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1.
Rev. chil. infectol ; 34(5): 458-466, oct. 2017. tab, graf
Article in Spanish | LILACS | ID: biblio-899743

ABSTRACT

Resumen Introducción: Los alimentos de origen animal frecuentemente están implicados en brotes de salmonelosis. Objetivo: Evaluar la frecuencia de Salmonella enterica en carnes molidas de pollo, res y cerdo (un total de 2.592 muestras) obtenidas de mercados sobre ruedas y supermercados de la Delegación Iztapalapa en la Ciudad de México, determinar la susceptibilidad antimicrobiana y efectuar ensayos de adherencia en las cepas aisladas. Métodos: El aislamiento de S. enterica se hizo de acuerdo a la BAM-FDA, la susceptibilidad antimicrobiana de acuerdo con CLSI y el ensayo de adherencia en células HEp-2 conforme a Baffone y cols., 2001. Resultados: Salmonella enterica fue aislada en 511 del total de muestras analizadas (19,7%), de las cuales 244 (47,7%), 152 (29,7%) y 115 (22,5%) correspondieron a carne molida de pollo, res y cerdo, respectivamente. La mayor frecuencia de resistencia de S. enterica a antimicrobianos fue a ampicilina y cloranfenicol en pollo, perfloxacina y ampicilina en res y carbenicilina, ampicilina, cloranfenicol, cefotaxima y perfloxacina en cerdo. Noventa por ciento de las cepas mostraron un patrón de adherencia agregativo. Conclusión: La frecuencia de S. enterica en productos cárnicos es alta, por lo que es importante la adecuada cocción de la carne para disminuir el riesgo de una salmonelosis.


Background: Food of animal origin is often involved in salmonellosis outbreaks. Aim: To evaluate the frequency of Salmonella enterica in chicken, beef and pork ground meat (a total of 2,592 samples) obtained from travelling markets and supermarkets at the Iztapalapa area of Mexico City, in order to determine the antimicrobial susceptibility and adherence capacity of isolated strains. Methods: Isolation of S. enterica was carried out according to the BAM-FDA, the microbial susceptibility according with CLSI and adherence assay on HEp-2 cell line according with Baffone et al., 2001. Results: S. enterica was isolated from 511 of all the analyzed samples (19.7%), from which 244 (47.7%), 152 (29.7%) and 115 (22.5%) corresponded to chicken, beef and pork ground meat, respectively. The highest frequency of resistance of S. enterica to antimicrobials was to ampicillin and chloramphenicol in chicken, perfloxacin and ampicillin in beef and carbenicillin, ampicillin, chloramphenicol, cefotaxime and perfloxacin in pork. Ninety percent of the strains showed an aggregative adherence pattern on HEp-2 cells. Conclusion: The frequency of S. enterica on meat products is high, which is the reason why a proper cooking of these ground meats is important in order to reduce the risk of acquiring salmonellosis.


Subject(s)
Animals , Poultry/microbiology , Bacterial Adhesion/physiology , Salmonella enterica/isolation & purification , Salmonella enterica/drug effects , Red Meat/microbiology , Anti-Bacterial Agents/pharmacology , Swine , Time Factors , Drug Resistance, Microbial , Cattle , Microbial Sensitivity Tests , Chickens , Cell Line, Tumor/microbiology , Serogroup , Food Microbiology , Mexico
2.
J. appl. oral sci ; 25(4): 436-441, July-Aug. 2017. tab, graf
Article in English | LILACS, BBO | ID: biblio-893638

ABSTRACT

Abstract Bacterial endotoxin (LPS) adhesion to orthodontic brackets is a known contributing factor to inflammation of the adjacent gingival tissues. Objective The aim of this study was to assess whether LPS adheres to orthodontic adhesive systems, comparing two commercial brands. Material and Methods Forty specimens were fabricated from Transbond XT and Light Bond composite and bonding agent components (n=10/component), then contaminated by immersion in a bacterial endotoxin solution. Contaminated and non-contaminated acrylic resin samples were used as positive and negative control groups, respectively. LPS quantification was performed by the Limulus Amebocyte Lysate QCL-1000™ test. Data obtained were scored and subjected to the Chi-square test using a significance level of 5%. Results There was endotoxin adhesion to all materials (p<0.05). No statistically significant difference was found between composites/bonding agents and acrylic resin (p>0.05). There was no significant difference (p>0.05) among commercial brands. Affinity of endotoxin was significantly greater for the bonding agents (p=0.0025). Conclusions LPS adhered to both orthodontic adhesive systems. Regardless of the brand, the endotoxin had higher affinity for the bonding agents than for the composites. There is no previous study assessing the affinity of LPS for orthodontic adhesive systems. This study revealed that LPS adheres to orthodontic adhesive systems. Therefore, additional care is recommended to orthodontic applications of these materials.


Subject(s)
Bacterial Adhesion/physiology , Lipopolysaccharides/physiology , Composite Resins/chemistry , Resin Cements/chemistry , Escherichia coli , Reference Values , Materials Testing , Enzyme-Linked Immunosorbent Assay , Lipopolysaccharides/isolation & purification
3.
Arq. gastroenterol ; 54(2): 130-134, Apr.-June 2017. tab, graf
Article in English | LILACS | ID: biblio-838836

ABSTRACT

ABSTRACT BACKGROUND The diarrheal syndrome is considered a serious public health problem all over the world and is considered a major cause of morbidity and mortality in developing countries. The high incidence of enteroaggregative Escherichia coli in diarrheal syndromes classified as an emerging pathogen of gastrointestinal infections. After decades of study, your pathogenesis remains uncertain and has been investigated mainly using in vitro models of adhesion in cellular lines. OBJECTIVE The present study investigated the interaction of enteroaggregative Escherichia coli strains isolated from childhood diarrhea with rabbit ileal and colonic mucosa ex vivo, using the in vitro organ culture model. METHODS The in vitro adhesion assays using cultured tissue were performed with the strains co-incubated with intestinal fragments of ileum and colon over a period of 6 hours. Each strain was tested with three intestinal fragments for each region. The fragments were analysed by scanning electron microscopy. RESULTS Through scanning electron microscopy we observed that all strains adhered to rabbit ileal and colonic mucosa, with the typical aggregative adherence pattern of “stacked bricks” on the epithelium. However, the highest degree of adherence was observed on colonic mucosa. Threadlike structures were found in greater numbers in the ileum compared to the colon. CONCLUSION These data showed that enteroaggregative Escherichia coli may have a high tropism for the human colon, which was ratified by the higher degree of adherence on the rabbit colonic mucosa. Finally, data indicated that in vitro organ culture of intestinal mucosa from rabbit may be used to elucidate the enteroaggregative Escherichia coli pathogenesis.


