Submitted by Renorbio (renorbioba@ufba.br) on 2017-08-15T14:32:14Z No. of bitstreams: 1 Tese_ Zulane Lima Sousa_ Atividades biológicas de extratos e frações das folhas de Persea americana e Syzygium malaccense_ 2016.pdf: 3239265 bytes, checksum: 5c62167b6ccb9fb8640e54efc7cd1af4 (MD5) / Approved for entry into archive by Delba Rosa (delba@ufba.br) on 2017-08-28T13:56:26Z (GMT) No. of bitstreams: 1 Tese_ Zulane Lima Sousa_ Atividades biológicas de extratos e frações das folhas de Persea americana e Syzygium malaccense_ 2016.pdf: 3239265 bytes, checksum: 5c62167b6ccb9fb8640e54efc7cd1af4 (MD5) / Made available in DSpace on 2017-08-28T13:56:26Z (GMT). No. of bitstreams: 1 Tese_ Zulane Lima Sousa_ Atividades biológicas de extratos e frações das folhas de Persea americana e Syzygium malaccense_ 2016.pdf: 3239265 bytes, checksum: 5c62167b6ccb9fb8640e54efc7cd1af4 (MD5) / Fundação de Amparo à Pesquisa do Estado da Bahia – FAPESB / Os produtos naturais são fonte de substâncias biologicamente ativas, especialmente as plantas, as quais passaram por anos de processos adaptativos contra fatores abióticos, como temperaturas extremas, falta de água, luz e nutrientes, assim como fatores bióticos, como insetos e micro-organismos. Esse processo adaptativo permitiu o desenvolvimento de metabólitos secundários que hoje são alvo para o estudo de compostos bioativos. Na área médica, a necessidade de novas drogas antimicrobianas ocorre devido ao crescente número de casos de micro-organismos resistentes às drogas de uso atual, além do que no caso dos antifúngicos, muitos são tóxicos, devido à similaridade de células eucariotas humanas e fúngicas. Na área agronômica, o problema de perda de produção pela ação de fitopatógenos é preocupante, principalmente para aquelas culturas que ainda não têm um método de controle eficaz e humanamente e ambientalmente seguros, como é o caso do cacaueiro (Theobroma cacao Linn.) infectado por Moniliophthora perniciosa (Stahel) Aime and Phillips-Mora, que causa a vassoura de bruxa. Com base nisto, este trabalho foi realizado com o objetivo de buscar substâncias bioativas em extratos das folhas de Persea americana Miller e Syzygium malaccense (L.) Merr. & Perry, plantas já utilizadas na medicina tradicional contra doenças infecciosas. A fração diclorometanólica de P. americana (PAD) apresentou atividade contra M. perniciosa com concentração de 500 µg/mL, apresentando 0,3% de sobrevivência quando comparado ao controle sem tratamento (p<0,05). Através dos estudos realizados, essa ação provavelmente é devida à presença de flavonoides nessa fração, os quais possivelmente agem sobre a membrana plasmática do fungo, levando-o à morte. Já o extrato bruto metanólico de S. malaccense (SMM) apresentou atividade contra Enterococcus faecalis resistente à vancomicina (ATCC 51299) e Staphylococcus aureus multirresistente (ATCC 43300), com concentração inibitória mínima (CIM) de 100 μg/mL. A fração aquosa de S. malaccense (SMA) mostrou atividade contra Candida albicans (CIM = 50 μg/mL) e Candida krusei (CIM = 6,25 μg/mL). A fração etanólica (SME) apresentou atividade contra Candida parapsilosis e C. albicans (CIMs = 100 μg/mL). Dessa maneira, tanto a fração PAD pode ser utilizada como uma alternativa para o controle da vassoura de bruxa, como as frações SMM, SMA e SME podem ser utilizadas para a elaboração de formulações farmacêuticas para o uso terapêutico. / Natural products are sources of biologically active substances, especially plants, which have gone through years of adaptive processes against abiotic factors such as extreme temperatures, and water, light and nutrients lack, as well as biotic factors such as insects and micro-organisms attacks. This adaptive process allowed the development of secondary metabolites that are now targeted for the study of bioactive compounds. In the medical field, the need for new antimicrobial drugs is due to the increasing number of cases of microorganisms resistant to currently useed drugs and in the case of antifungals many are toxic due to the similarity of human and fungal eukaryotic cells. In agronomy, the problem of lost production by plant pathogens action is worrying, especially for those crops that do not yet have an effective control method humanly and environmentally safe, as is the case of the cacao tree (Theobroma cacao Linn.) infected with Moniliophthora pernicious (Stahel) Aime and Phillips-Mora, which causes witches' broom. Based on this, this study was conducted with the objective of seeking bioactive compounds in extracts of Persea americana Miller and Syzygium malaccense (L.) Merr. & Perry leaves, plants already used in traditional medicine against infectious diseases. P. americana dichloromethane fraction (PAD) showed activity against M. perniciosa at concentration of 500 µg/mL, with 0.3% survival when compared to the untreated control (p <0.05). Through studies carried out, this action is probably due to the presence of flavonoids in this fraction, which possibly acts on the plasma membrane of the fungus, leading to death. S. malaccense crude methanol extract (SMM) showed activity against vancomycin-resistant Enterococcus faecalis (ATCC 51299) and multidrug-resistant Staphylococcus aureus (ATCC 43300) with minimum inhibitory concentration (MIC) of 100 µg/mL. The S. malaccense aqueous fraction (SMA) showed activity against Candida albicans (MIC = 50 µg/mL) and Candida krusei (MIC = 6.25 µg/mL). The S. malaccense ethanol fraction (SME) showed activity against Candida parapsilosis and C. albicans (MIC = 100 µg/mL). Thus, the PAD fraction may be used as an alternative for the control of witch's broom, as well as the SMM, SMA and SME fractions can be used for the preparation of pharmaceutical formulations for therapeutic use.

