Made available in DSpace on 2014-10-09T12:34:41Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:01:02Z (GMT). No. of bitstreams: 1 17900.pdf: 755420 bytes, checksum: b49381f0e220377f0173530306d2ddf1 (MD5) / Dissertacao (Mestrado Profissionalizante em Lasers em Odontologia) / IPEN/D-MPLO / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP; Faculdade de Odontologia, Universidade de São Paulo, São Paulo

Made available in DSpace on 2014-10-09T12:34:41Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:01:02Z (GMT). No. of bitstreams: 1 17900.pdf: 755420 bytes, checksum: b49381f0e220377f0173530306d2ddf1 (MD5) / Dissertacao (Mestrado Profissionalizante em Lasers em Odontologia) / IPEN/D-MPLO / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP; Faculdade de Odontologia, Universidade de São Paulo, São Paulo

Lentinula edodes é um cogumelo comestível que possui qualidades nutricionais, terapêuticas e medicinais. Além disso, muitos estudos na área médica têm comprovado que o cogumelo possui efeito antibiótico sobre microrganismos patogênicos ao homem. Na área agrícola, alguns trabalhos realizados com o cogumelo demonstraram possíveis efeitos no controle de fitopatógenos. O presente trabalho teve como objetivo demonstrar a produção de substâncias antimicrobianas por L. edodes ativas sobre Colletotrichum sublineolum, agente causal da antracnose em sorgo, Alternaria solani, responsável pela pinta preta do tomateiro, Xanthomonas axonopodis pv. passiflorae, agente causal da mancha bacteriana em maracujazeiro e Tobacco mosaic virus (TMV), causador de mosaico foliar em fumo. Para os testes com C. sublineolum e A. solani foram utilizados extratos aquosos de L. edodes, obtidos a partir de basidiocarpos desidratados em pó, dos isolados LE JAB-K, LE 96/22, LE 96/17 e LE 95/01. Os resultados evidenciaram que o extrato aquoso de basidiocarpos do isolado LE 96/22 inibiu o crescimento micelial in vitro e a formação de apressórios por C. sublineolum. Já os extratos dos isolados LE JAB-K e LE 95/01 apresentaram efeito inibitório na germinação de conídios e na formação de apressórios do patógeno. Em contrapartida, os extratos aquosos de basidiocarpos dos diferentes isolados de L. edodes não apresentaram efeito inibitório na germinação dos conídios e no crescimento micelial de A. solani. Por sua vez, os extratos aquosos de basidiocarpos a 20% (v/v) e o filtrado do crescimento micelial de L. edodes, misturados à suspensão de X. axonopodis pv. passiflorae, exibiram redução na multiplicação bacteriana. Todos os extratos aquosos de basidiocarpos dos diferentes isolados testados na multiplicação da bactéria mostraram-se termolábeis, quando autoclavados a 121 °C por 20 min. Em experimentos com plantas de fumo, os extratos aquosos de basidiocarpos dos isolados LE 96/17 e LE 96/22 adicionados à suspensão contendo partículas do TMV reduziram significativamente a ocorrência de lesões locais nas folhas. O extrato aquoso do isolado LE 96/22 apresentou compostos antivirais de natureza termoestável. Finalmente, o extrato aquoso de basidiocarpos do isolado LE 96/22, o qual apresentou a maior atividade antimicrobiana, foi purificado parcialmente por cromatografia de troca aniônica (CTA). O pico V apresentou efeito inibitório no crescimento micelial de C. sublineolum. Por sua vez, a multiplicação de X. axonopodis pv. passiflorae foi inibida pelos picos IV, V e VII. Já os picos I, II e III, obtidos em CTA por gradiente linear de NaCl e o pico I obtido em CTA pelo método “step wise”, reduziram significativamente a infectividade do TMV em plantas de fumo. Com base nesses resultados, evidencia-se a ação de preparações de L. edodes sobre fitopatógenos, o que demonstra o uso potencial do mesmo no controle