RESUMO CONTEXTO A síndrome diarréica é considerada um grave problema de saúde pública em todo o mundo e é considerada uma das principais causas de morbidade e mortalidade nos países em desenvolvimento. A elevada incidência de Escherichia coli enteroagregativa nas síndromes diarreicas a classificou como um patógeno emergente de infecções gastrintestinais. Depois de décadas de estudo, sua patogênese ainda é incerta e tem sido investigada usando principalmente modelos in vitro de adesão em linhagens celulares. OBJETIVO O presente estudo investigou a interação de cepas de Escherichia coli enteroagregativa isoladas de diarreia infantil com mucosa ileal e colônica de coelho ex vivo, utilizando o modelo de cultura de órgão in vitro. MÉTODOS Os ensaios de adesão in vitro utilizando tecido cultivado foram realizados com as cepas co-incubadas com fragmentos intestinais de íleo e de cólon durante um período de 6 horas. Cada cepa foi testada em três fragmentos intestinais para cada região. Os fragmentos foram analisados por microscopia eletrônica de varredura. RESULTADOS Através da microscopia eletrônica de varredura observamos que todas as cepas aderiram a mucosa ileal e colônica de coelho, com o padrão de aderência agregativo típico de “tijolos empilhados” no epitélio. Entretanto, o maior grau de adesão foi observado na mucosa do cólon. Estruturas filiformes foram encontradas em maior número no íleo em comparação com o cólon. CONCLUSÃO Esses dados mostraram que Escherichia coli enteroagregativa pode ter um maior tropismo para o cólon humano, o que foi ratificado pelo maior grau de aderência na mucosa do cólon de coelho. Finalmente, os dados indicaram que a cultura de órgão in vitro da mucosa intestinal de coelho pode ser utilizado para elucidar a patogênese de Escherichia coli enteroagregativa.


Subject(s)
Humans , Animals , Male , Bacterial Adhesion/physiology , Colon/microbiology , Escherichia coli/physiology , Escherichia coli Infections/microbiology , Ileum/microbiology , Intestinal Mucosa/microbiology , Phylogeny , Rabbits , Microscopy, Electron, Scanning , Colon/ultrastructure , Virulence Factors , Ileum/ultrastructure , Intestinal Mucosa/ultrastructure
4.
Braz. dent. j ; 27(3): 255-260, May-June 2016. tab, graf
Article in English | LILACS | ID: lil-782824

ABSTRACT

Abstract The periodontopathogen Aggregatibacter actinomycetemcomitans colonizes oral cavity by binding to and invading epithelial cells as well as by participating in biofilms formed on hard surfaces. Aae, an autotransporter protein, is implicated in bacterial adhesion to epithelial cells. Due to the multiple functions of bacterial autotransporter proteins, this study aimed to evaluate the role of aae in A. actinomycetemcomitans ability to adhere to both saliva-coated hydroxyapatite (SHA) and biofilm. An aae null mutant was constructed. Its hydrophobic properties as well as its ability to adhere to epithelial cells, SHA and to form biofilm were evaluated and compared with the parental strain, A. actinomycetemcomitans VT1169. The aae null mutant showed reduced hydrophobicity, as well as decreased binding to SHA and biofilm formation compared to the parental strain. These data suggest that aae mediates A. actinomycetemcomitans adhesion to epithelial cells and may be involved in biofilm formation and interaction with adsorbed salivary proteins.


Resumo O peridontopatógeno Aggregatibacter actinomycetemcomitans coloniza a cavidade oral aderindo e invadindo as células epiteliais e participando da formação de biofilme em superfícies duras. Aae, uma proteína autotransportadora está relacionada com a adesão bacteriana às células epiteliais. Devido às múltiplas funções desempenhadas por proteínas bacterianas autotransportadoras, este estudo teve como objetivo avaliar o papel de aae de A. actinomycetemcomitans tanto na capacidade de aderir à hidroxiapatita recoberta por saliva (SHA), quanto a de formar biofilme. Um mutante nulo aae foi construído. Suas propriedades hidrofóbicas, bem como a sia capacidade para aderir às células epiteliais, à SHA e para formar biofilme foram avaliadas e comparadas com a cepa -mãe, A. Actinomycetemcomitans VT1169. O mutante nulo aae apresentou redução de hidrofobicidade, assim como diminuição da adesão à SHA e na formação de biofilme, quando comparado à cepa parental. Estes dados sugerem que aae media a adesão de A. Actinomycetemcomitans às células epiteliais e pode também estar envolvida na formação de biofilme e na interação com proteínas salivares adsorvidas.


Subject(s)
Aggregatibacter actinomycetemcomitans/physiology , Bacterial Adhesion/physiology , Bacterial Proteins/physiology , Membrane Transport Proteins/physiology , Aggregatibacter actinomycetemcomitans/genetics , Bacterial Proteins/genetics , Biofilms , Gene Knockdown Techniques , Hydrophobic and Hydrophilic Interactions , Membrane Transport Proteins/genetics
5.
Biol. Res ; 49: 1-10, 2016. ilus, graf
Article in English | LILACS | ID: lil-774429

ABSTRACT

BACKGROUND: Vibrio parahaemolyticus (V. parahaemolyticus) is a Gram-negative, halophilic bacterium recognized as one of the most important foodborne pathogen. When ingested, V. parahaemolyticus causes a self-limiting illness (Vibriosis), characterized mainly by watery diarrhoea. Treatment is usually oral rehydration and/or antibiotics in complicated cases. Since 1996, the pathogenic and pandemic V. parahaemolyticus O3:K6 serotype has spread worldwide, increasing the reported number of vibriosis cases. Thus, the design of new strategies for pathogen control and illness prevention is necessary. Lactobacillus sp. grouped Gram positive innocuous bacteria, part of normal intestinal microbiota and usually used as oral vaccines for several diarrheic diseases. Recombinants strains of Lactobacillus (RL) expressing pathogen antigens can be used as part of an anti-adhesion strategy where RL block the pathogen union sites in host cells. Thus, we aimed to express MAM-7 V. parahaemolyticus adhesion protein in Lactobacillus sp. to generate an RL that prevents pathogen colonization RESULTS: We cloned the MAM-7 gene from V. parahaemolyticus RIMD 2210633 in Lactobacillus expression vectors. Recombinant strains (Lactobacillus rhamnosus pSEC-MAM7 and L. rhamnosus pCWA-MAM7) adhered to CaCo-2 cells and competed with the pathogen. However, the L. rhamnosus wild type strain showed the best capacity to inhibit pathogen colonization in vitro. In addition, LDH-assay showed that recombinant strains were cytotoxic compared with the wild type isogenic strain CONCLUSIONS: MAM-7 expression in lactobacilli reduces the intrinsic inhibitory capacity of L. rhamnosus against V. parahaemolyticus.


