Home About us Contact
|
Home About us Contact | ||
|
|
||
Bacterial Biofilms (bacterial + biofilm)
Selected AbstractsAdaptive responses to antimicrobial agents in biofilmsENVIRONMENTAL MICROBIOLOGY, Issue 8 2005Barbara Szomolay Summary Bacterial biofilms demonstrate adaptive resistance in response to antimicrobial stress more effectively than corresponding planktonic populations. We propose here that, in biofilms, reaction-diffusion limited penetration may result in only low levels of antimicrobial exposure to deeper regions of the biofilm. Sheltered cells are then able to enter an adapted resistant state if the local time scale for adaptation is faster than that for disinfection. This mechanism is not available to a planktonic population. A mathematical model is presented to illustrate. Results indicate that, for a sufficiently thick biofilm, cells in the biofilm implement adaptive responses more effectively than do freely suspended cells. Effective disinfection requires applied biocide concentration that increases quadratically or exponentially with biofilm thickness. [source] Quorum sensing controls biofilm formation in Vibrio choleraeMOLECULAR MICROBIOLOGY, Issue 1 2003Brian K. Hammer Summary Multiple quorum-sensing circuits function in parallel to control virulence and biofilm formation in Vibrio cholerae. In contrast to other bacterial pathogens that induce virulence factor production and/or biofilm formation at high cell density in the presence of quorum-sensing autoinducers, V. cholerae represses these behaviours at high cell density. Consistent with this, we show here that V. cholerae strains ,locked' in the regulatory state mimicking low cell density are enhanced for biofilm production whereas mutants ,locked' in the regulatory state mimicking high cell density are incapable of producing biofilms. The quorum-sensing cascade we have identified in V. cholerae regulates the transcription of genes involved in exopolysaccharide production (EPS), and variants that produce EPS and form biofilms arise at high frequency from non-EPS, non-biofilm producing strains. Our data show that spontaneous mutation of the transcriptional regulator hapR is responsible for this effect. Several toxigenic strains of V. cholerae possess a naturally occurring frameshift mutation in hapR. Thus, the distinct environments occupied by this aquatic pathogen presumably include niches where cell-cell communication is crucial, as well as ones where loss of quorum sensing via hapR mutation confers a selective advantage. Bacterial biofilms could represent a complex habitat where such differentiation occurs. [source] The resistance of maxillofacial reconstruction plates to biofilm formation in vitroTHE LARYNGOSCOPE, Issue 11 2003Brian E. Emery MD Abstract Objectives/Hypothesis: Bacterial biofilms, bacteria surrounded by a protective glycocalyx, have been demonstrated on bioimplants placed within and outside of the head and neck region. The presence of the biofilm often makes decontamination of an infected implant impossible, requiring removal of the implant. Infections attributable to biofilm formation within the facial skeleton after reconstruction with implants may result in delayed union, fibrous union, malunion, nonunion, and malocclusion. These complications often require removal of the implant and secondary surgery. Although the incidence of infections necessitating implant removal is relatively low, the increased numbers of implants being placed make this a growing problem. Previous work in the authors laboratory has demonstrated a resistance to biofilm formation on different types of pressure-equalizing tubes. The hypothesis evaluated in the study is that such resistance to biofilm formation is due to the inability of bacteria to adhere to the tubes because of the material's smoothness or surface charge. Study Design: A controlled observational study. Methods: Scanning electron microscopy was used to evaluate the formation of biofilms in vitro for a common strain of Staphylococcus aureus on four implantable materials. The implantable materials included titanium and polylactide resorbable plates. Results: Consistent with the authors' prior findings, they were able to produce bacterial biofilm reliably on a silicone pressure equalizing tube but were unable to demonstrate biofilm formation on the titanium or resorbable implants. Conclusion: The absence of biofilm formation on these implants can best be explained by the surface charge or polarity properties of these materials. These findings are consistent with the relatively low incidence of infections among patients receiving these implants in maxillofacial applications. [source] Candidate's Thesis: Direct Evidence of Bacterial Biofilms in Otitis Media,THE LARYNGOSCOPE, Issue 12 2001J. Christopher Post MD Abstract Objectives/Hypothesis Bacteriologic studies of otitis media with effusion (OME) using highly sensitive techniques of molecular biology such as the polymerase chain reaction have demonstrated that traditional culturing methods are inadequate to detect many viable bacteria present in OME. The presence of pathogens attached to the middle-ear mucosa as a bacterial