Orientadores: Pedro Luiz Rosalen, Marta Cristina Teixeira Duarte / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-24T14:33:59Z (GMT). No. of bitstreams: 1 Freires_IrlandeAlmeida_M.pdf: 5300507 bytes, checksum: c7e268d2d061cfb090db127a48e976f8 (MD5) Previous issue date: 2014 / Resumo: Introdução: A candidíase oral é uma infecção fúngica oportunista da cavidade oral, cujas taxas de prevalência e incidência vêm aumentando significativamente em todo o mundo. Assim, novas estratégias orientadas para gerir esta doença têm sido propostas, dentre as quais está o uso de óleos essenciais (OE) com propriedades antifúngicas. Evidências indicam que o OE de Coriandrum sativum L. (coentro) é um forte agente antifúngico contra Candida e, portanto, investigações devem dar continuidade ao conhecimento gerado. Objetivo: Este estudo buscou avaliar a atividade antifúngica e modo de ação do OE de C. sativum sobre Candida spp., e determinar os alvos moleculares afetados na expressão global do genoma humano. Material e Métodos: C. sativum foi obtido a partir do Banco de Germoplasmas do Centro Pluridisciplinar de Pesquisas Químicas, Biológicas e Agrícolas (Universidade Estadual de Campinas, SP, Brasil) cujo OE e fração ativa tiveram o perfil fitoquímico determinado por cromatografia gasosa acoplada a espectrometria de massa. Posteriormente, foram realizados testes com cinco cepas de referência de Candida: Determinação da Concentração Inibitória e Fungicida Mínima (CIM/CFM); modo de ação antifúngica (ensaio do sorbitol e ergosterol); Microscopia Eletrônica de Varredura (MEV) de biofilmes de Candida e testes de inibição de aderência em biofilme. Utilizou-se nistatina, anfotericina B ou caspofungina como controles positivos, além de controles negativos. Também, foi testado o efeito do OE sobre a atividade proteolítica de C. albicans. Por fim, um ensaio de farmacogenômica identificou quais alvos moleculares no genoma humano foram afetados pelo OE e fração ativa de C. sativum. Os testes foram realizados em triplicata de experimentos independentes e os dados foram tratados estatisticamente (ANOVA, pós-teste de Tukey, ?=0,05). Os dados da análise farmacogenômica foram processados nas plataformas GeneGo MetaCore® e David Bioinformatics Resources. Resultados: O perfil fitoquímico EO indicou monoterpenos (37,9%) e sesquiterpenos (62,1%) como compostos principais. Os valores de CIM/CFM para o OE variaram de 15,6 a 62,5 µg/mL. Quanto ao modo de ação, o OE de C. sativum parece se ligar ao ergosterol da membrana celular fúngica, aumentando a permeabilidade iônica e causando morte celular; entretanto, o OE não atua sobre vias de biossíntese da parede celular. Estes achados confirmam as alterações na integridade da morfologia do biofilme verificadas nas análises por MEV. Além disso, o OE apresentou atividade antiaderente em biofilme em baixas concentrações (15,6-62,5 µg/mL) contra as cepas testadas, bem como atividade contra proteases produzidas por C. albicans, sendo estatisticamente significante na CIM (p<0,05). Finalmente, o OE e sua fração ativa apresentaram baixa citotoxicidade em células humanas com CI30 de 359,8 e 366,7 µg/mL, respectivamente. As principais vias afetadas estão relacionadas com quimiocinas e MAP-quinase (apoptose, proliferação) bem como proteínas de adesão. Conclusões: O OE das folhas de C. sativum tem forte atividade antifúngica e antiaderente sobre Candida spp. e atividade anti-proteolítica sobre C. albicans, e atua aumentando a permeabilidade iônica da membrana celular, provavelmente devido ao efeito sinérgico de mono e sesquiterpenos. Análise farmacogenômica indicou baixa citotoxicidade do OE e sua fração ativa com alvos moleculares específicos afetados no genoma humano, o que incentiva o desenvolvimento de novas pesquisas pré-clínicas toxicológicas e clínicas nesta área / Abstract: Introduction: Oral candidiasis is a common opportunistic fungal infection of the oral cavity with increasingly significant worldwide prevalence and incidence rates. Accordingly, novel specific-targeted strategies to manage this ailment have been proposed, among which is the use of essential oils (EO) with antifungal properties. Coriandrum sativum L. (coriander) EO has proven antifungal activity against Candida species and thus deserves further investigation. Objective: This study aimed to evaluate the antifungal