Subject(s)
Humans , Adhesins, Bacterial/analysis , Bacterial Adhesion/physiology , Lactobacillus rhamnosus/physiology , Vibrio parahaemolyticus/pathogenicity , Biofilms/growth & development , Cell Line , Cytotoxicity, Immunologic , Electrophoresis, Polyacrylamide Gel , Gene Expression , Gentian Violet , Polymerase Chain Reaction , Vibrio Infections/prevention & control , Vibrio parahaemolyticus/growth & development , Vibrio parahaemolyticus/metabolism
6.
Rev. argent. microbiol ; 47(3): 261-273, set. 2015. ilus
Article in Spanish | LILACS | ID: biblio-1129897

ABSTRACT

With the aim of exploiting symbiotic nitrogen fixation, soybean crops are inoculated with selected strains of Bradyrhizobium japonicum, Bradyrhizobium diazoefficiens or Bradyrhizobium elkanii (collectively referred to as Bradyrhizobium spp.). The most common method of inoculation used is seed inoculation, whether performed immediately before sowing or using preinoculated seeds or pretreated seeds by the professional seed treatment. The methodology of inoculation should not only cover the seeds with living rhizobia, but must also optimize the chances of these rhizobia to infect the roots and nodulate. To this end, inoculated rhizobia must be in such an amount and condition that would allow them to overcome the competition exerted by the rhizobia of the allochthonous population of the soil, which are usually less effective for nitrogen fixation and thus dilute the effect of inoculation on yield. This optimization requires solving some queries related to the current knowledge of seed inoculation, which are addressed in this article. I conclude that the aspects that require further research are the adhesion and survival of rhizobia on seeds, the release of rhizobia once the seeds are deposited in the soil, and the movement of rhizobia from the vicinity of the seeds to the infection sites in the roots


Con el fin de aprovechar la fijación simbiótica de nitrógeno, el cultivo de soja se inocula con cepas seleccionadas de Bradyrhizobium japonicum, Bradyrhizobium diazoefficiens o Bradyrhizobium elkanii (conjuntamente referidas como Bradyrhizobium spp.). El método más común de hacerlo es la inoculación en semillas, ya sea que esta se realice en el momento previo a la siembra o que se utilicen semillas preinoculadas o pretratadas mediante el tratamiento profesional de semillas. La metodología de inoculación no debe limitarse a recubrir las semillas con rizobios vivos, sino que también debe optimizar las chances de esos rizobios para infectar las raíces y nodular. Para ello los rizobios inoculados deben estar en una cantidad y un estado tales que les permitan superar la competición ejercida por los rizobios de la población alóctona del suelo, los cuales usualmente son menos eficaces para la fijación de nitrógeno y así diluyen el efecto de la inoculación sobre el rendimiento. Esta optimización requiere resolver algunos interrogantes, que son abordados en el presente artículo. Concluyo que los aspectos que requieren más investigación son la adhesión y supervivencia de los rizobios en las semillas, la liberación de los rizobios una vez que las semillas se depositan en el suelo y el movimiento de los rizobios desde las inmediaciones de las semillas hasta los sitios de infección en las raíces


Subject(s)
Soybeans/growth & development , Soybeans/metabolism , Bradyrhizobium/growth & development , Bradyrhizobium/metabolism , Agricultural Inoculants/metabolism , Nitrogen Fixation , Bacterial Adhesion/physiology , Survival Analysis
7.
Mem. Inst. Oswaldo Cruz ; 109(4): 414-419, 03/07/2014. graf
Article in English | LILACS | ID: lil-716305

ABSTRACT

While the influence of water in Helicobacter pylori culturability and membrane integrity has been extensively studied, there are little data concerning the effect of this environment on virulence properties. Therefore, we studied the culturability of water-exposed H. pylori and determined whether there was any relation with the bacterium’s ability to adhere, produce functional components of pathogenicity and induce inflammation and alterations in apoptosis in an experimental model of human gastric epithelial cells. H. pylori partially retained the ability to adhere to epithelial cells even after complete loss of culturability. However, the microorganism is no longer effective in eliciting in vitro host cell inflammation and apoptosis, possibly due to the non-functionality of the cag type IV secretion system. These H. pylori-induced host cell responses, which are lost along with culturability, are known to increase epithelial cell turnover and, consequently, could have a deleterious effect on the initial H. pylori colonisation process. The fact that adhesion is maintained by H. pylori to the detriment of other factors involved in later infection stages appears to point to a modulation of the physiology of the pathogen after water exposure and might provide the microorganism with the necessary means to, at least transiently, colonise the human stomach.


