Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-09-20T18:10:22Z No. of bitstreams: 1 annelisafarahdasilva.pdf: 1005296 bytes, checksum: bd6fe0556315b033084ab37bb370e929 (MD5) / Approved for entry into archive by Diamantino Mayra (mayra.diamantino@ufjf.edu.br) on 2016-09-26T20:24:53Z (GMT) No. of bitstreams: 1 annelisafarahdasilva.pdf: 1005296 bytes, checksum: bd6fe0556315b033084ab37bb370e929 (MD5) / Made available in DSpace on 2016-09-26T20:24:53Z (GMT). No. of bitstreams: 1 annelisafarahdasilva.pdf: 1005296 bytes, checksum: bd6fe0556315b033084ab37bb370e929 (MD5) Previous issue date: 2010-07-05 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Recentes pesquisas demonstram que o xilitol, além de poder ser utilizado como um ingrediente alimentar, possui várias aplicações clínicas, sendo indicado para tratar diabetes, lesões renais e parenterais, bem como para prevenir otite, cáries dentárias, infecções pulmonares e osteoporose. Na literatura científica, há alguns relatos da atuação antiaderente do xilitol sobre diversas bactérias. Atualmente, com a emergência de microrganismos resistentes aos antimicrobianos convencionais, torna-se essencial a intensificação de pesquisas de novas estratégias para se prevenir e/ou tratar doenças ocasionadas pela presença destes patógenos. Uma abordagem promissora se baseia no conhecimento da aderência de microrganismos a diferentes superfícies, tanto no hospedeiro (in vivo) quanto em materiais médico-hospitalares, e de mecanismos que possam impedir essa colonização. O presente trabalho teve como objetivo avaliar a atividade antimicrobiana e a propriedade antiaderente do xilitol frente às cepas bacterianas Escherichia coli ATCC 8739, Escherichia coli clínica (EPEC), Salmonella enterica sorotipo Typhimurium clínica, Shigella flexneri ATCC 12022, Shigella flexneri clínica e Pseudomonas aeruginosa ATCC 9027. Foram realizados o teste de atividade antimicrobiana, para determinação da concentração inibitória mínima (CIM), e o teste de aderência, no qual os parâmetros: crescimento no meio de cultivo, número de unidades formadoras de colônias desprendidas e avaliação das lamínulas por microscopia eletrônica de varredura (MEV) foram analizados. Foi constatado que o xilitol, nas concentrações 0,5%; 2,5% e 5,0%, não possui atividade antimicrobiana sobre as cepas testadas. No entanto, foi demonstrada a inibição da aderência tanto pelas contagens dos bacilos desprendidos quanto pelas microfotografias obtidas por MEV. Para as concentrações de xilitol 0,5%; 2,5% e 5,0% foram desprendidos, respectivamente, 89 ± 6; 187 ± 12 e 257 ± 30 bacilos (E.coli ATCC 8739); 198 ± 28; 282 ± 20 e 311 ± 30 bacilos (E.coli clínica - EPEC); 65 ± 4; 23 ± 2 e 31 ± 1 bacilos (S. enterica sorotipo Typhimurium clínica); 131 ± 3; 120 ± 6 e 58 ± 6 bacilos (S. flexneri ATCC 12022); 779 ± 15; 146 ± 2 e 56 ± 6 bacilos (S. flexneri clínica); 2259 ± 130; 2990 ± 130 e 5330 ± 0 bacilos (P. aeruginosa ATCC 9027). Para todas as cepas testadas, os resultados revelaram que a inibição da aderência microbiana, observada nas lamínulas tratadas com xilitol, é provavelmente o mecanismo de ação deste composto. Este trabalho confere ao xilitol um maior valor agregado, torna-se uma biomolécula adequada para ser usada como adjuvante terapêutico para a prevenção e/ou tratamento de uma série de infecções recorrentes. / Recent researches show that xylitol, besides being used as a food ingredient, has many clinical applications, being used to treat diabetes, renal and parenteral lesions, as well as to prevent otitis media, caries, lung infections and osteoporosis. In the scientific literature, there are some reports on the anti-adherent action of xylitol on many bacteria. Currently, with the emergence of microorganisms resistant to