activity and mode of action of the EO from Coriandrum sativum L. leaves on Candida spp., and to determine the molecular targets affected in the whole-genome expression in human cells. Material and Methods: C. sativum was obtained from the Germoplasm Bank of the Research Center for Chemistry, Biology and Agriculture (University of Campinas, SP, Brazil) whose EO and active fraction had their phytochemical profile determined by Gas Chromatography coupled to Mass Spectrometry. Then, we carried out the following tests with five reference strains of Candida spp. (CBS): Minimum Inhibitory and Fungicidal Concentration (MIC/MFC); antifungal mode of action (sorbitol and ergosterol assays); Scanning Electron Microscopy (SEM) analysis of Candida biofilm and tests of inhibition of biofilm adherence. We used nystatin, amphotericin B or caspofungin as positive controls, in addition to negative controls. Also, we tested the effect of C. sativum EO on the proteolytic activity of C. albicans. Next, a pharmacogenomic assay identified which molecular targets in the human genome were affected by C. sativum EO and its active fraction. Tests were performed in triplicate of independent experiments and data were statistically treated (ANOVA, Tukey¿s post-test, ?=0.05). Pharmacogenomic data were processed on GeneGo MetaCore® and DAVID Bioinformatics Resources. Results: The EO phytochemical profile indicated monoterpenes (37.9 %) and sesquiterpenes (62.1 %) as major compounds. The MIC/MFC values for the EO ranged from 15.6 to 62.5 µg/ml. With regard to the mode of action, C. sativum EO may bind to membrane ergosterol, increasing ionic permeability and causing membrane damage to cell death, but it does not act on cell wall biosynthesis-related pathways. This mode of action confirms the changes in the integrity of the biofilm morphology as verified in the analyses by SEM. The EO showed anti-adherent activity at low concentrations (31.2 ¿ 62.5 µg/ml) against all strains tested, as well as activity against proteases produced by C. albicans, with statistical significance at MIC (P < 0.05). Finally, the EO and its active fraction had low cytotoxicity on human cells with IC30 of 359.8 and 366.7 µg/ml, respectively, affecting the pathways of chemokines and MAP-kinase (apoptosis, proliferation), as well as adhesion proteins. Conclusions: The EO from C. sativum leaves has strong antifungal and anti-adherent activity against Candida spp., as well as anti-proteolytic activity against C. albicans, and acts by increasing cell membrane ionic permeability, probably due to the synergistic effect of mono- and sesquiterpenes. Pharmacogenomic analyses revealed low cytotoxicity with specific targets affected in the human genome, which encourages further pre-clinical toxicological screening and clinical research in this field / Mestrado / Farmacologia, Anestesiologia e Terapeutica / Mestre em Odontologia

Orientador: Marta Cristina Teixeira Duarte / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-26T00:33:23Z (GMT). No. of bitstreams: 1 Gasperini_AlessandraMarcon_M.pdf: 1292764 bytes, checksum: fcbdfc6ab370ac6744e8002a57b4a69e (MD5) Previous issue date: 2014 / Resumo: O fungo Aspergillus flavus é responsável pela degradação de alimentos, além de produzir aflatoxinas que são reconhecidas como potentes carcinógenos. Atualmente, frente ao aumento acelerado de cepas resistentes aos antifúngicos sintéticos e à pressão dos consumidores pela substituição destes por produtos naturais, a busca de novos antimicrobianos a partir de óleos essenciais (OE) tem se tornado uma opção promissora. Neste contexto, o objetivo deste trabalho foi avaliar a capacidade de 13 OE, provenientes de plantas medicinais e aromáticas no controle do crescimento de A. flavus CCT 0614 e produção das aflatoxinas AFB1 e AFB2. Ensaios in vitro realizados pelo teste da microdiluição mostraram que os OE de Cinamommum burmanii (MIC 0,2 mg.mL-1) e Cymbopogon citratus, C. martinii, C. winterianus, Eugenia caryophyllata e T. vulgaris (MIC 0,5 mg.mL-1) apresentaram elevada ação anti ¿ A. flavus. Em relação à produção de biomassa seca pelo fungo na presença dos OE, nas concentrações correspondentes a de mínima concentração inibitória (MIC) e 2xMIC, houve diminuição significativa em relação ao controle (p > 0,05). A avaliação do crescimento radial de A. flavus quando submetido