A simple HPLC procedure for the quantitative determination of citral, the major fragrant component in the lemon grass, has been developed. The procedure involves a C-8 stationary phase using a 90:10 methanol: water pH 5 mobile phase containing 0.25% 1-octanesulfonic acid and an UV detector (set at 233 nm). The lemon grass leaves were harvested fresh at different times of the year and were soaked in methanol for 48 hours without any mechanical assistance to extract the citral and other methanol soluble components. The method showed good reproducibility with relative standard deviation of 2.8% and 10.8% for two different sets of samples. The method showed linearity in the range of 0.89 - 35.52 μg/mL. The average recovery was 104.8%. The amount of citral found as a percentage of the dried leaves are, 0.093, 0.27, 0.10, 0.13, 0.16, and 0.066 for fall, winter, three summer, and 1 commercial store samples, respectively.

Ce travail a été effectué dans le cadre d’une convention de cotutelle entre les Universités de Corse et d’Antananarivo. Il a contribué à la caractérisation de cinq Plantes de Madagascar à travers la composition chimique de leurs huiles essentielles (HE). Quatre plantes aromatiques et médicinales (Cymbopogon giganteus var. madagascariensis, Tana bojeriana, Croton kimosorum et Croton sp.) poussent à l’état spontané dans la région sub-aride du sud-ouest de Madagascar (Toliara) alors que l’espèce Vepris madagascarica est présente dans la forêt dense humide du nord-est (Vohémar). Ainsi, nous avons étudié par combinaison CPG (IR), CPG-SM et RMN 13C la composition chimique d’huiles essentielles obtenues par hydrodistillation (appareil type Clevenger) et par entraînement à la vapeur d’eau (Alambic).Les huiles essentielles de Cymbopogon giganteus Chiov. var. madagascariensis (A. Camus) ou Ahibero possèdent une composition chimique comparable avec celles des espèces Cymbopogon giganteus Chiov. africaines décrites dans la littérature. Elles sont dominées par les quatre menthadiénols (cis- et trans-p-mentha-1(7),8-dièn-2-ol, cis- et trans-p-mentha-2,8-dièn-1-ol) et le limonène. Nous avons aussi identifié quatre hydropéroxydes possédant le squelette p-menthane. Une monographie de l’huile essentielle d’Ahibero a été proposée à l’intention des utilisateurs de cette huile essentielle.La composition chimique de l’HE de Tana bojeriana Baker est caractérisée par la présence majoritaire de l’-phellandrène (30,4%), de la carvone (22,3%), du limonène (13,3%) et du dill éther (7,7%). Cette composition chimique apparait très proche de celle d’Anethum graveolens (Apiaceae). La présence du trans-p-menth-2-èn-1,6-diol et du p-mentha-1(7),2-dièn-6-ol lui confère une certaine originalité Elle pourrait donc constituer une alternative aux huiles essentielles qui sont bien implantées sur le marché des Plantes Aromatiques et Médicinales (PAM).La composition chimique de l’HE de feuilles de Vepris madagascarica (Baill.) H. Perrier est caractérisée par une très forte teneur en (E)-anéthole (78,2%). L’étude comparative de la composition chimique des huiles essentielles de feuilles (fraîches et séchées) et d’écorces de tronc confirme la prédominance du (E)-anéthole (69,5 à 84,6%). Compte tenu de la répartition restreinte de cette plante aromatique et en vue d’une gestion durable de cette espèce, son exploitation devrait être limitée aux huiles essentielles des feuilles qui pourraient ainsi constituer une source de (E)-anéthole, composé très utilisé en agro-alimentaire.Deux espèces de Croton endémiques de Madagascar ont été étudiées, Croton kimosorum Leandri (Zanapoly ou Napoly) et Croton sp. (Andriambolamena ou Riamena). Les huiles essentielles de ces deux plantes aromatiques et médicinales sont caractérisées par la présence du linalol (Zanapoly) et du limonène(Andriambolamena). La première HE est caractérisée par une majorité de composés oxygénés tandis que la seconde est dominée par les composés oléfiniques monoterpéniques. Dans les deux cas, de nombreux sesquiterpènes oxygénés ont été identifiés. Parmi eux, l’épi--bisabolol, jamais décrit dans une HE du genre Croton, est trouvé pour la première fois dans les deux espèces. / This study was conducted under an agreement of co-supervision between the Universities of Antananarivo (Madagascar) and Corsica (France). It contributed to the characterization of five malagasy plants through the chemical composition of their essential oils (EO). Four aromatic and medicinal plants: Cymbopogon giganteus var. madagascariensis, Tana bojeriana, Croton kimosorum and Croton sp. grow wild in the south-western region of Madagascar (Toliara) whereas the species Vepris madagascarica is widespread in the rainforest of the north-eastern region (Vohémar). The