ROCHA, Rafael dos Santos. Caracterização polifásica da comunidade bacteriana heterotrófica de ambiente de carcinicultura frente às tetraciclinas. 2016. 105 f. Tese (Doutorado em Engenharia de Pesca)-Universidade Federal do Ceará, Fortaleza, 2016. / Submitted by Vitor Campos (vitband@gmail.com) on 2016-10-05T22:40:10Z No. of bitstreams: 1 2016_tese_rsrocha.pdf: 2184416 bytes, checksum: 17a4ecb8bd02fce65742927d2927e854 (MD5) / Approved for entry into archive by Jairo Viana (jairo@ufc.br) on 2016-10-05T23:19:54Z (GMT) No. of bitstreams: 1 2016_tese_rsrocha.pdf: 2184416 bytes, checksum: 17a4ecb8bd02fce65742927d2927e854 (MD5) / Made available in DSpace on 2016-10-05T23:19:54Z (GMT). No. of bitstreams: 1 2016_tese_rsrocha.pdf: 2184416 bytes, checksum: 17a4ecb8bd02fce65742927d2927e854 (MD5) Previous issue date: 2016 / Antimicrobial agents of tetracycline class has been extensively used in aquaculture for disease control and growth promotion. Thus, the aim of this study was to evaluate the polyphase profile of the bacterial community in shrimp farming environment front antimicrobial classes of tetracyclines, focusing on Vibrio genus. Water and sediment samples were collected four (4) shrimp farms: two (2) in the state of Ceará and two (2) in the state of Rio Grande do Norte, of points: estuary, supply channel, pond and drainage channel. Cultivable heterotrophic bacteria (CHB), with resistance to tetracycline (CHB+TET) and oxytetracycline (CHB+OTC), total Vibrio, positive sucrose Vibrio and sucrose negative Vibrio were surveyed by Standard Plate Count method (SPC). The strains isolated from CHB, CHB+TET, CHB+OTC and Vibrio underwent phenotypic identification. The phenotypic profile of antimicrobial susceptibility of strains of Vibrio and other BHC was compared between the disk diffusion techniques and macrodilution by determining the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) for tetracycline and oxytetracycline antimicrobials. Resistance genes tet(M), tet(S) otr(A) and otr(B) were investigated in strains characterized as phenotypically resistant. Diversity of heterotrophic bacteria is influenced by wet and rainy seasons and also by the different collection points within the shrimp farms. Overall, the CHB, CHB+TET and CHB+OTC and Vibrio counts differ between the collection seasons, with higher measurements during the rainy season to the points of pond and drainage channel. 146 strains of CHB, 57 of CHB+TET and 45 of CHB+OTC were isolated, of which the most representative genera were Vibrio and Enterobacter. A total of 100 strains of Vibrio were isolated, which V. coralliilyticus was the most representative species. Antimicrobial profile of Vibrio and other CHB differed between disk-diffusion and macrodilution techniques, being the macrodilution the most efficient in the characterization of resistance among isolates. All strains of Vibrio and other CHB, phenotypically resistant, presented the tet(M) gene as a determinant of resistance determinat, but tet(S) otr(A) and otr(B) genes were not observed. Thus, it proves that Vibrio genus is most abundant among herotrotrophic bacteria with resistance to tetracyclines, with the bacterial diversity strongly influenced by seasons. / Antimicrobianos da classe das tetraciclinas têm sido extensivamente usados em aquicultura para o controle de doenças e como promotores de crescimento. Dessa forma, foi traçado como objetivo geral avaliar o perfil polifásico da comunidade bacteriana de ambiente de criação de camarão frente aos antimicrobianos das classes das tetraciclinas, com enfoque no gênero Vibrio. Amostras de água e sedimento foram coletadas de quatro (4) fazendas de carcinicultura: duas (2) no estado do Ceará e duas (2) no estado do Rio Grande do Norte, dos pontos: estuário, canal de abastecimento, viveiro e canal de drenagem. Foram pesquisadas bactérias heterotróficas cultiváveis (BHC), com resistência à tetraciclina (BHC+TET) e oxitetraciclina (BHC+OTC), Vibrio total, sacarose positivos e negativos pelo método de Contagem Padrão em Placas (CPP). As cepas isoladas de BHC, BHC+TET, BHC+OTC e Vibrio foram submetidas à identificação fenotípica. O perfil fenotípico de susceptibilidade antimicrobiana das cepas de Vibrio e demais BHC foi comparado entre as técnicas de disco-difusão e macrodiluição, pela determinação da Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM) para os antimicrobianos tetraciclina e oxitetraciclina. Os genes de resistência tet(M), tet(S), otr(A) e otr(B) foram pesquisados nas cepas fenotipicamente caracterizadas como resistentes. A diversidade de bactérias heterotróficas foi influenciada pelos períodos de estio e chuvoso e também pelos diferentes pontos de coleta dentro das fazendas de carcinicultura. No geral, as contagens de BHC, BHC+TET e BHC+OTC e Vibrio e demais BHC diferiram entre os períodos de coleta, com quantificações superiores no período chuvoso para os pontos de coleta viveiro e canal de drenagem. Foram isoladas 146 cepas de BHC, 57 de BHC+TET, 25 de BHC+OTC, das quais os gêneros mais representativos foram Vibrio e Enterobacter. Foi possível o isolamento de 100 cepas de Vibrio, das quais V. coralliilyticus foi a espécie mais representativa. O perfil antimicrobiano diferiu entre as cepas de Vibrio e demais BHC entre as técnicas de disco-difusão e macrodiluição, sendo a de macrodiluição a mais eficiente na caracterização de resistência dentre os isolados. Todas as cepas de Vibrio e demais BHC, fenotipicamente resistentes, apresentaram o gene tet(M) como determinante de resistência, no entanto os genes tet(S), otr(A) e otr(B) não foram verificados. Com isso, comprovou-se que Vibrio é gênero mais abundante dentre as bactérias heterotróficas com resistência às tetraciclinas, estando a diversidade bacteriana fortemente influenciada pelos períodos do ano.