à ação dos compostos voláteis (AV) do OE de C. burmanii a 0,2 mg.mL-1 mostrou inibição total até 336 h. Por outro lado, os OE de E. caryophyllata, T. vulgaris e C. citratus reduziram o crescimento radial do fungo quando incorporados ao meio de cultura e quando submetidos a ação dos voláteis. Os OE mais ativos foram analisados através de CG-EM, que permitiu identificar nos óleos compostos conhecidos pela ação antimicrobiana. Em seguida, o OE de C. citratus foi selecionado para estudo dos efeitos sobre a produção de aflatoxinas por A. flavus, quantificada por CLAE-FL. Os resultados mostraram que o OE de C. citratus foi capaz de reduzir a produção de AFB1 em 95% quando incorporado ao meio a 0,5 mg.mL-1, enquanto houve um aumento de cerca de 53% na produção da micotoxina a 0,25 mg.mL-1, após o mesmo período. Não foi detectada a presença da AFB2. Os resultados deste trabalho indicam que os OE estudados representam uma alternativa promissora no controle do crescimento A. flavus e o OE de C. citratus à produção de AFB1. A possibilidade de aplicação dos OE por AV é uma alternativa atrativa, pelo fato de não afetar as propriedades físicas e sensoriais do alimento / Abstract: The fungus Aspergillus flavus is responsible for damage in foods, and by aflatoxins production that are recognized as potent carcinogens. Currently, due to rapid increase of resistant strains to synthetics antifungal, and consumer pressure to replacing these synthetics compounds by natural products, the search for new antimicrobial from essential oils (EO) has become a promising option. This study aimed evaluate the capacity of 13 EO from medicinal and aromatic plants in the control of growth and aflatoxins B1 and B2 production by A. flavus CCT 0614. In vitro assays performed by microdilution method demonstrated that EO of Cinnamomum burmanii (MIC 0.2 mg.mL-1) and Cymbopogon citratus, C. martinii, C. winterianus, Eugenia caryophyllata and T. vulgaris (MIC 0.5 mg.mL-1) showed strong action anti - A. flavus. In relation to fungal biomass dry weight in the presence of OE in the concentrations corresponding to minimum inhibitory concentration (MIC) and 2xMIC, occurred a significant decrease compared to control (p< 0.05). The radial growth of A. flavus was inhibited totally until 336 h when submitted to the action of the volatile compounds (AV) of C. burmanii at 0.2 mg.mL-1. In contrast, the OE of E. caryophyllata, T. vulgaris and C. citratus reduced the radial growth of the mould when were incorporated to medium (IP) and when used in AV. EO that showed higher action were analysed by CG-MS this allowed identify the compounds known by antimicrobial action. The EO of C. citratus was selected to evaluation of the effects on the aflatoxins production by A. flavus, which was quantified by HPLC-FL. The results showed that C. citratus EO (0.5 mg.mL-1) was able to reduce 95% of AFB1 production, while the production increase about 53% with 0.25 mg.mL-1 after the same period. The presence of AFB2 was not detected. The most active EO were identified by GC-MS, which show the presence of compounds with known antimicrobial activity. The results of this study indicate that the EO studied represent a promising alternative to control the growth and AFB1 production by A. flavus. The possibility of application of EO by AV is an attractive alternative, in view of the fact that it does not affect the physical and sensory properties of food / Mestrado / Ciência de Alimentos / Mestra em Ciência de Alimentos

Orientadores: Glaucia Maria Pastore, Marta Cristina Teixeira Duarte / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-26T06:56:55Z (GMT). No. of bitstreams: 1 Simiqueli_AnaPaulaResende_D.pdf: 14118088 bytes, checksum: fde7134a6bda63a14dbd09e77af4109b (MD5) Previous issue date: 2014 / Resumo: Nos últimos anos, produtos naturais como os biossurfactantes e óleos essenciais têm sido objeto de intensa investigação científica. As suas atividades antimicrobianas e antiadesivas fazem com que eles apresentem potencial para serem utilizados para o controle de fungos fitopatógenos na agricultura e combate à micro-organismos multirresistentes e biofilmes na área médica e em indústrias de alimentos. Apesar da surfactina produzida por Bacillus subtilis possuir elevadas atividades superficiais e biológicas e ser um dos biossurfactantes mais estudados