Tesi: "Biofilms" – Grafiati

1

Bonsaglia, Erika Carolina Romão [UNESP]. "Produção de biofilme por Listeria monocytogenes isoladas de diferentes alimentos". Universidade Estadual Paulista (UNESP), 2012. http://hdl.handle.net/11449/87783.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Listeria monocytogenes é uma bactéria frequentemente encontrada em vários alimentos crus, como carne, queijo e legumes. Está amplamente distribuída no ambiente causando a contaminação de alimentos durante seu processamento. Atualmente, a contaminação por L. monocytogenes nas indústrias alimentícias é considerada um dos maiores problemas de segurança alimentar, pois uma vez presente na linha de processamento à bactéria dificilmente é eliminada pela grande capacidade em formar biofilme. A formação de biofilmes em equipamentos pode levar a sérios problemas de higiene e perdas econômicas devido à deterioração dos alimentos e deficiência dos equipamentos. Além disso, a contaminação por patógeno representa um grande risco para saúde. A incidência de listeriose é baixa, mas a taxa de letalidade é alta. Muitos dos focos principais de listeriose têm sido associados ao consumo de alimentos contaminados, especialmente os produtos lácteos. Assim, esse trabalho teve como objetivo analisar a capacidade de formação de biofilme, por 32 cepas de L. monocytogenes isoladas de diferentes alimentos, em diferentes materiais (aço inoxidável, vidro e poliestireno) e temperaturas (4º, 20º, 35ºC) observando a presença do gene luxS e também testar a eficácia de dois sanitizantes alcalino clorado, utilizados para sanitização de planta de processamento de alimentos. Os resultados demonstraram que entre as 32 cepas testadas 25 (78,1%) foram positivas para o gene luxS e a bactéria foi capaz de produzir biofilme em todos os materiais e temperaturas testadas, apresentando maior frequência de cepas produtoras nos materiais hidrofílicos (96,7% vidro e 95,6% inox) do que no hidrofóbico (28%). Os testes com sanitizantes mostraram que as concentrações recomendadas podem ser eficazes desde que utilizados em tempo mínimo...
Listeria monocytogenes is frequently found in many raw foods such as meat, cheese and vegetables, is widely distributed in the environment causing contamination of food during processing. Currently, contamination by L. monocytogenes in food is considered a major food security problems, because once present in the processing line, this bacteria is hardly eliminated by the great ability to form biofilm. The formation of biofilms on equipment can lead to serious health and economic losses due to spoilage and disability equipment. Furthermore, contamination pathogen is a great risk to health. The listeriosis incidence is low, but the fatality rate is high. Many of the main focuses of listeriosis have been linked to consumption of contaminated food, especially dairy products. Thus, this study aimed to analyze 32 strains of L. monocytogenes isolated from different foods to form biofilms on different materials (stainless steel, glass and polystyrene) and temperatures (4 º, 20 º, 35 º C) for the presence of the gene luxS and also test the effectiveness of two sanitizers alkaline chlorine used to sanitize the plant food processing. The results showed that the strains tested 25 (78.1%) were positive for the gene and the bacterium was able to produce biofilm in all materials and temperatures tested, with higher frequency of hydrophilic materials in producing strains (96.7% glass and 95.6% stainless) than in the hydrophobic (12.4%). The tests show that the sanitizing recommended concentrations may be effective if used in minimum time of 30 minutes for the product A and 15 for the product B has led to the elimination of 16 (50%) of the strains tested. We conclude that the gene luxS may be involved in the regulation of biofilm formation, but shouldn’t be the only one, once luxS-negative strains were also producers. We conclude... (Complete abstract click electronic access below)

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2

Leite, Andressa Rosa Perin [UNESP]. "Avaliação de aspectos relacionados á utilização de um adesivo para próteses totais convencionas: estudo in vitro". Universidade Estadual Paulista (UNESP), 2013. http://hdl.handle.net/11449/97326.

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O objetivo do presente estudo foi avaliar a influência da utilização de um adesivo para prótese (Ultra Corega creme) na formação de biofilme sobre a superfície interna de próteses totais e na microbiota bucal, no grau de satisfação, além da estimativa de custo médio diário do produto. Trinta pacientes receberam próteses totais novas, e foram divididos em dois protocolos: protocolo 1- utilização do adesivo durante os primeiros 15 dias de teste, seguida por não utilização de nenhum tipo de adesivo durante os próximos 15 dias; protocolo 2- não utilização de adesivo durante os primeiros 15 dias de teste, seguida por utilização do adesivo durante os próximos 15 dias. Após cada período de 15 dias, o biofilme formado na superfície interna das próteses totais foi corado e quantificado por meio de um método fotográfico com o auxílio de um software (Image Tool 3.00). Amostras de material da mucosa palatina e da superfície interna das próteses superiores foram plaqueadas em meios seletivos para Candida spp. e Streptococcus mutans e em um meio não seletivo. Ainda, foi aplicado um questionário para avaliação da satisfação com as próteses e o custo médio diário do produto foi estimado por meio de fórmulas matemáticas. Todas as análises foram realizadas com α = 0,05 e foram empregados testes apropriados à distribuição dos dados. Foi observada formação de biofilme semelhante com ou sem o uso do adesivo sobre as próteses superiores (Wilcoxon, p=0,255) e inferiores (Wilcoxon, p=0,433). Contagens de colônias semelhantes foram observadas com ou sem a utilização do adesivo na mucosa e na superfície interna da prótese total superior (p>0,05). O uso de adesivo proporcionou maior satisfação aos participantes (Wilcoxon, p=0,04). Em média, cada paciente utilizou 3,9 ± 0,3 gramas de adesivo por dia...
The aim of this study was to evaluate the influence of adhesive usage (Ultra Corega cream) on biofilm formation on the internal surface of dentures and oral microbiota, degree of satisfaction, and the estimated average daily cost of product. Thirty patients received new dentures, and have been divided into two protocols: Protocol 1-use of the adhesive during the first 15 days of the test, followed by not using adhesive over the next 15 days; Protocol 2- no use of adhesive for the first 15 days of the test, followed by use of adhesive over the next 15 days. After each period of 15 days, the internal surfaces of the dentures were stained and photographed and the areas (total internal surface and surface stained with biofilm) quantified (Image Tool 3.00). Samples of material from the palatal mucosa and the internal surface of the maxillary denture were plated on selective media for Candida spp. and Streptococcus mutans and a non-selective medium. A questionnaire was applied to evaluate satisfaction with the dentures and the average daily cost of the product was estimated by mathematical formulas. All analyzes were performed with α=.05 and appropriate tests were applied to the data distribution. Similar biofilm formation was found with or without adhesive usage for maxillary (Wilcoxon, p=.255) and mandibular dentures (Wilcoxon, p=.433). Similar colony counts were observed with or without adhesive for mucosa and the internal surface of dentures, irrespective of the culture medium (p>.05). The use of adhesive provided higher satisfaction (Wilcoxon, p=.04). On average, each patient used 3.9 ± 0.3 grams of adhesive per day, equivalent to R $ 4.02 for average daily cost. It is concluded that the use of the adhesive did not affect the quantification of the biofilm and the oral microbiota, besides providing greater overall satisfaction

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3

Klein, Marlise Inez. "Caracterização fisiologica e genetica de isolados clinicos de Streptococcus mutans". [s.n.], 2006. http://repositorio.unicamp.br/jspui/handle/REPOSIP/288642.

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Orientadores: Reginaldo Bruno Gonçalves, Renata de Oliveira Mattos-Graner
Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: Streptococcus mutans são os principais patógenos da cárie dentária, um importante problema de saúde pública no país. Para um melhor entendimento da biologia desta espécie bacteriana, o objetivo desta tese de doutorado foi avaliar características genéticas e fisiológicas relacionadas à virulência de isolados clínicos de S. mutans. Neste microrganismo, o sistema de indução da competência, "quorum-sensíng", é dependente da densidade celular e está envolvido na capacidade de crescimento em biofilme. Foram pesquisados, os genes de sistema "quorum-sensíng" e os genes envolvidos no processo de transformação da espécie S. mutans. Foi demonstrada a presença de todos os genes avaliados em S. mutans, bem como a presença de polimorfismo em alguns genes pesquisados, dados ainda inéditos na literatura. Posteriormente, genótipos clínicos de S. mutans foram submetidos a uma análise genética e fisiológica em que foram avaliadas a tolerância e adaptação ao pH ácido, a capacidade de formação de biofilme e curvas de crescimento sob diferentes condições (pHs e diferentes fontes de carbono). Os dados de expressão gênica confirmaram os resultados obtidos nas análises fisiológicas. As cepas clínicas apresentaram um comportamento heterogêneo frente aos mesmos desafios ambientais, o que pode favorecer a sobrevivência das mesmas na cavidade bucal, um ambiente com diversas condições de estresse. Os dados obtidos, comparados aos existentes na literatura, sugerem que além da atividade A TPase, os sistemas de transporte de açúcares fosfotransferase fosfoenolpiruvato:açúcar dependente (pep-PTS) também estariam envolvidos na tolerância de S. mutans ao estresse ácido. Em conjunto, as observações decorrentes das análises efetuadas, nesta tese, podem contribuir para a compreensão dos processos biológicos de S. mutans.
Abstract: Streptococcus mutans is considered the primary etiological agent of human dental caries, an important public health problem in Brazil. The purpose of this thesis was to evaluate genetic and physiologic properties related with virulence in clinical isolates of Streptococcus mutans. The quorum-sensing system, that induces and regulates genetic competence, depends on cellular density and also may play a role in biofilm growth and structure in S. mutans species. A screening of genes of the quorum-sensing system and genes involved with genetic transformation was performed. The analysis revealed that ali genes are widespread within the S. mutans species, and some genes presented polymorphisms. These data are new in the scientific literature. Furthermore, clinical genotypes of S. mutans were subjected to genetic and physiological analysis, including tolerance and acid adaptation to low pH, ability to form stable biofilm and growth kinetic under different conditions (pH and carbon source). The profile of gene expression confirms the data found in the physiologic assays. The studied strains shown heterogenic behavior at the same environmental challenge, that could favor these strains survive in the oral cavity, an environmental with several stress conditions. The data obtained here, and supported by scientific literature, suggest that besides ATPase activity' the sugar:phosphotransferase systems (PTS) could help to mount an adaptive acid tolerance response in S. mutans. Taken together all data obtained in this thesis may help to understand S. mutans biological processes.
Doutorado
Microbiologia e Imunologia
Doutor em Biologia Buco-Dental

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4

Bonsaglia, Erika Carolina Romão. "Produção de biofilme por Listeria monocytogenes isoladas de diferentes alimentos /". Botucatu, 2012. http://hdl.handle.net/11449/87783.

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Orientador: Vera Lúcia Mores Rall
Banca: José Paes de Almeida Nogueira
Banca: Ary Fernandes Junior
Resumo: Listeria monocytogenes é uma bactéria frequentemente encontrada em vários alimentos crus, como carne, queijo e legumes. Está amplamente distribuída no ambiente causando a contaminação de alimentos durante seu processamento. Atualmente, a contaminação por L. monocytogenes nas indústrias alimentícias é considerada um dos maiores problemas de segurança alimentar, pois uma vez presente na linha de processamento à bactéria dificilmente é eliminada pela grande capacidade em formar biofilme. A formação de biofilmes em equipamentos pode levar a sérios problemas de higiene e perdas econômicas devido à deterioração dos alimentos e deficiência dos equipamentos. Além disso, a contaminação por patógeno representa um grande risco para saúde. A incidência de listeriose é baixa, mas a taxa de letalidade é alta. Muitos dos focos principais de listeriose têm sido associados ao consumo de alimentos contaminados, especialmente os produtos lácteos. Assim, esse trabalho teve como objetivo analisar a capacidade de formação de biofilme, por 32 cepas de L. monocytogenes isoladas de diferentes alimentos, em diferentes materiais (aço inoxidável, vidro e poliestireno) e temperaturas (4º, 20º, 35ºC) observando a presença do gene luxS e também testar a eficácia de dois sanitizantes alcalino clorado, utilizados para sanitização de planta de processamento de alimentos. Os resultados demonstraram que entre as 32 cepas testadas 25 (78,1%) foram positivas para o gene luxS e a bactéria foi capaz de produzir biofilme em todos os materiais e temperaturas testadas, apresentando maior frequência de cepas produtoras nos materiais hidrofílicos (96,7% vidro e 95,6% inox) do que no hidrofóbico (28%). Os testes com sanitizantes mostraram que as concentrações recomendadas podem ser eficazes desde que utilizados em tempo mínimo... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Listeria monocytogenes is frequently found in many raw foods such as meat, cheese and vegetables, is widely distributed in the environment causing contamination of food during processing. Currently, contamination by L. monocytogenes in food is considered a major food security problems, because once present in the processing line, this bacteria is hardly eliminated by the great ability to form biofilm. The formation of biofilms on equipment can lead to serious health and economic losses due to spoilage and disability equipment. Furthermore, contamination pathogen is a great risk to health. The listeriosis incidence is low, but the fatality rate is high. Many of the main focuses of listeriosis have been linked to consumption of contaminated food, especially dairy products. Thus, this study aimed to analyze 32 strains of L. monocytogenes isolated from different foods to form biofilms on different materials (stainless steel, glass and polystyrene) and temperatures (4 º, 20 º, 35 º C) for the presence of the gene luxS and also test the effectiveness of two sanitizers alkaline chlorine used to sanitize the plant food processing. The results showed that the strains tested 25 (78.1%) were positive for the gene and the bacterium was able to produce biofilm in all materials and temperatures tested, with higher frequency of hydrophilic materials in producing strains (96.7% glass and 95.6% stainless) than in the hydrophobic (12.4%). The tests show that the sanitizing recommended concentrations may be effective if used in minimum time of 30 minutes for the product A and 15 for the product B has led to the elimination of 16 (50%) of the strains tested. We conclude that the gene luxS may be involved in the regulation of biofilm formation, but shouldn't be the only one, once luxS-negative strains were also producers. We conclude... (Complete abstract click electronic access below)
Mestre

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5

Leite, Andressa Rosa Perin. "Avaliação de aspectos relacionados á utilização de um adesivo para próteses totais convencionas: estudo in vitro /". Araraquara, 2013. http://hdl.handle.net/11449/97326.

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Orientador: Ana Carolina Pero
Banca: Marco Antonio Compagnoni
Banca: Raphael Freitas de Souza
Resumo: O objetivo do presente estudo foi avaliar a influência da utilização de um adesivo para prótese (Ultra Corega creme) na formação de biofilme sobre a superfície interna de próteses totais e na microbiota bucal, no grau de satisfação, além da estimativa de custo médio diário do produto. Trinta pacientes receberam próteses totais novas, e foram divididos em dois protocolos: protocolo 1- utilização do adesivo durante os primeiros 15 dias de teste, seguida por não utilização de nenhum tipo de adesivo durante os próximos 15 dias; protocolo 2- não utilização de adesivo durante os primeiros 15 dias de teste, seguida por utilização do adesivo durante os próximos 15 dias. Após cada período de 15 dias, o biofilme formado na superfície interna das próteses totais foi corado e quantificado por meio de um método fotográfico com o auxílio de um software (Image Tool 3.00). Amostras de material da mucosa palatina e da superfície interna das próteses superiores foram plaqueadas em meios seletivos para Candida spp. e Streptococcus mutans e em um meio não seletivo. Ainda, foi aplicado um questionário para avaliação da satisfação com as próteses e o custo médio diário do produto foi estimado por meio de fórmulas matemáticas. Todas as análises foram realizadas com α = 0,05 e foram empregados testes apropriados à distribuição dos dados. Foi observada formação de biofilme semelhante com ou sem o uso do adesivo sobre as próteses superiores (Wilcoxon, p=0,255) e inferiores (Wilcoxon, p=0,433). Contagens de colônias semelhantes foram observadas com ou sem a utilização do adesivo na mucosa e na superfície interna da prótese total superior (p>0,05). O uso de adesivo proporcionou maior satisfação aos participantes (Wilcoxon, p=0,04). Em média, cada paciente utilizou 3,9 ± 0,3 gramas de adesivo por dia... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The aim of this study was to evaluate the influence of adhesive usage (Ultra Corega cream) on biofilm formation on the internal surface of dentures and oral microbiota, degree of satisfaction, and the estimated average daily cost of product. Thirty patients received new dentures, and have been divided into two protocols: Protocol 1-use of the adhesive during the first 15 days of the test, followed by not using adhesive over the next 15 days; Protocol 2- no use of adhesive for the first 15 days of the test, followed by use of adhesive over the next 15 days. After each period of 15 days, the internal surfaces of the dentures were stained and photographed and the areas (total internal surface and surface stained with biofilm) quantified (Image Tool 3.00). Samples of material from the palatal mucosa and the internal surface of the maxillary denture were plated on selective media for Candida spp. and Streptococcus mutans and a non-selective medium. A questionnaire was applied to evaluate satisfaction with the dentures and the average daily cost of the product was estimated by mathematical formulas. All analyzes were performed with α=.05 and appropriate tests were applied to the data distribution. Similar biofilm formation was found with or without adhesive usage for maxillary (Wilcoxon, p=.255) and mandibular dentures (Wilcoxon, p=.433). Similar colony counts were observed with or without adhesive for mucosa and the internal surface of dentures, irrespective of the culture medium (p>.05). The use of adhesive provided higher satisfaction (Wilcoxon, p=.04). On average, each patient used 3.9 ± 0.3 grams of adhesive per day, equivalent to R $ 4.02 for average daily cost. It is concluded that the use of the adhesive did not affect the quantification of the biofilm and the oral microbiota, besides providing greater overall satisfaction
Doutor

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6

Brain, Stephen. "Monitoring microbial biofilms". Thesis, London South Bank University, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.337401.