conventional antibiotics, research for new strategies is necessary, aiming at preventing and/or treating diseases caused by the presence of such pathogens. A very promising approach is based on the awareness of the adherence of microorganisms to several surfaces, both in the host (in vivo) and in medical-hospital materials, and of the mechanisms which may prevent such colonization. The present study aimed at evaluating the antimicrobial activity and the anti-adherent property of xylitol on the bacterial strains Escherichia coli (ATCC 8739), clinical Escherichia coli (EPEC), clinical Salmonella enterica serotype Typhimurium, Shigella flexneri (ATCC 12022), clinical Shigella flexneri and Pseudomonas aeruginosa (ATCC 9027). The test of antimicrobial activity was realized to determinate a minimum inhibitory concentration (MIC) and the adhesion test were performed, according to which the parameters regarding growth in the culture medium, number of colony-forming units (CFUs) released and evaluation of slides using scanning electron microscopy (SEM) were analyzed. It was found that 0.5%, 2.5% and 5.0% concentrations of xylitol do not have antimicrobial activity on the strains tested. However, the inhibition of adherence was demonstrated both by counting the released bacilli and by SEM microphotographs. For 0.5%, 2.5% and 5.0% concentrations of xylitol, CFUs were released, respectively of 89 ± 6; 187 ± 12 and 257 ± 30 E.coli ATCC 8739 bacilli; 198 ± 28; 282 ± 20 and 311 ± 30 clinical E.coli – EPEC bacilli ; 65 ± 4; 23 ± 2 and 31 ± 1 clinical S. enterica sorotipo Typhimurium bacilli ; 131 ± 3; 120 ± 6 and 58 ± 6 S. flexneri ATCC 12022 bacilli; 779 ± 15; 146 ± 2 and 56 ± 6 clinical S. flexneri bacilli; 2259 ± 130; 2990 ± 130 and 5330 ± 0 P. aeruginosa ATCC 9027 bacilli. For all the strains tested, the results obtained show that the inhibition of microbial adhesion, observed in slides treated with xylitol, is probably the mechanism of action of this compound. This study gives xylitol a greater added value, making it an appropriate biomolecule to be used as adjuvant therapeutic for the prevention and/or treatment of a series of recurrent infections.

Submitted by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-24T12:20:42Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Anelise_Bravo.pdf: 334250 bytes, checksum: 202476b6e47db168114279f75ea259d0 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-24T13:56:04Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Anelise_Bravo.pdf: 334250 bytes, checksum: 202476b6e47db168114279f75ea259d0 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-24T13:56:12Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Anelise_Bravo.pdf: 334250 bytes, checksum: 202476b6e47db168114279f75ea259d0 (MD5) / Made available in DSpace on 2018-05-24T13:56:12Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Anelise_Bravo.pdf: 334250 bytes, checksum: 202476b6e47db168114279f75ea259d0 (MD5) Previous issue date: 2017-07-20 / Sem bolsa / A mussarela de leite de búfala, principal tipo de queijo obtido a partir desse leite no Brasil, é um produto novo no mercado, com alta aceitação pelos consumidores e excelentes perspectivas de comércio. O queijo é um alimento rico em nutrientes, o que favorece a proliferação de micro-organismos que podem provocar toxi-infecções ou intoxicações nos consumidores. A gastrenterite estafilocócica é causada pela ingestão de alimentos que contenham enterotoxinas produzidas por Staphylococcus coagulase positiva. Um problema que pode dificultar a eliminação de microorganismos indesejáveis na indústria de alimentos é a formação de biofilme. O objetivo deste estudo foi determinar o efeito da formação de biofilme por Staphylococcus coagulase positiva (SCP) isolados de queijo