The aim of this study was to determine propagation methodology of L. sericea taking into consideration the effect of season and storage. The response of tip and basal cuttings of L. sericea to three hormonal treatments and three growth media was investigated. Seed germination response to five constant temperatures and four pre-sowing treatments was also investigated. Using NMR spectroscopy, the effects of season and storage on metabolites in leaves of L. sericea were investigated. All treatments did not initiate or improve rooting of L sericea cuttings. Seed germination percentage was significantly affected by temperature and pre-sowing treatment compared to control. The optimum seed germination percentage was obtained at 15°C (53%) when treated with smoke water. Seasonal variation affected leaf metabolite profiles. Storage did not affect the antibacterial activity of L. sericea leaves and those harvested in winter showed an MIC value of ≤3.90μg/mL when tested on Propionibacterium acnes / Agriculture, Animal Health and Human Ecology / M. Sc. (Agriculture)

Made available in DSpace on 2014-08-13T14:50:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-02-14Bitstream added on 2014-08-13T18:00:04Z : No. of bitstreams: 1 000767277.pdf: 1576777 bytes, checksum: 8c82bd3957cc7f25dbc9fb4b5bb1b689 (MD5) / Antes da emergência da resistência à vancomicina, Enterococcus spp não eram considerados patógenos de grande risco para pacientes submetidos a cuidados em saúde. Estas bactérias raramente causavam infecções sistêmicas e eram isoladas principalmente de infecções urinárias. Atualmente, são considerados uma grande ameaça no ambiente hospitalar, devido à emergência dos enterococos resistentes à vancomicina (Vancomycinresistant enterococci, VRE). Nosso estudo pretendeu abordar de forma ampla a epidemiologia, os fatores de risco, a clonalidade e o prognóstico da aquisição de VRE em dois hospitais consorciados: O Hospital Estadual Bauru (HEB) e o Hospital Estadual Manuel de Abreu (HEMA), ambos pertencentes à Secretaria de Estado da Saúde de São Paulo. Para tanto, combinamos metodologias de epidemiologia clássica e molecular. O período de coleta de isolados foi de janeiro de 2010 a junho de 2012, correspondendo à época de emergência e disseminação dos VRE nos hospitais do estudo. Foram incluídos 122 sujeitos que adquiriram VRE em um dos hospitais, e 8 casos classificados como importados (isto é, procedentes de outros serviços de saúde). Os isolados eram procedentes de culturas clínicas (solicitadas pelos médicos responsáveis) ou de vigilância (swabs retais coletados sob orientação da Comissão de Controle de Infecção Hospitalar). Foram caracterizados por métodos usuais de identificação de espécie e antibiograma. Todos foram submetidos a tipagem molecular por Eletroforese de Campo Pulsado (Pulsed-Field Gel Electrophoresis, PFGE), utilizando a enzima de restrição smaI. Foram realizados estudos caso-controle (com análise multivariada), excluindo os sujeitos que adquiriram VRE em outros serviços. Para os 122 casos remanescentes, foram selecionados 244 controles, nos quais o VRE não foi identificado. Dentre os casos identificados nos hospitais do estudo, 113 (86,9%) foram caracterizados ... / In the decades that preceded the emergence of vancomycin resistance, Enterococcus spp. were not considered pathogens of greatest risk for patients undergoing health care. These bacteria rarely caused systemic infections and were isolated mainly from urinary tracr infections. Presently, they are considered a major threat in hospitals due to the emergence of vancomycin-resistant Enterococci (VRE). Our study sought to provide a wide evaluation of the epidemiology, risk factors, clonality and outcome of VRE colonization/infection in two hospitals from a public health consortium: Hospital Estadual Bauru (HEB) and Hospital Estadual Manuel de Abreu (HEMA) Hospital , both belonging to São Paulo State Health Secretariat. We combined classic epidemiological methods and molecular epidemiology. The study period extended from January 2010 through June 2012 , corresponding to the time of emergence and spread of VRE in hospitals in the study. We included 122 subjects who acquired VRE in one of the study hospitals, and 8 cases classified as imported (i.e., who acquired VRE in other health services) . The isolates were obtained from clinical cultures (requested by attending physicians) or surveillance cultures (rectal swabs collected under the guidance of the Committee on Hospital Infection Control). VRE were characterized by the usual methods of species identification and antibiotic susceptibility. All isolates were subjected to molecular typing using Pulsed-Field Gel Electrophoresis (PFGE) using the restriction enzyme SmaI . Case-control studies (with multivariable analysis) were performed excluding subjects who acquired VRE in other services. For the remaining 122 cases , 244 controls were selected , in which VRE was not identified. Among the cases from the study hospitals, 113 (86.9%) were identified as Enterococcus faecium and 17 (13.1%) and Enterococcus faecalis. Overall, 88 (67.7%) were isolated in HEB and 42 (32.7%) in HEMA . The autochthony was ...