identification of components of EOs produced by hydrodistillation (Clevenger type apparatus) and by steam distillation (Alembic) has been obtained by combination of GC (RI), GC-MS and 13C NMR.Cymbopogon giganteus Chiov. var. madagascariensis (A. Camus) EO or Ahibero EO of Madagascar has similar chemical composition with Cymbopogon giganteus Chiov. african species described in literature. Its chemical composition is dominated by four p-menthadienols (cis- and trans-p-mentha-1(7),8-dien-2-ol, cis- and trans-p-mentha-2,8-dien-1-ol) and limonene. Moreover, four peroxides derivatives bearing the p-menthane skeleton were also identified. A monography has been proposed to Ahibero EO users.Tana bojeriana Baker EO is characterized by the predominance of -phellandrene (30.4%), carvone (22.3%), limonene (13.3%) and dill ether (7.7%). The chemical composition of this EO was very close to that of Anetum graveolens (Apiaceae family), constituents. The presence of trans-p-menth-2-en-1,6-diol and p-mentha-1(7) ,2-dien-6-ol, gives some originality to Tana bojeriana EO. It could be therefore an alternative for essential oils that are already well established on the Medicinal and Aromatic Plants market.The chemical composition of Vepris madagascarica (Baill.) H. Perrier leaf EO is characterized by a high content of (E)-anethole (78.2%). The comparative study of the chemical composition of leaves (fresh and dried) and bark trunk EOs confirmed the predominance of (E)-anethole (69.5 to 84.6%). Taking account of the restricted distribution of this aromatic plant and the need for sustainable management of this species, its potential use would be limited to leaf oil and could be a source for this compound widely used in food industry.Two species of Croton endemic from Madagascar were studied, Croton kimosorum Leandri (Zanapoly or Napoly) and Croton sp. (Andriambolamena or Riamena). Both aromatic and medicinal plants EOs are characterized by the presence of linalool (Zanapoly) and limonene (Andriambolamena). Croton kimosorum is characterized by a majority of oxygenated compounds, while the second is dominated by hydrocarbon monoterpenes. In both cases, several oxygenated sesquiterpenes are identified. Among these, the epi--bisabolol, never reported in of Croton EOs, was found for the first time in both species.

Les huiles essentielles de citronnelle (Cymbopogon citratus), de géranium (Pelargonium graveolens) et de vétiver (Vetiveria zizanioides) sont utilisées partout dans le monde pour leur activité répulsive contre les principaux vecteurs (moustiques, tiques) de maladies infectieuses chez l'Homme (paludisme, chikungunya, …). L'application cutanée de ces produits naturels pour éviter le contact avec un vecteur n'avait pas été encore envisagée comme moyen de limiter les premiers stades de l’infection par l'agent pathogène transmis par le vecteur. Pour vérifier cette hypothèse, les travaux ont été consacrés à la mise en place d'un cadre structuré pour la réévaluation chimique et biologique des trois huiles essentielles sur le modèle du virus du Ross River (alphavirus) de la même famille que le virus du Chikungunya. La caractérisation chimique des huiles essentielles avec une technique de haute résolution (GC×GC/TOF-MS) a permis d'établir leur profil chémotypique précis. L'utilisation de marqueurs spécifiques (clones moléculaires du virus) a permis d'établir l'inhibition de la réplication virale en fonction des conditions d'application des huiles essentielles de géranium et citronnelle. Ces résultats suggèrent l'intérêt d’une huile essentielle répulsive dans les premiers stades d'une infection par un vecteur. À ce titre, l'étude comparative établit la haute valeur ajoutée de l'huile essentielle de géranium et oriente la recherche de nouveaux anti-infectieux naturels vers des complexes riches en monoterpènes. / Essential oils of citronella (Cymbopogon citratus), geranium (Pelargonium graveolens) and vetiver (Vetiveria zizanioides) are used worldwide as topical repellent against the main vectors (mosquitoes, ticks) of human infectious diseases (Malaria, chikungunya, …). Skin treatment with these natural products, initially to avoid contact with the vector had not yet been considered as a way to disrupt the early stages of infection when the repelling action fails. To check this hypothesis, a structured framework has been performed for the chemical and biological re-evaluation of the three essential oils. The latter was tested against Ross River virus (alphavirus) that belongs to the same family of Chikungunya virus. Analysis of essential oils using a high-resolution technique (GC × GC / TOF-MS) resulted in a more accurate chemotypical profile of the local production. The use of specific markers (molecular clones of the virus, Saclay CEA) allowed to establish the inhibition of viral replication depending of the conditions of geranium and citronella essential oils application. These results suggest the great interest of an essential oil topical repellent in the early stages of a vector infection. The comparative study established the high value of geranium essential oil and gave future direction to the discovery of new anti-infectious solutions from monoterpenes-rich natural complexes.