biofilm, rather than as free-floating organisms in a middle-ear effusion, has previously been suggested to explain these observations. The suggestion has been speculative, however, because no visual evidence of such biofilms on middle-ear mucosa has heretofore been collected. The hypotheses motivating the current study were: 1) biofilms of nontypable Hemophilus influenzae will form on the middle-ear mucosa of chinchillas in an experimental model of OME, 2) these biofilms will exhibit changes in density or structure over time, and 3) biofilms are also present on tympanostomy tubes in children with refractory post-tympanostomy otorrhea. The objective of this study was to collect visual evidence of the formation of bacterial biofilms in these situations. Study Design Laboratory study of bacteriology in an animal model and on medical devices removed from pediatric patients. Methods Experimental otitis media was induced in chinchillas by transbullar injection of nontypable H. influenzae. Animals were killed in a time series and the surface of the middle-ear mucosa was examined by scanning electron microscopy (SEM) for the presence of bacterial biofilms. Adult and fetal chinchilla uninfected controls were similarly examined for comparison. In addition, tympanostomy tubes that had been placed in children's ears to treat OME and removed after onset of refractory otorrhea or other problems were examined by SEM and by confocal scanning laser microscopy for bacterial biofilms, and compared with unused control tubes. Results Bacterial biofilms were visually detected by SEM on the middle-ear mucosa of multiple chinchillas in which H. influenzae otitis media had been induced. Qualitative evaluation indicated that the density and thickness of the biofilm might increase until at least 96 hours after injection. The appearance of the middle-ear mucosa of experimental animals contrasted with that of uninjected control animals. Robust bacterial biofilms were also visually detected on tympanostomy tubes removed from children's ears for clinical reasons, in contrast with unused control tubes. Conclusions Bacterial biofilms form on the middle-ear mucosa of chinchillas in experimentally induced H. influenzae otitis media and can form on tympanostomy tubes placed in children's ears. Such biofilms can be directly observed by microscopy. These results reinforce the hypothesis that the bacterial aggregates called biofilms, resistant to treatment by antibiotics and to detection by standard culture techniques, may play a major etiologic role in OME and in one of its frequent complications, post-tympanostomy otorrhea. [source] Exceptionally preserved tadpoles from the Miocene of Libros, Spain: ecomorphological reconstruction and the impact of ontogeny upon taphonomyLETHAIA, Issue 3 2010MARIA E. MCNAMARA McNamara, M.E., Orr, P.J., Kearns, S.L., Alcalá, L., Anadón, P. & Peñalver-Mollá, E. 2010: Exceptionally preserved tadpoles from the Miocene of Libros, Spain: ecomorphological reconstruction and the impact of ontogeny upon taphonomy. Lethaia, Vol. 43, pp. 290,306. The Libros exceptional biota from the Upper Miocene of NE Spain includes abundant frog tadpoles (Rana pueyoi) preserved in finely laminated lacustrine mudstones. The tadpoles exhibit a depressed body, short tail, low tail fins, dorso-laterally directed eyes and jaw sheaths; these features identify the Libros tadpoles as members of the benthic lentic ecomorphological guild. This, the first ecomorphological reconstruction of a fossil tadpole, supports phylogenetic evidence that this ecology is a conserved ranid feature. The soft-tissue features of the Libros tadpoles are characterized by several modes of preservation. The space occupied previously by the brain is defined by calcium carbonate, the nerve cord is defined by calcium phosphate, and jaw sheaths and bone marrow are preserved as organic remains. Gut contents (and coprolites adjacent to specimens) comprise ingested fine-grained sedimentary detritus and epiphyton. The body outline and the eyespots, nares, abdominal cavity, notochord, caudal myotomes and fins are defined by a carbonaceous bacterial biofilm. A similar biofilm in adult specimens of R. pueyoi from Libros defines only the body outline, not any internal anatomical features. In the adult frogs, but not in the tadpoles, calcium phosphate and calcium sulphate precipitated in association with integumentary tissues. These differences in the mode of preservation between the adult frogs and tadpoles reflect ontogenetic factors. ,Anuran, ecology, soft-tissue, tadpoles, taphonomy. [source] Microbial strengthening of loose sandLETTERS IN APPLIED MICROBIOLOGY, Issue 2 2010B.L. Banagan Abstract Aims:, To test whether the addition of Flavobacterium johnsoniae could increase the strength of saturated Ottawa 30 sand. Methods and Results:, A box model was built that simulates groundwater-like flow through a main sand compartment. Strength tests were performed at seven locations and at two depths, 10·8 and 20·3 cm below the top of the tank, using a vane shear device before and after the addition of bacteria. After the addition of Fl. johnsoniae, sand samples were