Subject(s)
Humans , Bacterial Adhesion/physiology , Epithelial Cells/microbiology , Helicobacter pylori/pathogenicity , Water , Antigens, Bacterial/physiology , Bacterial Secretion Systems , Bacterial Proteins/physiology , Gastric Mucosa/cytology , Host-Pathogen Interactions , Helicobacter pylori/growth & development , Virulence/physiology
8.
Dental press j. orthod. (Impr.) ; 19(1): 60-68, Jan-Feb/2014. tab, graf
Article in English | LILACS | ID: lil-709643

ABSTRACT

OBJECTIVE: To verify, by means of a systematic review, whether the design of brackets (conventional or self-ligating) influences adhesion and formation of Streptococcus mutans colonies. METHODS: Search strategy: four databases (Cochrane Central Register of Controlled Trials, Ovid ALL EMB Reviews, PubMed and BIREME) were selected to search relevant articles covering the period from January 1965 to December 2012. Selection Criteria: in first consensus by reading the title and abstract. The full text was obtained from publications that met the inclusion criteria. Data collection and analysis: Two reviewers independently extracted data using the keywords: conventional, self-ligating, biofilm, Streptococcus mutans, and systematic review; and independently evaluated the quality of the studies. In case of divergence, the technique of consensus was adopted. RESULTS: The search strategy resulted in 1,401 articles. The classification of scientific relevance revealed the high quality of the 6 eligible articles of which outcomes were not unanimous in reporting not only the influence of the design of the brackets (conventional or self-ligating) over adhesion and formation of colonies of Streptococcus mutans, but also that other factors such as the quality of the bracket type, the level of individual oral hygiene, bonding and age may have greater influence. Statistical analysis was not feasible because of the heterogeneous methodological design. CONCLUSIONS: Within the limitations of this study, it was concluded that there is no evidence for a possible influence of the design of the brackets (conventional or self-ligating) over colony formation and adhesion of Streptococcus mutans. .


OBJETIVO: verificar, por meio de uma revisão sistemática, se o design dos braquetes (convencionais ou autoligáveis) apresenta influência na aderência e formação de colônias de Streptococcus mutans. MÉTODOS: quatro bases de dados (Cochrane Central Register of Controlled Trials; Ovid ALL EMB Reviews; PubMed e BIREME) foram selecionadas para a busca por artigos relevantes, do período de janeiro de 1965 a dezembro de 2012. Os critérios de seleção foram inicialmente aplicados aos títulos e abstracts e o texto integral foi obtido de publicações que cumprira os critérios de inclusão. Dois revisores, de forma independente, extraíram os dados utilizando as palavras-chave "convencionais", "autoligados", "biofilme", "Streptococcus mutans" e "revisão sistemática" e avaliaram a qualidade metodológica dos estudos incluídos. No caso de divergência, foi adotada a técnica do consenso. RESULTADOS: a estratégia de busca resultou em 1.401 artigos. A classificação da relevância científica revelou alta qualidade dos 6 artigos elegíveis, cujos desfechos não foram unânimes em relatar a influência do design dos braquetes (convencionais ou autoligáveis) sobre a aderência e a formação de colônias de Streptococcus mutans, e que outros fatores como características dos tipos de braquetes, o nível de higiene bucal individual, colagem e idade dos indivíduos, podem ter maior influência. O tratamento estatístico foi inviável por causa do desenho metodológico heterogêneo. CONCLUSÕES: dentro das limitações do presente estudo, concluiu-se que não há evidência de uma possível influência do design dos braquetes (convencionais ou autoligáveis) sobre a aderência e a formação ...


Subject(s)
Humans , Biofilms/growth & development , Orthodontic Appliance Design , Orthodontic Brackets/microbiology , Streptococcus mutans/physiology , Bacterial Adhesion/physiology , Dental Plaque/microbiology
9.
Rev. méd. Chile ; 141(3): 291-297, mar. 2013. ilus
Article in Spanish | LILACS | ID: lil-677335

ABSTRACT

Background: Copper has a bactericidal activity against a series of bacterial strains. Aim: To measure resistance to bacterial adherence of copper (Cu) and stainless steel (SS) metal coupons. Material and Methods: Bacterial strains causing nosocomial infections in Chile were analyzed. Bacterial adherence was studied using apreviously described method based on a system of metal coupons that are immersed in culture media containing the bacteria ofinterest at room temperature. Results: Adherence to Cu and SS coupons was differentfor Methicillin-resistant Staphylococcus aureus (MRSA), Klebsiella pneumoniae and Acinetobacter baumannii strains. For these strains, no adherence to Cu coupons occurred during the 48 h observation period compared to a rapidly increasing adherence to SS coupons, with a final colony count of 1.00E + 07 cfu/mL. For two different Pseudomonas aeruginosa clinical strains, inhibition of adherence was not observed on Cu coupons, and colony counts were similar for Cu and SS using the standard inoculum (2-3 xlO7 cfu).Apartial decrease in adherence was observed for Cu but not for SS coupons, when a lower inoculum was used. Conclusions: Copper surfaces represent an interesting option to reduce bacterial contamination in the hospital environment due to its resistance to bacterial adhesión ofmost ofthe common nosocomial bacterial strains.


Subject(s)
Adult , Humans , Bacterial Adhesion/physiology , Copper , Cross Infection/microbiology , Stainless Steel , Acinetobacter baumannii/physiology , Colony Count, Microbial , Klebsiella pneumoniae/physiology , Methicillin-Resistant Staphylococcus aureus/physiology , Pseudomonas aeruginosa/physiology
10.
J. appl. oral sci ; 20(5): 544-549, Sept.-Oct. 2012. tab
Article in English | LILACS | ID: lil-654919

ABSTRACT

In Orthodontics, fixed appliances placed in the oral cavity are colonized by microorganisms. OBJECTIVE: The purpose of this study was to quantitatively determine the independent bacterial colonization of S. mutans and S. sobrinus in orthodontic composite resins. MATERIAL AND METHODS: Seven orthodontic composite adhesives for bonding brackets were selected and classified into 14 groups; (GIm, GIs) Enlight, (GIIm, GIIs) Grengloo, (GIIIm, GIIIs) Kurasper F, (GIVm, GIVs) BeautyOrtho Bond, (GVm, GVs) Transbond CC, (GVIm, GVIs) Turbo Bond II, (GVIIm, GVIIs) Blugloo. 60 blocks of 4x4x1 mm of each orthodontic composite resin were made (total 420 blocks), and gently polished with sand-paper and ultrasonically cleaned. S. mutans and S. sobrinus were independently cultivated. For the quantitative analysis, a radioactive marker was used to codify the bacteria (³H) adhered to the surface of the materials. The blocks were submerged in a solution with microorganisms previously radiolabeled and separated (210 blocks for S. mutans and 210 blocks for S. sobrinus) for 2 hours at 37ºC. Next, the blocks were placed in a combustion system, to capture the residues and measure the radiation. The statistical analysis was calculated with the ANOVA test (Sheffè post-hoc). RESULTS: Significant differences of bacterial adhesion were found amongst the groups. In the GIm and GIs the significant lowest scores for both microorganisms were shown; in contrast, the values of GVII for both bacteria were significantly the highest. CONCLUSIONS: This study showed that the orthodontic composite resin evaluated in the GIm and GIs, obtained the lowest adherence of S. mutans and S. sobrinus, which may reduce the enamel demineralization and the risk of white spot lesion formation.