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / Oxidative stress (OS) is involved in the development of several disorders involving the central nervous system, such as Alzheimer s disease (AD). Considering the limitations of current therapeutics for AD, there is still a great demand for discovery of new medical alternatives. In the first part of the present study it was investigated the antioxidant capacity and the chemical composition of three plants popularly used to treat neurological disorders: Melissa officinalis, Matricaria recutita and Cymbopogon citratus. The antioxidant effect of some phenolic compounds was also examined for comparative purposes. All plant extracts were effective in to combat OS, but Melissa officinalis presented the highest antioxidant effect. Among the purified compounds, quercetin had the highest antioxidant activity followed by gallic acid, quercitrin and rutin. In this work, we have demonstrated that the plant extracts could protect against OS induced by various pro-oxidant agents that induce lipid peroxidation by different process. Based on this study, which indicates that Melissa officinalis could be an effective agent in the prevention of various neurological diseases associated with OS. In the second part of this study, it was investigated the chemical composition and antioxidant activity of different fractions from Melissa officinalis extract. Furthermore, it was verified the effect of the most antioxidant fraction on acethylcholinesterase (AChE) activity and the effect of gallic acid on the matrix metalloproteinase-2 (MMP-2) activity. Ethyl acetate fraction presented the highest flavonoids content as well as antioxidant activity when compared with other tested fractions and also it caused AChE inhibition. Moreover, gallic acid inhibited MMP-2 activity. In conclusion, M. officinalis ethyl acetate fraction is suggested to be further investigated for its possible use in the treatment of AD, due its antioxidant, anticholinesterase activities and the MMP-2 inhibitory capacity of their phenolic compound, gallic acid. / O estresse oxidativo (EO) está envolvido no desenvolvimento de várias doenças relacionadas ao sistema nervoso central, tal como a Doença de Alzheimer (DA). Considerando as limitações da terapia existente para a DA, existe ainda a necessidade de novas alternativas médicas. Na primeira parte deste estudo, foi investigada a capacidade antioxidante e a composição química de três plantas usadas popularmente para tratar desordens neurológicas: Melissa officinalis, Matricaria recutita e Cymbopogon citratus. O efeito antioxidante de alguns compostos fenólicos também foi examinado para fins comparativos. Todos os extratos de plantas foram efetivos no combate ao EO induzido por diferentes agentes, mas o extrato aquoso da Melissa officinalis apresentou o maior efeito antioxidante. Dentre os compostos purificados, a quercetina teve a maior atividade antioxidante seguida pelo ácido gálico, quercitrina e rutina. Neste trabalho, foi mostrado que os extratos de plantas protegeram contra o EO induzido por vários agentes pró-oxidantes que induzem peroxidação lipídica por diferentes processos. Baseado nestes resultados, que indicam que a Melissa officinalis poderia ser um agente efetivo na prevenção de várias doenças neurológicas associadas ao EO, na segunda parte deste estudo, foram investigadas a composição química e a atividade antioxidante de diferentes frações do extrato de Melissa officinalis. Além disso, investigou-se o efeito da fração com maior potencial antioxidante na atividade da acetilcolinesterase (AChE) e o efeito do ácido gálico na atividade da metaloproteinase-2 da matriz (MMP-2). A fração acetato de etila apresentou maior conteúdo de flavonóides associada a uma maior atividade antioxidante quando comparado com as outras frações testadas e causou inibição da AChE. Além disso, o ácido gálico inibiu a atividade da MMP-2. Desta forma, sugere-se que a fração acetato de etila da M. officinalis poderia ser melhor investigada para o seu possível uso no tratamento da DA, devido às suas atividades antioxidante e anticolinesterase e a capacidade do ácido gálico, composto fenólico presente em sua composição, inibir a MMP-2.

