Submitted by JULIANA DA SILVA MENEZES AZOLA null (julianatutora23@yahoo.com.br) on 2017-01-05T18:20:16Z No. of bitstreams: 1 Tese_Juliana_da_Silva_Menezes_Azola.pdf: 1283526 bytes, checksum: 3710665b62f6ef126cbd8e2ca68a8629 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2017-01-10T17:16:20Z (GMT) No. of bitstreams: 1 azola_jsm_dr_jabo.pdf: 1283526 bytes, checksum: 3710665b62f6ef126cbd8e2ca68a8629 (MD5) / Made available in DSpace on 2017-01-10T17:16:20Z (GMT). No. of bitstreams: 1 azola_jsm_dr_jabo.pdf: 1283526 bytes, checksum: 3710665b62f6ef126cbd8e2ca68a8629 (MD5) Previous issue date: 2016-12-13 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A diarreia neonatal é uma das mais recorrentes enfermidades que acometem bezerros, acarretando prejuízos à pecuária leiteira. Nestes casos, um dos micro-organismos mais prevalentes é Escherichia coli, relacionada à contaminação fecal e a surtos alimentares. Para o tratamento, a crescente resistência bacteriana a antimicrobianos leva à busca por novas alternativas farmacológicas. O presente trabalho teve como objetivo geral testar a susceptibilidade de isolados de E. coli oriundos de bezerros neonatos diarreicos e sem diarreia a extratos vegetais. Os animais pertenceram a fazendas destinadas à produção leiteira no Estado de Minas Gerais, Brasil. As amostras foram submetidas ao isolamento microbiológico, à sorologia, à identificação genética por PCR e a testes antimicrobianos. Em relação aos antissoros polivalentes, houve isolados positivos a sorogrupos de importância clínica em humanos, como O26 e O111. Foram isoladas 36 estirpes oriundas de animais acometidos, positivas para pelo menos um gene testado, sendo eles stx1, stx2, eae, bfp e Sta. Relacionadas ao isolamento de animais sem diarreia, 9 estirpes foram carreadoras dos genes stx1, stx2 ou ambos, caracterizando os bovinos como reservatórios do patótipo STEC. O gene LT-II não foi encontrado. Quanto aos testes de susceptibilidade, encontrou-se isolados resistentes a diferentes classes de antimicrobianos. Entre os extratos testados, houve sensibilidade frente à planta Salvia officinalis L. (sálvia). Assim, esta vertente de estudo coloca-se como alternativa ao combate a patógenos bacterianos, válida para futuros testes quanto à descoberta de novos componentes químicos que possuam atividade antimicrobiana. / Neonatal diarrhea is one of the most recurrent illnesses that affect calves, resulting in losses to dairy farming. In these cases, one of the most prevalent microorganisms is Escherichia coli that is related to fecal contamination and food outbreaks. For the treatment, the increase bacterial resistance to antimicrobials leads to the search for new pharmacological alternatives. The main aim of this study was to test the susceptibility of E. coli isolated from neonatal calves with diarrhea and healthy to plant extracts. The animals were found in farms which are dairy-producing in the State of Minas Gerais, Brazil. The samples were subject to microbiological isolation, serology, PCR genetic identification and antimicrobial testing. In relation to polyvalent antiserum, there were positive isolates of clinical importance of serogroups in humans, such as O26 and O111. Thirty six strains were isolated from affected animals and they were positive to at least one gene tested, such as stx1, stx2, eae, bfp and Sta. Related to the healthy animals, 9 strains were carriers of the genes stx1, stx2 or both, characterizing the cattles as reservoirs of pathotype STEC. The gene LT-II was not found. In relation to susceptibility tests, resistant isolates to different classes of antimicrobials were found. Among the extracts that were tested, there was sensitivity to the Salvia officinalis L. plant (sálvia). Thus, this aspect of study can be alternative to combat the bacterial pathogens, valid for future tests as to the discovery of new chemical compounds with antimicrobial activity. / FAPESP: 14/06313-3