atualmente, os seus altos custos de produção e baixo rendimento tornam impraticável a sua produção em escala industrial. Deste modo, este trabalho estudou a produção contínua de surfactina e coprodução de amilases e proteases por B. subtilis utilizando manipueira como substrato em diferentes taxas de alimentação, visando à redução dos custos de processo e ao melhor aproveitamento do arsenal metabólico do micro-organismo. As suas atividades antimicrobiana e antibiofilme do biossurfactante produzido foram estudadas em conjunto com dez óleos essenciais extraídos plantas medicinais e aromáticas. Os valores de produção de biossurfactantes e atividades enzimáticas foram crescentes com o aumento da taxa de diluição empregada, encontrando-se os valores máximos de 2,9 mg.h-1, 938,3 U.mL-1 e 3,6 U.mL-1 para a produção de biossurfactantes e atividades amilolítica e proteolítica, respectivamente. O biossurfactante produzido demonstrou atividade antibacteriana contra Bacillus cereus, Rhodococcus equi e Listeria innocua e inibiu a formação de biofilmes de Staphylococcus aureus e Escherichia coli em até 92 e 78 %, respectivamente. Além disso, ele apresentou efeito sinérgico com os óleos essenciais de Aloysia gratissima e Cymbopogon winterianus para a inibição de R. equi, L. innocua e B. subtilis, com os óleos essenciais de Cinnamomum burmannii, Cymbopogon citratus e Lippia sidoides para a inibição dos fungos fitopatógenos Alternaria solani, Alternaria alternata e Mucor hiemalis e com o óleo essencial de C. winterianus para a inibição da formação de biofilme por E. coli / Abstract: In recent years, natural products such as biosurfactants and essential oils have been the subject of intense scientific research. Due to their antimicrobial and anti-adhesive activities, they show potential for controlling the growth of phytopathogenic fungi in agriculture and combating multi-drug resistant microorganisms and biofilms in medicine and food industries. Despite the high superficial and biological activities possessed by surfactin produced by Bacillus subtilis, their low yields and high production costs hamper their production on a commercial scale. Thus, this work studied the continuous production of surfactin and co-production of amylases and proteases by B. subtilis LB5a using cassava wastewater as substrate varying the dilution rate, in order to reduce production costs and improve the utilization of metabolic arsenal from the microorganism. The antimicrobial and anti-biofilm activities of the biossurfactant produced were studied in blends with ten essential oils extracted from medicinal and aromatic plants. Biossurfactant production values and enzymatic activities were higher with the increase of dilution rate, reaching the maximum values of 2,9 mg.h-1, 938,3 U.mL-1 and 3,6 U.mL-1 for the production of biossurfactants and amylolitic and proteolitic activities, respectively. The biossurfactant showed antibacterial activity against Bacillus cereus, Rhodococcus equi and Listeria innocua and inhibited up to 92 e 78 % of biofilm formation by Staphylococcus aureus and Escherichia, respectively. Moreover, it presented synergistic with the essential oils of Aloysia gratissima and Cymbopogon winterianus for the inhibition of R. equi, L. innocua and B. subtilis, with the essential oils of Cinnamomum burmannii, Cymbopogon citratus and Lippia sidoides for the inhibition of the phytopathogenic fungi Alternaria solani, Alternaria alternata and Mucor hiemalis and with the essential oil of C. winterianus for the prevention of biofilm formation by E. coli / Doutorado / Ciência de Alimentos / Doutora em Ciência de Alimentos

Submitted by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-18T22:31:24Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Carla_Daiane_Lubke_Ucker.pdf: 1869629 bytes, checksum: faefabb4e99813c029587cc3088d97be (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-18T22:44:22Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Carla_Daiane_Lubke_Ucker.pdf: 1869629 bytes, checksum: faefabb4e99813c029587cc3088d97be (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-18T22:44:29Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Carla_Daiane_Lubke_Ucker.pdf: 1869629 bytes, checksum: faefabb4e99813c029587cc3088d97be (MD5) / Made available in DSpace on 