obtained from multiple sampling ports on the vertical sides of the box model. The presence of a bacterial biofilm was confirmed by staining these sand samples with SYTO-9 and Alexa Fluor 633 and viewing with a confocal microscope. The average shear strength increases after the addition of Fl. johnsoniae were 15·2,87·5%, depending on the experimental conditions. Conclusions:,Flavobacterium johnsoniae caused a statistically significant increase in the strength of saturated Ottawa 30 sand. Significance and Impact of the Study:, Biofilm-forming bacteria can increase the shear strength of saturated sand. The addition of biofilm-forming bacteria to a building site may be an alternate method to mitigate the effects of liquefaction. [source] The resistance of maxillofacial reconstruction plates to biofilm formation in vitroTHE LARYNGOSCOPE, Issue 11 2003Brian E. Emery MD Abstract Objectives/Hypothesis: Bacterial biofilms, bacteria surrounded by a protective glycocalyx, have been demonstrated on bioimplants placed within and outside of the head and neck region. The presence of the biofilm often makes decontamination of an infected implant impossible, requiring removal of the implant. Infections attributable to biofilm formation within the facial skeleton after reconstruction with implants may result in delayed union, fibrous union, malunion, nonunion, and malocclusion. These complications often require removal of the implant and secondary surgery. Although the incidence of infections necessitating implant removal is relatively low, the increased numbers of implants being placed make this a growing problem. Previous work in the authors laboratory has demonstrated a resistance to biofilm formation on different types of pressure-equalizing tubes. The hypothesis evaluated in the study is that such resistance to biofilm formation is due to the inability of bacteria to adhere to the tubes because of the material's smoothness or surface charge. Study Design: A controlled observational study. Methods: Scanning electron microscopy was used to evaluate the formation of biofilms in vitro for a common strain of Staphylococcus aureus on four implantable materials. The implantable materials included titanium and polylactide resorbable plates. Results: Consistent with the authors' prior findings, they were able to produce bacterial biofilm reliably on a silicone pressure equalizing tube but were unable to demonstrate biofilm formation on the titanium or resorbable implants. Conclusion: The absence of biofilm formation on these implants can best be explained by the surface charge or polarity properties of these materials. These findings are consistent with the relatively low incidence of infections among patients receiving these implants in maxillofacial applications. [source] Candidate's Thesis: Direct Evidence of Bacterial Biofilms in Otitis Media,THE LARYNGOSCOPE, Issue 12 2001J. Christopher Post MD Abstract Objectives/Hypothesis Bacteriologic studies of otitis media with effusion (OME) using highly sensitive techniques of molecular biology such as the polymerase chain reaction have demonstrated that traditional culturing methods are inadequate to detect many viable bacteria present in OME. The presence of pathogens attached to the middle-ear mucosa as a bacterial biofilm, rather than as free-floating organisms in a middle-ear effusion, has previously been suggested to explain these observations. The suggestion has been speculative, however, because no visual evidence of such biofilms on middle-ear mucosa has heretofore been collected. The hypotheses motivating the current study were: 1) biofilms of nontypable Hemophilus influenzae will form on the middle-ear mucosa of chinchillas in an experimental model of OME, 2) these biofilms will exhibit changes in density or structure over time, and 3) biofilms are also present on tympanostomy tubes in children with refractory post-tympanostomy otorrhea. The objective of this study was to collect visual evidence of the formation of bacterial biofilms in these situations. Study Design Laboratory study of bacteriology in an animal model and on medical devices removed from pediatric patients. Methods Experimental otitis media was induced in chinchillas by transbullar injection of nontypable H. influenzae. Animals were killed in a time series and the surface of the middle-ear mucosa was examined by scanning electron microscopy (SEM) for the presence of bacterial biofilms. Adult and fetal chinchilla uninfected controls were similarly examined for comparison. In addition, tympanostomy tubes that had been placed in children's ears to treat OME and removed after onset of refractory otorrhea or other problems were examined by SEM and by confocal scanning laser microscopy for bacterial biofilms, and compared with unused control tubes. Results Bacterial biofilms were visually detected by SEM on the middle-ear mucosa of multiple chinchillas in which H. influenzae otitis media had been induced. Qualitative evaluation indicated that the density and thickness of the biofilm might increase until at least 96 hours after injection. The appearance of the middle-ear mucosa of experimental animals