O diabetes mellitus é uma doença que afeta o metabolismo de carboidratos, gorduras e proteínas e tem como aspectos característicos a hiperglicemia e a excreção da glicose pela urina. Estima-se que o número de casos da doença aumentará muito nos próximos anos, o que a torna preocupante para o sistema público de saúde dos países mais afetados. Além do uso de medicamentos alopáticos, tratamentos complementares como o uso de plantas medicinais pode contribuir para melhorar a qualidade de vida do paciente. Entre as ervas utilizadas, tanto no Brasil quanto em outras partes do mundo, encontram-se Bidens pilosa e Salvia officinalis. Os mecanismos responsáveis pela atividade antidiabética das plantas, geralmente estão associados aos seus metabólitos secundários, no entanto, a influência de micronutrientes não deve ser descartada. Estudos mostram que há uma correlação entre a concentração destes elementos com a presença e o desenvolvimento da doença. O objetivo deste trabalho foi estudar os elementos Cr, Fe, Mg, Mn, V e Zn em Bidens pilosa (picão) e Salvia officinalis (sálvia) cultivadas com tratamento normal (substrato comercial) e com adição dos elementos acima mencionados, e verificar a influência destes elementos na produção de metabólitos secundários que podem atuar como agentes hipoglicêmicos. A determinação e quantificação elementar foram feitas por meio da técnica de Análise por Ativação Neutrônica Instrumental, e para análise dos metabólitos secundários utilizou-se a técnica de Cromatografia Líquida de Alta Eficiência. O resultados indicaram que B. pilosa e S. officinalis podem ser usadas como fontes de Cr, Fe Mg, Mn, V e Zn. Foi observado que a B. pilosa absorveu maior quantidade de Fe no grupo tratamento, e que a S. officinalis foi capaz de acumular Zn nas folhas independente de ter ou não sido tratada. Quanto aos compostos do metabolismo secundário, os resultados indicaram que sua produção pelas plantas aparentemente não foi alterada pela adição da solução dos nutrientes. / Diabetes mellitus is a disease that affects the carbohydrates, lipids and proteins metabolism characterized by hyperglycemia and glucose excretion by urine. It is estimated that the number of cases of this disease will increase in the coming years, worrying the public health system of the most affected countries. Besides the use of allopathic medicine, complementary treatments such as the use of medicinal plants can contribute to improving the pacients quality of life. Among the herbs used, both in Brazil and in other countries, are Bidens pilosa and Salvia officinalis. The mechanisms responsible for antidiabetic activity of the plants, are usually associated with secondary metabolites, however, the influence of micronutrient content should not be discarded. Studies have shown that there is a correlation between the concentration of these elements and the presence and development of the disease. The objective of this work was to study the elements Cr, Fe, Mg, Mn, V and Zn concentration in Bidens pilosa (beggarticks) and Salvia officinalis (sage) cultivated with normal treatment (commercial substrate) and, with the addition of these elements, to verify their influence in the production of secondary metabolites that can act as hypoglycemic agents. The elemental determination and quantification were performed by means of Instrumental Neutron Activation Analysis technique. For the secondary metabolites analysis Liquid Chromatography High Performance technique was used. The results indicated that B. pilosa and S. officinalis may be used as sources of Cr, Fe, Mg, Mn, V and Zn. It was observed that B. pilosa absorbed a larger amount of Fe in the treatment group, and S. officinalis was able to accumulate Zn its in leaves whether treated or not. Considering the secondary metabolism compounds, the results indicated that its production by plants was apparently not altered by the addition of the nutrient solution.