mussarela de búfala sobre a sensibilidade a sanitizantes. A partir de contagens de SCP realizadas em 50 amostras de queijo mussarela de búfala foram obtidos isolados, que foram comparados entre si por rep-PCR e identificados bioquimicamente e por multiplex PCR. As cepas distintas foram testadas quanto a formação de biofilme em placas de microtitulação. As cepas forte formadoras e uma não formadora de biofilme foram testadas em superfícies de polietileno de alta densidade, aço inoxidável e vidro. Também foram testadas quanto à sensibilidade ao hipoclorito de sódio e ao iodo após a formação do biofilme. Vinte amostras de queijo albergavam SCP, entretanto as contagens estavam dentro dos limites estabelecidos pela legislação brasileira. A rep-PCR mostrou que cada uma das amostras que estavam contaminadas apresentava uma única cepa, as quais foram identificadas como S. aureus. Dois isolados foram classificados como forte formadores de biofilme, sete como moderados formadores, dez fracos formadores e um como não formador de biofilme. As duas cepas forte formadoras produziram biofilme nas três superfícies testadas. A aplicação dos sanitizantes hipoclorito de sódio e iodo promoveu uma redução das populações bacterianas de aproximadamente 2 log em todas as superfícies, tanto das cepas formadoras de biofilme como da não formadora. Embora as cepas formadoras de biofilme não sejam mais resistentes aos sanitizantes hipoclorito de sódio e iodo do que as não formadoras, elas atingem maiores concentrações no biofilme, o que resulta em maiores populações bacterianas remanescentes após a aplicação dos sanitizantes. / The Buffalo milk mozzarella cheese, main type of chesse obtained from this milk in Brazil, is a new product in the market, with high consumer acceptance and excellent prospects for trade. The cheese is rich in nutrients, which favors the proliferation of microorganisms that can cause food toxi-infections in the consumer. The staphylococcal gastro-enteritis is caused by ingestion of food containing enterotoxins produced by coagulase-positive Staphylococcus. One problem that may hinder the elimination of undesirable microorganisms in the food industry is the formation of biofilms. The objective of this study was to determine the effect of biofilm formation by coagulase-positive (CPS) Staphylococcus isolated from buffalo mozzarella cheese on sensitivity to sanitizers. From CPS counts carried out on 50 samples of buffalo mozzarella cheese were obtained isolates, which were compared by rep-PCR and biochemically identified and by multiplex PCR. The distinct strains were tested for biofilm formation in microtiter plates. Strong forming and non-biofilm forming strains were tested on high density polyethylene, stainless steel and glass surfaces. They were also tested for sensitivity to sodium hypochlorite and iodine after biofilm formation. Twenty samples of cheese harbor CPS, however the counts were within the limits established by Brazilian legislation. Rep-PCR showed that each of the samples that were contaminated had a single strain, which was identified as S. aureus. Two isolates were classified as strong biofilm formers, seven as moderate formers, ten weak formers and one as non-biofilm builder. The two strong forming strains produced biofilm on the three surfaces tested. The application of sodium hypochlorite and iodine sanitizers promoted a reduction of approximately 2 log bacterial populations on all surfaces of both the biofilm and non-forming strains. Although biofilm forming strains are no longer resistant to sanitizers sodium hypochlorite and iodine than non-forming sanitizers, they reach higher concentrations in the biofilm, resulting in larger bacterial populations remaining after application of the sanitizers.