contrasted with that of uninjected control animals. Robust bacterial biofilms were also visually detected on tympanostomy tubes removed from children's ears for clinical reasons, in contrast with unused control tubes. Conclusions Bacterial biofilms form on the middle-ear mucosa of chinchillas in experimentally induced H. influenzae otitis media and can form on tympanostomy tubes placed in children's ears. Such biofilms can be directly observed by microscopy. These results reinforce the hypothesis that the bacterial aggregates called biofilms, resistant to treatment by antibiotics and to detection by standard culture techniques, may play a major etiologic role in OME and in one of its frequent complications, post-tympanostomy otorrhea. [source] Use of episcopic differential interference contrast microscopy to identify bacterial biofilms on salad leaves and track colonization by Salmonella ThompsonENVIRONMENTAL MICROBIOLOGY, Issue 4 2008J. C. Warner Summary Zoonotic pathogens such as Salmonella can cause gastrointestinal illness if they are ingested with food. Foods such as salads pose a greater risk because they are consumed raw and have been the source of major outbreaks of disease from fresh produce. The novel light microscopy methods used in this study allow detailed, high resolution imaging of the leaf surface environment (the phyllosphere) and allow pathogen tracking. Episcopic differential interference contrast microscopy coupled with epifluorescence was used to view the natural microflora in situ on salad leaves and their topographical distribution. Fluorescent nucleic acid staining was used to differentiate between bacterial colonists and inorganic debris. Salmonella enterica serovar Thompson expressing green fluorescent protein was inoculated onto individual spinach leaves for 24 h at 22°C in order to observe spatial and temporal patterning of colonization on the two surfaces of each leaf under different osmotic conditions. The results obtained show that salad leaves are host to high numbers of bacteria, typically 105 per square millimetre. Cells are present in complex three-dimensional aggregations which often have a slimy appearance, suggesting the presence of biofilms. Washing of the leaves had little effect on the number of adherent pathogens, suggesting very strong attachment. Episcopic differential interference contrast microscopy is a rapid alternative to both scanning electron microscopy and confocal laser scanning microscopy for visualizing leaf topography and biofilm formation in the natural state. [source] Specificity in the settlement , modifying response of bacterial biofilms towards zoospores of the marine alga EnteromorphaENVIRONMENTAL MICROBIOLOGY, Issue 5 2003Pratixa Patel Summary Previous studies have shown that the rate of settlement of zoospores of the green alga Enteromorpha is stimulated by mixed microbial biofilms and that the number of zoospores settling is positively correlated with the number of bacteria in the biofilm. In the present study the specificity of this relationship has been investigated. Ninety-nine strains of marine bacteria were isolated from natural biofilms on rocks and the surface of Enteromorpha plants. Isolates were screened by denaturing gradient gel electrophoresis (DGGE) to eliminate replicates and 16S rDNA sequencing identified a total of 37 unique strains. Phylogenetic analysis revealed that the isolated bacterial strains belonged to three groups ,- Proteobacteria (28 strains), Cytophaga-Flavobacteria-Bacteroid (CFB) group (six strains) and ,- Proteobacteria (one strain). Two strains were unassigned, showing < 93% sequence similarity with the CFB group. The main genera of ,- Proteobacteria were Pseudoalteromonas (14 strains), Vibrio (five strains), Shewanella (five strains), Halomonas (three strains) and Pseudomonas (one strain). Spore settlement experiments were conducted on single-species biofilms, developed for different times on glass slides. The effect of correcting spore settlement values for biofilm density was evaluated. Results showed that the effect of bacterial strains on spore settlement was strain- but not taxon-specific and activity varied with the age of the biofilm. However, most of the strains belonging to genera Vibrio and Shewanella showed stimulation. Pseudoalteromonas strains showed a range of effects including settlement-inhibiting, paralysing and lysing activities. Spatial analysis of bacterial density in the presence and absence of spores revealed a range of different types of association between spores and bacteria. Overall, the spatial association between spores and bacteria appears to be independent of the overall quantitative influence of bacterial cells on spore settlement. [source] Protists with different feeding modes change biofilm morphologyFEMS MICROBIOLOGY ECOLOGY, Issue 2 2009Anne Böhme Abstract The effect of Dexiostoma (filter feeder), Vannella, Chilodonella (raptorial feeders), Spumella, and Neobodo (direct interception feeders) on the morphology of multispecies bacterial biofilms was investigated in small flow cells. The filter feeder Dexiostoma campylum did not alter biofilm volume and porosity but