O diabetes mellitus é uma doença que afeta o metabolismo de carboidratos, gorduras e proteínas e tem como aspectos característicos a hiperglicemia e a excreção da glicose pela urina. Estima-se que o número de casos da doença aumentará muito nos próximos anos, o que a torna preocupante para o sistema público de saúde dos países mais afetados. Além do uso de medicamentos alopáticos, tratamentos complementares como o uso de plantas medicinais pode contribuir para melhorar a qualidade de vida do paciente. Entre as ervas utilizadas, tanto no Brasil quanto em outras partes do mundo, encontram-se Bidens pilosa e Salvia officinalis. Os mecanismos responsáveis pela atividade antidiabética das plantas, geralmente estão associados aos seus metabólitos secundários, no entanto, a influência de micronutrientes não deve ser descartada. Estudos mostram que há uma correlação entre a concentração destes elementos com a presença e o desenvolvimento da doença. O objetivo deste trabalho foi estudar os elementos Cr, Fe, Mg, Mn, V e Zn em Bidens pilosa (picão) e Salvia officinalis (sálvia) cultivadas com tratamento normal (substrato comercial) e com adição dos elementos acima mencionados, e verificar a influência destes elementos na produção de metabólitos secundários que podem atuar como agentes hipoglicêmicos. A determinação e quantificação elementar foram feitas por meio da técnica de Análise por Ativação Neutrônica Instrumental, e para análise dos metabólitos secundários utilizou-se a técnica de Cromatografia Líquida de Alta Eficiência. O resultados indicaram que B. pilosa e S. officinalis podem ser usadas como fontes de Cr, Fe Mg, Mn, V e Zn. Foi observado que a B. pilosa absorveu maior quantidade de Fe no grupo tratamento, e que a S. officinalis foi capaz de acumular Zn nas folhas independente de ter ou não sido tratada. Quanto aos compostos do metabolismo secundário, os resultados indicaram que sua produção pelas plantas aparentemente não foi alterada pela adição da solução dos nutrientes. / Diabetes mellitus is a disease that affects the carbohydrates, lipids and proteins metabolism characterized by hyperglycemia and glucose excretion by urine. It is estimated that the number of cases of this disease will increase in the coming years, worrying the public health system of the most affected countries. Besides the use of allopathic medicine, complementary treatments such as the use of medicinal plants can contribute to improving the pacients quality of life. Among the herbs used, both in Brazil and in other countries, are Bidens pilosa and Salvia officinalis. The mechanisms responsible for antidiabetic activity of the plants, are usually associated with secondary metabolites, however, the influence of micronutrient content should not be discarded. Studies have shown that there is a correlation between the concentration of these elements and the presence and development of the disease. The objective of this work was to study the elements Cr, Fe, Mg, Mn, V and Zn concentration in Bidens pilosa (beggarticks) and Salvia officinalis (sage) cultivated with normal treatment (commercial substrate) and, with the addition of these elements, to verify their influence in the production of secondary metabolites that can act as hypoglycemic agents. The elemental determination and quantification were performed by means of Instrumental Neutron Activation Analysis technique. For the secondary metabolites analysis Liquid Chromatography High Performance technique was used. The results indicated that B. pilosa and S. officinalis may be used as sources of Cr, Fe, Mg, Mn, V and Zn. It was observed that B. pilosa absorbed a larger amount of Fe in the treatment group, and S. officinalis was able to accumulate Zn its in leaves whether treated or not. Considering the secondary metabolism compounds, the results indicated that its production by plants was apparently not altered by the addition of the nutrient solution.

<p>Glavni cilj istraživanja ove doktorske disertacije je bio valorizacija sporednog proizvoda žalfije (Salvia officinalis L.) iz fabrike čaja za dobijanje visoko-vrednih ekstrakata željenih osobina koji bi imali primenu u gotovim proizvodima prehrambene, kozmetičke i farmaceutske industrije.<br />U cilju ekstrakcije polifenolnih jedinjenja su primenjene konvencionalne (maceracija) i savremene (ultrazvučna ekstrakcija, mikrotalasna ekstrakcija i ekstrakcija subkritičnom vodom) ekstrakcione tehnike. U postupaku maceracije je optimizovana koncentracija etanola u ekstragensu, dok je za optimizaciju savremenih ekstrakcionih tehnika primenjen eksperimentalni dizajn i metoda odzivne povr&scaron;ine. Prinos polifenola (ukupnih fenola i ukupnih flavonoida) i antioksidativna aktivnost određena različitim in vitro testovima su bili ispitivani odzivi. Za svaki od navedenih ekstrakcionih postupaka je ispitivan uticaj glavnih ekstrakcionih parametara: ultrazvučna ekstrakcija (temperatura, vreme ekstrakcije i snaga ultrazvuka), mikrotalasna ekstrakcija (koncentracija etanola, vreme ekstrakcije i odnos rastvarač-droga) i ekstrakcija subkritičnom vodom (temperatura, vreme ekstrakcije i koncentracija HCl u ekstragensu). Tečni ekstrakti dobijeni ultrazvučnom, mikrotalasnom i subkritičnom ekstrakcijom su pokazali izuzetno visoku antioksidativnu aktivnost određenu &bdquo;hvatanjem&ldquo; DPPH i O2∙- radikala i kapacitetom redukcije Fe3+ jona i imali su znatno veći sadržaj polifenola u odnosu na ekstrakte dobijene maceracijom.