115 resultados para Formação de biofilme - Tese

em Universidade Federal do Rio Grande do Norte(UFRN)


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Photodynamic therapy (PDT) has been proposed as an alternative method for the treatment of biofilm-dependent oral diseases like dental caries. This therapy consists of simultaneous action of a visible light (L) and a photosensitizer (FS) in the presence of oxygen, which leads to production of different reactive oxygen species that can interact with the bacterial cell components, and promote cell death. This study aims to evaluate the antimicrobial action of PDT on oral bacteria in suspension, as well as the formation of mono and multi-species biofilms, in vitro, from a standard strain of Streptococcus mutans (ATCC 25175) and saliva samples, respectively. The dye methylene blue (MB) and toluidine blue (TB) were used at a concentration of 100 mg/ L and activated by halogen light (600 to 750 nm) from a modified hand held photopolymerizer (Ultralux ®, Dabi Atlante, Ribeirão Preto , São Paulo, Brazil.). Planktonic cultures were prepared and submitted to different experimental conditions: 1. PDT using TB 2. PDT using MB, 3. L+ FS- , 4. TB + L - ; 5. MB+ L-; 6. L- FS- (negative control) and 7. administration of 0.12% chlorhexidine digluconate (positive control) (Periogard ®, Colgate-Palmolive Company, New York, USA). The immediate and mediated action of PDT on bacterial suspensions, as well as its effect on biofilm formation were observed from the number of colony-forming units per milliliter (CFU/mL) and measures optical density (OD). The data were statistically analyzed using the Kruskal-Wallis and Mann-Whitney test for the significance level of 5%. According to the results, the PDT showed no antibacterial action on suspensions of S. mutans, regardless of the dye used. PDT with MB activated by halogen light was able to reduce 86.6% CFU/mL multi-species planktonic cultures, however, this reduction was not significant (p > 0.05). PDT showed antibacterial effect, mediate on multi-species planktonic cultures with TB (p < 0.001) and MB (p < 0.001), activated by halogen light. PDT was able to prevent the formation of multispecies biofilm, through the activation of TB by halogen light (p = 0.01). We conclude that activation of the dye toluidine blue and methylene blue, by halogen light (PDT) showed antimicrobial activity, compared to multi-species planktonic cultures prepared from saliva samples

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Candidiasis is a major oral manifestation in kidney transplant patients. Candida spp. possess essential virulence factors which contribute for the infectious process, including the ability to adhere to epithelial cells and biofilm formation. The extract obtained from the leaves of Eugenia uniflora [acetone: water (7:3, v/v)] has demonstrated antifungal activity against Candida spp. This study evaluated the influence of the extract of E. uniflora in adhesion to human buccal epithelial cells (HBEC) and biofilm formation of 42 strains of Candida spp. isolated from the oral cavity of kidney transplant patients. Candida spp. strains belonging to a culture collection were reactivated and phenotypically re-identified by classical and molecular methods (genotyping ABC and RAPD), when necessary, to complete the identification to the species level. For the virulence tests evaluated in vitro, yeasts were grown in the presence and absence of 1000 g/mL of the extract. A ratio of 10: 1 (Candida spp. cells x HBECs) was incubated for 1 hour at 37 ° C, 200 rpm, fixed with 10% formalin and the number of Candida cells adhered to 150 HBEC determined by optical microscope. Biofilms were formed on polystyrene microplates in the presence or absence of the extract. The quantification was performed with crystal violet staining at 570 nm. All isolates were viable and exhibited phenotypic characteristics suggestive of each species identified. Two strains presumptively identified as Candida dubliniensis belonged to this species as determined with genotyping ABC, while strains identified as belonging to the Candida parapsilosis species complex were differentiated by RAPD genotyping. Candida albicans was found to be the most adherent species to the buccal epithelia, while C. tropicalis showed remarkable biofilm formation.We could detect that the extract of E. uniflora was able to reduce adhesion to HBEC for both Candida albicans and non-Candida albicans Candida species. On the other hand, only 16 Candida spp. strains (36 %) showed reduced biofilm formation. However, two highly biofilm producer strains of C. tropicalis had an expressive reduction in biofilm formation. This study reinforces the idea that besides growth inhibition, E. uniflora may interfere with the expression of some virulence factors of Candida spp., and may be possibly applied in the future as a novel antifungal agent.