Subject(s)
Bacterial Adhesion/physiology , Composite Resins , Dental Cements , Orthodontic Brackets/microbiology , Streptococcus mutans/growth & development , Streptococcus sobrinus/growth & development , Acrylic Resins , Analysis of Variance , Bacterial Load , Bisphenol A-Glycidyl Methacrylate , Dental Polishing , Phosphoric Acids , Resin Cements , Surface Properties
11.
Mem. Inst. Oswaldo Cruz ; 107(4): 486-493, June 2012. ilus, graf
Article in English | LILACS | ID: lil-626442

ABSTRACT

Corynebacterium pseudodiphtheriticum is a well-known human pathogen that mainly causes respiratory disease and is associated with high mortality in compromised hosts. Little is known about the virulence factors and pathogenesis of C. pseudodiphtheriticum. In this study, cultured human epithelial (HEp-2) cells were used to analyse the adherence pattern, internalisation and intracellular survival of the ATCC 10700 type strain and two additional clinical isolates. These microorganisms exhibited an aggregative adherence-like pattern to HEp-2 cells characterised by clumps of bacteria with a "stacked-brick" appearance. The differences in the ability of these microorganisms to invade and survive within HEp-2 cells and replicate in the extracellular environment up to 24 h post infection were evaluated. The fluorescent actin staining test demonstrated that actin polymerisation is involved in the internalisation of the C. pseudodiphtheriticum strains. The depolymerisation of microfilaments by cytochalasin E significantly reduced the internalisation of C. pseudodiphtheriticum by HEp-2 cells. Bacterial internalisation and cytoskeletal rearrangement seemed to be partially triggered by the activation of tyrosine kinase activity. Although C. pseudodiphtheriticum strains did not demonstrate an ability to replicate intracellularly, HEp-2 cells were unable to fully clear the pathogen within 24 h. These characteristics may explain how some C. pseudodiphtheriticum strains cause severe infection in human patients.


Subject(s)
Humans , Bacterial Adhesion/physiology , Corynebacterium/pathogenicity , Epithelial Cells/microbiology , Corynebacterium/physiology , Virulence
12.
Braz. j. med. biol. res ; 45(5): 417-424, May 2012. ilus, tab
Article in English | LILACS | ID: lil-622765

ABSTRACT

The adhesins of extraintestinal pathogenic Escherichia coli are essential for mediating direct interactions between the microbes and the host cell surfaces that they infect. Using fluorescence microscopy and gentamycin protection assays, we observed that 49 sepsis-associated E. coli (SEPEC) strains isolated from human adults adhered to and invaded Vero cells in the presence of D-mannose (100%). In addition, bacteria concentrations of approximately 2 x 10(7) CFU/mL were recovered from Vero cells following an invasion assay. Furthermore, PCR analysis of adhesin genes showed that 98.0% of these SEPEC strains tested positive for fimH, 69.4% for flu, 53.1% for csgA, 38.8% for mat, and 32.7% for iha. Analysis of the invasin genes showed that 16.3% of the SEPEC strains were positive for tia, 12.3% for gimB, and 10.2% for ibeA. Therefore, these data suggest that SEPEC adhesion to cell surfaces occurs through non-fimH mechanisms. Scanning electron microscopy showed the formation of microcolonies on the Vero cell surface. SEPEC invasiveness was also confirmed by the presence of intracellular bacteria, and ultrastructural analysis using electron transmission microscopy revealed bacteria inside the Vero cells. Taken together, these results demonstrate that these SEPEC strains had the ability to adhere to and invade Vero cells. Moreover, these data support the theory that renal cells may be the predominant pathway through which SEPEC enters human blood vessels.


Subject(s)
Adult , Animals , Humans , Adhesins, Bacterial/physiology , Bacterial Adhesion/physiology , Epithelial Cells/microbiology , Escherichia coli Infections/microbiology , Escherichia coli/physiology , Sepsis/microbiology , Adhesins, Bacterial/genetics , Adhesins, Bacterial/ultrastructure , Bacterial Adhesion/genetics , Chlorocebus aethiops , Epithelial Cells/ultrastructure , Escherichia coli/genetics , Escherichia coli/ultrastructure , Genotype , Microscopy, Electron, Scanning , Microscopy, Fluorescence , Polymerase Chain Reaction , Vero Cells
13.
Biol. Res ; 45(1): 21-26, 2012. ilus, tab
Article in English | LILACS | ID: lil-626743

ABSTRACT

Shigella flexneri causes bacillary dysentery in humans. Essential to the establishment of the disease is the invasion of the colonic epithelial cells. Here we investigated the role of the lipopolysaccharide (LPS) O antigen in the ability of S. flexneri to adhere to and invade polarized Caco-2 cells. The S. flexneri 2a O antigen has two preferred chain lengths: a short O antigen (S-OAg) regulated by the WzzB protein and a very long O antigen (VL-OAg) regulated by Wzz pHS2. Mutants with defined deletions of the genes required for O-antigen assembly and polymerization were constructed and assayed for their abilities to adhere to and enter cultured epithelial cells. The results show that both VL- and S-OAg are required for invasion through the basolateral cell membrane. In contrast, the absence of O antigen does not impair adhesion. Purified LPS does not act as a competitor for the invasion of Caco-2 cells by the wild-type strain, suggesting that LPS is not directly involved in the internalization process by epithelial cells.