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7

Pinto, Geraldo Camilo de Souza [UNESP]. "Eficácia da terapia fotodinâmica antimicrobiana em biofilmes de Staphylococcus Aureus suscetível e resistente á meticilina". Universidade Estadual Paulista (UNESP), 2013. http://hdl.handle.net/11449/97310.

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A necessidade de superar o desafio criado pelos biofilmes resistentes aos tratamentos antimicrobianos convencionais tem levado à busca por tratamentos alternativos, como terapia fotodinâmica antimicrobiana (aPDT). Este estudo avaliou in vitro a eficácia da aPDT na inativação de biofilmes de Staphylococcus aureus suscetíveis e resistentes à meticilina (MRSA e MSSA), mediado pelos fotossensibilizadores (PSs) Curcumina (Cur) e Photodithazine® (PDZ). Biofilmes foram formados e tratados com diferentes concentrações de Cur (0, 20, 40 e 80 μM) e PDZ (0, 50 e 75 mg/L), e iluminados ou não por fonte de luz LED (Cur 455 ± 3 nm/ 5,28 J/cm2; PDZ 660 ± 3 nm/ 5,28 J/cm2 ou 50 J/cm²). Os grupos Controle Positivo (CP) não receberam nenhum PS e também não foram iluminados. A viabilidade dos micro-organismos após a aPDT foi avaliado pelo número de colônias viáveis, pelo ensaio de XTT e pela utilização do kit LIVE/DEAD® na Microscopia Confocal de Varredura à Laser (MCVL). Os resultados foram avaliados por análises de variância de dois fatores de efeitos fixos (ANOVA) e complementados por comparações múltiplas de médias pelo teste de Tukey. Para ambas as cepas, todas as concentrações de Cur e PDZ testadas reduziram significativamente a atividade metabólica e o UFC/mL para ambos micro-organismos quando comparado com os grupos CN (p0,05). Os resultados foram otimizados para a Cur quando utilizou-se a maior concentração (80 μM), para a PDZ, a maior redução nos micro-organismos foi observada quando associou-se a maior concentração de PDZ (75 mg/L) com a maior dose de luz (50 J/cm²). Os biofilmes submetidos a aPDT demostraram pela MCVL um maior número de células coradas em vermelho, indicando que a aPDT foi eficaz para promover danos ou morte às células bacterianas. Assim, a aPDT pode ser considerada promissora para atuar de forma sinérgica no tratamento de infecções bacterianas
The need to overcome the challenge created by biofilms regarding conventional antimicrobial approaches has lead to search of alternative treatments such as Antimicrobial Photodynamic Therapy (aPDT). This in vitro study evaluated the efficacy of aPDT using the photosensitizer (PS) Curcumin (Cur) and Photodithazine® (PDZ) in the inactivation of biofilms of methicillin susceptible and resistant S. aureus (MSSA and MRSA). Biofilms were treated with different Cur (0, 20, 40 or 80 μM of Cur) and PDZ concentrations (0, 50 or 75 mg/L) and illuminated or not with LED source (Cur 455 ± 3 nm/ 5.28 J/cm2; PDZ 660 ± 3 nm/ 5.28 J/cm2 or 50 J/cm²). Positive control samples were not exposed to PS or light. The microorganisms viability after aPDT were evaluated by counting the number of colonies, the XTT assay and LIVE/DEAD® staining using confocal laser scanning microscopy (CLSM). The results were evaluated by analysis of variance, two-factor fixed effects (ANOVA) and complemented by multiple comparisons by Tukey test. For both strains, all the tested Cur and PDZ concentrations reduced significantly both biofilm metabolic activity and CFU/mL compared to the negative control (p0.05). Moreover, the results were optimized for Cur when the higher concentration was used (80 μM); For PDZ, the best results were obtained when it was associated a higher concentration of PDZ (75 mg/L) with the higher dose of light (50 J/cm²). Biofilms submitted to aPDT showed a large number of red-stained colonies, indicating that this therapy was efficient in disrupting the bacterial membrane. It can be concluded that PS was efficient in reducing viable colonies of both S. aureus strains by damaging cell membrane and causing cell death. Thus, the aPDT is can be considered promising to act synergistically in the treatment of bacterial infections

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Quishida, Cristiane Campos Costa [UNESP]. "Estudo da eficácia da terapia fotodinâmica, mediada pelos fotossensibilizadores Photodithazine® e Curcumina, sobre biofilmes multi-espécies formados por Streptococcus mutans, Candida albicans e Candida glabrata". Universidade Estadual Paulista (UNESP), 2013. http://hdl.handle.net/11449/105516.

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A capacidade de aderência dos micro-organismos a diferentes superfícies, a resistência a medicamentos, bem como a interação entre as espécies visam garantir a sobrevivência e a proliferação dos mesmos, resultando em menor susceptibilidade dos patógenos aos tratamentos e aos procedimentos de desinfecção. Este estudo teve como objetivo avaliar in vitro a eficácia da Terapia Fotodinâmica (PDT) mediada pelos fotossensibilizadores (FSs) Photodithazine® (PDZ) e Curcumina (Cur) sobre biofilmes multi-espécies compostos por Streptococcus mutans, Candida albicans e Candida glabrata. Para isso, o estudo foi divido em três partes: 1- avaliação do efeito fotodinâmico das diferentes concentrações de PDZ (100, 150, 175, 200 e 250mg/L) associada à luz LED (660nm/ 37,5J/cm2) na inativação de biofilme multi-espécies formado em fundo de placa de 96 orifícios; 2- avaliação do efeito fotodinâmico quando realizadas uma e três aplicações sucessivas de PDT, mediada por PDZ (175 e 200mg/L) e luz LED (660nm/ 37,5J/cm2) na inativação de biofilmes multi-espécies, formado sobre corpos-de-prova de resina acrílica. 3- avaliação do efeito fotodinâmico de diferentes concentrações do FS Cur (80, 100 e 120μM) associada à luz LED (455nm/ 37,5J/cm2) sobre biofilmes multi-espécies cultivados por diferentes períodos (24 e 48h) sobre corpos-de-prova de resina acrílica. Adicionalmente para cada condição avaliada foi verificado o efeito da aplicação isolada de cada concentração dos FSs PDZ e Cur em contato com o biofilme na ausência de luz (P+L-) e o efeito isolado da luz (P-L+). Além disso, foi realizado o grupo controle positivo, no qual o biofilme não esteve em contato com o FS e não foram iluminados com luz LED (P-L-). Após a aplicação dos tratamentos propostos, a viabilidade dos micro-organismos foi avaliada por meio da contagem de colônias (UFC/mL), teste do XTT, determinação da...
The ability of adhesion of microrganisms to different surfaces, the drug resistance, as well as the interaction among the species help to ensure the survival and proliferation of these microrganisms and result in lower susceptibility of these pathogens to treatment and disinfection procedures. This study aimed to evaluate the in vitro efficacy of PDT mediated by photosensitizers (FSs) Photodithazine ® (PDZ) and Curcumin (Cur) against multispecies biofilms formed by Candida albicans, Candida glabrata and Streptococcus mutans. This study was divided into three parts: 1. evaluation of the effect of photodynamic therapy of different concentrations of PDZ (100, 150, 175, 200 and 250mg / L) associated with the LED light (660 nm / 37.5 J/cm2) in the inactivation of multspecies biofilms formed in 96-well microtiter plates; 2. evaluation of the photodynamic effect of one PDT applications only and three successive PDT applications mediated by PDZ (175 and 200 mg / L) and LED light (660 nm / 37.5 J/cm2) in the inactivation of multispecies biofilms, formed on acrylic resin samples; 3. evaluation of the photodynamic effect of different concentrations of Cur FS (80, 100, and 120μM) associated with LED light (455nm / 37.5 J/cm2) against multispecies biofilms grown during different periods (24 and 48h) on acrylic resin samples. Additional samples were treated either with FSs (PDZ or Cur) (P+L-) or LED light (P-L+) only. Positive control samples had neither light nor FSs (PDZ or Cur) (P-L-). After applying the proposed treatments, the viability of microrganisms was assessed by Colony Count (CFU / mL), the XTT assay, determination of the total biomass (CV) and confocal laser scanning microscopy (CLSM).The results were analyzed using ANOVA or Kruskal-Wallis test followed by Tukey or Dunn, respectively. For the biofilm formed in 96-well plate, the concentrations of 175 and 200mg/L showed greater reduction in cell...

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Monteiro, Douglas Roberto [UNESP]. "Análise da ação de nanopartículas de prata sobre o biofilme de espécies de Candida". Universidade Estadual Paulista (UNESP), 2012. http://hdl.handle.net/11449/105564.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
O objetivo deste estudo foi avaliar a atividade antifúngica de nanopartículas de prata (NP) contra os biofilmes de Candida albicans e Candida glabrata. NP foram sintetizadas por meio da redução do nitrato de prata com citrato de sódio e estabilizadas com amônia ou polivinilpirrolidona. Os testes de concentração inibitória mínima (CIM) das NP contra células de Candida foram baseados no método da microdiluição. NP foram aplicadas sobre os biofilmes de Candida (48 horas) e após 24 horas de contato sua atividade antifúngica foi determinada por meio da quantificação da biomassa total (coloração com violeta cristal (VC)) e por meio da enumeração das unidades formadoras de colônias (UFCs). Após o tratamento com NP, as matrizes dos biofilmes foram extraídas e analisadas em termos de proteínas, carboidratos e DNA, e a estrutura dos biofilmes foi analisada por meio da microscopia eletrônica de varredura e de epifluorescência. A atividade antibiofilme da combinação de NP com nistatina e clorexidina foi avaliada por meio dos ensaios de VC e UFCs. Leveduras viáveis foram recuperadas a partir dos biofilmes previamente tratados com NP e adicionadas às células epiteliais HeLa e aos poços de placas de poliestireno e, após 2 horas de contato, a adesão foi determinada usando VC. A eficácia de NP submetidas às variações de temperatura (50, 70 e 100ºC) e pH (5 e 9) também foi avaliada, assim como a susceptibilidade às NP dos biofilmes de Candida em diferentes fases de crescimento. Os resultados de CIM mostraram que NP foram fungicidas contra os isolados testados em concentrações baixas (0,4-3,3 μg/mL). NP foram mais efetivas na redução da biomassa para os biofilmes de C. glabrata (reduções de 90% na concentração de 108 μg/mL) do que para C. albicans, e promoveram reduções significativas no log10 do número de UFCs...
The aim of this study was to evaluate the antifungal activity of silver nanoparticles (SN) against Candida albicans and Candida glabrata biofilms. Colloidal suspensions of SN were synthesized by reducing silver nitrate with sodium citrate and stabilized with ammonia or polyvinylpyrrolidone. Minimal inhibitory concentrations (MIC) were performed for Candida cells grown in suspension following the microbroth dilution method. Candida biofilms (48 h) were treated with SN for 24 h and then the total biomass quantification (by crystal violet (CV) staining) and the colony forming units (CFUs) were determined. Also, after treating with SN, extracellular matrices were extracted from Candida biofilms and analyzed chemically in terms of proteins, carbohydrates and DNA. To investigate the biofilm structure, scanning electron microscopy and epifluorescence microscopy were carried out. The antibiofilm activity of SN in combination with nystatin and chlorhexidine was also assessed by CV and CFU. Moreover, viable yeasts were recovered from the biofilms pretreated with SN and added to HeLa epithelial cells or to empty wells of polystyrene plates and, after 2 h of contact, the adhesion capacity of the yeasts was determined by using CV staining. The antibiofilm efficacy of SN subjected to variations of temperature (50, 70 and 100ºC) and pH (5 and 9) was also evaluated. Finally, the susceptibility to SN of biofilms in different stages of growth was analyzed. MIC results showed that SN were fungicidal against the tested strains at very low concentrations (0.4- 3.3 μg/mL). SN were more effective in reducing the total biomass of C. glabrata (reductions around 90% at 108 μg/mL SN) than C. albicans biofilms, and provided significant log10 reduction of... (Complete abstract click electronic access below)

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Francisconi, Renata Serignoli [UNESP]. "Efeito do óleo essencial de Melaleuca alternifolia e de seu principal componente Terpinen-4-ol sobre isolados clínicos de Candida albicans resistentes". Universidade Estadual Paulista (UNESP), 2014. http://hdl.handle.net/11449/116010.

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O óleo essencial de Melaleuca alternifolia (TTO) é um extrato de ação antifúngica e preventiva em escala farmacêutica ou cosmética. O objetivo deste estudo foi avaliar a eficácia do TTO e Terpinen-4-ol sobre isolados clínicos de Candida albicans. Este estudo foi realizado em três fases: 1- Identificação da CIM (Concentração Inibitória Mínima) e CFM (Concentração Fungicida Mínima) do TTO (0,25 a 2%) e Terpinen-4- ol (0,11 a 0,95%) sobre C. albicans na forma planctônica. 2- Análise das diferentes concentrações do óleo sobre biofilme monoespécie de C. albicans por meio da contagem das UFC/mL e avaliação da atividade metabólica das células pelo método colorimétrico de XTT. 3- Análise da ação inibitória das soluções de TTO e Terpinen-4- ol sobre biofilmes de C.albicans (cepa padrão e isolados clínicos) formados em corpos de prova de resina acrílica termopolimerizável previamente cobertos com saliva humana, por meio do teste de XTT e por microscopia confocal de varredura à laser (MCVL). A nistatina (Sigma) foi utilizada como controle positivo. Os resultados mostraram que isolados de C. albicans planctônicos foram sensíveis ao TTO 1%, Terpinen-4-ol 0,47% e Nistatina 8 ?g/mL. As menores concentrações fungicidas para os isolados foram TTO 2 %, Terpinen-4-ol 0,95 % e Nistatina 16 ?g/mL. Quando analisado em biofilme através da quantificação (UFC/mL) e teste de XTT as concentrações de TTO 2 % e Terpinen-4-ol 0,95 % foram eficazes quando comparado ao controle, sendo que as amostras da genotipagem A2 e B1 foram as mais resistentes. Os resultados de MCVL mostraram que todos os biofilmes desenvolvidos em corpos de resina acrílica apresentaram-se semelhantes à ação da Nistatina. Os extratos avaliados apresentaram ação antifúngica para os isolados clínicos e podem ser considerados tratamento alternativo para paciente com candidíase.
The essential oil of Melaleuca alternifolia (TTO) is an extract of antifungal action and preventive in pharmaceutical or cosmetic scale. The aim of this study was to evaluate the efficacy of TTO and Terpinen-4-ol against clinical isolates of Candida albicans. This study was conducted in three phases: 1- Identification of MIC (Minimum Inhibitory Concentration) and CFM (Minimum Fungicidal Concentration) of TTO (0.25 to 2%) and Terpinen-4-ol (0.11 to 0.95 %) on C. albicans in planktonic form . 2- Analysis of different concentrations of oil on C. albicans biofilm single species by counting CFU/mL and evaluation of the metabolic activity of cells by XTT colorimetric method. 3- Analysis of the inhibitory action of TTO and Terpinen-4-ol solutions on C. albicans biofilms (standard strain and clinical isolates) formed in specimens of polymerizable resin acrylic microwave previously coated with human saliva , by means of the XTT test and confocal laser scanning microscopy (CLSM) . The nystatin (Sigma) was used as a positive control. The results showed that isolates of C. albicans planktonic were sensitive to TTO 1%, Terpinen-4-ol 0.47% and Nystatin 8 mg/mL. The smaller fungicidal concentrations for the isolates were TTO 2%, Terpinen-4-ol 0.95% and Nystatin 16 mg / mL. When analyzed by quantifying biofilm (CFU / ml) and XTT test, the concentrations TTO 2% and Terpinen-4-ol 0.95% were effective when compared to the control, and genotyping of samples A2 and B1 were more resistant. The CLSM results showed that all biofilms developed in bodies of acrylic resin were similar to the action of Nystatin. The extracts showed antifungal action for the clinical isolates and can be considered an alternative treatment for patients with candidiasis.

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11

Francisconi, Renata Serignoli. "Efeito do óleo essencial de Melaleuca alternifolia e de seu principal componente Terpinen-4-ol sobre isolados clínicos de Candida albicans resistentes /". Araraquara, 2014. http://hdl.handle.net/11449/116010.