A Bacteroides gingivalis adhesin mediating attachment to red blood cells and buccal epithelial cells was isolated, cloned and characterized. The isolation procedure involved gentle stirring of the cells followed by ammonium sulphate precipitation, ion-exchange and gel chromatography. The native molecule had a Mr in excess of 10⁶ kDa and was made up of subunits with an Mr of 43 kDa. Antisera raised to the adhesin and its subunits reacted with antigens on the surface of B. gingivalis cells. No reaction with fimbriae was seen. The IgG fractions from these antisera inhibited the adherence of B. gingivalis to host tissue. Proteolytic enzymes destroyed binding capability of whole cells and of the purified adhesin but the molecular weight of the haemagglutinin was not altered. A genomic library of B. gingivalis DNA was created in E. coli JM83. 5500 colonies were screened by a colony immunoassay with anti-S. gingivalis serum and by a direct haemagglutinating assay. 337 clones tested positive by the immunoassay and two clones, 1-3,and 1-49 tested positive for haemagglutinating activity. Both haemagglutinating positive recombinants had inserts of 3.2 kb. One clone, 1-49 was chosen for further characterization. E. coli 1-49 expressed a protein of 43 kDa that was not present in E. coli JM 83 control as seen by SDS-PAGE and Western blot analysis. Anti-1-49 serum inhibited the haemagglutinating activity of B. gingivalis and E. coli 1-49. This serum reacted with surface molecules on B. gingivalis and E. coli 1-49 as seen by immunogold electron microscopy and immunofluorescence, and to the purified haemagglutinin by Western blot analysis. Like the haemagglutinin on B. gingivalis, the haemagglutinating activity of E. coli 1-49 was destroyed by heating and proteolytic enzymes but the apparent size of the molecule as determined by SDS-PAGE was not affected. A bacterial coaggregating adhesin from B. gingivalis was isolated and characterized. The isolation procedure involved adsorption of the solubilized adhesin on S. mitis followed by elution with glycine buffer. SDS-PAGE of the boiled adhesin revealed a protein with an Mr of 46 kDa. Proteolytic digestion destroyed all bacterial aggregating activity and hydrolysed the 46 kd protein. Antisera raised to the 46 kDa protein reacted with surface molecules on all strains of B. gingivalis tested. This antiserum inhibited the coaggregation reaction between B. gingivalis and other bacteria. Vesicles produced by B. gingivalis were found to enhance the binding of S. sanguis to serum coated hydroxy apatite (SeHA). Maximum vesicle mediated binding took place at 37°C and was destroyed by heating. The lipopolysaccharide from several black pigmented bacteroides were isolated and characterized physically, chemically and immunologically. All of the LPS were of the smooth type and contained the sugars rhamnose, glucose, galactose, glucosamine and galactosamine; no KDO or heptose were found. / Science, Faculty of / Microbiology and Immunology, Department of / Graduate

Hipótese do Estudo: O presente estudo hipotetizou que superfícies tratadas de discos de titânio comercialmente puro (Ti c.p.) são mais susceptíveis a aderência bacteriana que as superfícies lisas. Objetivo: O objetivo deste trabalho foi avaliar, in situ, a aderência bacteriana em discos de Ti c.p. submetidos a dois diferentes tratamentos de superfície: duplo ataque ácido e anodização. Metodologia: Sessenta corpos-de-prova de Ti c.p. foram divididos em três grupos: Grupo 1 - discos de Ti c.p. liso (grupo controle); Grupo 2 - discos de Ti c.p. com superfície duplamente condicionada com ácido (Master Pours Implant, Conexão Sistemas Prótese LTDA, SP., Brasil) e o Grupo 3 - discos de Ti c.p. submetidos à irradiação laser (Vulcano Actives, Conexão Sistemas Prótese LTDA, SP., Brasil). Inicialmente foi avaliada a rugosidade superficial (Ra) de todos os corpos-de-prova. Para a avaliação in situ, foram confeccionadas dez moldeiras individuais e em cada uma delas fixados seis discos de titânio, sendo dois de cada grupo. Dez voluntários usaram, por 24 horas, a moldeira contendo os discos de Ti c.p. Após este período três discos de titânio de cada grupo foram levados ao Microscópio Eletrônico de Varredura (MEV) para