de agentes causais de doenças infecciosas em plantas. / Lentinula edodes is an edible mushroom that has nutritious, therapeutical and medicinal qualities. Moreover, many studies in the medical area have shown that the mushroom exhibits antibiotic effects on pathogenic microorganism to the man. In the agricultural area, work carried out with the mushroom has demonstrated its possible effects to control phytopathogens. The objective of the present work was to demonstrate the productionof antimicrobial substances of L. edodes active on Colletotrichum sublineolum, causal agent of anthracnose in sorghum, Alternaria solani, responsible for the black spot of the tomato plants, X. axonopodis pv. passiflorae, causal agent of the bacterial spot in passion fruit plants and on Tobacco mosaic virus, causal agent of the mosaic in tobacco plants. For the test with C. sublineolum and A. solani aqueous extracts were obtained from dehydrated fruiting bodies from the shiitake isolates LE JAB-K, LE 96/22, LE 96/17 and LE 95/01. The results showed that the fruiting body aqueous extract from isolate LE 96/22 inhibited micelial growth and appressorium formation by C. sublineolum. The aqueous extracts of isolates LE JAB-K and LE 95/01 exhibited inhibitory effect on conidium germination and on formation of appressorium by the patogen. On the other hand, the extracts of the different isolates of L. edodes did not exhibit inhibitory effect on conidium germination and micelial growth of A. solani. The aqueous extracts of fruiting bodies at 20% (v/v) concentration and filtrate of the micelial growth of L. edodes, when mixed to the suspension of X. axonopodis pv. passiflorae, exhibited decreased on bacterial multiplication. All the aqueous extracts of fruiting bodies tested from the different isolates in the bacterial multiplication were thermobile, when heated at 121 °C for 20 min. In experiments with tobacco plants, the aqueous extracts of fruiting bodies of isolates LE 96/17 and LE 96/22 when added to the suspension of TMV reduced the amount of local lesions on the leaves. When the aqueous extracts of LE 96/22 were heated the antiviral nature was not lost. Finally, the aqueous extract of fruiting bodies from isolate LE 96/22 that presented major antimicrobial activity was partially purified by anion exchange chromatography (AEC). The peak V exhibited inhibitory effect on micelial growth of C. sublineolum. Multiplication of X. axonopodis pv. passiflorae was inhibited by peaks IV, V and VII. Regarding TMV infectivity, peaks I, II and III, obtained in CTA through linear gradient of NaCl, and peak I also obtained through CTA by the method "step wise", significantly reduced virus infectivity in tobacco plants. Based upon these results, it is shown that preparations of L. edodes can interfere whith phytopathogen multiplication, demonstrating its potential to control plant diseases.

Master of Science / Department of Architectural Engineering and Construction Science / Julia A. Keen / Nanofibers are classified as fibers less than 1 micrometer in diameter. These fibers can be layered to form nanofibrous membranes, and these membranes offer great potential in the filtration industry. The membranes' smaller fiber diameters and pore sizes permit such filters to filter out more and smaller particulate. Additionally, antimicrobial agents can be incorporated into the membrane to inhibit fungal and bacterial growth on the membrane’s surface. This report evaluates nanofibrous membranes with antimicrobial agents and their potential in two specific locations: cleanrooms and protective environment rooms, where bacterial and fungal growth would have a detrimental effect on the process or occupant of the space.