stimulated the formation of larger microcolonies compared with ungrazed biofilms. In contrast, the raptorial feeder Vannella sp. efficiently grazed bacteria from the biofilm surface, leading to smaller microcolonies and lower maximal and basal layer thickness compared with ungrazed biofilms. Microcolony formation was not stimulated in the presence of the sessile Spumella sp. Chilodonella uncinata rasped bacteria from the outer surface leading to mushroom-shaped microcolonies. In the presence of C. uncinata and Spumella sp., the biofilm volume was 2.5,6.3 times lower compared with ungrazed biofilms. However, the biofilm porosity and the ratio of biofilm surface area to biofilm volume were 1.5,3.7 and 1.2,1.8 times higher, respectively. Thus, exchange of nutrients and gases between the biofilm and its surrounding fluid should also be improved in deeper biofilm layers, hence accelerating microbial growth. [source] Development of an ex vivo model for the study of microbial infection in human teethINTERNATIONAL ENDODONTIC JOURNAL, Issue 5 2007B. Patel Aims, (1) To infect human teeth artificially to mimic root canal and dentine infection, using the Constant Depth Film Fermenter (CDFF); (2) To verify the similarity of the infections to those found, in vivo, using culture and microscopy (SEM, LM and TEM). Methodology, Human teeth [n = 38 and n = 28, for phases I (preliminary) and II (definitive), respectively] were infected within the CDFF for a period of 28 days and at pre-selected time points were removed, externally decontaminated using validated protocols and subjected to either culture-dependent or microscopy protocols. The condition of the teeth was varied in phase I to establish the feasibility of the approach and identify optimal conditions. This informed the selection of optimal conditions for definitive test in phase II. For culture-dependent analysis in this phase, a dentine filing sample was obtained from the apical 5 mm of the root canal and cultured anaerobically to allow isolation of individual strains. Bacterial DNA was extracted from purified isolates, the 16S rRNA genes amplified by PCR and the amplicons sequenced for identity using sequence databases. Teeth assigned for microscopy were post-fixed in 3% gluteraldehyde after removal from the CDFF and then subjected to appropriate protocols prior to microscopic evaluation of the infection. Results, All three microscopy techniques and culture-dependent analysis confirmed infection of the human teeth using the CDFF, with root canal infections visually resembling closely those seen in vivo. Furthermore, partial 16S rRNA gene sequencing of DNA from cultured isolates confirmed a selective number of 7,9 genera/species in the apical portion of two teeth each at 7 and 28 days; these taxa are also commonly recovered from teeth with apical periodontitis, in vivo. There were no objective measures other than speciation and topographical evaluation to compare the artificial and real (in vivo) infections. Conclusions, The proposed ex vivo model has the potential for development into an investigative tool for studying the dynamics of bacterial ecology in infected root canals, both before and after treatment. Its advantage is the ability to control both the abiotic and biotic factors. There is a need for the development of objective measures to compare artificial and real bacterial biofilms. [source] Effect of conditioning films and a novel anti-adherent agent on bacterial adherence to dentineINTERNATIONAL ENDODONTIC JOURNAL, Issue 4 2001A. Maglad Aim,Adherence of bacteria to dentine is a prerequisite to infection of the root canal system, yet adherence of root canal bacteria to dentine is poorly understood. The aim of this study was to evaluate the effect of conditioning films and anti-adherent compounds on bacterial adherence to dentine. Methodology,Freshly extracted molar teeth were prepared and sectioned to give 225 discs of predetermined dimensions. The discs were allocated to two groups. Group 1 (n = 189) was divided into three subgroups (n = 63) and coated with one of three conditioning agents (artificial saliva, serum, or distilled water) prior to bacterial inoculation. Group 2 discs (n = 36) were treated with either a novel anti-adherent agent (PC1036, Biocompatibles) (n = 18) or distilled water (n = 18) prior to conditioning with artificial saliva. Monospecies bacterial biofilms were generated on the dentine discs by incubating them in brain heart infusion broth (37 gL,1) containing Streptococcus intermedius (Si), Enterococcus faecalis (Ef) or Lactobacillus fermentum (Lf) (originally isolated from infected root canals). The number of bacteria adhering to the discs in each of the groups was determined using standard serial dilution protocols. Additional discs were prepared under all conditions for scanning electron microscopy. Where appropriate, statistical analysis by one way anova, post hoc Bonferroni, and independent t -test were used. Results,Si adhered significantly better to dentine when conditioned with serum compared with artificial saliva (P = 0.005) or distilled water (P = 0.009). Conversely, Ef adhered significantly better to the