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Orientador: Denise Madalena Palomari Spolidorio
Banca: Cristiane Duque
Banca: Regina Maria Puppin-Rontani
Resumo: O óleo essencial de Melaleuca alternifolia (TTO) é um extrato de ação antifúngica e preventiva em escala farmacêutica ou cosmética. O objetivo deste estudo foi avaliar a eficácia do TTO e Terpinen-4-ol sobre isolados clínicos de Candida albicans. Este estudo foi realizado em três fases: 1- Identificação da CIM (Concentração Inibitória Mínima) e CFM (Concentração Fungicida Mínima) do TTO (0,25 a 2%) e Terpinen-4- ol (0,11 a 0,95%) sobre C. albicans na forma planctônica. 2- Análise das diferentes concentrações do óleo sobre biofilme monoespécie de C. albicans por meio da contagem das UFC/mL e avaliação da atividade metabólica das células pelo método colorimétrico de XTT. 3- Análise da ação inibitória das soluções de TTO e Terpinen-4- ol sobre biofilmes de C.albicans (cepa padrão e isolados clínicos) formados em corpos de prova de resina acrílica termopolimerizável previamente cobertos com saliva humana, por meio do teste de XTT e por microscopia confocal de varredura à laser (MCVL). A nistatina (Sigma) foi utilizada como controle positivo. Os resultados mostraram que isolados de C. albicans planctônicos foram sensíveis ao TTO 1%, Terpinen-4-ol 0,47% e Nistatina 8 μg/mL. As menores concentrações fungicidas para os isolados foram TTO 2 %, Terpinen-4-ol 0,95 % e Nistatina 16 μg/mL. Quando analisado em biofilme através da quantificação (UFC/mL) e teste de XTT as concentrações de TTO 2 % e Terpinen-4-ol 0,95 % foram eficazes quando comparado ao controle, sendo que as amostras da genotipagem A2 e B1 foram as mais resistentes. Os resultados de MCVL mostraram que todos os biofilmes desenvolvidos em corpos de resina acrílica apresentaram-se semelhantes à ação da Nistatina. Os extratos avaliados apresentaram ação antifúngica para os isolados clínicos e podem ser considerados tratamento alternativo para paciente com candidíase.
Abstract: The essential oil of Melaleuca alternifolia (TTO) is an extract of antifungal action and preventive in pharmaceutical or cosmetic scale. The aim of this study was to evaluate the efficacy of TTO and Terpinen-4-ol against clinical isolates of Candida albicans. This study was conducted in three phases: 1- Identification of MIC (Minimum Inhibitory Concentration) and CFM (Minimum Fungicidal Concentration) of TTO (0.25 to 2%) and Terpinen-4-ol (0.11 to 0.95 %) on C. albicans in planktonic form . 2- Analysis of different concentrations of oil on C. albicans biofilm single species by counting CFU/mL and evaluation of the metabolic activity of cells by XTT colorimetric method. 3- Analysis of the inhibitory action of TTO and Terpinen-4-ol solutions on C. albicans biofilms (standard strain and clinical isolates) formed in specimens of polymerizable resin acrylic microwave previously coated with human saliva , by means of the XTT test and confocal laser scanning microscopy (CLSM) . The nystatin (Sigma) was used as a positive control. The results showed that isolates of C. albicans planktonic were sensitive to TTO 1%, Terpinen-4-ol 0.47% and Nystatin 8 mg/mL. The smaller fungicidal concentrations for the isolates were TTO 2%, Terpinen-4-ol 0.95% and Nystatin 16 mg / mL. When analyzed by quantifying biofilm (CFU / ml) and XTT test, the concentrations TTO 2% and Terpinen-4-ol 0.95% were effective when compared to the control, and genotyping of samples A2 and B1 were more resistant. The CLSM results showed that all biofilms developed in bodies of acrylic resin were similar to the action of Nystatin. The extracts showed antifungal action for the clinical isolates and can be considered an alternative treatment for patients with candidiasis.
Mestre

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12

Monteiro, Douglas Roberto. "Análise da ação de nanopartículas de prata sobre o biofilme de espécies de Candida /". Araçatuba, 2012. http://hdl.handle.net/11449/105564.

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Orientador: Débora Barros Barbosa
Coorientador: Fernanda Lourenção Brighenti
Banca: Alberto Carlos Botazzo Delbem
Banca: Juliano Pelim Pessan
Banca: Alessandra Marçal Agostinho
Banca: Emerson Rodrigues de Camargo
Resumo: O objetivo deste estudo foi avaliar a atividade antifúngica de nanopartículas de prata (NP) contra os biofilmes de Candida albicans e Candida glabrata. NP foram sintetizadas por meio da redução do nitrato de prata com citrato de sódio e estabilizadas com amônia ou polivinilpirrolidona. Os testes de concentração inibitória mínima (CIM) das NP contra células de Candida foram baseados no método da microdiluição. NP foram aplicadas sobre os biofilmes de Candida (48 horas) e após 24 horas de contato sua atividade antifúngica foi determinada por meio da quantificação da biomassa total (coloração com violeta cristal (VC)) e por meio da enumeração das unidades formadoras de colônias (UFCs). Após o tratamento com NP, as matrizes dos biofilmes foram extraídas e analisadas em termos de proteínas, carboidratos e DNA, e a estrutura dos biofilmes foi analisada por meio da microscopia eletrônica de varredura e de epifluorescência. A atividade antibiofilme da combinação de NP com nistatina e clorexidina foi avaliada por meio dos ensaios de VC e UFCs. Leveduras viáveis foram recuperadas a partir dos biofilmes previamente tratados com NP e adicionadas às células epiteliais HeLa e aos poços de placas de poliestireno e, após 2 horas de contato, a adesão foi determinada usando VC. A eficácia de NP submetidas às variações de temperatura (50, 70 e 100ºC) e pH (5 e 9) também foi avaliada, assim como a susceptibilidade às NP dos biofilmes de Candida em diferentes fases de crescimento. Os resultados de CIM mostraram que NP foram fungicidas contra os isolados testados em concentrações baixas (0,4-3,3 μg/mL). NP foram mais efetivas na redução da biomassa para os biofilmes de C. glabrata (reduções de 90% na concentração de 108 μg/mL) do que para C. albicans, e promoveram reduções significativas no log10 do número de UFCs... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The aim of this study was to evaluate the antifungal activity of silver nanoparticles (SN) against Candida albicans and Candida glabrata biofilms. Colloidal suspensions of SN were synthesized by reducing silver nitrate with sodium citrate and stabilized with ammonia or polyvinylpyrrolidone. Minimal inhibitory concentrations (MIC) were performed for Candida cells grown in suspension following the microbroth dilution method. Candida biofilms (48 h) were treated with SN for 24 h and then the total biomass quantification (by crystal violet (CV) staining) and the colony forming units (CFUs) were determined. Also, after treating with SN, extracellular matrices were extracted from Candida biofilms and analyzed chemically in terms of proteins, carbohydrates and DNA. To investigate the biofilm structure, scanning electron microscopy and epifluorescence microscopy were carried out. The antibiofilm activity of SN in combination with nystatin and chlorhexidine was also assessed by CV and CFU. Moreover, viable yeasts were recovered from the biofilms pretreated with SN and added to HeLa epithelial cells or to empty wells of polystyrene plates and, after 2 h of contact, the adhesion capacity of the yeasts was determined by using CV staining. The antibiofilm efficacy of SN subjected to variations of temperature (50, 70 and 100ºC) and pH (5 and 9) was also evaluated. Finally, the susceptibility to SN of biofilms in different stages of growth was analyzed. MIC results showed that SN were fungicidal against the tested strains at very low concentrations (0.4- 3.3 μg/mL). SN were more effective in reducing the total biomass of C. glabrata (reductions around 90% at 108 μg/mL SN) than C. albicans biofilms, and provided significant log10 reduction of... (Complete abstract click electronic access below)
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Pinto, Geraldo Camilo de Souza. "Eficácia da terapia fotodinâmica antimicrobiana em biofilmes de Staphylococcus Aureus suscetível e resistente á meticilina /". Araraquara, 2013. http://hdl.handle.net/11449/97310.

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Abstract (sommario):

Orientador: Ana Claúdia Pavarina
Banca: Eunice Teresinha Giampaolo
Banca: Ana Paula Dias Ribeiro
Resumo: A necessidade de superar o desafio criado pelos biofilmes resistentes aos tratamentos antimicrobianos convencionais tem levado à busca por tratamentos alternativos, como terapia fotodinâmica antimicrobiana (aPDT). Este estudo avaliou in vitro a eficácia da aPDT na inativação de biofilmes de Staphylococcus aureus suscetíveis e resistentes à meticilina (MRSA e MSSA), mediado pelos fotossensibilizadores (PSs) Curcumina (Cur) e Photodithazine® (PDZ). Biofilmes foram formados e tratados com diferentes concentrações de Cur (0, 20, 40 e 80 μM) e PDZ (0, 50 e 75 mg/L), e iluminados ou não por fonte de luz LED (Cur 455 ± 3 nm/ 5,28 J/cm2; PDZ 660 ± 3 nm/ 5,28 J/cm2 ou 50 J/cm²). Os grupos Controle Positivo (CP) não receberam nenhum PS e também não foram iluminados. A viabilidade dos micro-organismos após a aPDT foi avaliado pelo número de colônias viáveis, pelo ensaio de XTT e pela utilização do kit LIVE/DEAD® na Microscopia Confocal de Varredura à Laser (MCVL). Os resultados foram avaliados por análises de variância de dois fatores de efeitos fixos (ANOVA) e complementados por comparações múltiplas de médias pelo teste de Tukey. Para ambas as cepas, todas as concentrações de Cur e PDZ testadas reduziram significativamente a atividade metabólica e o UFC/mL para ambos micro-organismos quando comparado com os grupos CN (p0,05). Os resultados foram otimizados para a Cur quando utilizou-se a maior concentração (80 μM), para a PDZ, a maior redução nos micro-organismos foi observada quando associou-se a maior concentração de PDZ (75 mg/L) com a maior dose de luz (50 J/cm²). Os biofilmes submetidos a aPDT demostraram pela MCVL um maior número de células coradas em vermelho, indicando que a aPDT foi eficaz para promover danos ou morte às células bacterianas. Assim, a aPDT pode ser considerada promissora para atuar de forma sinérgica no tratamento de infecções bacterianas
Abstract: The need to overcome the challenge created by biofilms regarding conventional antimicrobial approaches has lead to search of alternative treatments such as Antimicrobial Photodynamic Therapy (aPDT). This in vitro study evaluated the efficacy of aPDT using the photosensitizer (PS) Curcumin (Cur) and Photodithazine® (PDZ) in the inactivation of biofilms of methicillin susceptible and resistant S. aureus (MSSA and MRSA). Biofilms were treated with different Cur (0, 20, 40 or 80 μM of Cur) and PDZ concentrations (0, 50 or 75 mg/L) and illuminated or not with LED source (Cur 455 ± 3 nm/ 5.28 J/cm2; PDZ 660 ± 3 nm/ 5.28 J/cm2 or 50 J/cm²). Positive control samples were not exposed to PS or light. The microorganisms viability after aPDT were evaluated by counting the number of colonies, the XTT assay and LIVE/DEAD® staining using confocal laser scanning microscopy (CLSM). The results were evaluated by analysis of variance, two-factor fixed effects (ANOVA) and complemented by multiple comparisons by Tukey test. For both strains, all the tested Cur and PDZ concentrations reduced significantly both biofilm metabolic activity and CFU/mL compared to the negative control (p0.05). Moreover, the results were optimized for Cur when the higher concentration was used (80 μM); For PDZ, the best results were obtained when it was associated a higher concentration of PDZ (75 mg/L) with the higher dose of light (50 J/cm²). Biofilms submitted to aPDT showed a large number of red-stained colonies, indicating that this therapy was efficient in disrupting the bacterial membrane. It can be concluded that PS was efficient in reducing viable colonies of both S. aureus strains by damaging cell membrane and causing cell death. Thus, the aPDT is can be considered promising to act synergistically in the treatment of bacterial infections
Mestre

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Quishida, Cristiane Campos Costa. "Estudo da eficácia da terapia fotodinâmica, mediada pelos fotossensibilizadores Photodithazine® e Curcumina, sobre biofilmes multi-espécies formados por Streptococcus mutans, Candida albicans e Candida glabrata /". Araraquara : [s.n.], 2013. http://hdl.handle.net/11449/105516.

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Orientador: Ana Claúdia Pavarina
Banca: Juliana Campos Junqueira
Banca: Claúdia Helena Lovato da Silva
Banca: Ewerton Garcia de Oliveira Mima
Banca: Eunice Teresinha Giampaolo
Resumo: A capacidade de aderência dos micro-organismos a diferentes superfícies, a resistência a medicamentos, bem como a interação entre as espécies visam garantir a sobrevivência e a proliferação dos mesmos, resultando em menor susceptibilidade dos patógenos aos tratamentos e aos procedimentos de desinfecção. Este estudo teve como objetivo avaliar in vitro a eficácia da Terapia Fotodinâmica (PDT) mediada pelos fotossensibilizadores (FSs) Photodithazine® (PDZ) e Curcumina (Cur) sobre biofilmes multi-espécies compostos por Streptococcus mutans, Candida albicans e Candida glabrata. Para isso, o estudo foi divido em três partes: 1- avaliação do efeito fotodinâmico das diferentes concentrações de PDZ (100, 150, 175, 200 e 250mg/L) associada à luz LED (660nm/ 37,5J/cm2) na inativação de biofilme multi-espécies formado em fundo de placa de 96 orifícios; 2- avaliação do efeito fotodinâmico quando realizadas uma e três aplicações sucessivas de PDT, mediada por PDZ (175 e 200mg/L) e luz LED (660nm/ 37,5J/cm2) na inativação de biofilmes multi-espécies, formado sobre corpos-de-prova de resina acrílica. 3- avaliação do efeito fotodinâmico de diferentes concentrações do FS Cur (80, 100 e 120μM) associada à luz LED (455nm/ 37,5J/cm2) sobre biofilmes multi-espécies cultivados por diferentes períodos (24 e 48h) sobre corpos-de-prova de resina acrílica. Adicionalmente para cada condição avaliada foi verificado o efeito da aplicação isolada de cada concentração dos FSs PDZ e Cur em contato com o biofilme na ausência de luz (P+L-) e o efeito isolado da luz (P-L+). Além disso, foi realizado o grupo controle positivo, no qual o biofilme não esteve em contato com o FS e não foram iluminados com luz LED (P-L-). Após a aplicação dos tratamentos propostos, a viabilidade dos micro-organismos foi avaliada por meio da contagem de colônias (UFC/mL), teste do XTT, determinação da...
Abstract: The ability of adhesion of microrganisms to different surfaces, the drug resistance, as well as the interaction among the species help to ensure the survival and proliferation of these microrganisms and result in lower susceptibility of these pathogens to treatment and disinfection procedures. This study aimed to evaluate the in vitro efficacy of PDT mediated by photosensitizers (FSs) Photodithazine ® (PDZ) and Curcumin (Cur) against multispecies biofilms formed by Candida albicans, Candida glabrata and Streptococcus mutans. This study was divided into three parts: 1. evaluation of the effect of photodynamic therapy of different concentrations of PDZ (100, 150, 175, 200 and 250mg / L) associated with the LED light (660 nm / 37.5 J/cm2) in the inactivation of multspecies biofilms formed in 96-well microtiter plates; 2. evaluation of the photodynamic effect of one PDT applications only and three successive PDT applications mediated by PDZ (175 and 200 mg / L) and LED light (660 nm / 37.5 J/cm2) in the inactivation of multispecies biofilms, formed on acrylic resin samples; 3. evaluation of the photodynamic effect of different concentrations of Cur FS (80, 100, and 120μM) associated with LED light (455nm / 37.5 J/cm2) against multispecies biofilms grown during different periods (24 and 48h) on acrylic resin samples. Additional samples were treated either with FSs (PDZ or Cur) (P+L-) or LED light (P-L+) only. Positive control samples had neither light nor FSs (PDZ or Cur) (P-L-). After applying the proposed treatments, the viability of microrganisms was assessed by Colony Count (CFU / mL), the XTT assay, determination of the total biomass (CV) and confocal laser scanning microscopy (CLSM).The results were analyzed using ANOVA or Kruskal-Wallis test followed by Tukey or Dunn, respectively. For the biofilm formed in 96-well plate, the concentrations of 175 and 200mg/L showed greater reduction in cell...
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15

Al-Fattani, Mohammed A. A. "Role of the biofilm matrix in resistance of Candida biofilms to antifungal agents". Thesis, University of Glasgow, 2007. http://theses.gla.ac.uk/4076/.

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The aim of this project was to investigate the possible role of the biofilm matrix as a barrier to drug diffusion in Candida biofilms and in mixed species fungal-bacterial biofilms. The penetration of antifungal agents through single- and mixed-species biofilms containing Candida was investigated using a novel filter disk bioassay. Fluconazole permeated all single-species Candida biofilms more rapidly than flucytosine. Drug penetration was more extensive with C. albicans than with the other species and the rates of diffusion of either drug through biofilms of three strains of C. albicans were similar. In all cases, after 3 to 6h the drug concentration at the distal edge of the biofilm was very high (many times the MIC). Nevertheless, drug penetration failed to produce complete killing of biofilm cells. These results indicate that poor antifungal penetration is not a major drug resistance mechanism for Candida biofilms under these conditions. It has been reported that the production of extracellular matrix by Candida biofilms growing under static incubation conditions is relatively minimal, but increases dramatically when developing biofilms are subjected to a liquid flow. In this study, Candida biofilms were grown under flow conditions in a modified Robbins device (MRD). Biofilms of C. albicans grown in the MRD produced more matrix material than those grown statically, and were significantly more resistant (P<0.001) to amphotericin B. Biofilms of C. tropicalis synthesized large amounts of matrix material even when grown statically, and such biofilms were completely resistant to both amphotericin B and fluconazole. Mixed-species biofilms of C. albicans and S. epidermidis RP62A, when grown statically or in the MRD, were also completely resistant to amphotericin B and fluconazole. Mixed-species biofilms of C. albicans and S. epidermidis M7, on the other hand, were completely drug resistant only when grown under flow conditions. Overall, these findings demonstrate that the matrix can make a significant contribution to drug resistance in Candida biofilms, especially under conditions similar to those found in catheter infections in vivo, and that the composition of the matrix material is an important determinant in resistance.

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Saur, Thibaut. "Structuration morphologique et microbiologique des biofilms multi-espèces : de l’adhésion au biofilm mature". Thesis, Montpellier 2, 2014. http://www.theses.fr/2014MON20174/document.