Made available in DSpace on 2014-06-11T19:22:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-06Bitstream added on 2014-06-13T20:48:38Z : No. of bitstreams: 1 lorenzetti_er_me_botfca.pdf: 1497737 bytes, checksum: 301fad7ec025809e7f69e3df89b5a8b9 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Cymbopogon citratus (DC.) Stapf é uma gramínea perene, de grande importância para a indústria farmacêutica e alimentícia, fornecedora de óleo essencial. Um problema limitante para a produção comercial é a ferrugem, causada pelo patógeno Puccinia nakanishikii Dietel. O trabalho objetivou estudar a interação patógeno-hospedeiro e availar princípios ativos naturais no controle da doença. Para os estudos anatômicos foram elaborados cortes, objetivando analisar as estruturas lesionadas e o desenvolvimento do patógeno. Foram avaliadas as trocas gasosas nas diferentes severidades da doença. Realizaram-se testes in vitro, com diferentes agentes de controle sobre a germinação de esporos do patógeno, sendo estes: óleos essenciais de eucalipto, palma-rosa, cravo-da-índia, menta, limão, citronela e mil-folhas; extrato pirolenhoso de eucalipto e mentol cristalizado. A partir destes testes foram escolhidos quatro agentes para os testes em campo, óleo de citronela e de eucalipto, extrato pirolenhoso e mentol. A área foi preparada a partir de mudas, sendo manejadas até os seis meses. Quando todas as plantas apresentavam sintomas, iniciaram-se os tratamentos com aplicações semanais. Foi avaliada ainda a doença após um corte. Os sintomas iniciais manifestam-se como manchas amareladas evoluindo para a formação de pústulas bronzeadas alongadas. Quanto à avaliação anatômica, após a infecção houve o desenvolvimento das estruturas fúngicas, prosseguindo com o rompimento da cutícula, possibilitando a observação das estruturas do patógeno de forma clara sobre as células da epiderme. Nas regiões lignificadas dos feixes vasculares não há desenvolvimento dessas estruturas. Com o aumento da severidade da doença observou-se a redução da assimilação de gás carbônico. A concentração interna de gás carbônico foi maior em severidades mais altas. Tal fato relaciona-se... / Cymbopogon citratus (DC) Stapf is a perennial grass, that has a large importance to the pharmaceutical and food industry, supplier of essential oil. One limiting problem to the commercial production is the rust caused by pathogen Puccinia nakanishikii Dietel. This work aimed to investigate the pathogen-host interactions and contribute to the researches with alternative methods of disease control. For anatomical studies were prepared anatomical sections, with the objective to analyze the structures injured and the development of the pathogen with different corants. On cultivated plants were evaluated gas exchange in different severity levels. In vitro tests were performed with different control agents, to the germination of pathogen spores, which are: essential oil of eucalyptus, palmarosa, clove, peppermint, lemon, citronella and yarrow; pyroligneous extract of eucalyptus and crystallized menthol. From these tests four products were chosen for the tests in a camp experiment, citronella oil, eucalyptus oil, pyroligneous extract and menthol. The area was prepared from seedlings, which were managed by six months age. When all plants had symptoms the treatments begin, with weekly applications. One avaliation happened after the cut of the lemongrass. The initial symptoms manifest as yellow spots progressing to formation of elongated pustules. These symptoms beginning with the development of fungal structures, after occurred the disruption of the cuticle, so the structures of the pathogen may be clearly observed on epidermis parenchyma cells of the leaf abaxial surface, however, in regions of vascular bundles, no development of such structures, these regions that have lignified structures. With the increase of the disease severity was a significant reduction in the assimilation of carbonic gas. The internal concentration of carbon dioxide was major... (Complete abstract click electronic access below)

Ce travail de thèse, financé par l'Agence de Développement Economique de la Corse de la Collectivité Territoriale de Corse, a pour objectif la caractérisation de diverses plantes poussant à l'état spontané en Corse (genévriers, pin maritime) à travers la composition de leurs huiles essentielles et résines. Pour cela, différentes techniques chromatographiques et spectroscopiques ont été mises œuvre, incluant la RMN 13C des mélanges, méthode mise au point et développée au laboratoire. Ainsi, nous avons montré différents aspects de l'application de la méthode d'analyse par RMN 13C à travers i) l'étude d'huiles essentielles du genre Cymbopogon, ii) l'étude de la variabilité chimique des huiles essentielles d'hybrides entre la clémentine commune et la mandarine Willow Leaf et iii) la mise au point d'une méthode de quantification du dipropylene glycol. Ensuite, pour chacune des espèces du genre Juniperus (J. thurifera, J. oxycedrus ssp. macrocarpa, J. communis var. communis et J. communis ssp. alpina), nous avons réalisé un échantillonnage aussi important que possible en fonction de la disponibilité de la plante. Les huiles