control discs (distilled water) compared with serum conditioned discs (P = 0.016). The conditioning films had no effect on the adherence of Lf, which adhered to the dentine discs significantly less (P = 0.001) than either Si or Ef. The anti-adherent coating significantly reduced the number of Si adhering to the dentine compared with the control (P = 0.012). Conclusion,Given the importance of adherence in root canal infection it is conceivable that an anti-adherent compound, could be used to prevent bacterial recontamination of cavities or the root canal system. [source] Inhibition of biofilms associated with dentures and toothbrushes by tetrasodium EDTAJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2007D.A. Devine Abstract Aims:, We examined the efficacy of tetrasodium EDTA in eradicating biofilms derived from salivary inocula or pure cultures of Candida albicans on discs of polymethyl methacrylate (PMMA) denture base or on toothbrushes that had been used normally for 4,8 weeks. Its efficiency in virus neutralization was also determined. Methods and Results:, Overnight (16 h) treatment with 4% (w/v) tetrasodium EDTA solution reduced salivary and C. albicans biofilm viable counts by ,99%. Biofilm removal was confirmed using confocal laser scanning microscopy. Presence/absence of sucrose during biofilm formation had no effect on killing efficacy. Prolonged treatment of PMMA with tetrasodium EDTA did not influence subsequent formation of C. albicans biofilms or affect surface roughness of the PMMA, but it reduced subsequent biofilm formation from a salivary inoculum. Infectivities of herpes simplex virus and polio virus suspensions were reduced by >99·99% by treatment for 1 and 2 h, respectively. Conclusions:, Tetrasodium EDTA solution efficiently disinfected toothbrushes and PMMA discs, with the detachment of biofilms, and rapidly neutralized both nonenveloped and enveloped viruses. Significance and Impact of the Study:, Dentures and toothbrushes become contaminated by bacterial biofilms and by viruses. There is a need for disinfection methods that are rapidly effective, cost-effective, nontoxic and easily implemented. These studies indicate that tetrasodium EDTA solution has disinfection applications in the oral care field. [source] Antibacterial activity of dental composites containing zinc oxide nanoparticles,JOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 1 2010Berdan Aydin Sevinç Abstract The resin-based dental composites commonly used in restorations result in more plaque accumulation than other materials. Bacterial biofilm growth contributes to secondary caries and failure of resin-based dental composites. Methods to inhibit biofilm growth on dental composites have been sought for several decades. It is demonstrated here that zinc oxide nanoparticles (ZnO-NPs) blended at 10% (w/w) fraction into dental composites display antimicrobial activity and reduce growth of bacterial biofilms by roughly 80% for a single-species model dental biofilm. Antibacterial effectiveness of ZnO-NPs was assessed against Streptococcus sobrinus ATCC 27352 grown both planktonically and as biofilms on composites. Direct contact inhibition was observed by scanning electron microscopy and confocal laser scanning microscopy while biofilm formation was quantified by viable counts. An 80% reduction in bacterial counts was observed with 10% ZnO-NP-containing composites compared with their unmodified counterpart, indicating a statistically significant suppression of biofilm growth. Although, 20% of the bacterial population survived and could form a biofilm layer again, 10% ZnO-NP-containing composites maintained at least some inhibitory activity even after the third generation of biofilm growth. Microscopy demonstrated continuous biofilm formation for unmodified composites after 1-day growth, but only sparsely distributed biofilms formed on 10% ZnO-NP-containing composites. The minimum inhibitory concentration of ZnO-NPs suspended in S. sobrinus planktonic culture was 50 ,g mL,1. ZnO-NP-containing composites (10%) qualitatively showed less biofilm after 1-day-anaerobic growth of a three-species initial colonizer biofilm after being compared with unmodified composites, but did not significantly reduce growth after 3 days. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2010. [source] Laser desorption postionization for imaging MS of biological materialJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 2 2010Artem Akhmetov Abstract Vacuum ultraviolet single photon ionization (VUV SPI) is a soft ionization technique that has the potential to address many of the limitations of matrix-assisted laser desorption/ionization (MALDI) for imaging MS. Laser desorption postionization (LDPI) uses VUV SPI for postionization and is experimentally analogous to a MALDI instrument with the addition of a pulsed VUV light source. This review discusses progress in LDPI-MS over the last decade, with an emphasis on imaging MS of bacterial biofilms, analytes whose high salt environment make them particularly resistant to imaging by MALDI-MS. This review first considers fundamental aspects of VUV SPI including ionization mechanisms, cross