Subject(s)
Humans , Bacterial Adhesion/physiology , Bacterial Proteins/analysis , Dysentery, Bacillary/microbiology , O Antigens/chemistry , Shigella flexneri/pathogenicity , Dysentery, Bacillary/immunology , O Antigens/metabolism , Polymerization , Shigella flexneri/immunology
14.
Braz. j. med. biol. res ; 43(7): 611-618, July 2010. ilus
Article in English | LILACS | ID: lil-550737

ABSTRACT

Helicobacter pylori adhesion to gastric epithelial cells constitutes a key step in the establishment of a successful infection of the gastric mucosa. The high representation of outer membrane proteins in the bacterial genome suggests the relevance of those proteins in the establishment of profitable interactions with the host gastric cells. Gastric epithelial cells are protected by a mucous layer gel, mainly consisting of the MUC5AC and MUC6 mucins. In addition to this protective role, mucins harbor glycan-rich domains that constitute preferential binding sites of many pathogens. In this article we review the main players in the process of H. pylori adhesion to gastric epithelial cells, which contribute decisively to the high prevalence and chronicity of H. pylori infection. The BabA adhesin recognizes both H-type 1 and Lewis b blood-group antigens expressed on normal gastric mucosa of secretor individuals, contributing to the initial steps of infection. Upon colonization, persistent infection induces an inflammatory response with concomitant expression of sialylated antigens. The SabA adhesin mediates H. pylori binding to inflamed gastric mucosa by recognizing sialyl-Lewis a and sialyl-Lewis x antigens. The expression of the BabA and SabA adhesins is tightly regulated, permitting the bacteria to rapidly adapt to the changes of glycosylation of the host gastric mucosa that occur during infection, as well as to escape from the inflammatory response. The growing knowledge of the interactions between the bacterial adhesins and the host receptors will contribute to the design of alternative strategies for eradication of the infection.


Subject(s)
Animals , Humans , Antigens, Bacterial/metabolism , Bacterial Adhesion/physiology , Epithelial Cells/microbiology , Gastric Mucosa/microbiology , Helicobacter Infections/microbiology , Helicobacter pylori/physiology , Adhesins, Bacterial/metabolism , /metabolism , Helicobacter pylori/metabolism , Lewis Blood Group Antigens/metabolism
15.
Arch. latinoam. nutr ; 60(2): 175-178, jun. 2010. ilus, tab
Article in Spanish | LILACS | ID: lil-630314

ABSTRACT

Listeria monocytogenes, además de ser un género capaz de producir una enfermedad infecciosa grave en el hombre, puede formar biopelículas en distintas superficies relacionadas con el ambiente de producción alimentario. Éstas constituyen un serio problema debido a que son una fuente importante y constante de contaminación para los alimentos y el ambiente de producción, además de que las bacterias presentes en ellas poseen una aumentada resistencia hacia agentes físicos y químicos de uso frecuente. En el presente trabajo se estudió la capacidad de formación de biopelícula de cepas de L. monocytogenes previamente aisladas a partir de queso tierno bajo diferentes condiciones de temperatura y cultivo. Se utilizó una técnica de microplaca con diferentes medios de cultivo (CICC, CTS 1:20 y suero de queso) a diferentes temperaturas de incubación (refrigeración, ambiente y 35ºC). La capacidad de formación de biopelícula fue clasificada según la densidad óptica obtenida a 620 nm. Ninguna de las cepas evaluadas fue clasificada como formadora fuerte de biopelicula bajo ninguna de las variables estudiadas, sí se detectaron formadoras débiles y moderadas. Los resultados obtenidos ponen de manifiesto la influencia del contenido de nutrientes en el medio de cultivo sobre la formación de biopelícula, no obstante, el CICC fue el único medio que permitió la expresión de formadores moderados. Por el contrario, el suero de queso resultó poco favorecedor. La formación de biopelícula es un proceso multifactorial, donde el nivel de adsorción depende de gran cantidad de variables y cuyo estudio debe fomentarse, de manera que se desarrollen metodologías que permitan su reducción o eliminación, de manera que las industrias alimentarias aseguren productos inocuos y de buena calidad microbiológica.


Listeria monocytogenes is a bacteria associated with the production of severe infectious disease in human being, but also with the formation of biofilms in different surfaces related to the food production environment. Biofilm represents a serious problem in food industry, since it is a constant and important contamination source and also, bacteria present in it have an increased resistance towards physical and chemical agents of common use. The capacity of biofilm formation of L. monocytogenes strains previously isolated from soft cheese samples from Costa Rica was studied under different temperature and culture conditions. The microplate technique was performed using different culture media (BHIB, TSB 1:20 and cheese serum) and at different incubation temperatures (refrigeration, environmental and 35ºC). Biofilm formation capacity was classified according to the optical density obtained at 620nm. None of the strains evaluated was classified as strong biofilm former under any of the variables studied, nevertheless, weak and moderate formers were detected. The results obtained show the influence of the nutrient content of the culture media used over biofilm formation; BHIB was the only culture media that allowed the expression of moderate biofilm forms, contrary to cheese serum that did not promote biofilm production. Biofilm formation is a multifactorial process, where adsorption level depends on several variables and its study must be promoted in order to develop methodologies that allow its reduction or elimination, so food industries may offer safe food products to consumers.


Subject(s)
Biofilms/growth & development , Cheese/microbiology , Listeria monocytogenes/physiology , Temperature , Bacterial Adhesion/physiology , Colony Count, Microbial , Costa Rica , Listeria monocytogenes/isolation & purification , Time Factors
16.
Rev. Inst. Med. Trop. Säo Paulo ; 52(3): 139-143, May-June 2010. tab
Article in English | LILACS | ID: lil-550344

ABSTRACT

Phospholipase and proteinase production and the ability of adhesion to buccal epithelial cells (BEC) of 112 Candida isolates originated from oral cavity of HIV infected patients and from blood and catheter of intensive care unit patients were investigated. The proteinase production was detected by inoculation into bovine serum albumin (BSA) agar and the phospholipase activity was performed using egg yolk emulsion. A yeast suspension of each test strain was incubated with buccal epithelial cells and the number of adherence yeast to epithelial cells was counted. A percentage of 88.1 percent and 55.9 percent of Candida albicans and 69.8 percent and 37.7 percent of non-albicans Candida isolates produced proteinase and phospholipase, respectively. Non-albicans Candida isolated from catheter were more proteolytic than C. albicans isolates. Blood isolates were more proteolytic than catheter and oral cavity isolates while oral cavity isolates produced more phospholipase than those from blood and catheter. C. albicans isolates from oral cavity and from catheter were more adherent to BEC than non-albicans Candida isolates, but the adhesion was not different among the three sources analyzed. The results indicated differences in the production of phospholipase and proteinase and in the ability of adhesion to BEC among Candida spp. isolates from different sources. This study suggests that the pathogenicity of Candida can be correlated with the infected site.