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Les biofilms constituent un mode de vie microbien extrêmement répandu, aussi bien en milieu naturel que dans les environnements anthropisés. Dans ce dernier cas, les structures morphologique et microbiologique du biofilm vont conditionner son impact sur le système, que cet impact lui soit bénéfique ou préjudiciable. L'objectif de cette thèse est d'approfondir notre connaissance des phénomènes de structuration du biofilm afin, à terme, d'optimiser les performances de procédé. Afin de représenter au mieux les conditions industrielles, des régimes d'écoulement turbulents et des consortia microbiens complexes ont été utilisés. Une première partie se focalise sur l'impact des forces de cisaillement sur l'adhésion microbienne. Les résultats démontrent un changement progressif de la flore bactérienne fixée et de sa distribution spatiale. Dans un second temps, le projet s'est intéressé aux étapes de développement du biofilm et ont permis d'identifier un effet mémoire du biofilm mature. Il s'agit d'une conservation des structures morphologique et microbiologique au cours du temps en dépit d'un changement de régime hydrodynamique. Enfin la dernière partie a consisté en la mise au point d'une méthode de quantification des prédateurs mobiles dans les biofilms. Ces prédateurs participent à la structuration du biofilm et leur quantification peut s'avérer utile dans le contexte de l'épuration des eaux
Biofilms are a biological mode of life widely spread in both natural and engineered environments. In the last case, whether the biofilm is beneficial or detrimental for the process under consideration, both morphology and microbial community of the biofilm determine its impact. The objective of this thesis is to deepen our knowledge of biofilm structuring and, as a further goal, optimize a given process. Turbulent flows and multi-species consortia were used in order to better mimic industrial conditions. The first part of the project focused on the impact of shear stress on microbial adhesion. Results have demonstrated a gradual shift in bacterial communities with shear and a change in the spatial distribution of adhered microorganisms. Secondly, the work dealt with biofilm development. A memory effect, defined as the conservation of initial morphological and microbiological features despite a change in the environmental conditions, has been observed. Finally, a method for quantification of moving predators in mature biofilms has been developed. These predators actively shape the biofilm and their quantification is valuable, especially for wastewater treatment

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Salvin, Paule. "Etude des biofilms électroactifs issus des milieux humides de la Guyane Française : application aux piles à combustible microbiennes". Thesis, Antilles-Guyane, 2012. http://www.theses.fr/2012AGUY0560/document.

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Les biofilms électroactifs (EA) sont des consortia bactériens mono ou pluri-espèces, qui ont la faculté d’échanger des électrons provenant de leur métabolisme avec les surfaces solides conductrices des électrodes. Cette découverte est à l’origine d’un nouveau dispositif énergétique : la pile à combustible microbienne (PACM). Depuis les années 2000, la littérature scientifique sur les biofilms EA et sur les PACM explose, notamment grâce à la découverte de bactéries capables de transférer par voie directe – par des pili ou protéines transmembranaires – des électrons vers les électrodes. Plusieurs sources de bactéries EA ont été à ce jour découvertes, allant de cultures pures à des communautés bactériennes plus complexes. Ces dernières sont issues de milieux aqueux naturels (milieux marins ou d’eau douce), industriels ou urbains (effluents d’industrie, eaux usées domestiques). La plupart de ces sources bactériennes proviennent d’environnements de climat tempéré. Dans ce travail de thèse, plusieurs sols de milieux humides de la Guyane ont été identifiés comme étant de sources de bactéries EA. Les expériences menées sous potentiel d’électrode imposé et constant ont permis d’étudier l’adhésion à la surface d’électrode des biofilms EA issus de la flore endogène des milieux sélectionnés. La formation de bioanodes et de biocathodes a été possible en présence respectivement d’acétate et d’oxygène dans les milieux. Une étude par voltammétrie cyclique a mis en évidence les pics d’oxydo-réduction en lien avec les échanges électroniques du biofilm EA et de l’électrode. En optimisant la procédure de formation des biofilms EA par chronoampérométrie (surface d’électrode plus importante, apport en continu et progressif du substrat), une densité de courant maximale de 12 A/m2 et un rendement faradique de 24 % ont été obtenus. Une autre méthode pour former des biofilms EA à partir d’un milieu choisi, la mangrove, a consisté à utiliser deux prototypes de PACM : une pile à compartiment unique et à cathode à air, et une pile benthique. Dans les deux cas, les biofilms EA ont pu être formés et étudiés, complétant certaines observations faites sous potentiel imposé. La PACM benthique s’est avérée être la plus proche d’une application à grande échelle puisqu’elle a été complètement autonome : anode et cathode utilisant uniquement le milieu pour fonctionner. Elle a pu être étudiée en laboratoire comme sur le terrain
Electroactive biofilms (EA) are mono or multi-species bacterial consortia, which have the ability to exchange electrons from their metabolism with solid surfaces of conductive electrodes. This discovery is the basis for a device of energy production: microbial fuel cell (MFC). Since the 2000s, the scientific literature on EA biofilms and MFC explodes, thanks to the discovery of bacteria that are able to transfer directly – by pili or trans-membrane proteins – electrons to electrodes. Several sources of EA bacteria were discovered to date, ranging from pure cultures to more complex bacterial communities. Those last are from natural (marine and freshwater), industrial or urban (industrial effluents, domestic wastewater) aqueous environments. However, the vast majority of these are from temperate environments.In this thesis, several wetland soils of French Guiana have been identified as sources of EA bacteria. Experiments under poised and constant electrode potential were used to examine adherence to the electrode surface of EA biofilms from the endogenous flora of selected environments. Formation of bioanodes and biocathodes was possible in the presence respectively of acetate and oxygen in the media. A study by cyclic voltammetry showed the redox peaks related to electronic exchanges between EA biofilm and electrode. By optimizing the process of EA biofilm formation by chronoamperometry (larger electrode surface, providing continuous and progressive substrate), a maximum current density of 12 A/m2 and a coulombic efficiency of 24% were obtained.Another method to form EA biofilms from a chosen medium (mangrove) was to use two MFC prototypes: a single compartment and air cathode one, and a benthic one. In both cases, the EA biofilms have been trained and studied supplementing certain observations made under poised polarization. MFC benthic proved to be the closest to a wide application since it was completely autonomous, anode and cathode only using the medium to function. It has been studied in the laboratory and in the field

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18

Steinberg, Gregory. "Long-term Stationary Phase Behavior of Streptococcus pyogenes Biofilms". Master's thesis, Temple University Libraries, 2012. http://cdm16002.contentdm.oclc.org/cdm/ref/collection/p245801coll10/id/170151.

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Microbiology and Immunology
M.S.
Long-term Stationary Phase Behavior of Streptococcus pyogenes Biofilms Department of Microbiology and Immunology Streptococcus pyogenes is the etiological agent of many human diseases ranging from mild superficial skin infections and pharyngitis to life-threatening necrotizing fasciitis. There can be several complications as a result of S. pyogenes infection including post-streptococcal glomerulonephritis and rheumatic fever, which leads to rheumatic heart disease. Despite the significant virulence associated with the pathogen, the bacteria can also persist asymptomatically in human host carriers. S. pyogenes is characterized by significant strain-to-strain variation with many single nucleotide polymorphisms and differences in genetic content of up to 33% of the genome. Active infection is associated with the rapid growth of the pathogen, whereas survival or carriage is associated with slow growth. Our laboratory has demonstrated that during survival in long-term stationary phase cultures and in eukaryotic cells, S. pyogenes diversifies into a mixed population. Isolates from this population show diversification in their proteome, in metabolism, and in virulence factor transcription patterns. These are stable, heritable changes with unique mutations in global gene regulators in some isolates, suggesting that an accumulation of genetic mutations leads to diversification. There are two proposed modes of survival in the human host; by taking residence intracellularly in host cells and as biofilms. Previous studies showed that isolates surviving within eukaryotic cells acquire heritable changes in metabolism and virulence factor expression. Biofilms are highly organized structures formed by many bacteria, which provide resiliency to harsh environmental conditions. It has been demonstrated that S. pyogenes form biofilms in vivo and in vitro, and up to 90% of clinical isolates can form biofilms. Considering the resiliency of biofilms, and the organized roles played by individual cells in biofilms, we hypothesized that biofilms may provide S. pyogenes with a niche for persistence and diversification. Despite the capacity for survival of planktonic cells, we have found that viable cells could not be isolated from static biofilms after 10 days. No metabolic variants were found among biofilm isolates prior to loss of biofilm viability. Biofilm structure was examined using confocal microscopy to image cells after LiveDead® staining. These experiments revealed that the biofilms lost viability rapidly, and also appeared to disperse. Dispersion of 2-day old biofilms could be induced with culture supernatants collected from 7-day old planktonic cells. Overall, the results of these studies suggest that secreted factors from late stationary phase cultures induce biofilm dispersion and biofilms do not serve as a niche for long-term survival and diversification of S. pyogenes. Therefore, S. pyogenes biofilms may be more critical for initial colonization of the oropharynx. These studies may provide a valuable insight to the role of biofilms in S. pyogenes infections.
Temple University--Theses

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19

Pinheiro, Luiza [UNESP]. "Staphylococcus epidermis e Staphylococcus haemolycus: detecção de genes de biofilme, toxinas, resistência a antimicrobianos e tipagem clonal em isolados de hemoculturas". Universidade Estadual Paulista (UNESP), 2014. http://hdl.handle.net/11449/110357.

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Staphylococcus epidermidis e Staphylococcus haemolyticus são considerados patógenos oportunistas, cujas infecções estão principalmente associadas à produção de biofilme. As enterotoxinas, superantígenos relacionados a intoxicações alimentares, e as hemolisinas, moléculas capazes de causar lise de membranas celulares, parecem estar associadas à patogênese das infecções estafilocócicas, apesar de ainda ser questionado se elas constituem fatores de virulência importantes nesses organismos. O uso indiscriminado de antimicrobianos vem selecionando cepas resistentes à oxacilina com menor suscetibilidade à vancomicina. A resistência à oxacilina é codificada pelo gene mecA, contido em um elemento genético móvel, denominado SCCmec. Novos antimicrobianos, como a linezolida, tigeciclina, daptomicina e quinupristina/dalfopristina vêm sendo empregados no tratamento de infecções por estafilococos multirresistentes. Este estudo objetivou caracterizar S. epidermidis e S. haemolyticus isolados de hemoculturas quanto à presença de genes de hemolisinas, enterotoxinas, suscetibilidade aos antimicrobianos e perfil clonal. Cento e sessenta e nove isolados foram pesquisados quanto à presença do gene mecA por PCR em tempo real, estando presente em 100% dos S. haemolyticus e 92,9% dos S. epidermidis, cujos tipos de SCCmec mais frequentes, determinados por PCR Multiplex, foram, respectivamente, I e III. Os MICs (Concentração Inibitória Mínima) de linezolida, tigeciclina, daptomicina, quinupristina/dalfopristina e vancomicina foram obtidos por Etest® e revelaram 4,7% de isolados resistentes à tigeciclina, 1,2% resistentes ou com resistência intermediária à quinupristina/dalfopristina, sendo o MIC máximo de vancomicina igual a 3 μg/ml. A microdiluição em caldo para vancomicina mostrou MICs de 0,25 a 2 μg/ml, concordando em 88,2% com os resultados obtidos com Etest®. A tipagem por PFGE...
Staphylococcus epidermidis and Staphylococcus haemolyticus are opportunistic pathogens that cause infections which are mainly related to the formation of a biofilm. Enterotoxins, superantigens related to food poisoning and hemolysins, molecules able to lyse mammalian cells, seem to be involved in the pathogenesis of staphylococcal infections, although the question remains whether they represent important virulence factors in these organisms. The indiscriminate use of antimicrobial drugs has selected strains that are resistant to oxacillin, in addition to isolates with reduced vancomycin susceptibility. Oxacillin resistance is encoded by the mecA gene which is carried on a mobile genetic element, SCCmec. New antimicrobial drugs such as linezolid, tigecycline, daptomycin and quinupristin/dalfopristin are being used for the treatment of infections caused by multidrug-resistant staphylococci. The objective of this study was to characterize S. epidermidis and S. haemolyticus strains isolated from blood cultures regarding the presence of hemolysin and enterotoxin genes, antimicrobial susceptibility, and clonal profile. Investigation of the mecA gene by real-time PCR in 169 isolates revealed the presence of the gene in 100% of S. haemolyticus isolates and 92.9% of S. epidermidis. The most frequent SCCmec types determined by multiplex PCR were types I and III, respectively. The minimum inhibitory concentrations (MICs) of linezolid, tigecycline, daptomycin, quinupristin/dalfopristin and vancomycin determined by the Etest® showed that 4.7% of the isolates were resistant to tigecycline and 1.2% were resistant or intermediate resistant to quinupristin/dalfopristin. The maximum MIC for vancomycin was 3 μg/ml. Broth microdilution revealed vancomycin MICs of 0.25 to 2 μg/ml, showing 88.2% agreement with the Etest results. Pulsed-field gel electrophoresis (PFGE) typing revealed the presence of S. epidermidis ...

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Lobo, Carmélia Isabel Vitorino. "Interação de Streptococcus mutans e de Candida albicans em biofilme in vitro /". Araraquara, 2018. http://hdl.handle.net/11449/154397.

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Orientador: Marlise Inêz Klein
Resumo: O objetivo foi avaliar quais são os mecanismos de Streptococcus mutans e de Candida albicans que contribuem para aumentar a patogenicidade do biofilme misto. Biofilmes mistos e simples das cepas S. mutans UA159 (Sm) e C. albicans SC5314 (Ca) foram formados sobre discos de hidroxiapatita com película salivar, na presença de sacarose. O pH do meio de cultura foi aferido em diversas fases de desenvolvimento do biofilme. Após 43h de crescimento, foram realizadas análises bioquímicas (peso seco, proteinas, composição da matriz extracelular) e de população microbiana. A estrutura dos biofilmes foi avaliada por microscopia confocal em 19 e 43h. A expressão de genes de vias metabólicas de ambas espécies foi verificada em 28h. Os dados foram avaliados por métodos estatísticos considerando α=0,05. Verificou-se diferença do pH do meio para os três biofilmes nos tempos 19, 27 e 43h (p<0,001; ANOVA dois critérios, Tukey). Biofilmes de Sm e misto foram mais ácidos em 19 e 43h, enquanto biofilmes de Ca mantiveram o pH neutro (p>0,05). As quantidades do peso seco e de proteínas de biofilme misto foram maiores comparadas aos biofilmes simples, e menores para Ca (p=0,001; ANOVA um critério). Não houve diferença na quantidade de exopolissacarídeos solúveis entre biofilmes Sm e misto, porém o biofilme Ca apresentou menor quantidade (p<0,001; Kruskal-Wallis, Dunn). Houve maior quantidade de exopolissacarídeos insolúveis em biofilme misto (p=0,002). Verificou-se mesmo comportamento populacional pa... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The objective was to evaluate the mechanisms of Streptococcus mutans and Candida albicans that contribute to increasing the pathogenicity of the mixed-species biofilm. Mixed and single-species biofilms of the strains S. mutans UA159 (Sm) and C. albicans SC5314 (Ca) were formed on saliva-coated hydroxyapatite discs in the presence of sucrose. The pH values of the culture media were verified at distinct developmental phases of biofilms. After 43h of growth, the biofilms were subjected to distinct assays biochemical (dry weight, proteins, the composition of extracellular matrix) and microbiological (colony forming units), analyzes. The biofilm's structure was verified via confocal microscopy at 19 and 43h. Gene expression of metabolic ways from both species was evaluated at 28h. The data were assessed by statistical methods (α=0,05). There was a difference in the media pH for the three biofilms at times 19, 27 and 43h (p<0,001; two-way ANOVA, Tukey). Sm and mixed-species biofilms were acidic at 19 and 43h, while Ca biofilms maintained a neutral pH (p>0,05). The amounts of dry weight and proteins were higher for mixed-species biofilm compared to singlespecies biofilms, being lower for Ca (p=0,001; one-way ANOVA). The quantity of soluble exopolysaccharides was similar for Sm and mixed-species biofilms but Ca presented a lower amount than those biofilms (p<0,001; Kruskal-Wallis, Dunn). There was a higher amount of insoluble exopolysaccharides in mixed-species biofilm (p=0,002). There was no difference in Sm population in single- and mixed-species biofilms (p=0,404; Mann Whitney); however, the mixed-species biofilm presents a higher population of Ca versus the single-species biofilm (p<0,001; t-Test). The threedimensional structure analysis showed larger microcolonies in mixed-species biofilms versus Sm biofilm, and absence of these structures in Ca biofilm...(Complete abstract electronic access below)
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Turbiani, Franciele Rezende Barbosa. "Desenvolvimento e caracterização de filmes ativos de alginato de sodio reticulados com benzoato de calcio". [s.n.], 2007. http://repositorio.unicamp.br/jspui/handle/REPOSIP/266253.