<br />U cilju ekstrakcije terpenoidnih jedinjenja su primenjeni postupci Soxhlet ekstrakcije, destilacije vodenom parom i ekstrakcije superkritičnim CO2. Postupak superkritične ekstrakcije je optimizovan metodom odzivne povr&scaron;ine i optimalni uslovi ekstrakcije su bili pritisak 290 bar, temperatura 55˚C i protok CO2 od 0,4 kg/h, dok su predviđene i eksperimentalne vrednosti prinosa ekstrakcije bile 8,90 i 8,84%. Hemijskom analizom lipofilnih ekstrakata i etarskog ulja je utvrđeno da su oksidovani monoterpeni (&alpha;-tujon i kamfor), oksidovani seskviterpeni (viridiflorol) i polifenolni diterpeni (epirosmanol) dominantna jedinjenja prisutna u ovim uzorcima.<br />Tečni ekstrakti žalfije dobijeni konvencionalnim i savremenim ekstrakcionim tehnikama su spray drying tehnikom osu&scaron;eni u cilju prevođenja u stabilniju formu suvog ekstrakta. Suvim ekstraktima su određene fizičko-hemijske osobine (hemijski sastav, sadržaj vlage, higroskopnost, moć rehidratacije, WAI i WSI) i biolo&scaron;ka aktivnost (antimikrobna i antioksidativna aktivnost) i procenjena je njihova mogućnost primene u prehrambenoj i farmaceutskoj industriji.</p> / <p>The main aim of this dissertation was valorization of sage (Salvia officinalis L.) by-product from filter-tea factory for recovery of high-value extracts with desirable properties for application in food, cosmetics and pharmaceutical products.<br />Conventional (maceration) and novel (ultrasound-assisted, microwave-assisted and subcritical water extraction) extraction techniques were applied for polyphenols recovery. Ethanol concentration was optimized in maceration process, while experimental design and response surface methodology were applied for optimization of novel extraction techniques. Polyphenols yield (total phenols and total flavonoids) and antioxidant activity, determined by various in vitro assays, were investigated responses. Influence of main process parameters was determined for each technique: ultrasound-assisted extraction (temperature, extraction time and ultrasonic power), microwave-assisted extraction (ethanol concentration, extraction time and sample-solvent ratio) and subcritical water extraction (temperature, extraction time and HCl concentration in solvent). Liquid extracts obtained by novel extraction technique exhibited higher antioxidant activity determined by scavenging of DPPH and O2∙- radicals and reduction of Fe3+ and also provided higher polyphenols yield comparing to extracts obtained by maceration. Soxhlet extraction, hydrodistillation and supercritical fluid extraction were applied for terpenoids recovery. Supercritical fluid extraction process was optimized by response surface methodology and optimal conditions were pressure of 290 bar, temperature of 55˚C and CO2 flow rate of 0.4 kg/h, while predicted and experimentally obtained values of total extraction yield at these conditions were 8.90 i 8.84%, respectively. According to chemical analysis, the most abundant compounds in lipid extracts and essential oil were oxygenated monoterpenes (&alpha;-thujone and camphor), oxygenated sesquiterpenes (viridiflorol) and diterpene polyphenols (epirosmanol).<br />Sage liquid extracts obtained by conventional and novel extraction techniques were spray dried in order to obtain dry extract form. Physico-chemical properties (chemical profile, moisture content, hygroscopicity, rehydratation time, WAI and WSI) and biological activity (antimicrobial and antioxidant activity) were determined in dry extracts and their potential application in food and pharmaceutical formulations was discussed.</p>

Oleanolic and ursolic acid (OA and UA) are triterpenic acids with diverse biological activities that are of interest to the pharmaceutical industry. To investigate the scope for producing these compound using cell suspension cultures of Salvia species, calli from S. officinalis, S. virgata and S. fruticosa were induced using several plant growth regulator (PGR) combinations. Eleven lines were selected for suspension induction from a pool of calli. Six suspension cultures were established successfully and cultivated in the Respiration Activity MOnitoring System® (RAMOS®) to obtain online data on their growth kinetics and to establish appropriate sampling schedules for the determination of their OA and UA production. Based on their observed growth behaviour, OA and UA contents, and aggregation properties, one suspension culture from each studied Salvia species was selected for further optimisation. The μmax values for these suspension cultures ranged from 0.20 to 0.37°d-1, their OA and UA contents were greater than 1.3 and 1.2 mg g-1, respectively, and they afforded maximum volumetric yields of 21.0 mg l-1 for OA and 32.8 mg l-1 for UA. These results will be useful in the development of a refined Salvia suspension-based process for OA and UA production.