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Chromobacterium violaceum is a free-living bacillus, Gram-negative commonly found in water and sand of tropical and subtropical regions. One of its main characteristic it's the ability to produce the purple pigment named violacein, that shows countless biological activities. In 2003, the genome of this organism was totally sequenced and revealed important informations about the physiology of this bacteria. However, few post-genomics studies had been accomplished. This work evaluated the protein profile of C. violaceum cultivated in LB medium at 28ºC that allowed the identification and characterization of proteins related to a possible secretion system that wasn't identified and characterized yet in C. violaceum, to the quorum sensing system, to regulatory process of transcription and translation, stress adaptation and biotechnological potential. Moreover, the response of the bacteria to UVC radiation was evaluated. The comparison of the protein profile, analyzed through 2-D electrophoresis, of the control group versus the treatment group allowed the identification of 52 proteins that arose after stress induction. The obtained results enable the elaboration of a stress response pathway in C. violaceum generated by the UVC light. This pathway, that seems to be a general stress response, involves the expression of proteins related to cellular division, purine and pirimidine metabolism, heat chock or chaperones, energy supply, regulation of biofilm formation, transport, regulation of lytic cycle of bacteriophages, besides proteins that show undefined function. Despite the response present similarities with the classic SOS response of E. coli, we still cannot assert that C. violaceum shows a SOS-like response, mainly due to the absence of characterization of a LexA-like protein in this organism

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The studies in the education area have been defending the importance of forming autonomous teachers that they look for to understand the context in that they act, for that movement to make possible educational interventions with better conditions of the students' learning. In the extent of the teachers' formation, it has been discussed the importance of formative actions that they have as focus the analysis of the needs in the educational formation, as alternative to turn such more significant actions. Concerned with those subjects and with the school failure in the Brazilian education, especially in the children's of the public school literacy, we accomplished this work that aims at to investigate the needs of teachers' of the Fundamental Teaching of the public school formation, in the extent of their students' orthographic difficulties, building with those teachers knowledge on the process of teaching/learning spelling. The study was accomplished at a municipal school of the city of Parnamirim-RN, that offers the Fundamental Teaching I. the approach investigativa is of qualitative nature, where the case study and the research-action were used as methodologies. They participated in the research 7 teachers and 1 coordinator of the institution. For the construction of the data the following instruments were used: glimpsed semi-directing collective and individual, documental analysis, questionnaire and observation, that made possible the triangulation of the data for the content analysis. The main results pointed that the teachers' of that school initial needs were addressed not only to the teaching of the spelling, but also to the formation type that they would receive. We verified although the formative process was favorecedor of sockets of the teachers' conscience: related to the orthographic mistakes, to the strategies used at classroom and to the planning addressed to the teaching of the spelling. Finally, we concluded that the teachers' reflection researched about their own formation needs contributed for (trans)formations of their conceptions and practices related to the teaching of the spelling, even before some found difficulties, so much in the formation as in practice pedagogic developed by the participants

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Ce projet présente dans son premier chapitre une étude historique-bibliographique basée sur la Pédagogie Freinet en France, ainsi que sa formation républicaine, ses courants politiques et idéologiques qui ont moulés l organisation sociale et éducationnelle de ce pays. Ce travail montre le rôle des courants de gauche dans ce processus, cela se reflète aussi sur le jeune Freinet. Les changements opérés dans la décennie de 1950 dans le système scolaire, dans le propre Mouvement Freinet et les ruptures qui ont eues à l intérieur de ce mouvement. Cela se termine avec les directions prises dans les dernières décennies, son arrivée au Brésil et les perspectives actuelles. Le deuxième chapitre aborde la reconstitution historique du Brésil dont sa phase coloniale et impériale. Les événements historiques de la IIIe, IVe et Ve République sont mis en relief avec ceux du Mouvement Freinet. La phase militaire imposée au Brésil et les luttes pour la reconquête de la démocratie. Il se termine avec les considérations sur la LDBEN 9394/96 et l implantation du Curso Normal Superior; un bref historique de la Pédagogie Freinet au Brésil, des thèmes importants travaillés dans le troisième chapitre. Dans ce chapitre une recherche qualitative est décrite basée dans l Etnographie Critique et Multiréférentiellel et aussi dans la Recherche-Action. Elle est développée auprès d un groupe de cinq professeurs des premières classes de l école primaire au collège, inscrites dans le Curso Normal Superior. Les données ont été obtenues à partir de rencontres régulières avec le groupe la médiation didactique , des entretiens, des questionnaires, des observations dans les salles de classe, du travail final du cours, nommé Mémorial de Formation. À travers l ensemble des données, on recherche les apports et l actualité d une proposition pédagogique historiquement firmée pour le développement de la praxis éducative