abstract: Amazonia, inhabited and investigated for millennia, continues to astonish scientists with its cultural and natural diversity. Although Amazonia is rapidly changing, its vast and varied landscape still contains a complex natural pharmacopeia. The Amazonian tribes have accrued valuable environmental and geological knowledge that can be studied. This dissertation demonstrates that Indigenous Knowledge considered alongside Western Science can enhance our understanding of the relationship of people to geological materials and hydrological resources, and reveal mineral medicines with practical applications. I used methods from anthropology and geology to explore the geological knowledge of the Uitoto, a tribe of the Colombian Amazon. The Uitoto use two metaphors to describe Earth systems: 1. the earth is a body, and 2. the Amazon is a tree. I found that they classify surface-water systems according to observable characteristics and use mineral clays to treat various maladies. I argue that Uitoto knowledge about Amazonian mineral resources and surface water is practical, empirically–based and, in many cases, more nuanced than mainstream scientific knowledge. I studied the mode of action of a natural antibacterial clay from the Colombian Amazon (AMZ) to discover whether the Uitoto’s claims about the clay’s medicinal values was verifiable using the methods of Western Science. Natural antibacterial clays can inhibit the growth of human pathogens. Methods from microbiology and geochemistry were combined to evaluate the mineral-microbe interactions that inhibit growth of model Gram-negative (Escherichia coli) and Gram-positive (Bacillus subtilis) bacteria. The AMZ antibacterial clay contains 45 % kaolinites and 30 % smectites. Its high surface area maintains an acidic environment (pH 4.5) and releases high concentrations of aluminum. Aluminum accumulates in the outer membrane of E. coli by binding to phospholipids. Furthermore, the membrane’s permeability increases due to synergistic effects between aluminum and transition metals released from the AMZ (i.e. Fe, Cu). The changes in the membrane may compromise its function as a barrier. Understanding the antibacterial mechanism of AMZ is key for its safe use as a natural product. These findings can help us harness the capabilities of antibacterial clays more efficiently. Lastly, I integrated the results of this work in place-based, cross-cultural educational materials tailored for the tribal schools in the Colombian Amazon. The design of the units was informed by principles of curriculum design and successful pedagogic approaches for Native American students. The purpose of these educational materials is to return the results of research, enhance learning and participation of indigenous peoples in geosciences, and respond to the multicultural and plurilingual educational needs in countries such as Colombia. / Dissertation/Thesis / Doctoral Dissertation Geological Sciences 2016

Made available in DSpace on 2014-06-11T19:28:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-12-09Bitstream added on 2014-06-13T19:36:32Z : No. of bitstreams: 1 nogueira_lg_me_arafcf.pdf: 1061464 bytes, checksum: 0b8ec3ba710c3ab15f6b3d51c11fbadd (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O gênero Maytenus (Celastraceae), atualmente compreende cerca de 80 espécies distribuídas por todo território brasileiro. A espécie Maytenus ilicifolia é conhecida popularmente pelos nomes de espinheira-santa, espinheira-diva, salva-vidas, sombra de touro, cancerosa e coromilho do campo, e utilizada contra hiperacidez e ulcerações do estômago. Com o objetivo de caracterizar o potencial biológico desta planta, foi estudada a atividade antibacteriana e antioxidante do extrato diclorometânico (DCM), frações, subfrações e duas substâncias puras (maitenina e catequina) isoladas das cascas de raízes de M.ilicifolia, obtidas por maceração a frio. Foi utilizada a técnica de diluição em microplacas frente às bactérias Staphylococcus aureus ATCC 25923, Enterococcus faecalis ATCC 29212, Bacillus subtilis ATCC 19659, Escherichia coli ATCC 25922 e Pseudomonas aeruginosa ATCC 27853. A concentração inibitória mínima (CIM) para o extrato e as substâncias testadas, expresso como a menor concentração capaz de inibir crescimento bacteriano foi avaliada em microplacas contendo meio líquido aos quais foram adicionados extratos, frações, substâncias puras e antibióticos, sendo estes últimos usados como controles positivos. Para a atividade antioxidante foram realizados ensaios espectrofotométricos do 2,2'-Azinobis-3-etilbenzotiazolina-6-acido sulfônico (ABTS.