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Orientador: Theo Guenter Kieckbusch
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Quimica
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Resumo: Filmes biodegradáveis são produzidos a partir de polímeros naturais, principalmente polissacarídeos e proteínas, e tem potencial aplicação na área médica, farmacêutica e alimentícia. A incorporação de agentes ativos pode ampliar suas funções como embalagens antimicrobianas, por exemplo. Filmes foram confeccionados à base de alginato de sódio usando cloreto de cálcio como agente reticulante e glicerol como plastificante. O uso de benzoato de cálcio foi investigado como agente ativo (íons benzoato) e como auxiliar na reticulação (íons cálcio). Devido ao alto poder gelificante do Ca++, confeccionou-se, inicialmente, um filme de baixa reticulação a partir de soluções filmogênicas contendo até 0,54% de Ca++ (1º estágio). Esse filme sofreu uma reticulação complementar com excesso de Ca++ (2º estágio). Dentre os vários procedimentos avaliados (contato do filme com tecido e/ou esponja umidecidas, pincel ou rolo de pintura e imersão do filme em solução reticuladora), a simples imersão em solução contendo de 3 a 7% de CaCl2 no 2º estágio produziu filmes com alto grau de reticulação. O aumento da concentração de glicerol nessa solução melhora a manuseabilidade e plasticidade dos filmes, porém aumenta a solubilidade em água e o conteúdo de umidade dos mesmos e um adequado compromisso foi obtido usando 5% desse plastificante. Ensaios nos quais o CaCl2 foi substituído, total ou parcialmente, por benzoato de cálcio indicou que o mesmo não pode ser usado na solução do 2º estágio por favorecer a precipitação de cristais sobre o filme. Filmes ativos de 0,06 mm de espessura, pré-reticulados apenas com benzoato de cálcio e 0,7% de glicerol na solução do 1º estágio e imersos por 30 minutos em banho contendo 3% de CaCl2 e 5% de glicerol no 2º estágio, apresentaram baixa solubilidade em água (até 20% da matéria seca). Estes filmes têm baixo grau de intumescimento (< 50% da massa inicial), boa resistência mecânica à tração, mas baixa elasticidade. A permeabilidade ao vapor de água é moderada e os valores encontrados são típicos de biofilmes hidrofílicos. Ensaios de liberação de benzoato utilizando água como sorvedouro, apresentaram bom ajuste as soluções da 2ª Lei de Fick, com valores de difusividade efetiva do benzoato variando de 3 a 5.10-7 cm2/s. Os valores de difusividade diminuiram com o aumento da reticulação e aumentaram com o aumento da concentração de benzoato no filme
Abstract: Biodegradable films are produced from natural polymers, structurized mainly by polysaccharides or proteins, and have potential applications in the medical, pharmaceutical or food area. The incorporation of active agents can extend their application as antimicotic packaging, for instance. Films were manufactured with sodium alginate, using calcium chloride as cross-linking agent and glycerol as plasticizer. The use of calcium benzoate as active agent (benzoate ions) and as crosslinking agent (calcium ions) was investigated. Due to the strong gelling power of Ca++ ions, impeding smooth casting procedures, films with low reticulation are initially manufactured, using less than 0.54% Ca++ in the filmogenic solutions (1st stage). These films are further crosslinked with an excess of Ca++ by immersion in a solution of 3% to 7% of CaCl2 (2nd stage). Increasing the glycerol concentration in this solution improves the handling and plasticity of the films but increase water solubility and moisture content and an adequate compromise was obtained using 5% plasticizer. Tests conducted with partial or total substitution of CaCl2 by calcium benzoate indicated that the later could not be used in the 2nd stage solution since it promoted crystals precipitation on film surface. Active films, 0.06mm thick, pre-reticulated with calcium benzoate only and with 0.7% glycerol in the solution of the 1st stage, immersed for 30 minutes in a 3% CaCl2 and 5% glycerol solution (2nd stage) had around 17% moisture content and low water solubility (up to 20% of total dry mass). These films show low swelling degree (<50% of initial mass), good tension strength but low elongation ability. The water vapor permeability is moderate, typical of highly hydrophilic films. Benzoate liberation tests, using pure water as sink, presented good fit to solutions of Fick¿s 2nd law and effective diffusivities found varied from 4.2 to 6.3 × 10-7 cm2/s. The diffusivity values decreased with the degree of reticulation and increase with benzoate concentration in the film
Mestrado
Engenharia de Processos
Mestre em Engenharia Química

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22

Graziano, Talita Signoreti 1988. "Effects of statins in the bacterial viability and on biofilm of Staphylococcus aureus". [s.n.], 2014. http://repositorio.unicamp.br/jspui/handle/REPOSIP/289491.

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Orientadores: Karina Cogo Müller, Francisco Carlos Groppo
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: As estatinas são um grupo de fármacos que atuam como inibidores competitivos da enzima 3-Hidroxi-3-MetilGlutaril Coenzima-A Redutase (HMG-CoA redutase). Além de atuarem como importantes agentes hipolipemiantes, também apresentam outros efeitos, chamados de pleiotrópicos. Diversos estudos têm explorado um possível efeito protetor das estatinas atuando na redução na morbidade e mortalidade de várias doenças infecciosas. A atividade antimicrobiana das estatinas tem sido reportada por estudos in vivo e in vitro. O objetivo desse estudo foi avaliar os efeitos das estatinas sobre o crescimento e viabilidade de bactérias aeróbias patogênicas, e o efeito da sinvastatina sobre o biofilme de Staphylococcus aureus. Culturas das espécies de Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli e Enterococcus faecalis foram avaliadas na forma planctônica quanto à sensibilidade à atorvastatina, pravastatina e sinvastatina, através do teste de Concentração Inibitória Mínima (CIM). Além disso, diante da atividade apresentada pela sinvastatina contra S. aureus, foi determinada a ação dessa droga sobre a viabilidade celular através dos testes de Time-kill e Efeito pós-antibiótico (EPA). Também foi verificado um possível efeito sinérgico entre a sinvastatina e vancomicina. Por fim, a ação da sinvastatina foi avaliada contra biofilmes de S. aureus. Os valores de CIM da sinvastatina para o microrganismo S. aureus foram: 15,65 µg/ml (ATCC 29213) e 31,25 µg/ml (ATCC 33591, 43300, 14458 e 6538). A sinvastatina apresentou um perfil bacteriostático, e na concentração de 4xCIM seu EPA foi similar ao da vancomicina. Não foi encontrado nenhum tipo de interação entre a associação de sinvastatina e vancomicina. Entretanto, a sinvastatina foi capaz de reduzir a formação do biofilme nas concentrações entre 1/8CIM à 4xCIM. Além disso, na concentração 4xMIC foi capaz de diminuir a viabilidade, biomassa e a produção de polissacarídeos extracelulares e aumentar a produção de polissacarídeos intracelulares de biofilmes maduros de S. aureus. A produção de proteínas pelo biofilme não foi alterada. Em conclusão, os resultados encontrados mostram que a sinvastatina possui um grande potencial a ser explorado, principalmente em relação ao descobrimento de novos antimicrobianos
Abstract: Statins are drugs that competitively inhibit the enzyme 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA). Besides their important lipid-lowering action, they also are pleiotropic agents. Several studies have explored a possible protective effect of statins to reduce the morbidity and mortality of various infectious diseases. The antimicrobial activity of statins has been reported by in vivo and in vitro studies. The aim of this study was to evaluate the effects of statins on the growth, viability and biofilm formation of pathogenic aerobic bacteria. The Minimum Inhibitory Concentrations (MIC) of atorvastatin, pravastatin and simvastatin against planktonic cells of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Enterococcus faecalis strains were obtained. Since simvastatin showed activity against S. aureus, its effects on cell viability were evaluated in a time-kill and post-antibiotic effect (PAE) assays. A possible synergistic effect between simvastatin and vancomycin was also assessed. In addition, the effect of simvastatin against biofilms of S. aureus was tested. The MIC values of simvastatin for S. aureus were: 15.65 µg/ml (ATCC 29213) and 31.25 µg/ml (ATCC 33591, 43300, 14458 and 6538). Simvastatin showed a bacteriostatic profile, and in a 4x>MIC concentration the PAE was similar to vancomycin. No synergistic effect was found between simvastatin and vancomycin. Simvastatin was able to reduce the formation of biofilms in concentrations ranging from 1/8MIC to 4xMIC. In addition, the 4xMIC was able to decrease the viability, biomass and production of extracellular polysaccharides and increase the production of intracellular polysaccharides on mature biofilm of S. aureus. The protein production on biofilm was not altered in the presence of simvastatin . In conclusion, our results showed that simvastatin has a great potential to be explored, especially in relation to the development new antimicrobial agents
Mestrado
Farmacologia, Anestesiologia e Terapeutica
Mestra em Odontologia

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23

Domingues, Nádia [UNESP]. "Biofilmes multiespécies de Candida Albicans associados com streptococcus mitis e streptococcus sanguinis: estudo in vitro e in vivo". Universidade Estadual Paulista (UNESP), 2014. http://hdl.handle.net/11449/127663.

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Streptococcus sanguinis e Streptococcus mitis são colonizadores pioneiros do biofilme bucal e podem causar endocardite bacteriana. Candida albicans está presente na cavidade bucal de forma comensal, possui capacidade de formar biofilme e causar candidose bucal. O objetivo desse trabalho foi avaliar as interações entre C. albicans com S. sanguinis e S. mitis em biofilmes e suas influências in vivo em modelo experimental de invertebrado Galleria mellonella. Para o estudo da interação de C. albicans (ATCC 18804) com S. sanguinis (ATCC 7073) e S. mitis (ATCC 4945) foram formados biofilmes monoespécie e multiespécies de C. albicans com os micro-organismos (na concentração de 107 células/mL) em fundo de placa de 96 poços por 48 h, incubados em estufa a 37 °C com 5% CO2. Após crescimento, os biofilmes foram desprendidos e obtidas diluições decimais as quais foram semeadas em meios seletivos. Após incubação por 48 h/37 °C foi obtida a contagem de unidades formadoras de colônias por mililitro (UFC/mL). Em paralelo, foram realizados ensaios colorimétricos com o sal XTT, a fim de mensurar a atividade metabólica de C. albicans nos biofilmes isolados e associados, seguindo de leitura a 492 nm, e o resultado expresso em absorbância (Abs). No estudo in vivo, primeiro foram determinadas as concentrações subletais dos micro-organismos, e inoculadas em G. mellonella, e observado o efeito de C. albicans, S. mitis e S. sanguinis nas lagartas, avaliando se curva de sobrevivência. Avaliação morfológica dos biofilmes foi realizada por microscopia eletrônica de varredura (MEV). Os dados de contagem de UFC/mL e da atividade metabólica foram submetidos à análise de normalidade, e como foi observada distribuição normal, os resultados foram analisados estatisticamente pelos testes t de Student (para duas variáveis) e Tukey (para três ou mais variáveis), considerando-se diferença estatística quando p <0,05. Os dados obtidos na curva de..
Streptococcus sanguinis and Streptococcus mitis are pioneering colonizers of oral biofilm and can cause bacterial endocarditis. Candida albicans is present in the oral cavity in a commensal manner and also has the ability to form biofilm and cause oral candidiasis. The aim of this study was to evaluate the interactions between C. albicans with S. sanguinis and S. mitis biofilms and their influences in vivo on an experimental invertebrate model of Galleria mellonella. To study the interaction of C. albicans (ATCC 18804) with S. sanguinis (ATCC 7073) and S. mitis (ATCC 4945) monospecies and multispecies biofilms were formed of C. albicans and with micro-organisms (a concentration of 107 cells/ml) in the bottom of a 96-well plate for 48 h, incubated at 37 °C with 5% CO₂. After growth, biofilms were detached, and decimal dilutions were plated on selective media. After incubation for 48 h/37 °C the count of colony forming units per milliliter (CFU/mL) was obtained. Alongside, XTT salt colorimetric assays were carried in order to measure the metabolic activity of isolated and associated C. albicans biofilms, following reading at 492 nm, and the result expressed in absorbance (Abs). In the in vivo study, first the sublethal concentrations of microorganisms were determined, and inoculated into G. mellonella, and the effect of C. albicans, S. mitis and S. sanguinis in caterpillars was observed, evaluating the survival curve. Morphological evaluation of the biofilms was performed by scanning electron microscopy (SEM). The count of CFU/mL and metabolic activity were submitted to normality analisis, and as a normal distribution was observed, the results were statistically analyzed by Student's t test (for two variables) and Tukey (for three or more variables) considering statistical difference at p < 0.05. The G. mellonella survival curves data were analyzed by log-rank method. Streptococci influenced the reduction of biofilms of C. albicans and ....

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Pinheiro, Luiza. "Staphylococcus epidermis e Staphylococcus haemolycus : detecção de genes de biofilme, toxinas, resistência a antimicrobianos e tipagem clonal em isolados de hemoculturas /". Botucatu, 2014. http://hdl.handle.net/11449/110357.

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Orientador: Maria de Lourdes Ribeiro de Souza da Cunha
Banca: Elizabeth Pizzollito
Banca: Carlos Magno Castelo Branco Fortaleza
Resumo: Staphylococcus epidermidis e Staphylococcus haemolyticus são considerados patógenos oportunistas, cujas infecções estão principalmente associadas à produção de biofilme. As enterotoxinas, superantígenos relacionados a intoxicações alimentares, e as hemolisinas, moléculas capazes de causar lise de membranas celulares, parecem estar associadas à patogênese das infecções estafilocócicas, apesar de ainda ser questionado se elas constituem fatores de virulência importantes nesses organismos. O uso indiscriminado de antimicrobianos vem selecionando cepas resistentes à oxacilina com menor suscetibilidade à vancomicina. A resistência à oxacilina é codificada pelo gene mecA, contido em um elemento genético móvel, denominado SCCmec. Novos antimicrobianos, como a linezolida, tigeciclina, daptomicina e quinupristina/dalfopristina vêm sendo empregados no tratamento de infecções por estafilococos multirresistentes. Este estudo objetivou caracterizar S. epidermidis e S. haemolyticus isolados de hemoculturas quanto à presença de genes de hemolisinas, enterotoxinas, suscetibilidade aos antimicrobianos e perfil clonal. Cento e sessenta e nove isolados foram pesquisados quanto à presença do gene mecA por PCR em tempo real, estando presente em 100% dos S. haemolyticus e 92,9% dos S. epidermidis, cujos tipos de SCCmec mais frequentes, determinados por PCR Multiplex, foram, respectivamente, I e III. Os MICs (Concentração Inibitória Mínima) de linezolida, tigeciclina, daptomicina, quinupristina/dalfopristina e vancomicina foram obtidos por Etest® e revelaram 4,7% de isolados resistentes à tigeciclina, 1,2% resistentes ou com resistência intermediária à quinupristina/dalfopristina, sendo o MIC máximo de vancomicina igual a 3 μg/ml. A microdiluição em caldo para vancomicina mostrou MICs de 0,25 a 2 μg/ml, concordando em 88,2% com os resultados obtidos com Etest®. A tipagem por PFGE...
Abstract: Staphylococcus epidermidis and Staphylococcus haemolyticus are opportunistic pathogens that cause infections which are mainly related to the formation of a biofilm. Enterotoxins, superantigens related to food poisoning and hemolysins, molecules able to lyse mammalian cells, seem to be involved in the pathogenesis of staphylococcal infections, although the question remains whether they represent important virulence factors in these organisms. The indiscriminate use of antimicrobial drugs has selected strains that are resistant to oxacillin, in addition to isolates with reduced vancomycin susceptibility. Oxacillin resistance is encoded by the mecA gene which is carried on a mobile genetic element, SCCmec. New antimicrobial drugs such as linezolid, tigecycline, daptomycin and quinupristin/dalfopristin are being used for the treatment of infections caused by multidrug-resistant staphylococci. The objective of this study was to characterize S. epidermidis and S. haemolyticus strains isolated from blood cultures regarding the presence of hemolysin and enterotoxin genes, antimicrobial susceptibility, and clonal profile. Investigation of the mecA gene by real-time PCR in 169 isolates revealed the presence of the gene in 100% of S. haemolyticus isolates and 92.9% of S. epidermidis. The most frequent SCCmec types determined by multiplex PCR were types I and III, respectively. The minimum inhibitory concentrations (MICs) of linezolid, tigecycline, daptomycin, quinupristin/dalfopristin and vancomycin determined by the Etest® showed that 4.7% of the isolates were resistant to tigecycline and 1.2% were resistant or intermediate resistant to quinupristin/dalfopristin. The maximum MIC for vancomycin was 3 μg/ml. Broth microdilution revealed vancomycin MICs of 0.25 to 2 μg/ml, showing 88.2% agreement with the Etest results. Pulsed-field gel electrophoresis (PFGE) typing revealed the presence of S. epidermidis ...
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25

Domingues, Nádia. "Biofilmes multiespécies de Candida Albicans associados com streptococcus mitis e streptococcus sanguinis: estudo in vitro e in vivo /". São José dos Campos, 2014. http://hdl.handle.net/11449/127663.

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Orientador: Antonio Olavo Cardoso Jorge
Co-orientador: Cristiane Aparecida Pereira Correia
Banca: Silvia Maria Rodrigues Querido
Banca: Luciane Dias de Oliveira
Resumo: Streptococcus sanguinis e Streptococcus mitis são colonizadores pioneiros do biofilme bucal e podem causar endocardite bacteriana. Candida albicans está presente na cavidade bucal de forma comensal, possui capacidade de formar biofilme e causar candidose bucal. O objetivo desse trabalho foi avaliar as interações entre C. albicans com S. sanguinis e S. mitis em biofilmes e suas influências in vivo em modelo experimental de invertebrado Galleria mellonella. Para o estudo da interação de C. albicans (ATCC 18804) com S. sanguinis (ATCC 7073) e S. mitis (ATCC 4945) foram formados biofilmes monoespécie e multiespécies de C. albicans com os micro-organismos (na concentração de 107 células/mL) em fundo de placa de 96 poços por 48 h, incubados em estufa a 37 °C com 5% CO2. Após crescimento, os biofilmes foram desprendidos e obtidas diluições decimais as quais foram semeadas em meios seletivos. Após incubação por 48 h/37 °C foi obtida a contagem de unidades formadoras de colônias por mililitro (UFC/mL). Em paralelo, foram realizados ensaios colorimétricos com o sal XTT, a fim de mensurar a atividade metabólica de C. albicans nos biofilmes isolados e associados, seguindo de leitura a 492 nm, e o resultado expresso em absorbância (Abs). No estudo in vivo, primeiro foram determinadas as concentrações subletais dos micro-organismos, e inoculadas em G. mellonella, e observado o efeito de C. albicans, S. mitis e S. sanguinis nas lagartas, avaliando se curva de sobrevivência. Avaliação morfológica dos biofilmes foi realizada por microscopia eletrônica de varredura (MEV). Os dados de contagem de UFC/mL e da atividade metabólica foram submetidos à análise de normalidade, e como foi observada distribuição normal, os resultados foram analisados estatisticamente pelos testes t de Student (para duas variáveis) e Tukey (para três ou mais variáveis), considerando-se diferença estatística quando p <0,05. Os dados obtidos na curva de..
Abstract: Streptococcus sanguinis and Streptococcus mitis are pioneering colonizers of oral biofilm and can cause bacterial endocarditis. Candida albicans is present in the oral cavity in a commensal manner and also has the ability to form biofilm and cause oral candidiasis. The aim of this study was to evaluate the interactions between C. albicans with S. sanguinis and S. mitis biofilms and their influences in vivo on an experimental invertebrate model of Galleria mellonella. To study the interaction of C. albicans (ATCC 18804) with S. sanguinis (ATCC 7073) and S. mitis (ATCC 4945) monospecies and multispecies biofilms were formed of C. albicans and with micro-organisms (a concentration of 107 cells/ml) in the bottom of a 96-well plate for 48 h, incubated at 37 °C with 5% CO₂. After growth, biofilms were detached, and decimal dilutions were plated on selective media. After incubation for 48 h/37 °C the count of colony forming units per milliliter (CFU/mL) was obtained. Alongside, XTT salt colorimetric assays were carried in order to measure the metabolic activity of isolated and associated C. albicans biofilms, following reading at 492 nm, and the result expressed in absorbance (Abs). In the in vivo study, first the sublethal concentrations of microorganisms were determined, and inoculated into G. mellonella, and the effect of C. albicans, S. mitis and S. sanguinis in caterpillars was observed, evaluating the survival curve. Morphological evaluation of the biofilms was performed by scanning electron microscopy (SEM). The count of CFU/mL and metabolic activity were submitted to normality analisis, and as a normal distribution was observed, the results were statistically analyzed by Student's t test (for two variables) and Tukey (for three or more variables) considering statistical difference at p < 0.05. The G. mellonella survival curves data were analyzed by log-rank method. Streptococci influenced the reduction of biofilms of C. albicans and ....
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26

Tini, Ítalo Rigotti Pereira. "Osteogênese e formação de biofilmes em superfícies de titânio submetidas ao tratamento de implantação iônica por imersão em plasma de oxigênio /". São José dos Campos, 2019. http://hdl.handle.net/11449/183150.