Submitted by biblioteca unifenas (biblioteca@unifenas.br) on 2017-09-18T20:18:02Z No. of bitstreams: 1 Roberta Soares Terra Dissertação.pdf: 5586714 bytes, checksum: b90d918d9ac7cb880dcc55847cd8c5fd (MD5) / Made available in DSpace on 2017-09-18T20:18:02Z (GMT). No. of bitstreams: 1 Roberta Soares Terra Dissertação.pdf: 5586714 bytes, checksum: b90d918d9ac7cb880dcc55847cd8c5fd (MD5) Previous issue date: 2017-03-31 / Fundação de Amparo à Pesquisa do Estado de Minas Gerais - FAPEMIG / Salvia officinalis has been widely used in culinary and traditional medicine, and studies have identified numerous chemical compounds and potential therapeutic actions. This study evaluated the genotoxicity of lyophilized hydroalcoholic extract of S. officinalis L. (SO) leaves using the micronucleus assay in mouse bone marrow. The interaction between SO and the genotoxic effects induced by doxorubicin (DXR) was also analyzed – antigenotoxicity assay. Experimental groups consisting of male and female Swiss albinus mice (Unib: SW) were evaluated after 24-48h treatment with cyclophosphamide (CP; 50 mg/kg), DXR (5 mg/kg), NaCl 0.5-2 g/kg) and SO (0.5 g/kg) + DXR (5 mg/kg). The PCEMN analyzes showed differences (p0.05) between SO (1-2 g/kg) and NaCl treatments, regardless of gender and time. DXR induced EPCMNs significantly in both genders and treatment times. Groups of mice treated with DXR showed lower frequencies (p0.05) of PCEMNs when compared to CP control groups (50 mg/kg). Associative treatment (500 mg/kg SO + 5 mg/kg DXR) did not reduce the frequency of DXR-induced PCEMNs (p0.05). The PCE/NCE ratio between control (NaCl, CP and DXR) and experimental genotoxic and antigenotoxic (SO; SO + DXR) treatments were insignificant (p0.05). The results suggest moderately genotoxic effects (clastogeny and/or aneugeny) of S. officinalis L. leaves, dose-dependent (i.e., from 1 g/kg) and gender- and time-independent. However, S. officinalis has no systemic toxicity and antigenotoxic effects (SO + DXR) under the conditions established in the present micronucleus test in mouse bone marrow. / Salvia officinalis tem sido amplamente utilizada na culinária e na medicina tradicional, e estudos têm identificado inúmeros compostos químicos e potenciais ações terapêuticas. Esta pesquisa avaliou a genotoxicidade do extrato hidroalcoólico liofilizado de folhas de S. officinalis L. (SO) usando o ensaio do micronúcleo em medula óssea de camundongos. A interação entre SO e os efeitos genotóxicos induzidos pela doxorrubicina (DXR) também foi analisada – ensaio de antigenotoxicidade. Grupos experimentais constituídos de camundongos machos e fêmeas Swiss albinus (Unib: SW) foram avaliados após 24-48h de tratamento com ciclofosfamida (CP; 50 mg/Kg), DXR (5 mg/Kg), NaCl (150 mM), SO (0,5–2 g/Kg) e SO (0,5 g/Kg) + DXR (5 mg/Kg). As análises de EPCMNs mostraram diferenças (p0,05) entre os tratamentos de SO (1–2 g/Kg) e NaCl, independentemente do gênero e do tempo. DXR induziu EPCMNs significativamente em ambos os gêneros e tempos de tratamento. Grupos de camundongos tratados com DXR mostraram frequências menores (p<0,05) de EPCMNs quando comparados com os grupos controles CP (50 mg/Kg). O tratamento associativo (500 mg/Kg de SO + 5 mg/Kg de DXR) não reduziu a frequência de EPCMNs (p<0,05) induzida por DXR. As proporções de EPC/ENC entre tratamentos controles (NaCl, CP e DXR) e experimentais genotóxicos e antigenotóxicos (SO; SO + DXR) foram insignificantes (p0,05). Os resultados sugerem efeitos moderadamente genotóxicos (clastogenia e/ou aneugenia) de folhas de S. officinalis L., dose-dependente (i.e., a partir de 1 g/Kg) e gênero- e tempo-independentes. Contudo, S. officinalis não apresenta toxicidade sistêmica e efeitos antigenotóxicos (SO + DXR) nas condições estabelecidas no presente teste do micronúcleo em medula óssea de camundongos.