essentielles, préparées par hydrodistillation, ont été analysées par combinaison de différentes techniques (CC, CPG(Ir), CPG-SM, RMN 13C). Les résultats des analyses ont été soumis à un traitement statistique des données (ACP, k-means) quand le nombre d'échantillons était suffisant. Ainsi nous avons montré, dans deux cas, une variabilité chimique intraspécifique. Les échantillons de J. communis var. communis et de J. communis ssp. alpina se répartissent en deux groupes selon, dans le premier cas, leur teneur en -pinène et en sabinène et dans le second cas, leur teneur en limonène et en sabinène. Pour ce qui est des deux autres espèces, J. thurifera et J. oxycedrus ssp. macrocarpa, leurs huiles essentielles présentent comme composé majoritaire, respectivement le limonène et par l'-pinène, avec toutefois quelques compositions atypiques. Enfin, nous avons caractérisé l'huile essentielle de pin maritime (Pinus pinaster Ait.) de Corse, par l'analyse de 46 échantillons provenant d'aiguilles récoltées dans les différents massifs forestiers de l'île. Nous observons 3 types de compositions, dominées respectivement par des monoterpènes (-pinène), des sesquiterpènes (germacrène D) ou par des diterpènes (abiétadiène), sans qu'il soit possible pour autant de dégager des groupes bien définis. Nous avons également décrit la composition chimique de la fraction acide de 23 échantillons d'oléorésines de pin maritime, provenant de trois forêts de Corse. Sur l'ensemble de l'échantillonnage, nous avons identifié 11 acides résiniques. L'analyse statistique (ACP, K-means) de ces résultats suggère l'existence de deux groupes, sur la base des teneurs des acides lévopimarique et déhydroabiétique. Par ailleurs, le pinorésinol, produit à forte valeur ajoutée de la famille des lignanes, a été mis en évidence, pour la première fois à notre connaissance, dans des résines de pins.

Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2016-08-01T19:55:39Z No. of bitstreams: 3 Dissertação- Reginaldo Teixeira Nunes - 2014.pdf: 2255035 bytes, checksum: 70c145eb933df8fd6b89f116a91cade4 (MD5) Ata da Defesa de Mestrado. Reginaldo Texeira Nunes.jpg: 2607204 bytes, checksum: fb38ba2b64baafd566c2d99c81304371 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-02T12:26:59Z (GMT) No. of bitstreams: 3 Dissertação- Reginaldo Teixeira Nunes - 2014.pdf: 2255035 bytes, checksum: 70c145eb933df8fd6b89f116a91cade4 (MD5) Ata da Defesa de Mestrado. Reginaldo Texeira Nunes.jpg: 2607204 bytes, checksum: fb38ba2b64baafd566c2d99c81304371 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-08-02T12:26:59Z (GMT). No. of bitstreams: 3 Dissertação- Reginaldo Teixeira Nunes - 2014.pdf: 2255035 bytes, checksum: 70c145eb933df8fd6b89f116a91cade4 (MD5) Ata da Defesa de Mestrado. Reginaldo Texeira Nunes.jpg: 2607204 bytes, checksum: fb38ba2b64baafd566c2d99c81304371 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2014-07-04 / The basidiomycete Cryptococcus neoformans is a pathogenic yeast responsible for causing meningoencephalitis in patients with a compromised immune system, leading to high mortality rates worldwide. Presently available antifungal drugs for treating this disease are not always efficient and they may lead to serious side effects and sometimes to fungal resistance. This situation has caused an increasing search for natural products derived from plants with antifungal action. Cymbopogon winterianus Jowitt (Poaceae), popularly known as citronella and Thymus vulgaris L. (Labiatae), known as thyme, are two plants commonly used in folk medicine. This study aimed at evaluating the biological activity of both C. winterianus and T. vulgaris essential oils (EOs) on C. neoformans clinical isolates. We evaluated the in vitro susceptibility of C. neoformans to each EO by the broth microdilution method, the in vitro interaction between EOs and fluconazole by the checkerboard method, their toxicity on human erythrocytes and their effects on the mitochondrial metabolism of fungal cells. Both EOs showed to possess antifungal activity against the tested isolates, with minimum inhibitory concentrations (MICs) ranging from 32 to 256 μg/mL for C. winterianus and from 32 to 128 μg/mL for T. vulgaris. No synergism between the EOs and fluconazole was observed. C. winterianus and T. vulgaris EOs did not cause hemolysis on human erythrocytes at concentrations corresponding to MIC values, indicating, therefore, low toxicity. The MTT assays showed no alterations on mitochondrial metabolism of fungal cells by C. winterianus at concentrations ≤ 128μg/mL, as well as no alterations at any concentrations corresponding to MIC values by T. vulgaris, suggesting that the main mechanism of action of both EOs is not by interfering with mitochondrial activity. Results show that C. winterianus and T. vulgaris EOs may be promising on the production of new antifungal drugs. / O basidiomiceto Cryptococcus neoformans é uma levedura patogênica causadora de meningoencefalite em pacientes imunocomprometidos, levando a altas taxas de mortalidade em todo o mundo. Os antifúngicos atualmente disponíveis para o tratamento dessa enfermidade nem sempre são eficazes, podendo causar efeitos colaterais graves e levar a casos de resistência. Diante dessa situação, a