visualização do biofilme formado. O biofilme foi removido dos 17 discos restantes e, em sete deles, foi realizado o ensaio de MTT para quantificar as bactérias viáveis e em dez a Reação em Cadeia de Polimerase em Tempo Real (PCR Real Time, quantitativo) para quantificação total das bactérias aderidas e do Streptococcus oralis. Resultados: Os dados obtidos foram estatisticamente analisados utilizando Análise de Variância (ANOVA) One Way, seguido do pós-teste de Tamhane. Com relação a Ra, a diferença observada entres os Grupos 3 e 1 foi de 0,652 0,098μm e entre os Grupos 3 e 2 de 0,688 0,099μm. Nas micrografias foi observado biofilme aderido em todos os discos de titânio. O ensaio de MTT não evidenciou diferença significativa entre os grupos Grupo 1 (0,1190,048), 2 (0,1360,079) e 3 (0,1210,074), p=0,89. Os resultados da PCR Real Time não evidenciaram diferença significativa quanto ao número de bactérias entre os grupos estudados, tanto para o Universal (UN), Grupos 1- 372.477 (87.556;889.408), Grupo 2- 294.834 (51.742;648.674) e Grupo 3- 497.393 (150.596;2.333.567) com valor de p=0,88, quanto para o Streptococcus oralis (SO), Grupos 1-3.623(262;31.603), Grupo 2- 477(154;42.292) e Grupo 3- 9.002(1.053;147.154) com valor de p=0,42. Conclusões: Os discos do grupo 1 apresentaram rugosidade superficial igual aos do grupo 2 e ambos uma rugosidade superficial menor que os discos do grupo 3. A adesão bacteriana não foi influenciada pela rugosidade superficial dos discos de titânio. Não foi observada diferença significante na adesão bacteriana entre os grupos estudados, tanto pelo ensaio de MTT, quanto pela PCR real time. / Hypothesis of the Study: This study hypothesized that treated surfaces of discs commercially pure titanium (CP Ti) discs are more susceptible to bacterial adhesion than smooth surfaces. Objective: The objective of this study was to evaluate in situ the bacterial adherence in cp Ti discs subjected to two different surface treatments: double etching and anodizing. Methods: Sixty Ti specimens were divided into three groups: Group 1 - smooth cp Ti discs (control group), Group 2 - cp double Ti discs with surface acid etching (Master Pours Implant , Connection Prosthesis Systems LTD, SP., Brazil) and Group 3 - laser irradiated cp Ti discs subjected (Vulcan Actives , Connection Implant Systems LTD, SP., Brazil). Initially we evaluated the surface roughness (Ra) of all specimens. To assess in situ, individual trays were prepared and ten in each set of six disc titanium, two in each group. Ten volunteers wore what for 24 hours, the tray containing discs cpTi After this period three titanium disks from each group were taken to the Scanning Electron Microscope (SEM) to visualize the biofilm. The biofilm was removed from the 17 remaining disks, and seven of them, was held by the colorimetric MTT method to quantify living cells and ten to Real Time Polymerase Chain Reaction (Real Time PCR) for quantification of adhered bacteria and Streptococcus oralis. Results: Data were statistically analyzed using analysis of variance (ANOVA) One Way, followed by post-test Tamhane. With respect to Ra, the difference observed between the groups 3:01 was 0.652 0.098 m and between Groups 3 and 2 of 0.688 0.099 micrometers. In micrographs biofilm attached was observed in all titanium disks. The MTT method showed no significant difference among groups Group 1 (0.119 0.048), 2 (0.136 0.079) and 3 (0.121 0.074), p = 0.89. The Real Time PCR results showed no significant difference in number bacterial between the groups for both the Universal (UN), Group 1- 372.477 (87.556;889.408), Group 2- 294.834 (51.742;648.674) and Group 3- 497.393 (150.596;2.333.567) with p = 0, 88, as for Streptococcus oralis (SO), Group 1- 3.623(262;31.603), Group 2- 477(154;42.292) and Group 3- 9.002(1.053;147.154) with p = 0.42. Conclusions: The disks of group 1 showed surface roughness equal to Group 2 and both had a surface roughness smaller than the disks of group 3. The bacterial adhesion was not influenced by the surface roughness of the titanium disks. There was no significant difference in bacterial adhesion between the groups, both by MTT method, and by real time PCR.

No description available.

No description available.