2018-05-18T22:44:29Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Carla_Daiane_Lubke_Ucker.pdf: 1869629 bytes, checksum: faefabb4e99813c029587cc3088d97be (MD5) Previous issue date: 2016-10-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Óleos essenciais obtidos de plantas podem apresentar propriedades antimicrobianas e antioxidantes, possibilitando sua utilização em alimentos e medicamentos. O jambolão (Syzygium cumini) e a Melaleuca alternifolia, ambos pertencentes à família Myrtaceae, podem ser utilizados na obtenção de óleo essencial. O forte odor característico e a insolubilidade dos óleos essenciais são fatores limitantes para sua aplicação, devido a isso uma alternativa para ampliar o uso é o processo de desodorização. Dessa forma o objetivo do presente trabalho foi avaliar as características do óleo essencial de jambolão e óleo desodorizado de Melaleuca alternifolia, microencapsular o óleo desodorizado de Melaleuca alternifolia e avaliar suas propriedades. O óleo essencial de jambolão, extraído das sementes, apresentou como principal constituinte o β-cariofileno e não apresentou atividade antioxidante no método DPPH. Os óleos essenciais das folhas e sementes de jambolão apresentaram atividade antimicrobiana frente a Escherichia coli, Staphylococcus aureus, Salmonella Typhimurium, Listeria monocytogenes, Trichoderma spp. e Rizhopus spp. O óleo desodorizado de Melaleuca alternifolia apresentou baixo teor de compostos fenólicos e ausência de terpenos. Entretanto, apresentou atividade antibacteriana frente a Escherichia coli, Staphylococcus aureus, Salmonella Typhimurium, Listeria monocytogenes e ausência de atividade antifúngica. Obteve-se alta eficiência de encapsulação do óleo desodorizado de Melaleuca alternifolia pela técnica de coacervação simples utilizando quitosana e ácido tânico, os resultados foram confirmados pela técnica de Calorimetria Diferencial de Varredura. As micropartículas apresentaram formato irregular, tamanho distinto e inibição frente aos microrganismos avaliados, devido à propriedade antimicrobiana da quitosana, porém sem atividade antioxidante. Não foi observada influência quanto a presença do ácido tânico nas micropartículas. O óleo desodorizado de Melaleuca alternifolia apresenta potencial de aplicação em hambúrguer em função de ter apresentado atividade antimicrobiana in situ frente à Escherichia coli, já os óleos essenciais de jambolão não puderam ser analisados in situ devido ao baixo rendimento. / Essential oils obtained from plants may have antimicrobial and antioxidant properties, allowing its use in food and medicine. The jambolan (Syzygium cumini) and Melaleuca alternifolia, both belonging to the Myrtaceae family can be used to obtain an essential oil. The strong characteristic odor and the insolubility of the essential oils are limiting factors for its use due to it an alternative to extend the use is the deodorization process. Thus the objective of this study was to evaluate the essential oil characteristics jambolan and deodorized oil Melaleuca alternifolia, microencapsulating the deodorized oil Melaleuca alternifolia and evaluate their properties. The essential oil jambolan, extracted from the seeds, presented as the main constituent the β-caryophyllene and did not show antioxidant activity in the DPPH method. Essential oils from the leaves and seeds jambolan showed antimicrobial activity against Escherichia coli, Staphylococcus aureus, Salmonella Typhimurium, Listeria monocytogenes, Trichoderma spp. and Rizhopus spp. The deodorized oil Melaleuca alternifolia showed low content of phenolic compounds and absence of terpenes. However, the oil showed antibacterial activity and no antifungal activity. It was obtained high encapsulation efficiency of deodorized oil Melaleuca alternifolia by simple coacervation technique using chitosan and tannic acid, the results were confirmed by Calorimetry Differential Scanning technique. The microparticles presented irregular size and distinctive shape. The microparticles showed potent inhibition compared to the microorganisms evaluated due to the antimicrobial property of chitosan, but no antioxidant activity. No significant differences were observed for the presence of tannic acid in the microparticles. The deodorized oil Melaleuca alternifolia has potential application in burger due to present antimicrobial activity against the Escherichia coli.