sections, quantum yields of ionization, dissociation and potential mass limits. The most common sources of pulsed VUV radiation are then described along with a newly constructed LDPI-MS instrument with imaging capabilities. Next, the detection and imaging of small molecules within intact biofilms is demonstrated by LDPI-MS using 7.87 eV (157.6 nm) VUV photons from a molecular fluorine excimer laser, followed by the use of aromatic tags for detection of selected species within the biofilm. The final section considers the future prospects for imaging intact biological samples by LDPI-MS. Copyright © 2010 John Wiley & Sons, Ltd. [source] Antimicrobial photodynamic therapy combined with conventional endodontic treatment to eliminate root canal biofilm infectionLASERS IN SURGERY AND MEDICINE, Issue 1 2007Aguinaldo S. Garcez DDS Abstract Background and Objective To compare the effectiveness of antimicrobial photodynamic therapy (PDT), standard endodontic treatment and the combined treatment to eliminate bacterial biofilms present in infected root canals. Study Design/Materials and Methods Ten single-rooted freshly extracted human teeth were inoculated with stable bioluminescent Gram-negative bacteria, Proteus mirabilis and Pseudomonas aeruginosa to form 3-day biofilms in prepared root canals. Bioluminescence imaging was used to serially quantify bacterial burdens. PDT employed a conjugate between polyethylenimine and chlorin(e6) as the photosensitizer (PS) and 660-nm diode laser light delivered into the root canal via a 200-µ fiber, and this was compared and combined with standard endodontic treatment using mechanical debridement and antiseptic irrigation. Results Endodontic therapy alone reduced bacterial bioluminescence by 90% while PDT alone reduced bioluminescence by 95%. The combination reduced bioluminescence by >98%, and importantly the bacterial regrowth observed 24 hours after treatment was much less for the combination (P<0.0005) than for either single treatment. Conclusions Bioluminescence imaging is an efficient way to monitor endodontic therapy. Antimicrobial PDT may have a role to play in optimized endodontic therapy. Lasers Surg. Med. © 2006 Wiley-Liss, Inc. [source] Disruption of fungal and bacterial biofilms by lauroyl glucoseLETTERS IN APPLIED MICROBIOLOGY, Issue 5 2008D.H. Dusane Abstract Aim:, The ability of enzymatically synthesized lauroyl glucose to disrupt fungal (Candida albicans, Candida lipolytica) and bacterial (Pseudomonas aeruginosa PAO1, Pseudomonas aureofaciens) biofilms was investigated. Methods and Results:, Preformed biofilms of C. albicans and C. lipolytica in polystyrene microtitre plates were disrupted upto 45% and 65%, respectively, while P. aeruginosa and P. aureofaciens biofilms were disrupted by 51% and 57%. Precoating of the microtitre wells with lauroyl glucose affected cell attachment and biofilm growth of all the cultures to a lesser extent. With C. albicans and C. lipolytica, there was 11% and 32% decrease in the development of biofilms, respectively. With P. aeruginosa and P. aureofaciens, the reduction was 21% and 12% after 48 h. Lauroyl glucose effectively inhibited the formation of biofilms on glass slide surfaces when added along with the inoculum. Analysis by confocal laser scanning microscopy showed that the growth of the biofilms was lesser as compared with the control experiments. Lauroyl glucose displayed minimum inhibitory concentration values >500 ,g ml,1 for the test cultures and was comparable to that obtained with acetyl salicylate. Conclusion:, Lauroyl glucose reduces biofilm growth of all the four test cultures on polystyrene and glass surfaces. Significance and Impact of the Study:, This report is a novel application of the enzymatically synthesized, environmental-friendly nonionic surfactant. [source] Ion transport in roots: measurement of fluxes using ion-selective microelectrodes to characterize transporter functionPLANT CELL & ENVIRONMENT, Issue 1 2001I. A. Newman ABSTRACT The transport of mineral ions into and out of tissues and cells is central to the life of plants. Ion transport and the plasma membrane transporters themselves have been studied using a variety of techniques. In the last 15 years, measurement of specific ion fluxes has contributed to the characterization of transport systems. Progress in molecular genetics is allowing gene identification and controlled expression of transporter molecules. However the molecular expression of transporter gene products must be characterized at the functional level. The ion-selective microelectrode technique to measure specific ion fluxes non-invasively is ideally suited to this purpose. This technique, its theory, its links with others and its application and prospects in plant science, are discussed. Ions studied include hydrogen, potassium, sodium, ammonium, calcium, chloride and nitrate. Applications discussed include: solute ion uptake by roots; gravitropism and other processes in the root cap, meristematic and elongation zones; Nod factor effect on root hairs; osmotic and salt stresses; oscillations; the effects of light and temperature. Studies have included intact