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This paper gives clues for the educative action in nutrition subjects. It deals about the professors experiences deployments lived in 2003 e 2004, both in Nutrition undergraduate course of the Federal University of the Rio Grande do Norte (UFRN), in Nutrition Education and Supervised Internship in Social Nutrition academic disciplines, as well as in the II Update Course in Nutrition Practices for Health Basic Care , offered to the supervisors nutritionists of internship, in this same department, being able to be characterized as an action-research, with interventionism purpose. The study stands out the importance of a new point of view about the nutritionist formation to overcome the limits imposed by the scientism, and the adoption of a complex and reflexive reference about the educational practice in this area. The corpus is made up by 81 undergraduate students alimentary autobiography (source of generating subjects for interventions with the nutritionists), 17 questionnaires and 05 interviews, being 03 of them biography (the start up for an initial dialogue with the nutritionists). The data found and the professors experiences allied to a theoretic reference, by the light of the education proposals for the XXI century were used as establishment elements for the proposition of five guidance axles used to build a complex and reflexive nutrition education, which are: 1) Take the cookery and the culture of eating together as significant elements for the human being integral formation; 2) Conceive the religion manifestations associated to feeding process as relevant elements of the human food culture; 3) Discuss the rupture nature/culture aiming the preservation of live in earth; 4) Search for the overcoming of the identity conflicts by a higher inclosing conscience degrees of being part of this process. 5) Face the limits of fragmented formation. The presented thesis stands that the autobiography method, allied to the freirean pedagogy and to a complex reference, could be taken as an important tool to the health educative subjects, contributing to the formation of reflective individuals able to transform themselves and the world.

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Chromobacterium violaceum is a free-living bacillus, Gram-negative commonly found in water and sand of tropical and subtropical regions. One of its main characteristic it's the ability to produce the purple pigment named violacein, that shows countless biological activities. In 2003, the genome of this organism was totally sequenced and revealed important informations about the physiology of this bacteria. However, few post-genomics studies had been accomplished. This work evaluated the protein profile of C. violaceum cultivated in LB medium at 28ºC that allowed the identification and characterization of proteins related to a possible secretion system that wasn't identified and characterized yet in C. violaceum, to the quorum sensing system, to regulatory process of transcription and translation, stress adaptation and biotechnological potential. Moreover, the response of the bacteria to UVC radiation was evaluated. The comparison of the protein profile, analyzed through 2-D electrophoresis, of the control group versus the treatment group allowed the identification of 52 proteins that arose after stress induction. The obtained results enable the elaboration of a stress response pathway in C. violaceum generated by the UVC light. This pathway, that seems to be a general stress response, involves the expression of proteins related to cellular division, purine and pirimidine metabolism, heat chock or chaperones, energy supply, regulation of biofilm formation, transport, regulation of lytic cycle of bacteriophages, besides proteins that show undefined function. Despite the response present similarities with the classic SOS response of E. coli, we still cannot assert that C. violaceum shows a SOS-like response, mainly due to the absence of characterization of a LexA-like protein in this organism