Syzygium cordatum Hoscht ex. C Krauss, also known as water berry, is normally used by the people of South Africa for respiratory ailments including tuberculosis, stomach complaints, treatment of wounds and as emetics. An extract of the leaves can be used as a purgative for diarrhoea treatment. The leaves of Syzygium cordatum Myrtaceae were obtained from the Eastern Cape Province of South Africa, air dried and sequential solvent extraction was done to obtain various non volatile crude extracts. The volatile extract, that is the essential oil was extracted from the leaves using hydrodistillation and analysis of compounds was done by GC/MS for composition. 32 compounds were obtained from the fresh leaves and 18 compounds were obtained from the dry leaves. The fresh oil contains caryophyllene (11.8 percent) and caryophyllene oxide (11.1 percent) as the main sesquiterpene component. α-Pinene(5.0 percent) was the only monoterpene compound identified in the fresh oil in substantial amount. The dry leaves oil had copanene (17.0 percent), β-Caryophellene (26.0 percent), cubenol (6.5 percent) and caryophellene oxide (14.2 percent) as the dominant constituent of the oil. Summary of the classes of compounds in the oil revealed that the chemical profile of both oils were dominated by sesquiterpenoid compounds. This is the first time that terpenoids compounds are being identified in both the fresh and dry leaf oil of S. cordatum. Hexane leaf extract was selected due to the interest in the terpenoid compounds. Column chromatography of the hexane crude gave five (5) of which two are fully reported. The isolates were fully elucidated using spectroscopic methods to be β-Sitosterol (HC3) and Friedela-3-one (HC1A/HC1D). Cytotoxicity analysis was carried out on the crude using the Brine shrimps assay. Isolates 1C and1D showed significant lethality using the brine shrimps assay with lethality values (LC50) of 4.105mg/ml for HC1C and 4.11mg/ml for 1D/1A respectively.

SFME and HD for the extraction of essential oil in Lavandula officinalis in Alice have been reported. A total of 59 compounds were identified with the major compound being 1,8-cineole, an oxygenated monoterpene, with 46.89% and 44.84% yield obtained for HD and SFME respectively. Charge transfer (CT) complexes formed between α-pinene, 1,8-cineole and camphor as electron donors with iodine as the electron acceptor have been studied spectrophotometrically in methylene chloride solution. The Benesi- Hildebrand equation has been applied to estimate the formation constant (Kf) and molecular extinction coefficient (εCT). The value of Kf is the highest in camphor-I2 complex compared to the other two complexes. Antibacterial assessment was carried out on the various reagents, determining the MIC of individual reagents and in combination. The results show an improvement, on combination of the various reagents than when tested alone.

The common maize weevil, Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), a well known pest of stored-maize grain in most parts of the world, was identified as one of the major constraints of harvested maize grains in the Eastern Cape Province, South Africa. The use of plants or their products is one of the recent methods being investigated for insect pest control worldwide. Therefore, the main aim of the present study was to identify available plants in the Eastern Cape that could be used to combat the problem of Sitophilus zeamais in stored-maize grains. For the realization of the aims of this research, the following studies were carried out; a preliminary survey was conducted to obtain baseline information on the farmers’ knowledge and experience of indigenous insect pest control methods in the Eastern Cape. Also, studies on the insecticidal potential of the essential oils of some plants were investigated against the maize weevil. The quality parameters of maize grains treated with the essential oils was also studied and, using a rat model, the toxicity of the essential oils was investigated. The outcome from this study revealed that there is awareness amongst the farmers in the Eastern Cape on the use of plants or their products to control insect pests. Unfortunately, such methods are currently being neglected and the knowledge of their application was found to be eroding. Among the various essential oils screened were those from Mentha longifolia L. and Tagetes minuta L. which evoked an appreciable level of contact, fumigant and repellent toxicity on the maize weevil. Further work done to determine the effects of the oils on maize stored over a period of three months revealed that the two oils had no adverse effect on the proximate compositions and some quality parameters of the stored maize. However, the toxicological study conducted on rats showed that the oils at tested concentrations exhibited some level of toxicity. It is, therefore, suggested that the essential oils of M. longifolia and T. minuta should not be used to treat maize grains intended for human consumption.