roots, leaf mesophyll and other tissues, protoplasts and bacterial biofilms. A multi-ion capability of the technique will greatly assist functional genomics, particularly when coupled with imaging techniques, patch clamping and the use of suitable mutants. [source] Candidate's Thesis: Direct Evidence of Bacterial Biofilms in Otitis Media,THE LARYNGOSCOPE, Issue 12 2001J. Christopher Post MD Abstract Objectives/Hypothesis Bacteriologic studies of otitis media with effusion (OME) using highly sensitive techniques of molecular biology such as the polymerase chain reaction have demonstrated that traditional culturing methods are inadequate to detect many viable bacteria present in OME. The presence of pathogens attached to the middle-ear mucosa as a bacterial biofilm, rather than as free-floating organisms in a middle-ear effusion, has previously been suggested to explain these observations. The suggestion has been speculative, however, because no visual evidence of such biofilms on middle-ear mucosa has heretofore been collected. The hypotheses motivating the current study were: 1) biofilms of nontypable Hemophilus influenzae will form on the middle-ear mucosa of chinchillas in an experimental model of OME, 2) these biofilms will exhibit changes in density or structure over time, and 3) biofilms are also present on tympanostomy tubes in children with refractory post-tympanostomy otorrhea. The objective of this study was to collect visual evidence of the formation of bacterial biofilms in these situations. Study Design Laboratory study of bacteriology in an animal model and on medical devices removed from pediatric patients. Methods Experimental otitis media was induced in chinchillas by transbullar injection of nontypable H. influenzae. Animals were killed in a time series and the surface of the middle-ear mucosa was examined by scanning electron microscopy (SEM) for the presence of bacterial biofilms. Adult and fetal chinchilla uninfected controls were similarly examined for comparison. In addition, tympanostomy tubes that had been placed in children's ears to treat OME and removed after onset of refractory otorrhea or other problems were examined by SEM and by confocal scanning laser microscopy for bacterial biofilms, and compared with unused control tubes. Results Bacterial biofilms were visually detected by SEM on the middle-ear mucosa of multiple chinchillas in which H. influenzae otitis media had been induced. Qualitative evaluation indicated that the density and thickness of the biofilm might increase until at least 96 hours after injection. The appearance of the middle-ear mucosa of experimental animals contrasted with that of uninjected control animals. Robust bacterial biofilms were also visually detected on tympanostomy tubes removed from children's ears for clinical reasons, in contrast with unused control tubes. Conclusions Bacterial biofilms form on the middle-ear mucosa of chinchillas in experimentally induced H. influenzae otitis media and can form on tympanostomy tubes placed in children's ears. Such biofilms can be directly observed by microscopy. These results reinforce the hypothesis that the bacterial aggregates called biofilms, resistant to treatment by antibiotics and to detection by standard culture techniques, may play a major etiologic role in OME and in one of its frequent complications, post-tympanostomy otorrhea. [source] An in vitro model of bacterial infections in wounds and other soft tissuesAPMIS, Issue 2 2010MARIA WERTHÉN Werthén M, Henriksson L, Jensen PØ, Sternberg C, Givskov M, Bjarnsholt T. An in vitro model of bacterial infections in wounds and other soft tissues. APMIS 2010; 118: 156,64. There is growing evidence that bacteria play a crucial role in the persistence of chronic wounds. These bacteria are most probably present in polymer-embedded aggregates that represent the biofilm mode of growth. Much work has been carried out to study the development of biofilms in vitro, in particular in attachment to solid surfaces. The observations from the chronic wounds indicate that the bacteria are not attached to a solid surface. Consequently, a new in vitro model is required to investigate biofilms in more wound-like settings. This study describes such a novel in vitro model, with bacteria growing as biofilm aggregates in a collagen gel matrix with serum protein mimicking the wound bed of chronic wounds. The model was verified to comprise important hallmarks of biofilms such as the bacterial embedment in a matrix and increased antibiotic tolerance. Furthermore, we have verified the relevance of the model by comparing the organization of the bacteria in the model with the organization of the bacteria in a real chronic wound. We believe that we have developed an important new model for investigating bacterial biofilms in chronic wounds. This model may be used to study biofilm development in chronic wounds and to develop novel diagnostic tools as well as treatment strategies. [source] How strong are bacterial biofilms?BIOTECHNOLOGY & BIOENGINEERING, Issue 5 2010Article first published online: 19 FEB 2010 No abstract is available for this article. [source] | ||||||||||||||||