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Orientador: Luana Marotta Reis de Vasconcellos
Coorientador: Adriano Gonçalves dos Reis
Banca: Marianne Spalding
Banca: Newton Soares da Silva
Banca: Alexandre Luiz Souto Borges
Banca: Jonatas Rafael de Oliveira
Resumo: Neste estudo foram caracterizadas superfícies de titânio (Ti) obtidas após utilização de diferentes temperaturas pela técnica de implantação iônica por imersão em plasma de oxigênio (O-IIIP), bem como correlacionado o efeito deste tratamento com a osteogênese e formação de biofilmes microbianos monotípicos. As amostras foram caracterizadas por meio de análises de química de superfície, rugosidade e textura da superfície, molhabilidade e resistência à corrosão. Além disso, análises de biocompatibilidade por meio de interação e viabilidade celular, conteúdo de proteína total, atividade de fosfatase alcalina e quantificação de nódulos de mineralização foram realizadas sobre a linhagem celular MG-63 (osteoblato humano). Análise de formação de biofilmes de Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus mutans e Candida albicans sobre as superfícies também foi realizada. Os dados foram estatisticamente analisados por teste ANOVA e Tukey (p<0,05, p<0,001 e p<0,0001). Os resultados das análises de química de superfície demonstraram um aumento proporcional da quantidade de O conforme aumento da temperatura utilizada na técnica de O-IIIP, verificando ainda a presença de TiO2 nos grupos tratados a 500ºC e 600ºC. Foi observado que, em escala nanométrica, houve um aumento significativo da rugosidade e da área superficial nas amostras tratadas com O-IIIP conforme aumento da temperatura utilizada, apresentando ainda, um aumento significativo da hidrofobicidade e resistência à ... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: In this study, titanium surfaces were produced by the ion implantation technique, immersing samples in oxygen plasma (O-IIIP), at different temperatures. Therapeutic effects of the surface modification were evaluated for osteogenesis and formation of monotypic microbial biofilms. Roughness, texture, wettability, corrosion resistance and chemical composition of the samples were characterized. Moreover, biocompatibility of the produced materials was verified by cell interaction and viability, total protein content, alkaline phosphatase activity, and quantification of mineralization nodules assays were performed on MG-63 (human osteoblate) cells. Biofilm formation of Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus mutans and Candida albicans on surfaces was also evaluated. Data were statistically analyzed by ANOVA and Tukey test. A proportional intensification in the amount of oxygen was observed as the temperature used in the O-IIIP technique raised, also, TiO2 was observed in the groups treated at 500 ºC and 600 ºC. At nanoscale, there was a statistic increase in both roughness and surface area in samples treated with O-IIIP as a result of the increase of the temperature used. Hydrophobicity and corrosion resistance were also higher in samples treated with OIIIP. According to the performed biocompatibility analyzes, cell viability, total protein production, alkaline phosphatase activity and the formation of mineralized nodules were stimulated and increased in the group treated with O-IIIP at 600 ºC, compared to the other groups. In the assays performed with monotypic microbial biofilms, a statistic reduction of microorganisms was observed especially in the groups submitted to O-IIIP treatment at 500 ºC and 600 ºC. Therefore, we demonstrated here that O-IIIP technique was able to chemically and physically modify surfaces,... (Complete abstract click electronic access below)
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27

Norwood, David Edmund. "Listeria monocytogenes in biofilms". Thesis, Queen's University Belfast, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.314102.

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28

Jones, Steven Michael. "Nosocomial pathogens within biofilms". Thesis, University of Exeter, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.370017.

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29

Skillman, Lucy C. L. C. "Enterobacterial mixed species biofilms". Thesis, University of Edinburgh, 1998. http://hdl.handle.net/1842/12962.

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Green fluorescent protein (GFP) was used as a tool to examine the interactions between pairs of bacterial species. A plasmid encoding GFP from Aequorea victoria was transformed into strains of Enterobacter agglomerans (EntGFP) and E. coli ATCC 11229 (E. coliGFP). UV illumination of plates enabled species to be identified in a mixture; their spatial distribution was examined by UV microscopy; and fluorescence measurements were used to quantify adhesion and inhibition of adhesion of the strains to other cells or cell components. Cooperation between EntGFP and Klebsiella pneumoniae G1 (KlebG1) resulted in enhanced biofilm formation and alteration of dual species biofilm properties. E. coliGFP and Serratia marcescens 87b (Serr87b) stably coexisted in biofilms but did not affect the growth of each other. The other bacterial partnerships examined were competitive, with the end result that one species dominated the biofilm. Microscopic examination of EntGFP and KlebGI dual species biofilms showed that the two species were often closely juxtaposed in microcolonies, suggesting the interactions involved surface associated macromolecules. They appeared to directly interact through adhesin/receptor interactions and proteins were isolated which could form the basis of their specific interactions. In addition, EPS affected coadhesion non-specifically. Compared to single species biofilms, both species in cooperative dual species biofilms had increased resistance to disinfectants and antibiotics. Neutral dual species biofilms of E. coliGFP and Serr87b did not show increased resistance to disinfection. Therefore, the enhanced resistance was related to the specific interactions and disruption of the cooperative partnership though the integration of a third species, Serr87b, led to a decrease in resistance. The methods developed provide a convenient technique for the examination of mixed species biofilm communities where the unique interactions between species determine the true properties and resistance of the resultant biofilms.

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Samanta, Priyankar. "Gene Regulation in Biofilms". Thesis, North Dakota State University, 2011. https://hdl.handle.net/10365/29330.

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Sessile bacterial communities which form on the solid surface or solid-liquid interface are known as biofilms. Both single species and multispecies biofilms are characterized by an extracellular matrix of polymeric substances which gives them several hundred times more antibiotic resistances than a planktonic bacterial culture. Though bacteria are the most common causative agent of various diseases, because of the high antibiotic resistance, biofilms cause complications of various diseases like cystic fibrosis, prosthetic valve endocarditis, chronic pulmonary diseases, catheter-associated urinary tract infections and several other diseases. From past studies, quorum sensing has been established as a novel target mechanism against biofilms; in this study, the two-component signal transduction systems (2CSTSs) have been focused. Once better understood, 2CSTSs can serve as a novel drug target and prevention mechanism for biofilm associated diseases. According to prior high-throughput experiments and phenotype microarray experiments by our lab, several 2CSTSs like OmpR-EnvZ, RcsCDB along with the global regulator FlhD/FlhC were hypothesized to have an important effect on various developmental stages of biofilm formation. From that past study, we postulated that acetate metabolism may be an important aspect for biofilm formation. In this study, we tested and confirmed this hypothesis. We observed biofilms formed by several mutants in 2CSTS, as well as mutants in acetate metabolism, using Scanning Electron Microscopy (SEM). We found quantitative and qualitative differences in the biofilm of the acetate mutants when compared to their isogenic parental Escherichia coli strain. An additional mutation in rcsB with acetate mutant strains forms less clumpy biofilms whereas an additional mutation in dcuR results in the formation of less biofilms. So the structural and the quantitative differences of acetate mutant biofilms depend on additional mutations in rcsB and dcuR. Though a number of studies have been done on the temporal gene expression within biofilms, spatial gene expression of the mature biofilm is a big gap of knowledge. The future aim of this study is to study the temporal as well as the spatial gene expression of different 2CSTSs in the biofilm. In my MS thesis, I have constructed selected promoter fused GFP /RFP plasmids and some other fusion plasmids were purchased from the promoter collections from Open Biosystems, lastly E. coli AJW678 bacterial strains were transformed with these GFP /RFP fused plasmids. A 96 well microtiter plate assay was performed to study the temporal expression from the promoters by quantifying the fluorescence intensity in the planktonic culture. According to this experiment, the highest expression of flhD was after 20 hours whereas, the expression of ompR increases up to 7 days, which indicates that the flhD expresses earlier than ompR. The decreasing phase of flhD expression was paralleled by the sharpest increase in ompR expression as phosphorylated OmpR is an inhibitor of flhD expression.
National Institutes of Health (NIH grant 1R15AI089403)
United States. Animal and Plant Health Inspection Service

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31

Galy, Olivier. "Microrhéologie des biofilms bactériens". Paris 6, 2011. http://www.theses.fr/2011PA066295.

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Cette thèse traite de l'hétérogénéité mécanique des biofilms bactériens. La première partie de cette étude est dédiée développement d'un nouveau montage expérimental qui permet de mesurer l'hétérogénéité mécanique au sein d'un biofilm bactérien. A l'aide d'un électroaimant, on peut appliquer des forces sur des billes magnétiques micrométriques insérées à l'intérieur d'un biofilm. La mesure du déplacement occasionné permet d'extraire la compliance et la viscosité locale du matériau. Sur l'exemple d'un biofilm cultivé à partir d'une souche d'Escherichia coli surexprimant les pili de conjugaison de type F, nous avons pu mettre en évidence deux régions distinctes mécaniquement au sein d'un biofilm. La deuxième partie de ce projet a permis d'étudier l'influence de divers facteurs sur les propriétés viscoélastiques locales des biofilms. Ainsi, il a été possible de mettre en exergue l'influence d'un facteur moléculaire sur les propriétés mécaniques d'un biofilm et nous avons montré que l'expression des curli donne une forte rigidité et une apparente homogénéité au biofilm. Par ailleurs, nous avons aussi pu relier la vitesse du flux de croissance d'un biofilm et ses propriétés mécaniques locales. Enfin, le problème de la résistance des biofilms aux antibiotiques, à travers la réponse mécanique d'un biofilm soumis à un traitement, a été abordé. Ce nouveau montage expérimental nous a permis de faire un premier pas vers la compréhension de la dynamique locale des biofilms.

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Pires, Regina Helena [UNESP]. "Formação de biofilmes e resistência a antifúngico e biocidas em Candida parapsilosis e C. orthopsilosis isoladas de águas usadas para hemodiálise". Universidade Estadual Paulista (UNESP), 2010. http://hdl.handle.net/11449/103857.

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Unidades de hemodiálise utilizam enormes quantidades de água potável para o funcionamento de rins artificiais”. Problemas microbiológicos com sistemas de distribuição de água potável são geralmente associados à presença de biofilmes formados por adesão e crescimento de microrganismos em superfícies. O patógeno humano Candida é capaz de crescer como biofilme na maioria dos aparatos médicos de uso corrente. Nos sistemas de hemodiálise, a formação de biofilme é relevante devido à contínua liberação de microrganismos que o compõe, os quais mostram fenótipo diverso daquele encontrado em seus homólogos de vida livre (forma planctônica de crescimento) com relação à transcrição gênica, velocidade de crescimento e habilidade de resistir a antimicrobianos e biocidas. Candida parapsilosis, atualmente dividida em três espécies distintas, tem sido associada às infecções decorrentes do uso de dispositivos médicos constituídos de material plástico e também de proliferar em soluções ricas em glicose, sendo que, tipicamente, tais infecções são relacionadas à formação de biofilmes. Assim, este trabalho teve como principais objetivos caracterizar molecularmente isolados identificados como C. parapsilosis advindos do circuito hídrico de uma Unidade de Hemodiálise coletados entre 2004 e 2006; verificar a capacidade de formação de biofilmes pelos mesmos; o efeito de produtos sintéticos e naturais, bem como a sensibilidade aos agentes antimicrobianos de uso clínico: anfotericina, fluconazol e voriconazol nos isolados. Por outro lado, técnicas proteômicas foram usadas para comparar o perfil de proteínas em isolados de C. parapsilosis capazes de crescerem no modo biofilme e em isolados que não apresentaram essa propriedade. Assim, 100 isolados fenotipicamente classificados como C. parapsilosis foram reclassificados por técnicas moleculares. Biofilmes foram...
Hemodialysis units use large amounts of water for the operation of artificial kidneys. Microbiological problems with distribution systems of drinking water are usually associated with the presence of biofilms formed by adhesion and growth of microorganisms on surfaces. The human pathogen Candida is able to grow as biofilm in most medical devices in current use. In hemodialysis systems, biofilm formation is relevant because of the continuous release of organisms that compose it, which have a phenotype different from their counterparts found in free-living (planktonic growth form) with respect to gene transcription, growth rate and ability to resist antibiotics and disinfectants. Candida parapsilosis, currently divided into three distinct species, has been linked to infections resulting from the use of medical devices made of plastic material and also to proliferate in glucose-rich solutions, and, typically, such infections are related to the formation of biofilms. The goals of the present study were to molecularly characterize isolates identified as C. parapsilosis coming from the water circuit of a hemodialysis unit between 2004 and 2006, assess at ability to form biofilms by these species, and to monitor the effect of synthetic and natural, as well the susceptibilities to antifungal agents in clinical use: amphotericin, fluconazole and voriconazole. Moreover, proteomic techniques were used to compare the profile of proteins isolated from C. parapsilosis able to grow in the biofilm mode and isolates that did not have this property. Thus, 100 strains classified as C. parapsilosis were identified by molecular techniques. Candida biofilms were formed in microplates and monitored by using colorimetric assay of tetrazolium salts reducing method (MTT), by the enumeration of colony forming units (CFU/mL), and by confocal scanning laser microscopy. The susceptibility to antifungal drugs... (Complete abstract click electronic access below)

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33

Li, Yung-Hua. "The influence of the environment on biofilms of oral bacteria". Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1998. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp03/NQ32000.pdf.

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34

McWilliams, Leon. "In vitro antimicrobial susceptibility of S. aureus in suspensions, biofilms, and resuspended biofilms /". Available to subscribers only, 2009. http://proquest.umi.com/pqdweb?did=1796356191&sid=7&Fmt=2&clientId=1509&RQT=309&VName=PQD.

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35

McWilliams, Leon Albert. "IN VITRO ANTIMICROBIAL SUSCEPTIBILITY OF S. AUREUS IN SUSPENSIONS, BIOFILMS, AND RESUSPENDED BIOFILMS". OpenSIUC, 2009. https://opensiuc.lib.siu.edu/theses/489.

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AN ABSTRACT OF THE THESIS OF Leon McWilliams, for the Masters of Science Degree in Medical Microbiology and Immunology, presented March 5, 2009 at Southern Illinois University. IN VITRO ANTIMICROBIAL SUSCEPTIBILITY OF S. AUREUS IN SUSPENSIONS, BIOFILMS, AND RESUSPENDED BIOFILMS MAJOR PROFESSOR: Dr. Kounosuke Watabe Whether in nature, a laboratory, or in a hospital, microorganisms exist by attaching themselves to living and nonliving objects. The range of objects microorganisms can adhere to vary from soil, internal medical devices, living tissues such tooth enamel and the lungs (1-4). As these microorganisms grow on the various objects, they develop polysaccharide layers (slime) that aid in attachment and development of a matrix (1). The development of the microorganism colony, and subsequent slime layer, is called a biofilm. The study of biofilms is of such clinical significance because of the microorganisms' altered physiological state in the biofilm. Previous studies have reported that microorganisms have significantly higher resistance in biofilm form as opposed to floating cells (1-2,5). Theories for this phenomenon include slower growth rates of cells, enzymes deactivating the antimicrobials before they can completely diffuse through the biofilm, and the very sub-population of cells that form a biofilm form a highly protected phenotypic state (1,2,3). Researchers continue to debate whether this altered physiological state of microorganisms in biofilms is due to intrinsic or extrinsic factors. The increased resistance to antimicrobials makes biofilms a very significance and expensive problem in the medical field. According to the Center for Disease Control (CDC), the medical community spends over 6 billion dollars in its battle against antibiotic-resistant microorganisms (3,38). Many indwelling medical devices such as catheters and pacemakers are susceptible to development of biofilms. Once a biofilm has been established on an indwelling medical device, clumps of cells periodically break off from the biofilm. These clumps of cells can travel in the blood stream, causing reoccurring systemic infections in the host. Due to biofilms' extraordinary resistance to antimicrobials and the occasional breakage of clumps from the primary structure, an individual infected by a biofilm may have to fight off its ill effects for months, years, even the rest of his or her life ( 3). In our research, we will study of Staphylococcus aureus. This microorganism is recognized as a key pathogen in human diseases. S. aureus is a frequent cause of community-acquired infections such as endocarditis, osteomyelitis, pneumonia, and septic arthritis (2,4). According to the CDC, the medical community invests 160 million dollars annually to combat emerging antibiotic-resistant strains of S. aureus (34). S. aureus's wide spectrum of diseases is due to its great ability to adhere to many inert and biological surfaces (4). For our project, we will use the antimicrobials vancomycin, oxacillin, and synercid. We will study the antimicrobial susceptibility of S. aureus by using suspensions, biofilms, and resuspended biofilms.