A produção de proteinase e fosfolipase e habilidade de adesão à célula epitelial bucal de 112 isolados de Candida originadas da cavidade bucal de pacientes infectados pelo HIV e de sangue e cateter de pacientes hospitalizados foram investigados. A produção de proteinase foi detectada por inoculação em ágar soro albumina bovina e a atividade de fosfolipase foi realizada usando emulsão de gema de ovo. A suspensão de levedura de cada isolado foi incubada com célula epitelial e o número de leveduras aderidas a célula epitelial foi contada. Uma porcentagem de 88,1 e 55,9 por cento de C. albicans e 69,8 e 37,7 por cento de isolados de Candida não albicans produziram proteinase e fosfolipase, respectivamente. Candida não albicans obtidas do cateter foram mais proteolíticos que isolados de Candida albicans (p < 0,001). Isolados do sangue foram mais proteolíticos do que isolados do cateter e cavidade bucal, enquanto isolados da cavidade bucal produziram mais fosfolipase do que aqueles isolados do sangue e cateter. C. albicans isoladas da cavidade bucal e do cateter foram mais aderentes à célula epitelial bucal do que isolados de Candida não albicans, mas não houve diferença na adesão entre os três locais analisados. Os resultados indicaram diferenças na produção de fosfolipase e proteinase e na habilidade de adesão à célula epitelial bucal entre os isolados de Candida das diferentes fontes. Este estudo sugere que a patogenicidade de Candida spp pode estar correlacionada ao local infectado.


Subject(s)
Humans , Aspartic Acid Proteases/biosynthesis , Bacterial Adhesion/physiology , Candida/enzymology , Candida/physiology , Phospholipases/biosynthesis , Candida/isolation & purification , Catheters, Indwelling/microbiology , Epithelial Cells/microbiology , HIV Infections/microbiology , Mouth/microbiology
17.
Braz. j. med. biol. res ; 41(11): 986-991, Nov. 2008. ilus, tab
Article in English | LILACS | ID: lil-500362

ABSTRACT

Invasive diseases caused by Corynebacterium diphtheriae have been described increasingly. Several reports indicate the destructive feature of endocarditis attributable to nontoxigenic strains. However, few reports have dealt with the pathogenicity of invasive strains. The present investigation demonstrates a phenotypic trait that may be used to identify potentially invasive strains. The study also draws attention to clinical and microbiological aspects observed in 5 cases of endocarditis due to C. diphtheriae that occurred outside Europe. Four cases occurred in female school-age children (7-14 years) treated at different hospitals in Rio de Janeiro, Brazil. All patients developed other complications including septicemia, renal failure and/or arthritis. Surgical treatment was performed on 2 patients for valve replacement. Lethality was observed in 40 percent of the cases. Microorganisms isolated from 5 blood samples and identified as C. diphtheriae subsp mitis (N = 4) and C. diphtheriae subsp gravis (N = 1) displayed an aggregative adherence pattern to HEp-2 cells and identical one-dimensional SDS-PAGE protein profiles. Aggregative-adhering invasive strains of C. diphtheriae showed 5 distinct RAPD profiles. Despite the clonal diversity, all 5 C. diphtheriae invasive isolates seemed to display special bacterial adhesive properties that may favor blood-barrier disruption and systemic dissemination of bacteria. In conclusion, blood isolates from patients with endocarditis exhibited a unique adhering pattern, suggesting a pathogenic role of aggregative-adhering C. diphtheriae of different clones in endocarditis. Accordingly, the aggregative-adherence pattern may be used as an indication of some invasive potential of C. diphtheriae strains.


Subject(s)
Adolescent , Child , Female , Humans , Bacterial Adhesion/physiology , Corynebacterium diphtheriae/pathogenicity , Endocarditis, Bacterial/microbiology , Bacterial Typing Techniques , Cells, Cultured , Corynebacterium diphtheriae/genetics , Corynebacterium diphtheriae/isolation & purification , Electrophoresis, Polyacrylamide Gel , Genotype , Phenotype , Random Amplified Polymorphic DNA Technique , Species Specificity
18.
Int. j. odontostomatol. (Print) ; 2(1): 95-99, jul. 2008.
Article in English | LILACS | ID: lil-545859

ABSTRACT

Tongue mucosae of one day postnatal rat was examined by transmission electron microscopic and HRSEMmethods.The specimens were fixed using Karnovsky solution and embedded in Spurr resin for transmission electron microscopy. For HRSEM methods, the samples were fixed in 2 percent osmiun tetroxide, dehydrated in alcohol, critical point dried and coated with gold-palladium. The results demonstrated that the surface of tongue present the filiform and fungiform papillae covered by numerous keratinized epithelial cells. The bacteriae are attached to the surfaces of microplicae at random, demonstrating in their three-dimensional HRSEM images. At high magnification, the transmission electron microscopic images revealed the adhesion of bacteriae to the cell membrane by glycocalyx. The fibrillar structures surrounding the bacteriae are clearly seen.


Un día después del nacimiento, la mucosa de la lengua una rata fue examinada por el microscopio electrónico de transmisión y método de ARMEB. Los especímenes fueron fijados mediante uan solución Karnovsky y embebido en resina Spurr para microscopía electrónica de transmisión. Para el método ARMEB, las muestras fueron fijadas en tetróxido de osmio 2 por ciento, deshidratados en alcohol, secados al punto crítico y recubierto con oro-paladio. Los resultados demostraron que la superficie de la lengua presentaba papilas filiformes y fungiformes cubiertas por numerosas células epiteliales queratinizadas. Las bacterias se unen a las superficies de las microplicas al azar, lo que se demuestra en sus tres dimensiones las imágenes en ARMEB. A gran aumento, las imágenes del microscopio electrónico de transmisión revelan la adhesión de bacterias a la membrana celular por el glicocalix. Las estructuras fibrilares que rodean a las bacterias son claramente visibles.