busca por produtos naturais, com ação antifúngica, oriundos de plantas tem crescido consideravelmente. Cymbopogon winterianus Jowitt (Poaceae), conhecida popularmente como citronela e Thymus vulgaris L. (Labiatae), conhecida como tomilho-branco, são duas plantas comumente utilizadas na medicina popular. Este trabalho avaliou a atividade biológica dos óleos essenciais (OEs) de ambas, em isolados clínicos de C. neoformans. Foram avaliados a suscetibilidade in vitro destes isolados ao OE de cada planta pelo método de microdiluição em caldo, a interação in vitro entre os OEs e fluconazol pelo método de checkerboard, sua toxicidade em eritrócitos humanos e seus efeitos no metabolismo mitocondrial de células fúngicas. Ambos os OEs apresentaram atividade antifúngica, com concentrações inibitórias mínimas (CIMs) variando de 32 a 256μg/mL, para C. winterianus, e de 32 a 128μg/mL para T. vulgaris. Não houve sinergismo entre os OEs e fluconazol. Os OEs de C. winterianus e T. vulgaris não causaram hemólise em eritrócitos humanos nas concentrações correspondentes às CIMs, mostrando-se de baixa toxicidade. Nos ensaios com MTT, observou-se que não houve alterações no metabolismo mitocondrial das células fúngicas por C. winterianus em concentrações iguais ou inferiores a 128μg/mL, e em nenhum dos valores correspondentes às CIMs por T. vulgaris, sugerindo que o principal mecanismo de ação antifúngica desses OEs nas CIMs não ocorre por interferência na atividade mitocondrial. Os resultados obtidos demonstram que OEs de C. winterianus e T. vulgaris podem ser promissores para a produção de novos antifúngicos.

Orientador: Ary Fernandes Junior / Resumo: Os óleos essenciais (OE) são produtos do metabolismo secundário da planta e são conhecidos pelas suas propriedades antimicrobianas. O principal componente do OE da palmarosa é o geraniol (70-80%). A incorporação de compostos da biodiversidade agrega valor as formulações dermocosméticas. OEs são uma boa opção devido à semelhança estrutural com o equilíbrio hidro-lipídico da pele. Assim, o objetivo do estudo foi avaliar a atividade antibacteriana desses compostos e o desenvolvimento de protótipos de dermocosméticos com nanopartículas para o tratamento da acne vulgar. Nanoemulsões com OE e geraniol foram preparadas de acordo com os valores do equilíbrio hidrofílico-lipofílico (EHL) e a caracterização das nanoemulsões seguiram os parâmetros da ANVISA (2004) para testes preliminares de estabilidade. Foram preparadas dez formulações. Nanoemulsões especiais contendo nanopartículas de prata (AgNP) foram preparadas e testadas contra C. acnes. Os ensaios de microtitulação com resazurina (REMA) foram realizados para determinar a Concentração Inibitória Mínima (CIM) e a Concentração Bactericida Mínima (CBM) para o OE de palmarosa e geraniol contra cepas American Type Culture Collection (ATCC) de Cutibacterium acnes 11827, 11828 e 6919. A curva de sobrevivência (CS) foi realizada para analisar o sinergismo entre os compostos e a clindamicina contra C. acnes, cepa 6919. As formulações foram estáveis ao pH, condutividade elétrica, densidade, centrifugação e análise de stress térmico. Anális... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Essential oils (EO) are secondary metabolism products of plant and they are known for antimicrobial properties.The major component of palmarosa EO is geraniol (70-80%). Incorporation of compounds of biodiversity guarantee value to dermocosmetic formulations. EOs are good option due to structural similarity with skin hydro-lipid balance. Thus, the desire was to evaluate the antibacterial activity to these compounds and the development of dermocosmetic prototypes with nanoparticles to treat acne vulgaris. Nanoemulsion with EO and geraniol were prepared according to values of lipophilic-hydrophilic balance (LHB) and nanoemulsions characterization followed the ANVISA (2004) parameters for preliminary stability tests. Ten formulations were prepared. Special nanoemulsion containing silver nanoparticles (AgNP) was prepared and tested against C. acnes. Resazurin Microtiter Assays (REMA) was performed to determine the Minimum inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) for palmarosa EO and geraniol against American Type Culture Collection (ATCC) strains of Cutibacterium acnes 11827, 11828 and 6919. The target of Time Kill Curve (TKC) test was determinate the synergism between compounds and clindamycin to C. acnes strain 6919. The formulations were stable to pH, electrical conductivity, density, centrifugation and thermal stress analysis. Analysis of size, zeta potential (ZP) and polydispersity index (PDI) were between 60 and 165 d.nm; -16.6 and -34.8 mv... (Complete abstract click electronic access below) / Mestre