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This laboratory study involves the participation of a group with professionals from different areas that had contributed to the construction of a multidisciplinary knowledge, about biological response of titanium surfaces modified through thermochemical treatment by plasma. Thus, the crystalline phase was previously characterized in relation to the topography, roughness, molhability and nitrogen concentration in the samples surface. It s indispensable that materials implanted can influence in a good cellular response as well as promotes a bacteria action. Surfaces modified by plasma were exposed to different cultures such as: cellular (human osteoblastic) and bacteria (Staphylococcus epidermidis ATCC35984 and Pseudomonas aeruginosa ATCC 27853) in order to evaluate the biological response. It was evaluated the adhesion, proliferation, morphology and cellular preference of human ostheoblastic cells (HOST), as well as the formation of a biofilm and bacteria proliferation. It was still analyzed the bacteria selectivity ability in relation to the surfaces. The software Image Pro Plus was used to the counting of cells and bacteria adhered to the surface of disks. The results were submitted to the variance analysis (ANOVA), and then, by the Kruskal-Wallis test, using GraphPad Instat ® software, version 3.5 to Windows. The nitrided samples in spite of show a higher roughness and molhability showed a smaller bacteria growing and higher cellular proliferation, when compared to non treated samples, indicating that the treated material present a high efficiency to biomedical implants

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Despite Candida species are often human commensals isolated from various oral sites such as: tongue, cheek and palatal mucosa plus subgingival region, there are some properties linked to the organism commonly known as virulence factors which confer them the ability to produce disease. Oral candidiasis is one of the main oral manifestations reported in literature related to kidney transplant patients. The objectives of the present study were to identify and investigate virulence factors of yeasts isolated from the oral cavity of kidney transplant recipients admitted at the Hospital Universitário Onofre Lopes, in Natal RN. Seventy Candida species isolated from 111 kidney transplant recipients were investigated in this study. Identification of the isolates was performed by using the evidence of germ tube formation, hypertonic broth, tolerance to grow at 42°C, micromorphology and biochemical profiles. We observed a high rate of isolation of yeasts from the oral cavity of kidney transplant recipients (63.1%) being C. albicans was the most prevalent species. Oral candidiasis was diagnosed in 14.4% of transplant recipients. We evaluated virulence properties of the isolates regarding to: biofilm formation on polystyrene microplates as well as XTT reduction, adherence to acrylic resin and human buccal epithelial cells and proteinase activity. Most isolates were able to form biofilm by the method of adhesion to polystyrene. All isolates of Candida spp. remained viable during biofilm formation when analyzed by the method of XTT reduction. The number of CFU attached to the acrylic resin suggested high adherence for C. parapsilosis. C. albicans isolates showed higher median adherence to human buccal epithelial cells than non-C. albicans Candida isolates. Nevertheless, this difference was not statistically significant. C. dubliniensis showed low ability to adhere to plastic and epithelial cells and biofilm formation. Proteolytic activity was observed for all the isolates investigated, including the unique isolate of C. dubliniensis. There was a statistically significant association between proteinase production and the presence of oral candidiasis. Studies related to oral candidiasis in renal transplant recipients are limited to clinical and epidemiological data, but investigations concerning Candida spp. virulence factor for this group of individuals are still scarce. We emphasize the importance of studies related to virulence factors of yeasts isolated from this population to contribute to the knowledge of microbiological aspects of oral candidiasis