A presença de bactérias no cultivo in vitro de plantas é normalmente relacionada à contaminação, acarretando o descarte de microplantas de alto valor comercial. No entanto, essa presença pode ser caracterizada como \"manifestações\", decorrentes de possíveis estresses físicos ou nutricionais, da microbiota endofítica, que coloniza ativamente os tecidos vegetais, e que não necessariamente, afeta o desenvolvimento da planta. Sendo assim, o objetivo deste trabalho foi controlar a manifestação bacteriana, preservando as microcepas de Eucalyptus cloeziana em desenvolvimento in vitro, adicionando-se ao meio de cultura, os antibióticos gentamicina, ciprofloxacina, rifampicina, timentin (isolados ou combinados entre si) e o biocida clorometil-isotiazolinona + metil-isotiazolinona. O monitoramento do trabalho foi realizado por meio de avaliações semanais, visando verificar possíveis influências dos agentes antimicrobianos na morfofisiologia do vegetal e a eficiência no controle das manifestações bacterianas. Para tanto, foram avaliados parâmetros referentes ao incremento de massa seca, número de brotações, teor de acúmulo de nutrientes, análises histológicas e histoquímicas das microcepas. Para verificar possíveis flutuações das comunidades bacterianas endofíticas nas microcepas, utilizou-se a técnica de DGGE. Todas as análises foram realizadas aos 15 e 30 dias após a adição dos agentes antimicrobianos no meio de cultura, exceto a avaliação nutricional que foi feita apenas no final do experimento (30 dias). Os resultados obtidos permitiram inferir que ao longo dos 30 dias de cultivo, o antibiótico gentamicina, ocasionou queda no desenvolvimento das microcepas e não foi eficiente no controle das manifestações bacterianas. Os antibióticos rifampicina e ciprofloxacina apesar de não serem efetivos no controle da manifestação bacteriana, não afetaram o desenvolvimento das microcepas. Conclui-se que, o biocida clorometil-isotiazolinona + metil-isotiazolinona efetivamente controlou as manifestações bacterianas, sem alterar a morfofisiologia das microcepas, bem como o timentin, que além de controlar as manifestações, proporcionou maior vigor das microcepas em função da melhor assimilação dos nutrientes. / The in vitro bacterial presence in plant cultivation, usually is related to contamination, resulting in the elimination of the microplants with high commercial value. However, this presence can be characterized as \"manifestations\" arising from possible physical or nutritional stresses, from microbial endophytic that actively colonize plant tissues, and do not necessarily affect plant development. Therefore, the purpose of this study was to control the bacterial manifestation, preserving in vitro developing of Eucalyptus cloeziana microstumps, by adding to the culture medium, the antibiotics gentamicin, ciprofloxacin, rifampicin, timentin (alone or combined) and biocide + chloromethyl isothiazolinone methyl-isothiazolinone. The monitoring work was conducted through weekly evaluations, to verify the possible influence of antimicrobial agents in the plant morphophysiology and the efficient control of bacterial manifestations. Therefore, were evaluated parameters related to the increase in dry weight, number of buds, content of nutrient accumulation, histological and histochemical the microstumps. DGGE technique was used to check possible fluctuations of endophytic bacterial communities in microstumps. All analyzes were performed at 15 and 30 days after the addition of antimicrobial agents in the culture medium, except for the nutritional assessment that was made only at the end of the experiment (30 days). The results allow to infer that over the 30 days of culture, the antibiotic gentamicin, resulted in the microstumps development decrease and was not effective in controlling bacterial manifestations. The antibiotics rifampicin and ciprofloxacin although were not effective in controlling the bacterial manifestation, affected the development of microstumps. It was concluded that the biocide chloromethyl-methyl-Isothiazolinone + Isothiazolinone effectively controlled the bacterial manifestations without modifying the morphophysiology of the microstumps, well as the timentin, which besides controlling the manifestations, provided greater vigor of microstumps due to better assimilation of nutrients.