+); do íon hipoclorito (OCl-) e da taurina-cloramina (via oxidação do 3,3’,5,5’, tetrametilbenzidina), utilizando como padrões quercetina e cisteína. Foi demonstrada forte atividade da maitenina com CIM de 0,48 μg/mL para as bactérias Gram positivas, e a catequina não apresentou atividades contra as bactérias testadas. Na avaliação da atividade antioxidante o extrato DCM e a maitenina não foram tão eficazes quanto a atividade da catequina que... / The genus Maytenus (Celastraceae) currently includes 80 species distributed throughout the Brazilian territory. The species Maytenus ilicifolia popularly known by the names of “espinheira-santa”, “espinheira-diva”, “salva-vidas”, “sombra de touro”, “cancerosa” and “coromilho do campo”, with activity against hyperacidity and gastric ulcers. In order to characterize the biological potential of this plant, was studied the antibacterial activity of the dichloromethane extract (DCM), fractions, subfractions and two substances (maitenin and catechin) isolated from the roots of M. ilicifolia obtained by cold maceration. To evaluation of the antimicrobial potential was used the microplates dilution technique using the bacteria Staphylococcus aureus ATCC 25923, Enterococcus faecalis ATCC 29212, Bacillus subtilis ATCC 19659, Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853. Minimum inhibitory concentration (MIC) for the extract and the substances tested, expressed as the lowest concentration able to inhibit bacterial growth, was evaluated in microplates containing liquid medium to which were added extracts, fractions, subfractions, pures substances and antibiotics, the latter being used as positive controls. For the antioxidant activity tests were performed spectrophotometric: 2,2 '-Azinobis-3-ethylbenzothiazoline-6- sulfonic acid (ABTS.+); of OCl- and taurinecloramine (via oxidation of 3,3', 5,5 ' , tetramethylbenzidine), using quercetin and cysteine as standards. It was demonstrated a strong activity to maitenin, with MIC of 0,48 μg/mL for Gram positive bacteria and catechin did not show activity against the bacteria tested. In the evaluation of antioxidant activity the extract DCM and maitenin... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:23:04Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-02-24Bitstream added on 2014-06-13T19:49:40Z : No. of bitstreams: 1 machado_bfmt_me_botib.pdf: 809978 bytes, checksum: dfad13979717e24ed1396c50d10ae323 (MD5) / Esta dissertação encontra-se dividida em introdução geral e 3 capítulos, sendo estes no formato de manuscritos a serem enviados para publicação em 3 periódicos distintos. Considerando a crescente utilização dos produtos naturais, especialmente dos obtidos a partir de plantas, objetivou-se estudar a ação antibacteriana de 27 óleos essenciais de uso em aromaterapia, sobre linhagens de Staphylococcus aureus (n=10), Escherichia coli (n=9), e Pseudomonas aeruginosa (n=9), isoladas de casos clínicos humanos, utilizando a metodologia dos discos (difusão) e determinação da concentração inibitória mínima (CIM) (diluição) em Mueller Hinton Ágar (Capítulo I). Verificou-se que as linhagens de S. aureus foram mais susceptíveis que as de Gram negativas, sendo que os valores de CIM90% foi de 0,21mg/mL para os óleos de pimenta negra (Piper nigrum) e tea tree (Melaleuca alternifolia) e 26,52mg/mL para o óleo de copaíba (Copaíba officinalis). Para E. coli, o óleo de canela (Cinnamomum cassia) foi o mais efetivo, com 2,0 mg/mL para CIM90% enquanto para P. aeruginosa o valor foi de 8,29 mg/mL com cravo da índia (Syzigium aromaticum). Utilizando valores de CIM obtidos in vitro foram selecionados 5 óleos (cravo da índia-Syzygium aromaticum, gerânio-Pelargonium graveolens, lavanda -Lavandula angustifolia, palmarosa-Cymbopogon martini e tea tree-Melaleuca alternifolia) e novamente sobre linhagens de Escherichia coli, Staphylococcus aureus e Pseudomonas aeruginosa, foi verificada a ação antibacteriana através da diluição individual de cada óleo em água e salina visando redução na contagem bacteriana em função do tempo (Capítulo II). Tanto nos ensaios em água quanto salina, verificou-se que o perfil de sensibilidade das linhagens bacterianas aos óleos essenciais foram próximos entre si, porém significativamente distintos comparados aos ensaios controles... / This thesis is divided into general introduction and three chapters, which are in the format of manuscripts to be submitted for publication in three separate journals. Considering the increasing use of natural products, especially those obtained from plants, aimed to study the antibacterial activity of 27 essential oils used in aromatherapy, on Staphylococcus aureus (n=10), Escherichia coli (n=9), and Pseudomonas aeruginosa (n=9), isolated from human clinical cases, using the methodology of the disks (diffusion) and determination of minimum inhibitory concentration (MIC) (dilution) on Mueller Hinton Agar (Chapter I). It was found that the strains of S. aureus were more susceptible than the Gram negative, and the values of MIC90% was 0.21 mg/mL for the oils of black pepper (Piper nigrum) and tea tree (Melaleuca alternifolia) and 26.52 mg/mL for oil Copaiba (Copaiba officinalis). For E. coli strains, the oil of cinnamon (Cinnamomum cassia) was the most effective, with 2.0 