Made available in DSpace on 2014-06-11T19:32:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-09-01Bitstream added on 2014-06-13T21:03:35Z : No. of bitstreams: 1 povh_ja_dr_botib.pdf: 514154 bytes, checksum: 54ccc04cdb9b8c8830d238e77200844a (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Este estudo objetivou avaliar o efeito de reguladores vegetais e bioestimulantes no desenvolvimento, teor e composição de óleo essencial, teores de flavonóides e fenóis totais, atividade antioxidante e atividade das enzimas peroxidase e polifenoloxidade de plantas de Salvia officinalis L. Para tanto, instalou-se experimento em casa de vegetação, do Departamento de Botânica, Instituto de Biociências, da Universidade Estadual Paulisa - UNESP, Botucatu, SP. O delineamento experimental foi inteiramente casualizado com 6 tratamentos contendo três repetições. As plantas foram tratadas com as seguintes substâncias: 1- controle (água); 2- IBA a 100 mg L-1; 3- GA3 a 70 mg L-1; 4-benzilaminopurina (BAP) a 70 mg L-1; 5- GA3+cinetina+IBA - Stimulate® a 1% e 5- GA4+7+N-(fenilmetil)-1-purina-6- amino - Promalin® a 100 mg L-1. As avaliações fisiológicas foram realizadas em 7 coletas, pelas seguintes características: altura da planta, área foliar, massa seca de caule, folhas e da parte aérea da planta. Para as avaliações fitoquímicas, foram avaliadas a produção de massa seca da parte aérea e rendimento do óleo essencial, aos 90 e 120 D.A.E. As análises bioquímicas foram determinadas nas sete coletas, determinando-se: os teores de flavonóides e fenóis totais, atividade das enzimas peroxidase e polifenoloxidase e atividade antioxidante (CE50). Analisando-se os resultados obtidos foi possível verificar que plantas tratadas com IBA e GA3 promoveram maiores incrementos na produtividade vegetal, principalmente, na formação da parte aérea. A produção de óleo essencial foi incrementada pela aplicação de IBA nas plantas, mas também tratamentos com GA3 e o bioestimulante Stimulate® também influenciaram positivamente no acúmulo de óleo essencial. O maior rendimento de óleo essencial foi obtido na coleta aos 120 D.A.E. Os... / This study aimed at evaluating the effect of plant growth regulators and biostimulants on development, essential oil content and composition, total flavonoid and phenol levels, besides peroxidase, polyphenol oxidase and antioxidant activities in Salvia officinalis L plants. The experiment was carried out in a greenhouse from Department of Botany, Institute of Biosciences, São Paulo State University - UNESP, Botucatu, São Paulo State, Brazil. Experimental design was completely randomized with six treatments and three replicates. Plants were subjected to the following substances: 1- control (water); 2- IBA 100 mg L-1; 3- GA3 70 mg L-1; 4- benzylaminopurine (BAP) 70 mg L-1; 5- GA3+kinetin+IBA - Stimulate® 1%, and 5- GA4+7+ N-(phenylmethyl)-1H-purine-6-amine - Promalin® 100 mg L- 1. Physiological evaluations were performed in seven harvestings and consisted of plant height, leaf area, and dry matter of stem, leaves and shoot. As regards phytochemical evaluations, shoot dry matter and essential oil yield were assessed at 90 and 120 D.A.E. Also, the following biochemical evaluations were performed: total flavonoid and phenol levels, besides peroxidase, polyphenol oxidase and antioxidant (CE50) activities. IBA- and GA3- treated plants had higher plant productivity, mainly concerning shoot formation. IBA application led to an increase in essential oil yield, which was also positively influenced by GA3 and Stimulate®, a biostimulant. The highest essential oil yield was detected at 120 D.A.E. Plant growth regulators, biostimulants and harvesting times did not lead to alterations in the essential oil chemical composition. IBA-treated plants presented the highest total phenol and flavonoid levels. Plants subjected to IBA, GA3 and Promalin® had higher antioxidant activity. Considering enzymatic activities, there was an increase in peroxidases when plants were treated with GA3... (Complete abstract click electronic access below)