Os processos industriais de produção de álcool existentes no Brasil reutilizam o fermento em ciclos fermentativos consecutivos. Paralelamente, o excedente produzido pela multiplicação das células de levedura durante esse processo é seco e comercializado, principalmente no mercado externo, como ingrediente para ração animal. As práticas usualmente utilizadas nas indústrias para reduzir a contaminação bacteriana são o tratamento ácido do creme de levedura e a aplicação de antibióticos. Porém, desde que foram detectados altos níveis de resíduos de antibióticos na levedura destinada à ração animal, seu uso tem sido condenado pela comunidade internacional. Desde então as indústrias brasileiras têm buscado alternativas aos antibióticos para o controle da contaminação bacteriana. O objetivo deste trabalho foi avaliar a atividade antimicrobiana do extrato da própolis sobre as bactérias do gênero Lactobacillus fermentum e Bacillus subtillis, que são alguns dos contaminantes da fermentação alcoólica. O maior pico de produção de compostos fenólicos totais foi com uso de etanol 76% como solvente, à 58°C por 50 minutos, apresentando 68,95 mg de CFT/g de própolis e um halo de inibição de 6 e 5mm para Bacillus e Lactobacillus, respectivamente. As faixas das variáveis que maximizaram o teor alcoólico foi o processo fermentativo conduzido a 32°C, com 41 g de células de leveduras/L, em um meio contento 18,5°Brix. Apesar do antimicrobiano comercial ter apresentado maior eficiência na redução da contaminação (94,46% e 97,40% para Lactobacillus e Bacillus, respectivamente), o extrato de própolis tem potencial para ser utilizado no controle dos contaminantes bacterianos presentes nas fermentações etanólicas, sendo responsável por redução de 54,24% e 67,02% para Lactobacillus e Bacillus, respectivamente. / The industrial processes of ethanol production in Brazil reuse yeast fermentation in consecutive cycles. In parallel, the surplus produced by the multiplication of yeast cells during this process is dried and sold, mainly in foreign markets as an ingredient for animal feed. The practices commonly used in industry to reduce bacterial contamination are the acid treatment of the yeast cream and the application of antibiotics. However, since they were detected high levels of antibiotic residues in yeast destined for animal feed, its use has been condemned by the international community. Since then Brazilian industries have sought alternatives to antibiotics to control bacterial contamination. The aim of this study was to evaluate the antimicrobial activity of the extract of propolis on bacteria of the genus Lactobacillus and Bacillus, which are some of the contaminants of fermentation. The highest peak production of phenolic compounds was using 76% ethanol as solvent at 58°C for 50 minutes, with 68,95 mg of CFT/g of propolis and a halo of inhibition of 6 and 5mm for Bacillus and Lactobacillus, respectively. The ranges of variables that maximized the alcoholic fermentation was conducted at 32°C with 41g of yeast cells/L in media containing a 18,5°Brix. Despite having larger commercial antimicrobial effectiveness in reducing contamination (94,46% and 97,40% for Lactobacillus and Bacillus, respectively), the extract of propolis has potential to be used in the control of bacterial contaminants present in the ethanolic fermentation, and responsible for reduction of 54,24% and 67,02% for Lactobacillus and Bacillus, respectively.

Face à l’émergence de pathogènes multi-résistants aux antibiotiques classiques, et au développement des armes biologiques, la découverte de nouveaux agents antimicrobiens reste un enjeu majeur de santé public. Dans ce contexte, la conception d’oligomères peptidomimétiques, capables de mimer le caractère amphiphile et la structure en hélice des peptides antimicrobiens naturels, effecteurs clés de l’immunité innée, offre d’intéressantes perspectives. Il a été établi que des foldamères à base d’urées amphipathiques, structurés en hélice-2,5, possédaient une forte activité bactéricide contre Bacillus anthracis, bactérie considérée comme une arme biologique potentielle. En vue d’optimiser l’activité anthracidale et la sélectivité in vitro de la première génération de composés, une étude relation structure-activité a été initiée en réalisant une série de modifications (i.e. séquence primaire, longueur et squelette de l’oligourée). Des oligomères originaux possédant des motifs isostères de type thiourée et guanidine ont ainsi été préparés en