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36

Larousse, Marie. "Étude de l’interaction entre Phytophthora parasitica et le microbiote rhizosphérique à la surface de la plante hôte Solanum lycopersicum". Thesis, Nice, 2016. http://www.theses.fr/2016NICE4038.

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Les oomycètes phytopathogènes ont co-évolué avec les microbiotes des plantes hôtes. Il en résulte la formation de biofilms et des réseaux complexes d’interactions dont nous commençons juste à comprendre l’incidence sur la biologie et la virulence des oomycètes. Déterminer la nature de ces interactions et leur rôle dans le contexte d’une infection est aujourd’hui un enjeu cognitif qui concerne la caractérisation des mécanismes moléculaires et communautaires sous-jacents. C’est également une opportunité en termes d’innovation pour élaborer des méthodes de lutte alternative à l’usage de fongicides. Dans ce contexte, ce travail de thèse a consisté à étudier, d’une part, la formation de biofilm chez Phytophthora parasitica, un oomycète polyphage et tellurique, ainsi que, d’autre part, les interactions au sein de ce biofilm entre P. parasitica et le microbiote procaryote rhizosphérique de la plante hôte Solanum lycopersicum. L’analyse du génome de P. parasitica et du transcriptome du biofilm a conduit à la caractérisation d’une nouvelle famille de mucines restreinte à la lignée des oomycètes, les protéines MUCL. Ces 315 protéines sécrétées (25-30 kDa) sont réparties en 15 groupes et possèdent deux domaines : un domaine hautement conservé de fonction inconnue ; un domaine caractéristique des mucines, riche en résidus Sérine et Thréonine avec de très nombreux sites putatifs d’O-glycosylation. Chez P. parasitica, les 3 gènes PPMUCL1/2/3 sont exprimés et co-régulés spécifiquement au stade biofilm
The interactions between a pathogen and the host surface resident microbiota are critical to disease outbreak. These interactions shape the distribution, the density and the genetic diversity of inoculum. However for most plant pathogens how each of these interactions acts on disease as a single one or as a member of a functional network remains to be specified. This issue is addressed here through the analysis of two types of interactions involving the polyphagous oomycete P.parasitica : (i) the intraspecific interaction that leads to monospecific biofilm formation by P. parasitica zoospores on plant surface; (ii) the interspecific interactions that occur between P. parasitica biofilm and the prokaryotic microbiota of Solanum lycopersicum rhizosphere. The biology of monospecific biofilm is investigated through the characterization of MUCL, a new oomycete-specific Mucin-like Protein family. Gene profiling, biochemical and immunohistological analyses define the extent of this family and lead to identify three members, PPMUCL1/2/3, as residing in P. parasitica biofilm. The Phytophthora parasitica-Microbiota interaction is explored using first a metagenomic approach. Two microbial metagenomes derived from a soil of a tomato greenhouse is defined and compared after 16S RNA gene sequencing: M1 which corresponds to the sub-rhizospheric microbiota able to colonize the roots of axenic tomato seedlings; M2, the sub-microbiota able to colonize the tomato seedling roots previously coated with P. parasitica monospecific biofilm. A representative collection of microorganisms from M2 were also obtained through in vitro selection on a medium prepared from P. parasitica extract

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Guilhen, Cyril. "Caractérisations transcriptionnelle et phénotypique de bactéries dispersées de biofilm à Klebsiella pneumoniae". Thesis, Université Clermont Auvergne‎ (2017-2020), 2017. http://www.theses.fr/2017CLFAS015.

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Le développement d’un biofilm est un processus complexe qui se décline en plusieurs étapes. L’adhésion initiale des bactéries au support est suivie de la formation de microcolonies, puis de la synthèse d’une matrice extracellulaire qui englobe les bactéries donnant lieu à un biofilm mature. La dernière étape, sans doute la moins bien décrite jusqu’à présent, consiste en la dispersion du biofilm. Ce phénomène, contrôlé génétiquement, est déclenché en réponse à de nombreux signaux physiques et chimiques, telles que la température, la disponibilité en nutriments ou l’accumulation de molécules signal (peptides autoinducteurs du quorum sensing, monoxyde d’azote...). Les bactéries sessiles synthétisent alors une panoplie d’effecteurs qui permettent à certaines cellules de quitter le biofilm pour coloniser de nouveaux environnements. Dans le cadre d’infections liées aux biofilms, ce processus de dispersion est un mécanisme clé permettant la libération dans l’organisme de bactéries à partir de biofilms souvent constitués sur des matériels médicaux invasifs. Cependant, la physiologie des cellules ainsi dispersées reste relativement méconnue mais doit probablement jouer un rôle majeur dans la colonisation ultérieure et l’infection chez l’hôte. L’objectif de ce travail était de caractériser au niveau transcriptionnel et phénotypique les cellules dispersées de biofilms formés par le pathogène opportuniste Klebsiella pneumoniae, et ceci comparativement aux formes planctoniques et sessiles. Une analyse transcriptionnelle a été réalisée à partir des formes planctoniques (exponentielles et stationnaires), sessiles et dispersées, en combinant un modèle expérimental de culture en flux (flow-cell) et un séquençage global des ARN (RNAseq). Les données obtenues ont indiqué que les bactéries dispersées affichaient un profil transcriptionnel unique et distinct des autres formes bactériennes. De plus, cette analyse transcriptionnelle des formes planctoniques, sessiles et dispersées de biofilm a permis de dresser une carte exhaustive du transcriptome de K. pneumoniae au cours des différentes phases de croissance et de dégager des gènes « signatures » de chacune d’elle. Le profil transcriptionnel unique des bactéries dispersées suggère qu’elles possèdent une physiologie spécifique leur permettant de coloniser efficacement de nouveaux environnements. L’analyse des propriétés des bactéries dispersées a ainsi constitué la deuxième partie du projet, en se focalisant sur les mécanismes clés dans la physiopathologie des infections nosocomiales à K. pneumoniae, à savoir les capacités d’adhésion, de colonisation et de virulence. Le profilage phénotypique a montré que les bactéries dispersées de biofilm colonisaient significativement mieux les surfaces biotiques et abiotiques que des bactéries planctoniques. Cette capacité de colonisation accrue ne s’expliquait ni par une meilleure adhésion initiale des bactéries, ni par une activité métabolique supérieure, mais par une capacité des bactéries dispersées à former très rapidement des microcolonies. À ceci s’ajoutait un pouvoir pathogène augmenté des bactéries dispersées comparativement à la forme planctonique, avec notamment une meilleure résistance à l’activité bactéricide des macrophages. La mise au point d’un modèle murin d’infection pulmonaire adapté a été initiée ; il permettra d’analyser la virulence de ces bactéries dispersées. L’ensemble des résultats indique que les bactéries dispersées de biofilm se placent dans un état unique du cycle de vie bactérien, en étant transcriptionnellement différentes des autres formes bactériennes, et avec des propriétés mixtes, se rapprochant à la fois de la forme planctonique (activité métabolique et taux de croissance élevée) et de la forme sessile (fort pouvoir de colonisation et meilleure résistance à la phagocytose). L’ensemble de ces propriétés confère certainement un rôle déterminant dans le déclenchement d’infections liées aux biofilms
Biofilm development is a complex process involving several steps. The bacterial adhesion to the surface is followed by formation of microcolonies and synthesis of extracellular matrix, which encased bacteria giving rise to mature biofilm. The last step, probably the most poorly described, corresponds to biofilm dispersal. This process is genetically controlled and triggered in response to a wide range of physical and chemical signals, such as temperature, nutrients availability or the accumulation of signal molecules (quorum sensing autoinducers, nitric oxide...). In response to these signals, sessile bacteria synthesize various effectors, which allow a subset of cells to leave the biofilm and colonize new environments. In the context of biofilm-related infections, the biofilm dispersal process is a key mechanism for the release of bacteria from biofilms formed on invasive medicals devices. Little is known about the properties of the resulting biofilm-dispersed bacteria, but they probably play a major role in the subsequent colonization and infection processes within the host. The objective of this work was to characterize the transcriptome and the phenotype of biofilm-dispersed bacteria of the opportunist pathogen Klebsiella pneumoniae comparatively to the planktonic and sessile forms. A transcriptional analysis was carried out using planktonic (both exponential and stationary forms), sessile and biofilm-dispersed bacteria by combining a flow-cell experimental model with global RNA sequencing (RNAseq). Results indicated that biofilm- dispersed bacteria displayed a unique transcriptional pattern in the bacterial lifecycle. Furthermore, analysis of the whole transcriptome of planktonic, sessile and biofilm-dispersed bacteria allowed to emphasize the transcriptional changes occurring in the course of K. pneumoniae lifestyle transitions and to select transcriptional signatures genes for the five bacterial physiological states. The unique transcriptional pattern of biofilm-dispersed bacteria suggests that they display specific physiological characteristics required to colonize new environments. The second part of this work consisted in analyzing the properties of biofilm-dispersed bacteria, by focusing on key mechanisms involved in the physiopathology of K. pneumoniae: adhesion, colonization and virulence. Phenotypic profiling showed that biofilm-dispersed bacteria colonized significantly more biotic and abiotic surfaces than their planktonic counterparts. This increased colonization capacity was not due to an enhanced adhesion or a higher metabolic activity but rather to the intrinsic capacity of the biofilm-dispersed bacteria to form rapidly microcolonies. Besides, biofilm-dispersed bacteria were more pathogenic than their planktonic counterparts showing a better resistance to the bactericidal activity of macrophages. The development of a murine pulmonary infection model is in progress and will enable to assess the virulence of biofilm-dispersed bacteria. Our results indicate that biofilm-dispersed bacteria are placed in a unique state in the bacterial lifecycle, being transcriptionally different from other bacterial forms, and with mixed properties, approaching both the planktonic form (elevated metabolic activity and growth rate) and the sessile form (strong colonization power and high resistance to phagocytosis). These properties certainly play a decisive role in the initiation of biofilm-related infections

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Wolf, Kleber Luís [UNESP]. "Propriedades físico-químicas e mecânicas de biofilmes elaborados a partir de fibra e pó de colágeno". Universidade Estadual Paulista (UNESP), 2007. http://hdl.handle.net/11449/90777.

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O desenvolvimento de novos materiais é uma área de pesquisas que ganha cada vez mais importância nos meios acadêmicos e industriais. Essa área de pesquisa é vital para a adequação dos materiais existentes às necessidades de aplicação, pois estas estão em constante mudança. O desenvolvimento de biofilmes exige a disponibilidade de um composto - normalmente um hidrocolóide - que possua a propriedade de formar uma matriz filmogênica. O colágeno de origem bovina possui tal propriedade e, por ser abundante no Brasil, é uma matéria-prima interessante para a produção de biofilmes. Por outro lado, para melhorar as características mecânicas e de barreira à umidade dos filmes finais, além do hidrocolóide principal, em geral são adicionadas outras substâncias à matriz filmogênica. O objetivo desse trabalho foi caracterizar física e quimicamente as fibras e pó de colágeno, obtidos a partir de pele bovina, como matérias-primas potencialmente formadoras de filme, bem como avaliar a influência das proporções de fibra e pó de colágeno usadas nas propriedades dos filmes. Nesse sentido, para as matérias-primas, foram realizadas as seguintes análises: composição centesimal, solubilidade em água, isotermas de sorção de água, aminograma, granulometria, calorimetria diferencial de varredura, espectroscopia de infravermelho e reologia da solução filmogênica. A fibra e o pó de colágeno apresentam, praticamente, o mesmo teor de proteína em sua composição e a diferença entre eles, além do formato, aparece na solubilidade em água, que se supõe ser, em parte, devida à diferença de tamanho entre as partículas. Na caracterização dos filmes, realizaram-se testes de tração, solubilidade e permeabilidade ao vapor de água, determinação de cor e opacidade e microscopia.
The development of new materials is a research area that has received increasing importance in the academic and industrial. This research area is vital for adaptation of existent materials to application needs, since they are in constant change. The biofilms development demands the availability of a material - usually a hydrocolloid - that possesses the property of forming a filmogenic matrix. The collagen of bovine origin possesses such property and, for being abundant in Brazil, it is an interesting raw material for edible and biodegradable films production. On the other hand, in order to improve the mechanical and water vapor barrier characteristics of the final films, in addition to the main hydrocolloid, other substances are generally added to the film forming solution. The objective of this work was to determine the physical and chemical characteristic of collagen fibers and powder, obtained from bovine skin, as potentially film forming raw materials. The following analyses were accomplished: centesimal composition, solubility in water, water sorption isotherms, aminogram, size distribution, differential scanning calorimetric, FTIR and rheology of the filmogenic solution. The collagen fiber and powder present practically the same protein content and in addition to particles shape the main difference between them, appears in the solubility in water that is supposed to be, partly, due to the size difference between fiber and powder particles. The characterization was accomplished by traction tests and the measurement of solubility and water vapor permeability, color and opacity as well as by microscopy. The objective was evaluating the influence of collagen fiber and powder proportions on the films. In this case, fibers could act as reinforcement composite to the film forming matrix, with the peculiarity of being a system where the polymer and the composite ...(Complete abstract click eletronic access below)

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Borges, Aline Chiodi [UNESP]. "Atividade antifúngica e citotoxicidade do jato de plasma frio sob pressão atmosférica". Universidade Estadual Paulista (UNESP), 2016. http://hdl.handle.net/11449/148694.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
As doenças fúngicas representam grande desafio para a área médica e odontológica devido à crescente prevalência e aumento da resistência aos antifúngicos. O plasma frio sob pressão atmosférica (PFPA) é uma mistura gasosa contendo partículas carregadas, radicais livres e radiação. Sua potencial aplicação em doenças infecciosas foi relatada, contudo a literatura carece de estudo sistemático sobre o efeito antifúngico, mecanismos de ação e potencial citotóxico. Neste trabalho foi avaliado o efeito antifúngico do PFPA sobre Candida albicans através de ensaios em células planctônicas e biofilmes, efeitos sobre a integridade de parede celular e membrana plasmática, morfogênese, produção de exoenzimas, aderência às células epiteliais e efeito no tratamento in vivo de lesões de candidose oral induzida em modelo murino. Ainda, avaliou-se o efeito do PFPA sobre culturas de Trichophyton rubrum, além de efeitos sobre capacidade de adesão de conídios. Ainda, o potencial citotóxico foi investigado usando células epiteliais. PFPA em modo de tensão contínuo (MC) foi capaz de reduzir a aderência de C. albicans às células epiteliais, modular a transição levedura-hifa na cepa SC 5314 e comprometer a viabilidade de biofilmes. PFPA-MC se mostrou citotóxico em parâmetros efetivos frente a biofilmes de C. albicans. Porém, não foi observado efeito citotóxico quando o PFPA foi utilizado em modo de tensão pulsada (MP). A exposição ao PFPA-MP reduziu a invasão de C. albicans no epitélio in vivo. O PFPA-MP foi capaz de afetar o crescimento de T. rubrum a partir de 10 minutos e de afetar a sua capacidade de aderência. Assim, conclui-se que PFPA apresenta efeito antifúngico contra C. albicans e T. rubrum e é capaz de interferir em fatores de virulência de ambos os micro-organismos.
Fungal diseases represent a great challenge to the medical and dental areas, due to the increasing prevalence and antifungal resistance. Atmospheric pressure plasma jet (APPJ) is a gaseous mixture containing charged particles, free radicals and radiation. Its potential application in infectious disease has been reported, however there is still a lack of a systematic study on the antifungal effect, mechanism of action and citotoxicc potential. The general aim of this project was to evaluate the antifungal effect of APPJ on Candida albicans in planktonic and biofilm cultures, effects on cell wall and cell membrane integrity, morphogenesis, exoenzymes production, adherence to epithelial cells and in vivo effect in the treatment of oral candidosis in murine model will be performed. The effect of APPJ on Trichophyton rubrum cultures and on adherence capability were also evaluated. The cytotoxic potential was evaluated in vitro. APPJ in continuous tension mode (CM) was able to reduce the adherence and yeast-hyphae transition in C. albicans SC 5314 and to decrease biofilm viability. APPJ-CM showed cytotoxic effect in the parameters effective to C. albicans biofilm. Conversely, no cytotoxic effect on epithelial cells were observed when pulsed (PM) plasma jet was used. In vivo tests showed that APPJ-PM was able to prevent C. albicans invasion to the epithelium. T. rubrum cultures were affectd by APPJ-PM after 15 minutes of exposure and conidia adherence was impaired by 10 minutes exposure. In conclusion, APPJ showed antifungal effect against C. albicans and T. rubrum and can also impair virulence factors in both microorganisms.
FAPESP: 2014/02354-7

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Albuquerque, Maria Tereza Pedrosa de. "Ação de medicações intracanal sobre biofilme microbiano localizado em áreas de reabsorção apical de dentes humanos extraídos /". São José dos Campos :, 2012. http://hdl.handle.net/11449/90386.