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Vulvovaginal candidiasis (VVC) is one of the most common causes of vaginitis and affects about 75% of women of reproductive age. The majority of cases (80 to 90%) are due to C. albicans, the most virulent species of the genus Candida. Virulence attributes are scarcely investigated and the source of infection remains uncertain. Objective: This study aimed to evaluate the virulence factors and genotypes of clinical isolates of C. albicans sequentially obtained from the anus and vagina of patients with sporadic and recurrent VVC. Materials and methods: We analyzed 62 clinical isolates of C. albicans (36 vaginal and 26 anal strains). Direct examination of vaginal and anal samples and colony forming units (CFU) counts were performed. Yeasts were identified using the chromogenic media CHROMagar Candida® and by classical methodology, and phenotypically characterized regarding to virulence factors, including the ability to adhere to epithelial cells, proteinase activity, morphogenesis and biofilm formation. The genotypes of the strains were investigated with ABC genotyping, microsatellite genotyping with primer M13 and RAPD. Results: We found 100% agreement between direct examination and culture of vaginal samples. Filamentous forms were present in most of the samples of vaginal secretion, which presented CFU counts significantly higher than the samples of anal secretion. There was no statistically significant difference between virulence factors of infecting vaginal isolates and those presented by colonizing anal isolates; as well as for the comparison of the vaginal isolates from patients with different clinical conditions (sporadic or recurrent VVC). There was a decrease in the ability to adhere to HBEC, morphogenesis and biofilm formation of the vaginal isolates during the progress of infection. There was an association between the ability to express different virulence factors and the clinical manifestations presented by the patients. Genotype A was the most prevalent (93.6%), followed by genotype C (6.4%). We found maintenance of the same ABC genotype and greater prevalence of microevolution for the vaginal strains of C. albicans sequentially obtained. Vaginal and anal isolates of C. albicans obtained simultaneously from the same patient presented the same ABC genotype and high genetic relatedness. Conclusion: It is noteworthy that the proliferation of yeast and bud-to-hypha transition are important for the establishment of CVV. The expression of virulence factors is important for the pathogenesis of VVC, although it does not seem to be determinant in the transition from colonization to infection or to the installation of recurrent condition. Genotype A seems to be dominant over the others in both vaginal and anal isolates of patients with VVC. The most common scenario was microevolution of the strains of C. albicans in the vaginal environment. It is suggested that the anal reservoir constituted a possible source of vaginal infection, in most cases assessed

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Periodontal infections consist of a group of inflammatory conditions caused by microorganisms that colonize the tooth surface through the formation of dental biofilm. Chronic infections such as periodontitis have been associated to the development and progression of atherosclerosis. AIM: Detect cultivatable and non-cultivatable periodontopathogenic bacteria in atheromatous plaques; search for factors associated to the presence of these bacteria in the atheromatous plaques and characterize the presence of cultivatable and non-cultivatable bacteria in these plaques. METHODOLOGY: A cross-sectional study was performed with a sample of 30 patients diagnosed with atherosclerosis in the carotid, coronary or femoral arteries and surgically treated with angioplasty and stent implant, bypass or endarterectomy. The plaques were collected during surgery and analyzed using the PCR molecular technique for the presence of the DNA of the cultivatable bacteria Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Treponema denticola and of the non-cultivatable Synergistes phylotypes. The patients were examined in the infirmary, after the surgery, where they also responded to a questionnaire aimed at determining factors associated to the presence of periodontopathogenic bacteria in the atheromatous plaques. RESULTS: All patients with tooth (66,7%) possessed disease periodontal, being 95% severe and 65% widespread. No periodontopathogenic bacteria were found in the atheromatous plaques. However, four samples (13.3%) were positive for the presence of bacteria. Of these, three participants were dentate, being two carriers of widespread severe chronic periodontite and one of located severe chronic periodontitis. None of them told the accomplishment of procedures associated to possible bacteremia episodes, as treatment endodontic, extraction the last six months or some procedure surgical dental. CONCLUSION: The periodontopathogenic bacteria studied were not found in the atheromatous plaques, making it impossible to establish the prevalence of these pathogens or the factors associated to their presence in plaques, the detection of positive samples for bacteria suggests that other periodontal and non-periodontal pathogens be studied in an attempt at discovering the association or not between periodontal disease and/or others infections and atherosclerosis, from the presence of these bacteria in atheromas

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Fucans, sulphated polysaccharides that contain L-fucose in its constitution, obtained from species of Phaeophyceae of the Sargassum kind, display several biological activities. Heterofucans from Sargassum filipendula are bioactive molecules that contain strong antiproliferative and antioxidant activity. However, their immunomodulatory and antimicrobial activities have not yet been examined. In this context, the aim of this research was to evaluate the heterofucans as for their immunomodulatory capacity and antimicrobial action against Leishmania infantum, Trichomonas vaginalis, Staphylococcus epidermidis and Klebsiella pneumonia (KPC). The five heterofucans obtained from S. filipendula show activities that are distant as stimulants of the immune system and microbial agent. The SF0.5V, SF0.7V amd SF1.0V heterofucans were capable of acting in the activation of murine and human macrophages. In addition to that, SF0.5V has shown antibiofilm activity of S. epidermides and SF0.7V and 1.0V almost completely inhibited the survival of the protozoan T. vaginalis. Results such as this one, reflect the broad range of action of the sulphated polysaccharides obtained from seaweeds, especially from the species S.filipendula