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-02-17T13:13:31Z No. of bitstreams: 1 Antonio Alves Pimenta Neto.pdf: 891078 bytes, checksum: 3c3c0666af1521e887fa16686d4d993c (MD5) / Made available in DSpace on 2017-02-17T13:13:31Z (GMT). No. of bitstreams: 1 Antonio Alves Pimenta Neto.pdf: 891078 bytes, checksum: 3c3c0666af1521e887fa16686d4d993c (MD5) Previous issue date: 2012-02-29 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / This study aimed to evaluate the effectiveness of extracts and vegetable oils in controlling diseases of Solanaceae caused by Phytophthora nicotianae and its effects on the isolates of the pathogen. The effect of means and methods of cultivation in the sporulation of the pathogen was first studied to enable the inoculations and tests with oils and plant extracts. The first study was to evaluate the influence of various liquid and solid culture media, obtained from plant host tissues and/or indicated in the literature for the cultivation of Phytophthora spp, as well as different light regimes: constant dark (D), 12h alternating light (LD), and constant light (L) and the hydrogen potential in growth and sporulation of P. nicotianae. The mycelial growth was obtained at 10 and 15 days of culture, through the quantification of biomass. The development of the colonies in the agar media was monitored through daily measurement of radial growth. The spore (zoospores/mL) was obtained in different ways in the 10th and 15th day in a Neubauer chamber, and the data transformed to √ x +1. In the agar media, (D) showed the largest growth increases, however means differed in all light regimes, especially those of cassava, eggplant, tomato and vegetable juice (V8), the first two with the largest increases and the last two with the lowest. The light regimes did not significantly affect growth in liquid media with 10 days of incubation. In some ways the presence of light is inversely proportional to the vegetative growth, but was an essential factor for sporulation, since it showed the presence of zoospores only (L) and (LD), except for the V8 medium, where it obtained the highest sporulation in the absence of light (D). Strains, media, and the presence of agar, promoted statistical difference in the number of zoospores/mL in (L) and (LD). More acids culture media induced more sporulation of P. nicotianae and less mycelial growth. The second study was divided in "in vitro" and "in vivo” bioassays aiming to analyse the antifungal and biocontrole effect of vegetable oils and extracts of Syzygium aromaticum and Cymbopogon nardus on fruit and tomato and eggplant seedlings inoculated with P. nicotianae. It was found that the products inhibited the germination of the mycelial growth and zoospores obtained from S. aromaticum at 0,5 μL/ ml and 10% concentrations CAE and EO, respectively. Treatments with C. nardus EO and CAE at 1,0 μL/ ml and 20% respectively, delayed progression of disease in fruit and seedlings compared to inoculated control. It can be inferred that the products obtained from S. aromaticum and C. nardus, have the potential to reduce the attack of this pathogen on tomato and eggplant. / O presente trabalho teve como objetivo avaliar a eficiência de extratos e óleos vegetais no controle de doenças causadas por Phytophthora nicotianae em tomateiro e berinjela, bem como seus efeitos sobre os isolados do patógeno. A esporulação do patógeno foi inicialmente estudado para viabilizar as inoculações e os testes com os óleos e extratos vegetais. O primeiro estudo consistiu em avaliar a influência de diferentes meios de cultura líquidos e agarisados, obtidos de órgão vegetais de hospedeiros e/ou indicados na literatura para o cultivo de Phytophthora spp; além de diferentes regimes de luminosidade: ausência de luz (E), alternância luminosa de 12h (LE), e luz constante (L); e potencial hidrogeniônico no crescimento e esporulação de P. nicotianae. Nos meios agarisados, (E) proporcionou os maiores crescimentos, entretanto em todos os regimes de luz os meios se diferenciaram, destacando-se dos demais os de mandioca, berinjela, tomate, e suco de vegetais (V8), os dois primeiros com os maiores crescimentos e os dois últimos com os menores. Os regimes de luz não influenciaram significativamente o crescimento nos meios líquidos com 10 dias de incubação. Em alguns meios a presença de luz foi inversamente proporcional ao crescimento vegetativo, mas foi um fator essencial para a esporulação, pois se verificou a presença de zoósporos somente em (L) e (LE), a exceção do meio V8, no qual se obteve a mais alta esporulação na ausência de luz (E). Em (L) e (LE), os isolados, meios e a presença ou não de ágar, promoveram diferenças estatísticas quanto ao número de zoósporos/mL. Verificou-se que meios mais ácidos proporcionam um menor crescimento, mas uma maior esporulação para P. nicotianae. O segundo estudo foi dividido em bioensaios “in vitro” e “in vivo”, analisado o efeito fungitóxico e biocontrolador de óleos e extratos vegetais de Syzygium aromaticum e Cymbopogon nardus em frutos e plântulas de tomateiro e berinjela inoculados com P. nicotianae. Constatou-se que os produtos que mais inibiram o crescimento micelial e a germinação dos zoósporos, foram obtidos de S. aromaticum, a partir das concentrações de 0,5 μL/mL e 10% do OE e EBA, respectivamente. Enquanto que o tratamento que mais retardou a evolução da doença em frutos e plântulas quando comparado com a testemunha inoculada, foi o OE e EBA de C. nardus nas concentrações de 1,0 μL/mL e 20%, respectivamente. Com isso podemos inferir que os produtos obtidos de S. aromaticum e C. nardus, têm potencial para reduzir o ataque deste patógeno em plantas de tomate e berinjela.