mg/mL for MIC 90% while for P. aeruginosa, the value was 8.29 mg/mL with clove (Syzigium aromaticum). Using MIC values obtained in vitro were selected five oils (cloves-Syzygium aromaticum, geranium-Pelargonium graveolens, lavender- Lavandula angustifolia, palmarosa-Cymbopogon martini and tea tree-Melaleuca alternifolia) and the bacteria Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa, antibacterial activity was checked by dilution of individual oil and saline water in order to reduce the bacterial count in function of time (Chapter II). Both tests in saline and water, it was found that the sensitivity of bacterial strains to essential oils were close together, but were significantly different compared to control tests. Essential oils have demonstrated the potential inhibitor of the three bacterial strains, and the results were different from those obtained in vitro-ágar with Gram negative... (Complete abstract click electronic access below)

Modern medicine is challenged continuously by the increasing prevalence of multi-drug resistant bacteria. Therefore, the development of alternatives to traditional antibiotics is an urgent necessity. Cationic antimicrobial peptides (CAMPs) are components of the innate immune defense system. Histones, generally known as proteins that package and regulate the transcription of DNA, share all of the essential antimicrobial traits of CAMPs, and could be promising alternatives to antibiotics. In this study, I investigated the antimicrobial properties of nucleated-erythrocyte-specific linker histone H5 and its derived peptides. Histone H5 was extracted and purified from chicken erythrocytes using an acid extraction followed by ion exchange chromatography using a step salt gradient; the purity (>95%) was verified by densitometry and proteomics analysis. Purified histone H5 demonstrated potent antimicrobial activity against various Gram-positive and Gram-negative planktonic bacteria, including resistant strains such as methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE), as well as anti-biofilm activity against Listeria monocytogenes and Pseudomonas aeruginosa. Furthermore, scanning electron microscopy (SEM) revealed significant damage to L. monocytogenes and P. aeruginosa bacterial cell surfaces after histone H5 treatment. The potential for histone toxicity towards mammalian cells was investigated with a hemolytic assay which determined that even at the highest concentration tested (1 mg/mL), histone H5 was non-hemolytic. An in silico analysis determined the predicted antimicrobial domain of histone H5 of which six histone H5-derived peptides with potential antimicrobial activity were identified. These six histone H5-derived peptides were synthesized and tested against bacterial pathogens to determine their antimicrobial properties. Although the H5-derived peptides were identified within the predicted antimicrobial domain of histone H5, they did not possess more potent antimicrobial activity than the full length protein. Overall, this study demonstrates that histone H5 and histone H5-derived peptides could be promising candidates in the development of novel anti-infective therapeutics.

CoordenaÃÃo de AperfeÃoamento de Pessoal de NÃvel Superior / Antimicrobial agents of tetracycline class has been extensively used in aquaculture for disease control and growth promotion. Thus, the aim of this study was to evaluate the polyphase profile of the bacterial community in shrimp farming environment front antimicrobial classes of tetracyclines, focusing on Vibrio genus. Water and sediment samples were collected four (4) shrimp farms: two (2) in the state of Cearà and two (2) in the state of Rio Grande do Norte, of points: estuary, supply channel, pond and drainage channel. Cultivable heterotrophic bacteria (CHB), with resistance to tetracycline (CHB+TET) and oxytetracycline (CHB+OTC), total Vibrio, positive sucrose Vibrio and sucrose negative Vibrio were surveyed by Standard Plate Count method (SPC). The strains isolated from CHB, CHB+TET, CHB+OTC and Vibrio underwent phenotypic identification. The phenotypic profile of antimicrobial susceptibility of strains of Vibrio and other BHC was compared between the disk diffusion techniques and macrodilution by determining the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) for tetracycline and oxytetracycline antimicrobials. Resistance genes tet(M), tet(S) otr(A) and otr(B) were investigated in strains characterized as phenotypically resistant. Diversity of heterotrophic bacteria is influenced by wet and rainy seasons and also by the different collection points within the shrimp farms. Overall, the CHB, CHB+TET and CHB+OTC and Vibrio counts differ between the collection seasons, with higher measurements during the rainy season to the points of pond and drainage channel. 