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The central interest of this thesis is to comprehend how the public action impels the formation and transformation of the tourist destinies. The research was based on the premise that the public actions are the result of the mediation process of state and non-state actors considered important in a section, which interact aiming for prevailing their interests and world visions above the others. The case of Porto de Galinhas beach, in Pernambuco, locus of the investigation of this thesis, allowed the analysis of a multiplicity of actors on the formation and implementation of local actions toward the development of the tourism between the years 1970 and 2010, as well as permitted the comprehension of the construction of the referential on the interventions made. This thesis, of a qualitative nature, has as theoretical support the cognitive approach of analysis of the public policies developed in France, and it has as main exponents the authors Bruno Jobert and Pierre Muller. This choice was made by the emphasis on the cognitive and normative factors of the politics, which aspects are not very explored in the studies of public policies in Brazil. As the source of the data collection, documental, bibliographic and field researches were utilized to the (re)constitution of the formation and transformation in the site concerned. The analysis techniques applied were the content and the documental analysis. To trace the public action referential, it started by the characterization of the touristic section frontiers and the creation of images by the main international body: the World Tourism Organization, of which analysis of the minutes of the meetings underscored guidelines to the member countries, including Brazil, which compounds the global-sectorial reference of the section. As from the analysis of the evolution of the tourism in the country, was identified that public policies in Brazil passed by transformations in their organization over the years, indicating changes in the referential that guided the interventions. These guidelines and transformations were identified in the construction of the tourist destination of Porto de Galinhas, of which data was systematized and presented in four historical periods, in which were discussed the values, the standard, the algorithms, the images and the important mediators. It has been revealed that the State worked in different roles in the decades analyzed in local tourism. From the 1990s, however, new actors were inserted in the formulation and implementation of policies developed, especially for local hotelkeepers. These, through their association, establishes a leadership relation in the local touristic section, thereby, they could set their hegemony and spread their own interest. The leadership acquired by a group of actors, in the case of Porto de Galinhas, does not mean that trade within the industry were neutralized, but that there is a cognitive framework that confronts the actors involved. In spite of the advances achieved by the work of the mediators in the last decades, that resulted in an amplification and diversification of the activity in the area, as well as the consolidation at the beach, as a tourist destiny of national standout, the position of the place is instable, concerned to the competitiveness, once that there is an situation of social and environmental unsustainability

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In this study, it has been investigated the influence of silver film deposition onto 100% polyester woven and non-woven, on the survival of Escherichia coli and Staphylococcus aureus in contact with these surfaces. The treatment was performedin a chamber containing the working gas at low pressure (~ 10-2 mbar). Some process parameters such as as voltage: 470 V; pressure: 10-2 mbar; current : 0.40 A and gas flow: 6 and 10 cm3/min were kept constant. For the treatments with purêargon plasma using a flow of 6 and 10 cm3/min, different treatment times were evaluated, such as, 10 , 20, 30, 40, 50 and 60 minutes. Contact angle (sessile drop), measurements were used to determine the surface tension of the treated fabrics and its influence on the bacteria grow as weel as the possibilities of a biofilm formation. The formation of a silver film, as well as the amount of this element was verified byEDX technique. The topography was observed through scanning electron microscopy (SEM) to determine the size of silver grains formed on the surfaces of the fabric and assess homogeneity of treatment. The X-ray diffraction (XRD) was used to analyze the structure of silver film deposition. The woven fabric treatments enabled the formation of silver particulate films with particle size larger than the non-woven fabrics. With respect to bacterial growth, all fabrics were shown to be bactericidal for Staphylococcus aureus (S. aureus), while for the Escherichia coli (E. coli), the best results were found for the non-woven fabric (TNT) treated with a flow of 10 cm3/min to both bacteria