Orientador: Elizabeth Orika Ono / Resumo: Este estudo objetivou avaliar o efeito de reguladores vegetais e bioestimulantes no desenvolvimento, teor e composição de óleo essencial, teores de flavonóides e fenóis totais, atividade antioxidante e atividade das enzimas peroxidase e polifenoloxidade de plantas de Salvia officinalis L. Para tanto, instalou-se experimento em casa de vegetação, do Departamento de Botânica, Instituto de Biociências, da Universidade Estadual Paulisa - UNESP, Botucatu, SP. O delineamento experimental foi inteiramente casualizado com 6 tratamentos contendo três repetições. As plantas foram tratadas com as seguintes substâncias: 1- controle (água); 2- IBA a 100 mg L-1; 3- GA3 a 70 mg L-1; 4-benzilaminopurina (BAP) a 70 mg L-1; 5- GA3+cinetina+IBA - Stimulate® a 1% e 5- GA4+7+N-(fenilmetil)-1-purina-6- amino - Promalin® a 100 mg L-1. As avaliações fisiológicas foram realizadas em 7 coletas, pelas seguintes características: altura da planta, área foliar, massa seca de caule, folhas e da parte aérea da planta. Para as avaliações fitoquímicas, foram avaliadas a produção de massa seca da parte aérea e rendimento do óleo essencial, aos 90 e 120 D.A.E. As análises bioquímicas foram determinadas nas sete coletas, determinando-se: os teores de flavonóides e fenóis totais, atividade das enzimas peroxidase e polifenoloxidase e atividade antioxidante (CE50). Analisando-se os resultados obtidos foi possível verificar que plantas tratadas com IBA e GA3 promoveram maiores incrementos na produtividade vegetal, principalmente, na formação da parte aérea. A produção de óleo essencial foi incrementada pela aplicação de IBA nas plantas, mas também tratamentos com GA3 e o bioestimulante Stimulate® também influenciaram positivamente no acúmulo de óleo essencial. O maior rendimento de óleo essencial foi obtido na coleta aos 120 D.A.E. Os... (Resumo completo, clicar acesso eletrôncio abaixo) / Abstract: This study aimed at evaluating the effect of plant growth regulators and biostimulants on development, essential oil content and composition, total flavonoid and phenol levels, besides peroxidase, polyphenol oxidase and antioxidant activities in Salvia officinalis L plants. The experiment was carried out in a greenhouse from Department of Botany, Institute of Biosciences, São Paulo State University - UNESP, Botucatu, São Paulo State, Brazil. Experimental design was completely randomized with six treatments and three replicates. Plants were subjected to the following substances: 1- control (water); 2- IBA 100 mg L-1; 3- GA3 70 mg L-1; 4- benzylaminopurine (BAP) 70 mg L-1; 5- GA3+kinetin+IBA - Stimulate® 1%, and 5- GA4+7+ N-(phenylmethyl)-1H-purine-6-amine - Promalin® 100 mg L- 1. Physiological evaluations were performed in seven harvestings and consisted of plant height, leaf area, and dry matter of stem, leaves and shoot. As regards phytochemical evaluations, shoot dry matter and essential oil yield were assessed at 90 and 120 D.A.E. Also, the following biochemical evaluations were performed: total flavonoid and phenol levels, besides peroxidase, polyphenol oxidase and antioxidant (CE50) activities. IBA- and GA3- treated plants had higher plant productivity, mainly concerning shoot formation. IBA application led to an increase in essential oil yield, which was also positively influenced by GA3 and Stimulate®, a biostimulant. The highest essential oil yield was detected at 120 D.A.E. Plant growth regulators, biostimulants and harvesting times did not lead to alterations in the essential oil chemical composition. IBA-treated plants presented the highest total phenol and flavonoid levels. Plants subjected to IBA, GA3 and Promalin® had higher antioxidant activity. Considering enzymatic activities, there was an increase in peroxidases when plants were treated with GA3... (Complete abstract click electronic access below) / Doutor

En esta tesis se estudió el fraccionamiento de aceites esenciales obtenidos de plantas aromáticas comunes en el país mediante el uso de dióxido de carbono supercrítico, con el objetivo de obtener compuestos o fracciones con actividad biocida. Se seleccionaron tres plantas: Tagetes minuta, Salvia officinalis y Mentha piperita. La extracción del aceite esencial se realizó mediante hidrodestilación, determinándose su composición mediante cromatografía de gases. Como parte de esta tesis, se construyó un equipo experimental de alta presión, con el cual se realizaron mediciones de solubilidad y composición de la fase supercrítica para cada sistema [CO2 + aceite esencial] en distintas condiciones de presión y temperatura (70–120 bar, 313–323 K), utilizando el método dinámico o de gas saturado. Se realizaron también ensayos de fraccionamiento semicontinuo para determinar la influencia de la composición en el grado de separación. Con los resultados experimentales obtenidos se validó un modelo termodinámico predictivo basado en la ecuación de estado a contribución grupal (GC-EOS), previo ajuste de los parámetros necesarios con datos provenientes de otras fuentes. Utilizando dicho modelo como base, y reduciendo la complejidad de los sistemas en términos de dos o tres pseudo-componentes, se realizaron simulaciones de columnas de fraccionamiento de múltiples etapas en contra-corriente, analizando la influencia de las variables operativas (presión, temperatura, relación solvente/ alimentación, relación de reflujo, etc.) con el objetivo de hallar condiciones recomendables de operación para separar los compuestos biocidas de interés. En el análisis, se utilizaron herramientas y consideraciones de la Ingeniería del Equilibrio de Fases. En el caso del aceite de T. minuta se procuró separar la fracción de cetonas insaturadas (ocimenonas); en el caso del aceite de S. officinalis se planteó la purificación de la fracción de monoterpenos oxigenados, separándolos a la vez de los hidrocarburos y de los sesquiterpenos; en el caso del aceite de M. piperita se planteó la separación de mentona y mentol. La actividad biocida de las fracciones obtenidas experimentalmente a partir del aceite de T. minuta fueron ensayadas frente a dos parásitos: el gorgojo del maíz y el nemátodo del nudo de la raíz. / The fractionation of essential oils from local aromatic species using supercritical carbon dioxide was studied, in order to obtain compounds or fractions with biocidal activity. Three species were chosen: Tagetes minuta, Salvia officinalis y Mentha piperita. The essential oil was extracted by hydrodistillation and its composition determined by gas chromatography. As a part of this project, a lab-scale high-pressure experimental apparatus was built. With this equipment, measurements of solubility and composition in the supercritical phase were performed for each [CO2 + essential oil] system at different pressure and temperature conditions (70–120 bar, 313–323 K), using a dynamic or gas saturated method. Semi-continuous fractionation experiments were also carried out in order to determine the effect of composition in the separation performance. With the experimental data a predictive thermodynamic model based on the group contribution equation of state (GC-EOS) was validated, after fitting the necessary parameters with data from other sources. With this model as background, and reducing the systems complexity in terms of two or three pseudo-components, simulations of multistage counter-current columns were carried out, analyzing the influence of the operation parameters (pressure, temperature, solvent-to-feed ratio, reflux ratio, etc.), in order to find recommended conditions for the separation of the biocidal components. In this analysis, phase equilibrium engineering tools and considerations were applied. In the case of T. minuta, the goal was to isolate the unsaturated ketones fraction (ocimenones); in the case of S. officinalis the goal was the separation of the oxygenated monoterpenes fraction from both the hydrocarbon and the sesquiterpene fraction; finally in the case of M. piperita the separation between menthone and menthol was proposed. The biocidal activity of the fractions obtained experimentally from T. minuta oil was tested against the corn weevil and the root knot nemathod.

Oleanolic and ursolic acid (OA and UA) are triterpenic acids with diverse biological activities that are of interest to the pharmaceutical industry. To investigate the scope for producing these compound using cell suspension cultures of Salvia species, calli from S. officinalis, S. virgata and S. fruticosa were induced using several plant growth regulator (PGR) combinations. Eleven lines were selected for suspension induction from a pool of calli. Six suspension cultures were established successfully and cultivated in the Respiration Activity MOnitoring System® (RAMOS®) to obtain online data on their growth kinetics and to establish appropriate sampling schedules for the determination of their OA and UA production. Based on their observed growth behaviour, OA and UA contents, and aggregation properties, one suspension culture from each studied Salvia species was selected for further optimisation. The μmax values for these suspension cultures ranged from 0.20 to 0.37°d-1, their OA and UA contents were greater than 1.3 and 1.2 mg g-1, respectively, and they afforded maximum volumetric yields of 21.0 mg l-1 for OA and 32.8 mg l-1 for UA. These results will be useful in the development of a refined Salvia suspension-based process for OA and UA production.