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Orientador: Marcia Carneiro Varela
Banca: Claudio Antônio Talge Carvalho
Banca: Caio Casar Randi Ferraz
Resumo: O objetivo desse estudo foi avaliar a ação de medicações intracanal sobre biofilme microbiano localizado em áreas de reabsorção externa de dentes humanos extraídos. Para isto, 90 raízes de dentes humanos extraídos apresentando reabsorção apical externa foram submetidas à instrumentação padronizada. Em seguida foram divididas aleatoriamente em 6 grupos (n=15). As raízes foram expostas a cepa de E. faecalis (ATCC 29212) por 10 dias para induzir a formação de biofilme microbiano nas áreas de reabsorção externa apical. Após a formação do biofilme, foram inseridas medicações intracanal de acordo com os grupos. Grupo (HC): hidróxido de cálcio com solução fisiológica; Grupo (HC-CHX): hidróxido de cálcio com Clorexidina gel 2%; Grupo (HC-GEN) hidróxido de cálcio com extrato glicólico de gengibre 20%; Grupo (CHX): clorexidina gel 2%; Grupo (GEN): extrato glicólico de gengibre; Grupo controle (CC): solução salina fisiológica. Cada grupo permaneceu com medicação no interior dos canais radiculares por um período de 15 (quinze) dias. Ao final desse período as raízes foram avaliadas através do cálculo de unidades formadoras de colônias por superfície do espécime (UFC/ml) e através de análise em Microscópio Eletrônico de Varredura (MEV). Os resultados foram analisados pelos testes estatísticos de ANOVA e Tukey. Verificouse que nenhuma medicação intracanal eliminou completamente o biofilme, mas houve redução dos microrganismos pela contagem de UFC/mL no grupo CHX, seguido pelo grupo HC. Os demais grupos apresentaram resultados semelhantes ao grupo controle. As imagens por MEV confirmaram a permanência do biofilme nas áreas de reabsorção apical. Conclui-se que as medicações intracanal não foram capazes de eliminar completamente o biofilme apical de E. faecalis in vitro(AU)
Abstract: The aim of this study is to evaluate the effects of intracanal medication on microbial biofilm located in apical external root resorptions of extracted human teeth. 90 roots of extracted human teeth with apical external root surface were submitted to standardized instrumentation. The roots will be randomly divided into 6 groups of 15 roots each (n=15).The roots were exposed to E.faecalis strains (ATCC 29212) for 10 days to induce microbial biofilm formation in resorptions of apical external root surface. After biofilm formation, intracanal medications were inserted in each group. Group (HC): calcium hydroxide with sterile saline solution; Group (HC-CHX): calcium hydroxide with Chlorhexidine gel 2%; Group (HCGEN): calcium hydroxide with glycolic extract of ginger 20%; Group (CHX): chlorhexidine gel 2%; Group (GEN): extract of ginger 20%; Control group (CC): sterile saline solution. Each group remained with medication in the root canal for fifteen (15) days. After this period the roots were evaluated by calculating the colony-forming units per surface of the specimen (CFU / ml) and by analysis of Scanning Electron Microscope. The results will be analyzed using statistical tests of ANOVA and Tukey. It was found that no dressing completely eliminated biofilm, but it was a decrease in microorganisms number according to CFU / mL in the CHX group, followed by HC group. The other groups showed similar results to the control group. The SEM images confirmed the permanence of biofilm in areas of apical resorption. It is concluded that the dressing is not able to completely eliminate the biofilm apical E. faecalis in vitro(AU)
Mestre

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Fernandes, Renan Aparecido. "Fitossíntese de nanopartículas de prata a partir de extrato de cascas de romã e desenvolvimento de formulação para tratamento de feridas : avaliação antimicrobiana, citotóxica e potencial cicatrizante em um modelo in vivo /". Araçatuba, 2017. http://hdl.handle.net/11449/151982.

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Orientador: Debora de Barros Barbosa
Coorientadora: Andresa Aparecida Berretta e Silva
Banca: Alberto Carlos Botazzo Delbem
Banca: Denise Pedrini Ostini
Banca: Luiz Fernando Gorup
Banca: Elaine Cristina Guerbach Conti
Resumo: O objetivo deste estudo foi avaliar a capacidade de produção de nanopartículas de prata através do extrato da casca de romã, e produzir formulações contendo estas nanopartículas para uso em feridas. Elas foram testadas quanto à ação antimicrobiana, citotóxica e potencial cicatrizante. Para a produção das nanopartículas de prata propôs-se uma síntese utilizando-se como base duas metodologias já estabelecidas na literatura, utilizando-se para reação carboximetilcelulose, propilenoglicol, nitrato de prata, água e extrato da casca de romã como agente redutor. O extrato da casca de romã foi caracterizado em parâmetros como pH, massa seca e quantidade de taninos bioativos (ácido elágico e totais fenólicos expressos em ácido gálico). Os totais fenólicos do extrato foram também dosados após seu aquecimento nas diferentes condições de tempo e temperatura propostos para as sínteses das nanopartículas de prata (12 minutos, 1 hora e 2 horas, à 50ºC e 100ºC). As nanopartículas de prata produzidas foram, então, adicionadas a uma solução contendo compostos para produção de formulações para serem utilizadas no tratamento de feridas. Elas foram caracterizadas através de espectroscopia UV-Visível, microscopia eletrônica de varredura (MEV), potencial zeta e dosagem de íons remanescentes após as reações. A atividade antimicrobiana tanto das nanopartículas como de suas formulações contra Candida albicans SC 5314 e Staphylococcus aureus ATCC 25923 foi avaliada por meio do método da microdiluição. ... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The aim of this study was to investigate the production of silver nanoparticles through peel extract of pomegranate, and produce formulations containing these particles to be used in wound healings. Its antimicrobial action, cytotoxicity and healing potential were tested. The synthesis of silver nanoparticles were based on two methods proposed in the literature with some modifications, which were used carboxymethylcellulose, propylene glycol, silver nitrate, water and peel extract of pomegranate as reducing agent. The peel extract was characterized by pH, dry mass and bioactive tannins (elagic acid and total phenols expressed as galic acid). The total phenols were also quantified after being heated at 50ºC and 100ºC for 12 minutes, 1 hour and 2 hours. Then, silver nanoparticles were added in a solution containing products to develop a formulation to be tested in wound healing. They were characterized by UV-Vis spectroscopy, scanning electron microscopy (SEM), zeta potential and the quantification of remaining silver ions after the synthesis reaction. The antimicrobial activity of the nanoparticles and formulations were tested against Candida albicans (SC 5314) e Staphylococcus aureus (ATCC 25923) by microdilution method. After submitting the peel extract to different conditions of temperature and times (50ºC and 100ºC for 12 minutes, 1 hour and 2 hours), it was noted that the values of the minimum inhibitory concentration was not affected and were 391 μg/ml and 781 μg/ml for S. aureus and C. albicans. The formation of silver nanoparticles was confirmed through the formation of characteristic peaks in the UV-Vis spectroscopy and SEM images, and it was observed that the reaction at 50ºC for 12 min produced silver nanoparticles with regular forms and better dispersed in the formulation. The synthesis proposed promoted...
Doutor

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Karthikeyan, Subramanian. "The emergence of degradative biofilms". Thesis, National Library of Canada = Bibliothèque nationale du Canada, 2000. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape2/PQDD_0029/NQ63886.pdf.

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Baker, Jonathan Paul. "Physiological gradients in oral biofilms". Thesis, Cardiff University, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.582603.

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44

Cairns, Scott. "Biofilms : biomaterials and chronic wounds". Thesis, Cardiff University, 2012. http://orca.cf.ac.uk/50559/.

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Healthcare associated infections (HCAIs) are a large and growing problem. Bacterial infections of patients and on the medical devices used to treat them represent a significant source of morbidity and mortality. There is also a significant economical impact to the healthcare system attributed to HCAIs. While bacterial infections per se are not a novel problem, the discovery of an adherent polymicrobial phenotype called a biofilm is. A biofilm is defined by its structure and the community of bacteria therein. This study investigated bacteria biofilms in a number of pertinent clinical scenarios. To achieve this, samples were taken from five different but related clinical areas where biofilms are known to infect or are suspected to, namely endotracheal tubes, tracheostomy tubes, burn wounds, chronic wounds and chronic wound dressings. Samples were analysed using microbiological and molecular analysis techniques, the latter included polymerase chain reactions, species-specific PCR and denaturing gradient gel electrophoresis to assess microbial diversity. Fluorescent in-situ hybridization was used subsequently to analyse species orientation and biofilm structure within the biofilm. This study showed a diverse bacterial population in all the samples, with the presence of oral biota in the ETT specimens, changing to commensal bacteria over time. Large threedimensional biofilm structures were present in the specimens confirming the presence of biofilms, and within one of the chronic wound dressings where a complex biofilm was visible within the matrix of the dressing itself. These findings have considerable significance clinically, not only in demonstrating the need for biofilm targeted diagnostic techniques, but also in highlighting the need for specific biofilm treatment modalities in critical care, burn services and chronic wound management.

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Hughes, Kevin A. "Bacterial biofilms and their exopolysaccharides". Thesis, University of Edinburgh, 1997. http://hdl.handle.net/1842/15053.

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Bacterial biofilms are formed when bacteria in a liquid environment adhere to a surface, multiply to form microcolonies and synthesis a protective glycocalyx composed mainly of hydrated exopolysaccharide (EPS). Bacterial biofilms form in natural, medical and industrial environments and are usually highly heterogeneous. Biofilms can be single or multi-species and their characteristics are dictated by the environment in which they develop. The biofilm bacteria analysed were isolated from a factory environment and all were members of the Enterobacteriaceae. The composition of their extracellular polysaccharides was examined using standard biochemical assays, HPLC and paper chromatography. Most were anionic due to uronic acids, except for the EPS of two strains, 53b and Ent (both Enterobacter agglomerans), which contained only neutral sugars and were highly insoluble. Periodate oxidation revealed a high degree of 1→3 linkages. Bacteriophages which possessed polysaccharide depolymerase enzymes specific for the EPS of strains 53b and Ent were isolated from sewage from a number of sources. The glycanase of one phage (SF153b) was highly specific, had optima over a wide range of temperature and pH, and activity was increased by Ca²⁺ ions. Degradation of 53b and Ent EPS by the depolymerase produced oligosaccharide repeat units. HPLC and size exclusion chromatography gave an estimation of DP for 53b and Ent repeat units of 8-9 and 7-8 respectively. Polysaccharide samples derived from 53b biofilm and planktonic cells were both degraded by the same phage glycanase suggesting that biofilm formation does not stimulate production of a biofilm specific EPS.

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Athukorala, Arachchi Seneviratne Chaminda Jayampath. "Molecular microbiology of candida biofilms". Thesis, Click to view the E-thesis via HKUTO, 2008. http://sunzi.lib.hku.hk/hkuto/record/B4068751X.

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Ryder, Victoria Joanne. "The mutability of Staphylococcal biofilms". Thesis, University of Leeds, 2010. http://etheses.whiterose.ac.uk/2049/.

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In their natural environment, bacteria are often found as sessile populations, known as biofilms, typified by surface adherence and extracellular matrix production. This growth phase confers broad spectrum antibiotic recalcitrance, through undefined mechanisms. This work sought to investigate a role for increased mutability ofstaphylococci in biofilms, as a contributing mechanism in biofilm antibiotic recalcitrance. Initially, a novel biofilm model for use with staphylococci was established that utilised cellulose disks and incorporated human plasma to promote bacterial surface adherence. This system was validated by demonstrating antibiotic recalcitrance, dissemination by D-amino acids, and similarities with transcriptional profiles of other biofilm models. Using this novel biofilm model, and the Sorbarod flow system, mutation frequencies (MFs) were determined for Staphylococcus aureus and Staphylococcus epidermidis in biofilms and compared with those in planktonic cultures. This revealed increases in MF of up to 68-fold compared with planktonic cultures. The role of oxidative stress in biofilm mutability was investigated by addition of antioxidants to biofilms. Here, MFs were reduced up to 5-fold, suggesting a role for oxidative damage. Transcriptional profiling of biofilms revealed upregulation of several genes involved in DNA repair, compared with planktonic cultures. Genes encoding antioxidant activity were also investigated; of these only sodA was upregulated. Staphyloxanthin biosysthetic genes, however, were downregulated. During these studies, S. aureus biofilms were found to generate phenotypic variants. White variants (WVs) and large pale variants (LPVs) were mostly found amongst cells shed from biofilms. WVs had lost biofilm forming capacity and had mutations in the alternative sigma factor, SigB. LPVs retained biofilm forming capacity, but the genetic basis of their variation remains undefined. In summary, I have shown that staphylococci exhibit enhanced mutability in biofilms, accelerating the emergence of antibiotic resistance and morphological variants. This may result from oxidative stress, causing the enhanced accumulation of mutations within the genome.

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Castro, Pedro Coimbra de Almeida Osório de. "Biofilmes em endodontia". Master's thesis, [s.n.], 2014. http://hdl.handle.net/10284/4386.

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Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Medicina Dentária
Introdução: Um biofilme é uma comunidade estruturada de células bacterianas envolvidas numa matriz constituída por substâncias poliméricas e aderido a uma superfície sólida. Esta comunidade permite um modo protegido de crescimento que permite a sobrevivência dos constituintes celulares em ambientes hostis e fornece uma tolerância aumentada a agentes antimicrobianos. Objectivo: Tentar compreender a forma como os constituintes celulares se organizam e ter um melhor conhecimento das formas de resistência antimicrobiana características da organização em biofilmes, como também de rever os métodos actualmente usados para a desinfecção no tratamento endodôntico e abordando métodos alternativos ainda em estudo. Materiais e métodos: Para tal realizou-se uma pesquisa bibliográfica nos principais motores de busca: Pubmed, B-On, SciELO, Science Direct, como também no repositório da Universidade Fernando Pessoa e da Faculdade de Medicina Dentária do Porto utilizando as palavras-chave “Biofilms”, “Apical Periodontitis”, “Enterococcus faecalis” e “Biofilm Treatment “ que foram associadas de várias formas. Desta pesquisa efectuada, entre Junho de 2014 e Julho de 2014, foram escolhidos 117 artigos em Português e Inglês dos quais foram usados 89. Resultados: A forma como actualmente procedemos à desinfecção do sistema de canais radiculares, passando pela instrumentação mecânica e irrigação química não é totalmente satisfatória no que toca a uma total erradicação dos microorganismos devido a várias limitações como a complexidade anatómica dos canais e a ecologia presente no interior dos mesmos. Conclusões: De futuro, terão que ser desenvolvidas outras estratégias antimicrobianas para suplementar as existentes. Embora estas pareçam promissoras in vitro elas carecem de estudos in vivo, os quais serão necessários no futuro para ultrapassar as várias limitações presentes no sistema de canais radiculares. Introduction: A biofilm is a structured bacterial cell community enveloped in a matrix composed of polymeric substances and attached to a solid surface. This community allows for a protected way of growth that permits the survival of the cellular components in hostile environments and provides a higher tolerance to antimicrobial agents. Objective: Trying to understand the way cellular components are organized and have a better knowledge of the antimicrobial resistances that are characteristic of the way biofilms are structured, as well as to review the currently used methods for disinfection in an endodontic treatment and to address alternative methods still in study. Materials and Methods: To this end a bibliographic research was performed on the main search engines: Pubmed, B-On, SciELO, Science Direct, and also on Universidade Fernando Pessoa and Faculdade de Medicina Dentária do Porto’s repository using “Biofilms”, “Apical Periodontitis”, “Enterococcus faecalis” and “Biofilm Treatment“ as key-words that were associated in many forms. From this research performed between June 2014 and July 2014, were selected 117 articles in Portuguese and English and from those, 89 were used. Results: The way that we currently proceed regarding the disinfection of the root canal system using mechanical instrumentation and chemical irrigation is not fully satisfactory, when it comes to the total eradication of the microorganisms present due to several limitations like the complexity of the root canal anatomy and the ecology present inside the root canal. Conclusions: In the future, other antimicrobial strategies will have to be developed to supplement the currently used ones. Although these look promising in vitro they lack in vivo studies, that will be necessary in the future to overcome the several limitations present in the root canal system.

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Alamri, Hadi M. "Antimicrobial efficacy of different calcium hydroxide containing preparations against biofilms at different stages of biofilm development". Thesis, University of British Columbia, 2015. http://hdl.handle.net/2429/55988.

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Objective: To quantify and assess the antibacterial effect of different medicaments on young and aged biofilms, and to modify the medicaments in order to increase their antibacterial effect. Hypotheses: Microbes in aged biofilms grown from a mixture of oral bacteria are more resistant to the antimicrobial activity of calcium hydroxide than microbes in young biofilms. Biofilms are less resistant to calcium hydroxide combined with other antimicrobial agents than to pure calcium hydroxide. Methodology: Collagen coated hydroxyapatite disks were immersed in plaque suspension solution and incubated for one and three weeks to grow young and aged biofilms, respectively. The tested medicaments were calcium hydroxide, iodine potassium iodide, cetrimide, and the following combinations: iodine potassium iodide + cetrimide, calcium hydroxide, calcium hydroxide + iodine potassium iodide, calcium hydroxide + cetrimide, calcium hydroxide + iodine potassium iodide + cetrimide. After exposure to the medicaments for one day, one week, and two weeks, biofilms on disks were stained with a LIVE/DEAD viability stain and imaged using confocal laser scanning microscopy. The three-dimensional reconstructions of the images were done and proportions of green and red fluorescence were measured and statistically analyzed. Results: Aged biofilms were thicker than the young biofilms. All tested medicaments showed reduced antibacterial activity on the aged biofilms compared to young biofilms. Combining iodine potassium iodide to cetrimide had an additive effect and mixed with calcium hydroxide showed stronger antibacterial effect than calcium hydroxide alone. Conclusions: Aged biofilms are more resistant to antibacterial agents than young biofilms. Combining iodine potassium iodide and cetrimide to calcium hydroxide resulted in an antibacterial effect that was stronger than using calcium hydroxide alone.
Dentistry, Faculty of
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Tuck, Benjamin. "Investigating Multispecies Biofilms on Steel Surfaces in Seawater and Biofilm Inhibition by a Novel, Multifunctional Inhibitor". Thesis, Curtin University, 2022. http://hdl.handle.net/20.500.11937/89066.

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Biofilm formation is a global, $multi-billion phenomenon spanning a plethora of stakeholders. This thesis investigates critical fundamental aspects of biofilm formation on steel and evaluates the efficacy of a novel, environmentally sustainable and multifunctional inhibitor compound developed through a broader Australian Research Council Discovery Project collaboration. Focused on sustainable and effective biofilm disruption, results from this thesis are used to expand fundamental knowledge and generate a targeted approach to biofilm mitigation that improves biocide function.

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Tesi sul tema "Biofilms"

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