solution puis sur support solide. Des études conformationnelles approfondies soulignèrent que seule l’insertion de lien thiourée à proximité du dipôle négatif était bien tolérée par l’hélice-2,5. Parallèlement, les études in vivo ont montré une forte stabilité des oligourées avec une accumulation sélective dans le rein ainsi qu’une protection partielle des souris contre l’infection systémique par Bacillus anthracis. Enfin l’étude de l’interaction de ces oligourées avec des membranes lipidiques modèles a confirmé leur capacité à perturber les membranes et a mis en avant des mécanismes d’action différents selon le type de lipides utilisés. / The increasing antibiotic resistance among pathogens and the emergence of biological weapons have highlighted the urgent need of new antimicrobial agents. In this context, the design of peptidomimetics as urea-based foldamers, capable of mimicking the amphiphilic character and conformation of natural antimicrobial peptides, key effector molecules of innate immunity, offers new prospects. It has been previously established that amphipathic oligourea 2.5-helices have a strong bactericidal activity against Bacillus anthracis, bacteria considered as a potential biological warfare agent. Based on these results and with the aim of optimizing the potency and selectivity in vitro of the first generation of compounds, a structure-activity relationship study was carried out by performing series of modifications on a lead compound (i.e. side-chain replacement, size and backbone modifications). Among them, new series oligomers incorporating isosteric substitutions such as thiourea and guanidine moieties were prepared in solution then on solid support. Interestingly, the conformational studies revealed that only the insertion of thiourea linkage near the negative end of the helix dipole was well-tolerated by the 2.5-helix. Concurrently, in vivo studies highlighted a strong stability of the lead oligourea with a selective accumulation in the kidneys as well as a partial protection of the mice after systemic infection by Bacillus anthracis. Finally, biophysical interaction studies of selected oligoureas with model membranes confirmed their capacity to disturb membranes and showed different mechanisms of action depending on the lipid composition.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / O ambiente estuarino à reconhecidamente um local de aporte de Ãguas contaminadas de diferentes fontes. O gÃnero Vibrio à encontrado, naturalmente, nesses locais, sendo relatadas, em inÃmeros trabalhos cientÃficos, cepas resistentes a antimicrobianos utilizados, rotineiramente, no tratamento mÃdico. Foram selecionadas setenta cepas de Vibrio oriundas de amostras de Ãgua e sedimento do estuÃrio do Rio AcaraÃ, litoral Oeste do CearÃ, Brasil para o experimento. O antibiograma foi proposto em duas baterias, sendo uma com Ãgar Mueller-Hinton solubilizado em Ãgua destilada (MH+AD) e a outra em Ãgua do mar (MH+AM), alÃm de ser testada a presenÃa dos fatores de virulÃncia proteases (caseinase, elastase e gelatinase), lipases, fosfolipases, DNases, amilases, urease e hemolisinas (β-hemÃlise pelo teste de Kanagawa em Ãgar Wagatsuma). Os antimicrobianos testados foram NAL, AMP, ATM, CFL, CIP, CLO, CTX, EST, GEN, OTC, PEN, SUT e TET. Em MH+AD, o perfil de resistÃncia foi verificado para 47 (67,1%) cepas a PEN, 26 (37,1%) cepas a AMP e CFL, 8 (11,4%) a OTC, 6 (8,6%) a TET, 3 (4,3%) a ATM, e 1 (1,4%) a CLO, enquanto para MH+AM, 68 (97,1%) cepas foram resistentes a OTC, 67 (95,7%) a TET, 57 (81,4%) a PEN, 34 (48,6%) a CFL, 29 (41,4%) a AMP, 21 (30,0%) a EST, 20 (28,5%) a GEN, 5 (7,2%) a ATM e NAL, 3 (4,3%) a SUT e 1 (1,4%) a CIP. As cepas resistentes em MH+AD foram submetidas à cura do plasmÃdio pelo agente curagÃnico acridine Orange. Foi verificada resistÃncia plasmidial em 36,21% das cepas resistentes a AMP, 5,17% a ATM, 37,93% a CFL, 13,79% a OTC, 53,45% a PEN e 6,90% a TET. Por ordem de recorrÃncia, 67 (91,42%) cepas de Vibrio spp. analisadas apresentam atividade da enzima urease, seguidas de 57 (81,42%) para lipase, 54 (77,14%) a amilase, 53 (75,72%) a gelatinase, 43 (61,42%) a caseinase, 26 (37,14%) a fosfolipase, 17 (24,28%) a DNase, 16 (22,86%) a elastase e 13 (18,57%) a β-hemÃlise (Kanagawa). A Ãgua do mar influenciou, significativamente, a caracterizaÃÃo da resistÃncia das cepas de Vibrio spp. analisadas, principalmente, sobre os antimicrobianos das