Variations in the yield, chemical composition, antibacterial, and antioxidant properties of the essential oils of Rosmarinus officinalis L. cultivated in Alice, Eastern Cape of South Africa over a period of 12 months using the solvent-free microwave extraction and traditional hydrodistillation methods were evaluated. The GC-MS analyses of the essential oils revealed the presence of 33 compounds with 1,8-cineole, a-pinene, camphor, verbenone, bornyl acetate and camphene constituting about 80 percent of the oils throughout the period of investigation, with the solvent-free microwave extraction method generally yielding more of the major components than the hydrodistillation method. Each of the major components of the oils varied in quantity and quality of yield at different periods of the year. The method of extraction and time of harvest are of importance to the quantity and quality of essential oil of Rosmarinus officinalis. Higher amounts of oxygenated monoterpenes such as borneol, camphor, terpene- 4-ol, linalool, a-terpeneol were present in the oil of SFME in comparison with HD. However, HD oil contained more monoterpene hydrocarbons such as a-pinene, camphene, β-pinene, myrcene, a-phellanderene, 1,8-cineole, trans- β-ocimene, γ-teprinene, and cis-sabinene hydrate than SFME extracted oil. Accumulation of monoterpene alcohols and ketones was observed during maturation process of Rosmarinus leaves. Quantitative evaluation of antibacterial activity, minimum inhibitory concentration values were determined using a serial microplate dilution method. The essential oils obtained using both methods of extraction were active against all the bacteria tested at a concentration of 10 mg mL-1. The minimum inhibitory concentrations for the SFME extracted oils ranged between 0.23 and 1.88 mg mL-1, while those of the HD extracted oils varied between 0.94 and 7.5 mg mL-1, thus suggesting that the oil obtained by solvent free microwave extraction was more active against bacteria than the oil obtained through hydrodistillation. The antioxidant and free radical scavenging activity of the obtained oils were tested by means of 1,1-diphenyl-2-picrylhydrazyl radical (DPPH+) assay and β- carotene bleaching test. In the DPPH+ assay, while the free radical scavenging activity of the oil obtained by SFME method showed percentage inhibitions of between 48.8 percent and 67 percent, the HD derived oil showed inhibitions of between 52.2 percent and 65.30 percent at concentrations of 0.33, 0.50 and 1.0 mg mL-1, respectively. In the β-carotene bleaching assay, the percentage inhibition increased with increasing concentration of both oils with a higher antioxidant activity of the oil obtained through the SFME than the HD method. Thin layer chromatography (TLC) was used to analyze the chemical composition of the extracts using three eluent solvent systems of varying polarities i. e. CEF, BEA and EMW and sprayed with vanillin-sulfuric acid. The chemical composition of the different extracts was similar with the exception of methanol and water extracts which had only one or two visible compounds after treating with vanillin-spray reagent. To evaluate the number of antibacterial compounds present in the fractions, bioautography was used against two most important nosocomial microorganisms. S. aureus (Gram positive) and E. coli (Gram negative). Nearly all the crude serial extraction fractions contained compounds that inhibited the growth of E. coli. The hexane extract had the most lines of inhibition followed by ethyl acetate. Bioassay-guided fractionation against E. coli was used to isolate antibacterial compounds. The largest number of antibacterial compounds occurred in the hexane fraction. Furthermore we tried to complete the characterization by extracting and studying other biologically important plant metabolites such as phenolic compounds to evaluate the antioxidant capacity of Rosmarinus extracts.

Orientador: Miriam Dupas Hubinger / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-22T05:48:31Z (GMT). No. of bitstreams: 1 Oriani_VivianBoesso_M.pdf: 12741078 bytes, checksum: 5f41652e5d2d38885c526edf6c2e0cf7 (MD5) Previous issue date: 2013 / Resumo: Coberturas comestíveis foram desenvolvidas e caracterizadas pela combinação de fécula de mandioca (2 e 3% p/v) com óleos essenciais de casca de canela (Cinnamomum zeylanicum) (0,05 a 0,30% v / v) ou de erva doce (Foeniculum vulgare L.) (0,05-0,30 % v / v). A adição do óleo teve como propósito desenvolver uma cobertura com propriedades antimicrobianas, antioxidantes e com capacidade de barreira ao vapor de água, já que os óleos essenciais têm característica hidrofóbica. Foram avaliados estabilidade física, tamanho de gota e viscosidade das coberturas comestíveis emulsionadas e os filmes foram caracterizados através das propriedades mecânicas, solubilidade e microscopia de força atômica. As coberturas emulsionadas com 2% de fécula de mandioca mostraram-se estáveis e as de 3% de fécula apresentaram separação da água do sistema emulsionado. A cobertura foi feitas em fatias de maçãs para o teste de resistência ao vapor de água e medida da taxa de respiração. A formulação com a concentração máxima dos dois óleos essenciais, em ambas as concentrações de fécula de mandioca, resultou em aumento da resistência ao vapor de água. A taxa respiratória das fatias de maçãs com cobertura mostrou efeito de barreira ao CO2 em relação aos frutos sem cobertura. As