146 strains of CHB, 57 of CHB+TET and 45 of CHB+OTC were isolated, of which the most representative genera were Vibrio and Enterobacter. A total of 100 strains of Vibrio were isolated, which V. coralliilyticus was the most representative species. Antimicrobial profile of Vibrio and other CHB differed between disk-diffusion and macrodilution techniques, being the macrodilution the most efficient in the characterization of resistance among isolates. All strains of Vibrio and other CHB, phenotypically resistant, presented the tet(M) gene as a determinant of resistance determinat, but tet(S) otr(A) and otr(B) genes were not observed. Thus, it proves that Vibrio genus is most abundant among herotrotrophic bacteria with resistance to tetracyclines, with the bacterial diversity strongly influenced by seasons. / Antimicrobianos da classe das tetraciclinas tÃm sido extensivamente usados em aquicultura para o controle de doenÃas e como promotores de crescimento. Dessa forma, foi traÃado como objetivo geral avaliar o perfil polifÃsico da comunidade bacteriana de ambiente de criaÃÃo de camarÃo frente aos antimicrobianos das classes das tetraciclinas, com enfoque no gÃnero Vibrio. Amostras de Ãgua e sedimento foram coletadas de quatro (4) fazendas de carcinicultura: duas (2) no estado do Cearà e duas (2) no estado do Rio Grande do Norte, dos pontos: estuÃrio, canal de abastecimento, viveiro e canal de drenagem. Foram pesquisadas bactÃrias heterotrÃficas cultivÃveis (BHC), com resistÃncia à tetraciclina (BHC+TET) e oxitetraciclina (BHC+OTC), Vibrio total, sacarose positivos e negativos pelo mÃtodo de Contagem PadrÃo em Placas (CPP). As cepas isoladas de BHC, BHC+TET, BHC+OTC e Vibrio foram submetidas à identificaÃÃo fenotÃpica. O perfil fenotÃpico de susceptibilidade antimicrobiana das cepas de Vibrio e demais BHC foi comparado entre as tÃcnicas de disco-difusÃo e macrodiluiÃÃo, pela determinaÃÃo da ConcentraÃÃo InibitÃria MÃnima (CIM) e ConcentraÃÃo Bactericida MÃnima (CBM) para os antimicrobianos tetraciclina e oxitetraciclina. Os genes de resistÃncia tet(M), tet(S), otr(A) e otr(B) foram pesquisados nas cepas fenotipicamente caracterizadas como resistentes. A diversidade de bactÃrias heterotrÃficas foi influenciada pelos perÃodos de estio e chuvoso e tambÃm pelos diferentes pontos de coleta dentro das fazendas de carcinicultura. No geral, as contagens de BHC, BHC+TET e BHC+OTC e Vibrio e demais BHC diferiram entre os perÃodos de coleta, com quantificaÃÃes superiores no perÃodo chuvoso para os pontos de coleta viveiro e canal de drenagem. Foram isoladas 146 cepas de BHC, 57 de BHC+TET, 25 de BHC+OTC, das quais os gÃneros mais representativos foram Vibrio e Enterobacter. Foi possÃvel o isolamento de 100 cepas de Vibrio, das quais V. coralliilyticus foi a espÃcie mais representativa. O perfil antimicrobiano diferiu entre as cepas de Vibrio e demais BHC entre as tÃcnicas de disco-difusÃo e macrodiluiÃÃo, sendo a de macrodiluiÃÃo a mais eficiente na caracterizaÃÃo de resistÃncia dentre os isolados. Todas as cepas de Vibrio e demais BHC, fenotipicamente resistentes, apresentaram o gene tet(M) como determinante de resistÃncia, no entanto os genes tet(S), otr(A) e otr(B) nÃo foram verificados. Com isso, comprovou-se que Vibrio à gÃnero mais abundante dentre as bactÃrias heterotrÃficas com resistÃncia Ãs tetraciclinas, estando a diversidade bacteriana fortemente influenciada pelos perÃodos do ano.

O objetivo deste trabalho foi produzir hidroxiapatita (HA) contendo diferentes tipos de própolis de origem brasileira e avaliar a possível atividade antimicrobiana dos materiais. Os extratos etanólicos de própolis (EEP) vermelha, verde e marrom foram obtidos em solução alcoólica 80%. EEP verde e vermelha (8 mg/mL e 20 mg/mL) foram incorporados ao material a 10% (m/v) via atomização (spray drying), obtendo-se HA-GP8, HA-GP20, HA-RP8 e HA-RP20. Os EEP e os materiais foram caracterizados quanto ao conteúdo de compostos fenólicos e flavonoides totais. A atividade antimicrobiana dos EEP foi avaliada por difusão em ágar, concentração inibitória mínima (CIM) e bactericida mínima (CBM) frente à Staphylococcus aureus (S. aureus), Staphylococcus epidermidis (S. epidermidis) e Escherichia coli (E. coli). Os pós de HA incorporados com própolis foram avaliados frente à S. aureus por contagem de colônias bacterianas em placa, CIM e CBM. A caracterização dos materiais foi realizada por difração de raios X (DRX), espectroscopia na região do infravermelho com transformada de Fourier (FTIR) e microscopia eletrônica de varredura (MEV). A citotoxicidade foi determinada pelo cálculo da viabilidade celular, realizada pelo método de incorporação do vermelho neutro. Para os extratos, o conteúdo de fenólicos variou entre 278,3 e 325,6 mg EAG/g ES. EEP vermelha mostrou maior conteúdo de flavonoides (112,4 mg EQ/g ES) em relação aos outros EEP (48,4 e 52,3 mg EQ/g ES), apresentando maior atividade inibitória (CIM) frente à S. aureus e S. Epidermidis (12,5 &micro;g/mL), comparado ao EEP verde (100 e 200 &micro;g/mL) e marrom (200 &micro;g/mL). Para CBM, os EEP