classes das tetraciclinas, alÃm de ter sido detectada mÃltipla resistÃncia associada à cepas com fatores preliminares de virulÃncia. / The estuarine environment is recognized as an intake of contaminated water from different sources. The genus Vibrio is found in those places, being reported in many papers resistant strains to antibiotics used routinely in medical treatment. Seventy strains of Vibrio were selected originated from samples of water and sediment from the Acaraà estuary, West coast of CearÃ, Brazil to experiment. The antibiogram was proposed in two batteries, one with Mueller Hinton agar dissolved in distilled water (MH+DW) and the other in seawater (MH+SW), and is tested the presence of virulence factors protease (caseinase, elastase and gelatinase), lipases, phospholipases, DNases, amylase, urease, and hemolysin (β-hemolysis test by Kanagawa Wagatsuma agar). The antimicrobials tested were NAL, AMP, ATM, CFL, CIP, CHL, CTX, STP, GEN, OTC, PEN, TCY and STX. In MH+DW, the resistance profile was observed for 47 (67.1%) strains to PEN, 26 (37.1%) to AMP and CFL, 8 (11.4%) to OTC, 6 (8.6%) to TCY, 3 (4.3%) to ATM and 1 (1.4%) to CHL, while for MH+SW, 68 (97.1%) strains were resistant to OTC, 67 (95.7%) to TCY, 57 (81.4%) to PEN, 34 (48.6%) to CFL, 29 (41.4%) to AMP , 21 (30.0%) to STP, 20 (28.5%) to GEN, 5 (7.2%) to ATM and NAL, 3 (4.3%) to STX and 1 (1.4%) to CIP. The resistant strains in MH+DW were subjected to plasmid curing by acridine orange agent. Resistance plasmid was observed in 36.21% of resistant strains to AMP, 5.17% to ATM, 37.93% to CFL, 13.79% to OTC, 53.45% to PEN and 6.90% to TCY. By order of recurrence, 67 (91.42%) strains of Vibrio spp. have analyzed the enzyme urease, followed by 57 (81.42%) to lipase, 54 (77.14%) to amylase, 53 (75.72%) to gelatinase, 43 (61.42%) to caseinase, 26 (37.14%) to phospholipase, 17 (24.28%) to DNase, 16 (22.86%) to elastase and 13 (18.57%) to β-hemolysis (Kanagawa). Seawater influence significantly the characterization of the resistance of strains of Vibrio spp. analyzed mainly on the drugs of the tetracycline class, and has been detected multidrug resistant strains associated with preliminary virulence factors.

Infecções hospitalares são um problema de saúde pública. Quatorze linhagens de fungos isolados de mangue foram avaliadas quanto ao potencial de síntese de nanopartículas de prata (NP Ag) por método biológico com atividade antimicrobiana, sendo dois fungos selecionados. A caracterização físico-química das NP foi realizada por espectrofotometria, microscopia eletrônica e espectroscopia de correlação de fótons. A banda de plasmon evidenciou a formação de NP entre 10-30 nm. A ação antimicrobiana foi avaliada em espécies de Candida e bactérias. As NP foram mais efetivas nas espécies de C. guilhermondi, C. parapsilosis, P. aeruginosa e S. aureus com concentrações inibitórias variando de 0,015 a 0,132 µg/mL. A ação antifúngica de tecidos impregnados com NP Ag foi testada em C. albicans, C. glabrata e C. parapsilosis com inibição do crescimento na faixa de 68,41 97,91 %. Em E. coli e S. aureus a inibição do crescimento foi de 100% na concentração de 2,2 µg/mL. Os fungos foram identificados como Bionectra ochroleuca e Aspergillus tubingensis, respectivamente. / Hospital infections are a public health problem. Fourteen strains of fungi isolated from the mangrove were evaluated regarded to their capacity to synthesize silver nanoparticles (Ag NP) by a biological method with antimicrobial activity and two fungi were selected. The physicochemical characterization was performed by spectrophotometry, electron microscopy and photon correlation spectroscopy. The plasmon band evidenced the formation of NP from 10-30 nm. The antimicrobial activity was evaluated against Candida species and bacteria. The NP were more effective against the C. guilhermondi, C. parapsilosis, P. aeruginosa e S. aureus especies with inhibitory concentrations from 0.015 to 0.132 µg/mL (1.3 a 12 µM). The antifungal activity of the fabrics impregnated with Ag NP was tested against C. albicans, C. glabrata and C. parapsilosis, presenting inhibition from 68.41 97.91 %. Against E. coli and S. aureus the growth inhibition was of 100% in the concentration of 2.2 µg/mL. The fungi were identified as Bionectra ochroleuca and Aspergillus tubingensis, respectively.

Thesis (M.Sc. (Pharmacology)) -- University of Limpopo, 2018. / Refer to the document