"The effect of bacterial surface biopolymers on bacterial adhesion to surfaces was studied through experiments and modeling. Atomic Force Microscopy (AFM) provided the tool to measure the interaction forces between different bacterial cells and silicon nitride tips under different chemical conditions at a nanoscopic level. Two bacterial strains were considered: Pseudomonas putida KT2442 and Escherichia coli K-12 JM109. This study addressed the following issues: 1) the effect of solution ionic strength and solvent polarity on adhesion between Pseudomonas putida KT2442 and the silicon nitride AFM tip, 2) role of heterogeneity of bacterial surface biopolymers on bacterial adhesion, 3) role of lipopolysaccharides (LPS) on adhesion at three different scales: continuous, batch, and nanoscale, and 4) nature of interactions between E. coli JM109 and a model surface (silicon nitride tip). To address the first issue, formamide, water, and methanol were used to investigate the effect of polarity on surface characteristics of biopolymers on the bacterial surface while a range of salt concentrations between that of water to 1 M KCl were used to study the effect of ionic strength. The adhesion increased with decreasing polarity of the solvent, indicating that the polymers on the bacterial surface are hydrophilic in nature. The adhesion was slightly affected by ionic strength variations up to a concentration of 0.1 M KCl; this may have been due to the fact that the ionic concentration in the solution did not counterbalance the ionic concentration in the biopolymer brush on the bacterial surface. However, a dramatic increase in the adhesion magnitude was observed when the salt concentration increased above 0.1 M KCl. This transition in adhesion with ionic strength from a low to high value induced a transition in the elasticity of the bacterial surface biopolymers. The biopolymer brush layer did change from rigid to soft with increasing the ionic strength. The elasticity was quantified mainly by the use of the freely jointed chain (FJC) model. Our interest in investigating the role of heterogeneity on adhesion developed from the results of the first study. The bacterial surface polymers were thought to be different in their chemical and physical nature since they were found to span a range of segment lengths. Analyzing the adhesion forces for P. putida KT2442 showed that the bacterial surface is heterogeneous. The heterogeneity was evident on the same cell surface and between different cells from the same population. To resolve the third issue, approximately, 80% of the surface LPS of E. coli K-12 JM109 were removed by treating the cells with 100 mM ethylenediaminetetraacetic acid (EDTA). The effect of LPS removal on the adhesion of the cells to the silicon nitride tip was studied in water and phosphate buffered silane (PBS). The adhesion results from the AFM experiments were compared to batch retention experiments with glass as the substratum and column attachment experiments with columns packed with quartz sand. LPS controlled bacterial adhesion to the different surfaces in the study at three scales: batch, continuous, and nano-scale. Finally, the nature of interactions between E. coli JM109 and a model surface (silicon nitride tip) were investigated in solvents of varying polarity (formamide, water, and methanol). The Young’s modulus of elasticity for the bacterial surface was estimated by fitting of the Hertzian model to the force-indentation curves. Young’s modulus values increased as the solvent polarity decreased, indicating a stiffer bacterial surface in lower polarity solvents. The average adhesion force in each solvent was negatively correlated with the dielectric constant of the solvent, suggesting hydrophilic biopolymers. Specific and non-specific interaction forces between the AFM tip and the biopolymers were further characterized by applying a Poisson statistical analysis to the discrete adhesion data. The specific and non-specific interaction forces were the highest in methanol (-4 and -1.48 nN respectively). These values are in accordance with the high adhesion magnitude values measured with AFM in methanol. The results of my different studies emphasized the important role of AFM in studying biological interactions to different surfaces and in characterizing bacterial surface biopolymers."

Thesis--University of Turku, Finland, 2003. / Includes bibliographical references.

Thesis--University of Turku, Finland, 2003. / Includes bibliographical references.