Subject(s)
Animals , Rats , Bacteria/ultrastructure , Epithelial Cells/microbiology , Epithelial Cells/ultrastructure , Tongue/microbiology , Tongue/ultrastructure , Bacterial Adhesion/physiology , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission
19.
Braz. dent. j ; 18(4): 320-323, 2007. ilus
Article in English | LILACS | ID: lil-474472

ABSTRACT

Togue mucosa surface of 3-day postnatal rats was examined under transmission electron microscopy (TEM) and high-resolution scanning electron microscopy (HRSEM). For HRSEM analysis, the specimens were fixed in the same solution for 24 h, postfixed in 2 percent osmiun tetroxide, critical-point dried and coated with platinum-palladium. For TEM analysis, the specimens were fixed using modified Karnovsky solution and embedded in Spurr resin. The results revealed the presence of numerous microplicae in the membrane surface of keratinized epithelial cells to which groups of bacteria were attached. These bacteria were staphylococcus and coccus organized either in rows or at random, which were visualized in three-dimensional HRSEM images. At high magnification, the TEM images revealed the adhesion of bacteria to the cell membrane through numerous filamentous structures comprising the glycocalyx. The fine fibrillar structures rising from each bacterium and from cell membrane were clearly seen. These characteristics on bacteria structure may be used for future control or prevention of bacterial diseases and for installation of the oral native flora.


A superfície lingual de ratos de três dias de idade foi examinada em microscópia eletrônica de transmissão (MET) e em microscópia eletrônica varredura de alta resolução (MEVAR). Para o método de MEVAR, os espécimes foram fixados na mesma solução por 24 h, pós fixados em solução de tetróxido de ósmio a 2 por cento, secos em ponto crítico e cobertos com platina- paládio. Para análise em MET, os espécimes foram fixados utilizando-se solução de Karnovsky modificada e emblocadas em resina Spurr. Os resultados mostraram a presença de numerosas micropregas na membrana superficial das células epiteliais queratinizadas, nas quais estavam aderidos grupos de bactérias. Estas bactérias eram estafilococos e cocos, organizados em fileiras ou a esmo, e puderam ser observadas em imagens tri-dimensionais em MEVAR. Em maiores aumentos, as imagens em MET revelaram a adesão de bactérias nas células por meio de numerosas estruturas filamentares compondo o glicocálice. As delicadas estruturas filamentares na periferia das bactérias e das células foram nitidamente identificadas. Estas características da estrutura bacteriana podem ser utilizadas, no futuro, para controle e prevenção de doenças bacteriana, bem como para a instalação da flora oral nativa.


Subject(s)
Animals , Rats , Bacteria/ultrastructure , Bacterial Adhesion/physiology , Mouth Mucosa/microbiology , Tongue/microbiology , Animals, Newborn , Cell Membrane/microbiology , Cell Membrane/ultrastructure , Epithelial Cells/microbiology , Epithelial Cells/ultrastructure , Glycocalyx/microbiology , Glycocalyx/ultrastructure , Imaging, Three-Dimensional , Keratins/ultrastructure , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Mouth Mucosa/ultrastructure , Rats, Wistar , Staphylococcus/ultrastructure , Taste Buds/microbiology , Taste Buds/ultrastructure , Tongue/ultrastructure
20.
Braz. oral res ; 20(4): 307-311, Oct.-Dec. 2006. ilus, tab
Article in English | LILACS | ID: lil-440177

ABSTRACT

The aim of this study was to evaluate surface free energy (SFE), surface roughness (SR) and surface hardness (SH) of two commercially available materials for fabricating dental implant abutments. In addition, the specimens were investigated by scanning electron microscopy (SEM) to determine the surface morphology. Twenty five discs (5 x 2 mm) of Ti-6Al-4V and Tilite (Ni-Cr-Ti) alloys were used in this study. Surface free energy was determined by the contact angle formed between a drop of distilled, deionized water and the surface of the specimen of each material. The surface roughness was measured with a mechanical profilometer and the surface hardness was evaluated by means of the Vickers hardness micro indentation test. SFE, SR and SH data were analyzed using one-way ANOVA (p < 0.05). Statistical differences (p < 0.05) were found between Ti-6Al-4V (36.2 erg.cm-2; 0.2 æm) and Tilite (30.9 erg.cm-2; 0.16 æm) for SFE and SR. However, the differences between the surface hardness values of Ti-6Al-4V (325.0 kg/mm²) and Tilite (324.3 kg/mm²) were not statistically significant (p > 0.05). Evaluations by SEM revealed different surface morphology. Within the limits of this study, it can be concluded that the Ti-6Al-4V and Tilite alloys showed differences in surface properties, except for surface hardness, suggesting that both alloys may be considered appropriate for producing abutments. Further studies are, however, necessary to elucidate the biological responses to implant abutments made with these alloys.


O objetivo deste estudo foi avaliar a energia livre de superfície (ELS), rugosidade superficial (RS) e dureza de superfície (DS) de dois materiais disponíveis comercialmente para fabricação de "abutments" de implante. Em acréscimo, os espécimes foram investigados por microscopia eletrônica de varredura (MEV) para determinar a morfologia de superfície. Vinte e cinco discos de ligas de Ti-6Al-4V e Tilite (Ni-Cr-Ti) (5 x 2 mm) foram usados neste estudo. A energia livre de superfície foi determinada pela mensuração do ângulo de contato formado entre uma gota de água destilada e deionizada e a superfície do espécime para cada material. A rugosidade superficial foi mensurada com uso de um rugosímetro e a dureza de superfície foi avaliada por meio do teste de microdureza Vickers. Os dados foram analisados usando Análise de variância (P < 0,05). Foram encontradas diferenças significantes (P < 0,05) para os valores de ELS e RS entre Ti-6Al-4V (36,2 erg.cm-2; 0,2 æm) e Tilite (30,9 erg.cm-2; 0,16 æm). Entretanto, os valores de dureza de superfície não foram diferentes para ambos os materiais (Ti-6Al-4V - 325,0 kg/mm² e Tilite - 324,3 kg/mm²) (P > 0,05). A análise topográfica dos materiais pela MEV revelou superfícies diferentes. Dentro das limitações deste estudo, concluiu-se que as ligas de Ti-6Al-4V e Tilite mostraram diferenças nas propriedades de superfícies, exceto dureza, sugerindo que ambas as ligas são adequadas para a confecção de "abutments". Entretanto, estudos são necessários para evidenciar as respostas biológicas aos "abutments" de implantes confeccionados com essas ligas.


Subject(s)
Bacterial Adhesion/physiology , Dental Abutments/microbiology , Dental Alloys/chemistry , Surface Properties , Titanium/chemistry , Analysis of Variance , Biocompatible Materials , Hardness , Materials Testing , Microscopy, Electron, Scanning
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