Orientador: Luis Vitor Silva do Sacramento / Banca: João Domingos Rodrigues / Banca: Celso Dornellas Fernandes / Resumo: Cymbopogon citratus (DC.) Stapf é uma gramínea perene, de grande importância para a indústria farmacêutica e alimentícia, fornecedora de óleo essencial. Um problema limitante para a produção comercial é a ferrugem, causada pelo patógeno Puccinia nakanishikii Dietel. O trabalho objetivou estudar a interação patógeno-hospedeiro e availar princípios ativos naturais no controle da doença. Para os estudos anatômicos foram elaborados cortes, objetivando analisar as estruturas lesionadas e o desenvolvimento do patógeno. Foram avaliadas as trocas gasosas nas diferentes severidades da doença. Realizaram-se testes in vitro, com diferentes agentes de controle sobre a germinação de esporos do patógeno, sendo estes: óleos essenciais de eucalipto, palma-rosa, cravo-da-índia, menta, limão, citronela e mil-folhas; extrato pirolenhoso de eucalipto e mentol cristalizado. A partir destes testes foram escolhidos quatro agentes para os testes em campo, óleo de citronela e de eucalipto, extrato pirolenhoso e mentol. A área foi preparada a partir de mudas, sendo manejadas até os seis meses. Quando todas as plantas apresentavam sintomas, iniciaram-se os tratamentos com aplicações semanais. Foi avaliada ainda a doença após um corte. Os sintomas iniciais manifestam-se como manchas amareladas evoluindo para a formação de pústulas bronzeadas alongadas. Quanto à avaliação anatômica, após a infecção houve o desenvolvimento das estruturas fúngicas, prosseguindo com o rompimento da cutícula, possibilitando a observação das estruturas do patógeno de forma clara sobre as células da epiderme. Nas regiões lignificadas dos feixes vasculares não há desenvolvimento dessas estruturas. Com o aumento da severidade da doença observou-se a redução da assimilação de gás carbônico. A concentração interna de gás carbônico foi maior em severidades mais altas. Tal fato relaciona-se... (Resumo completo clicar acesso eletrônico abaixo) / Abstract: Cymbopogon citratus (DC) Stapf is a perennial grass, that has a large importance to the pharmaceutical and food industry, supplier of essential oil. One limiting problem to the commercial production is the rust caused by pathogen Puccinia nakanishikii Dietel. This work aimed to investigate the pathogen-host interactions and contribute to the researches with alternative methods of disease control. For anatomical studies were prepared anatomical sections, with the objective to analyze the structures injured and the development of the pathogen with different corants. On cultivated plants were evaluated gas exchange in different severity levels. In vitro tests were performed with different control agents, to the germination of pathogen spores, which are: essential oil of eucalyptus, palmarosa, clove, peppermint, lemon, citronella and yarrow; pyroligneous extract of eucalyptus and crystallized menthol. From these tests four products were chosen for the tests in a camp experiment, citronella oil, eucalyptus oil, pyroligneous extract and menthol. The area was prepared from seedlings, which were managed by six months age. When all plants had symptoms the treatments begin, with weekly applications. One avaliation happened after the cut of the lemongrass. The initial symptoms manifest as yellow spots progressing to formation of elongated pustules. These symptoms beginning with the development of fungal structures, after occurred the disruption of the cuticle, so the structures of the pathogen may be clearly observed on epidermis parenchyma cells of the leaf abaxial surface, however, in regions of vascular bundles, no development of such structures, these regions that have lignified structures. With the increase of the disease severity was a significant reduction in the assimilation of carbonic gas. The internal concentration of carbon dioxide was major... (Complete abstract click electronic access below) / Mestre