vermelha e verde foram mais efetivos (800 &micro;g/mL) comparado ao EEP marrom (1600 &micro;g/mL), frente às mesmas bactérias. Entretanto, não foi observada atividade frente à E coli. A caracterização dos pós incorporados ou não com própolis apresentou estrutura cristalina e morfologia aparentemente esférica, indicando diminuição no grau de aglomeração com a adição de própolis. FTIR indicou a presença de grupos funcionais característicos para HA e própolis. Os materiais apresentaram alta liberação de fenólicos (228,3 a 327,6 mg EAG/g ES) e menores quantidades de flavonoides (14,0 a 35,8 mg EQ/g ES), sendo as maiores quantidades de flavonoides atribuída à HA contendo própolis vermelha. Foi verificado efeito bactericida a partir de 0,5 h (HA-RP20 e HA-GP20) e 1 h (HA-RP8 e HA-GP8) e menor atividade inibitória (CIM) para HA-GP20 e HA-GP8 (175,4 e 182,0 &micro;g/mL) e para HA-RP20 e HA-RP8 (51,7 e 66,8 &micro;g/mL), comparado aos EEP. Entretanto, observou-se maior atividade bactericida (CBM) para HA-GP20 e HA-GP8 (701,5 e 728,0 &micro;g/mL) e para HA-RP20 e HA-RP8 (206,5 e 267,0 &micro;g/mL), em relação aos EEP. O ensaio de citotoxicidade mostrou valores para IC50 (concentração que reflete 50% da viabilidade celular) de 387,1 e 84,8 &micro;g/mL para as amostras HA-GP8 e HA-RP8, respectivamente. Considerando os resultados obtidos neste trabalho, sugere-se que a HA incorporada com própolis (HA-GP8 e HA-RP8) pode ser utilizada como possível agente antimicrobiano, inibindo o crescimento de S. aureus, respeitando-se os valores máximos para IC50. Entretanto, não poderia ser utilizada como agente bactericida, uma vez que nestas condições os materiais apresentaram efeito citotóxico. / The aim of this study was to produce hydroxyapatite (HA) containing different types of propolis of Brazilian origin and to evaluate the possible antimicrobial activity materials. The ethanolic extracts of red, green and brown propolis (EEP) were obtained in alcoholic solution 80%. Green and red EEP (8 mg / mL and 20 mg / mL) were incorporated into the 10% (m/v) via atomization (spray drying), obtaining HA-GP8, HA-GP20, HA-RP8 and HA -RP20. EEP and materials were characterized regarding the content of phenolic compounds and total flavonoids. The antimicrobial activity of the EPS was evaluated by diffusion in agar, minimum inhibitory concentration (MIC) and minimum bactericidal (MBC) against Staphylococcus aureus (S. aureus), Staphylococcus epidermidis (S. epidermidis) and Escherichia coli (E. coli). The HA powders incorporated with propolis were evaluated against S. aureus by plate colony counting , CIM and CBM. Materials characterization was made by X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM). The cytotoxicity of the materials was determined by cell viability calculation, which was made by the neutral red incorporation method. For extracts, phenolic content ranged from 278.3 to 325.6 mg EAG/g ES. The red EEP showed a higher flavonoid content (112.4 mg EQ/g ES) than the other EEP (48.4 and 52.3 mg EQ/g ES), showing a higher inhibitory activity (MIC) against S. aureus and S. epidermidis (12.5 &micro;g/mL) compared to green EEP (100 and 200 &micro;g/mL) and brown (200 &micro;g/mL). For CBM, red and green EEP were more effective (800 &micro;g/mL) compared to brown EEP (1600 &micro;g/mL) against the same bacteria. However, no activity was observed against E coli. The characterization of the powders incorporated or not with propolis presented crystalline structure and apparently spherical morphology, indicating a decrease in the degree of agglomeration with the addition of propolis. FTIR indicated the presence of functional groups characteristic for HA and propolis. The materials presented high phenolic release (228.3 to 327.6 mg EAG/g ES) and lower amounts of flavonoids (14.0 to 35.8 mg EQ/g ES), with the highest amounts of flavonoids attributed to HA containing red propolis. The bactericidal effect for all materials was observed within the interval of 0.5 (HA-RP20 and HA-GP20) to 1 hour (HA-RP8 and HA-GP8). The materials showed lower inhibitory activity (MIC) for HA-GP20 and HA-GP8 (175, 4 and 182.0 &micro;g/mL) and for HA-RP20 and HA-RP8 (51.7 and 66.8 &micro;g/mL) compared to EEP. However, higher bactericidal activity (MBC) was observed for HA-GP20 and HA-GP8 (701.5 and 728.0 &micro;g/mL) and for HA-RP20 and HA-RP8 (206.5 and 267.0 &micro;g/mL) when compared to the EEP. The cytotoxicity assay showed values for IC50 (concentration that reflects 50% of cellular viability) of 387.1 and 84.8 &micro;g/mL for HA-GP8 and HA-RP8 samples, respectively. Considering the results obtained in this work, it is suggested that HA incorporated with propolis (HA-GP8 and HA-RP8) can be used as a possible antimicrobial agent, inhibiting the growth of S. aureus, respecting the maximum values for IC50. However, it could not be used as a bactericidal agent, since under these conditions the materials had a cytotoxic effect.