análises de propriedades mecânicas e microscopia de força atômica dos filmes mostraram que as películas com óleo essencial de casca de canela podem ter apresentado descontinuidade da matriz formadora de filmes. As coberturas com óleo essencial de casca de canela apresentaram efeito significativo para a atividade antioxidante. O teste de halo de inibição dos microrganismos mostrou que a formulação com 0,3% de óleo essencial de casca de canela apresenta uma pequena inibição para Staphylococcus aureus e Salmonella choleraesius / Abstract: Edible coatings formed by a combination of cassava starch (2 and 3% w / v) with essential oil of cinnamon bark (Cinnamomum zeylanicum) (0.05 to 0.30% v / v) or fennel (Foeniculum vulgare L.) (0.05 to 0.30% v / v) were developed and characterized. The addition of oil had the objective of developing a coating with antimicrobial and antioxidant properties and capacity to form a better water vapor barrier, due to the aggregation of the hydrophobic characteristics of essential oil. Physical stability, particle size and viscosity of the edible coatings were analyzed and films were characterized by mechanical properties, solubility and atomic force microscopy measures. Coatings emulsified with 2% cassava starch were stable and the 3% starch concentration with added essential oil showed water separation from the emulsified system. The coatings were applied to apple slices for water vapor resistance and respiration rate tests. The formulation with maximum concentration of both essential oils, at both concentrations of cassava starch, resulted in increased water vapor resistance, and respiration rates of coated apple slices showed a CO2 barrier effect in relation to fruit without coating. Mechanical properties analysis and atomic force microscopy showed that films added with cinnamon bark essential oil presented discontinuity on its matrix-forming capacity. Coating with cinnamon bark essential oil had significant antioxidant capacity. Inhibition of microorganisms tests proved that the formulation with 0.3% essential oil of cinnamon bark had a small inhibiting activity against Staphylococcus aureus and Salmonella choleraesius / Mestrado / Engenharia de Alimentos / Mestra em Engenharia de Alimentos

Orientadores: Lauro Euclides Soares Barata, Fabio Augusto / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-18T13:25:10Z (GMT). No. of bitstreams: 1 Souza_RitadeCassiaZacardide_M.pdf: 1828503 bytes, checksum: c564b2bcd8fc2c55c0690d12a6c9dc53 (MD5) Previous issue date: 2011 / Resumo: O presente trabalho é apresentado em dois capítulos: o primeiro compreende o desenvolvimento de uma metodologia de diferenciação entre duas espécies de Aniba frequentemente confundidas, Aniba parviflora e Aniba rosaeodora e o segundo compreende o estudo da composição química dos óleos essenciais de Aniba parviflora através de cromatografia gasosa acoplada a espectrometria de massas (CG-EM), cromatografia gasosa com detector de ionização de chama (CG-DIC) e cromatografia gasosa bidimensional abrangente acoplada à espectrometria de massas (CG × CGEM). Para a diferenciação das espécies foi utilizada a avaliação das suas frações voláteis (headspace) através da SPME-CG (Microextração em Fase Sólida acoplada a Cromatografia Gasosa). Foram analisadas um total de 100 amostras de folhas das duas espécies e a diferenciação foi feita através do tratamento quimiométrico dos dados obtidos. Foram analisados ainda óleos essenciais das folhas e da madeira de A. parviflora obtidos por arraste a vapor. O estudo destes forneceu além da composição química, a análise quantitativa dos seus componentes. Por fim foi realizada a comparação da técnica de CG-EM com a inovadora técnica de CGxCG-EM que possibilitou um aumento de 50% na identificação dos componentes e aproximadamente de 80% na separação e detecção. Os resultados obtidos permitem dizer que e possível diferenciar duas espécies botanicamente vizinhas através de técnicas analíticas de maneira objetiva com respaldo estatístico / Abstract: This work consists of two main parts: The first part involves the development of a differentiation methodology between two Aniba species that are often confused with one another, they are Aniba parviflora and Aniba rosaeodora. The second part involves the chemical composition study of Aniba parviflora essential oils through gas chromatography coupled to mass spectrometry (GC-MS), gas chromatography with flame ionization detection (GC-FID) and comprehensive two-dimensional gas chromatography (GC × GC- MS). In the first part, the species differentiation was performed using SPME-GC and chemometric tools for the assessment of their volatile fraction (headspace). A total of 100 different leaf samples coming from distinct individuals of these two species were examined and the differentiation was made based on the SPME-GC chromatograms data that were treated using chemometric tools. In the second part the essential oils of A. parviflora coming both from leaves and wood were also analyzed and its study resulted in the oils chemical composition and in the quantitative analyses of its components. This work provided also a comparison between two techniques, GC-MS and the innovative GCxGC-MS. The later technique allowed an increase of at least 80 % of total components separation and detection and an increase of more than 50 % of the total identified components / Mestrado / Quimica Organica / Mestre em Química