Les matériaux en contact avec des fluides biologiques peuvent être colonisés par de nombreux microorganismes (bactéries, levures…) et des macromolécules telles que les protéines. Lorsqu’il s’agit de bactéries pathogènes, l’adhésion bactérienne devient un problème, en particulier dans les milieux agroalimentaire et biomédical, car elle se poursuit jusqu’à la formation de biofilms bactériens, des bio-structures plus résistantes à l’action des antibiotiques que les bactéries isolées. Malgré une littérature abondante sur les processus d’adhésion bactérienne, l’interface surface – bactérie est encore mal comprise principalement à cause du manque de caractérisation à l’échelle moléculaire. Dans ce travail, nous utilisons une technique d’optique non linéaire du second ordre, la spectroscopie vibrationnelle de Génération de Fréquence Somme (SFG) à large bande, pour sonder spécifiquement des interfaces ordonnées à l’échelle moléculaire. Le principe consiste à envoyer un faisceau picoseconde visible et un faisceau femtoseconde infrarouge (accordé aux longueurs d’onde des vibrations des molécules de la surface) sur le substrat en contact avec des biomolécules en milieu aqueux. L’optimisation de la déconvolution et la modélisation du spectre SFG expérimental, réalisées dans le cadre de travail, permettent d’obtenir quantitativement la conformation des molécules de la surface du substrat. Nous nous sommes intéressés à la colonisation d’une surface structurée en « brosse » composée de monocouches autoassemblées (SAM) hydrophobes d’OctaDécaneThiol (ODT) par des bactéries Lactococcus lactis. Afin de reproduire les conditions naturelles de la colonisation bactérienne, nous avons aussi étudié le rôle de la présence de protéines, en l’occurrence l’albumine de sérum bovin. L’étude par spectroscopie SFG couplée avec des mesures de microscopie confocale de fluorescence a permis de proposer un mécanisme de l’adhésion bactérienne sur la SAM d’ODT, qui dépend de la présence des protéines. Nous avons démontré que les bactéries seules en suspension avaient un impact sur la conformation du support pouvant conduire à une augmentation ou à une diminution de la colonisation bactérienne selon le caractère hydrophobe / hydrophile de la paroi bactérienne. La présence des protéines avant ou pendant la colonisation bactérienne conduit à de nouveaux changements structuraux de la SAM d’ODT et à une importante diminution de l’adhésion bactérienne et du biofilm résultant (indépendamment du caractère hydrophobe / hydrophile de la paroi bactérienne). Cette étude démontre d’une part la faisabilité et l’intérêt de la spectroscopie vibrationnelle SFG pour l’étude de l’adhésion bactérienne in situ, et d’autre part que l’effet des bactéries et des protéines sur la conformation des surfaces est à prendre en compte lors de l’ingénierie de nouveaux matériaux à effet antiadhésif et/ou bactéricide. / Materials exposed to a fluid can be contaminated by microorganisms and macromolecules such as proteins. In food industry or biomedicine, surface colonization by pathogenic bacteria is harmful because it leads to biofilm formation, a microbial consortium more resistant to antiobiotics than planktonic bacteria. Despite of an abundant literature on bacterial adhesion, bacteria-surface interactions still require more studies, in particular at the molecular level. In this work, Broad Band Sum Frequency Generation (BBSFG) spectroscopy was employed to investigate specifically well-ordered interfaces at the molecular level. BBSFG consists in overlapping in time and in space a picosecond visible beam and a femtosecond infrared beam onto the sample in contact with bacteria in water. The IR beam is tuned to the wavelength range of suitable vibrational transitions of adsorbed molecules. Optimisation of the deconvolution procedure of SFG spectra and modeling were performed to derive quantitatively the molecular conformational changes of the interface in response to bacterial adhesion. We have studied the colonization of a well-defined “brush” surface composed of hydrophobic OctaDecaneThiols (ODT) Self-Assembled Monolayer (SAM) by Lactococcus lactis bacteria. In order to mimic natural conditions of bacterial adhesion, where bacteria and proteins coexist, we have also studied the role of Bovin Serum Albumin (BSA) proteins. SFG data and fluorescence confocal microscopy measurements led us to propose a mechanism of bacterial adhesion onto the ODT SAM which depends on the presence of BSA. We have demonstrated that adhesion of bacteria in distilled water induces measurable effects on the ODT SAM conformation and on the bacterial adhesion strength, depending on the hydrophobic / hydrophilic character of the bacterial cell wall. Different behaviors of the ODT were observed when BSA proteins were present before or during bacterial colonization. In particular, BSA leads to a marked antimicrobial effect (independent of the hydrophobic / hydrophilic character of the bacterial surface). This study demonstrates the potential of BBSFG to study in situ bacterial adhesion. It also shows that the modification of surface properties by bacterial adhesion must be taken into account for the design of materials suitable to control or eradicate biofilm formation.