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Vitro Biofilms (vitro + biofilm)
Selected AbstractsThe characterization of functions involved in the establishment and maturation of Klebsiella pneumoniae in vitro biofilm reveals dual roles for surface exopolysaccharidesENVIRONMENTAL MICROBIOLOGY, Issue 3 2008Damien Balestrino Summary The ability to form biofilm is seen as an increasingly important colonization strategy among both pathogenic and environmental Klebsiella pneumoniae strains. The aim of the present study was to identify abiotic surface colonization factors of K. pneumoniae using different models at different phases of biofilm development. A 2200 K. pneumoniae mutant library previously obtained by signature-tagged mutagenesis was screened in static and dynamic culture models to detect clones impaired at early and/or mature stages of biofilm formation. A total of 28 mutants were affected during late phases of biofilm formation, whereas 16 mutants displayed early adhesion defect. These mutants corresponded to genes involved in potential cellular and DNA metabolism pathways and to membrane transport functions. Eight mutants were deficient in capsule or LPS production. Gene disruption and microscopic analyses showed that LPS is involved in initial adhesion on both glass and polyvinyl-chloride and the capsule required for the appropriate initial coverage of substratum and the construction of mature biofilm architecture. These results give new insight into the bacterial factors sequentially associated with the ability to colonize an abiotic surface and reveal the dual roles played by surface exopolysaccharides during K. pneumoniae biofilm formation. [source] Cariogenicity of soluble starch in oral in vitro biofilm and experimental rat caries studies: a comparisonJOURNAL OF APPLIED MICROBIOLOGY, Issue 3 2008T. Thurnheer Abstract Aims:, Common belief suggests that starch is less cariogenic than sugar; however, the related literature is quite controversial. We aimed to compare cariogenic and microbiological effects of soluble starch in both a standard animal model and an oral biofilm system, and to assess the possible substitution of the animal model. Methods and Results:, Six-species biofilms were grown anaerobically on enamel discs in saliva and medium with glucose/sucrose, starch (average molecular weight of 5000, average polymerization grade of 31), or mixtures thereof. After 64·5 h of biofilm formation, the microbiota were quantitated by cultivation and demineralization was measured by quantitative light-induced fluorescence. To assess caries incidence in rats, the same microbiota as in the biofilm experiments were applied. The animals were fed diets containing either glucose, glucose/sucrose, glucose/sucrose/starch or starch alone. Results with both models show that demineralization was significantly smaller with starch than sucrose. Conclusions:, The data demonstrate that soluble starch is substantially less cariogenic than glucose/sucrose. Significance and Impact of the Study:, By leading to the same scientific evidence as its in vivo counterpart, the described in vitro biofilm system provides an interesting and valuable tool in the quest to reduce experimentation with animals. [source] Biofilm formation and cellulose expression among diverse environmental Pseudomonas isolatesENVIRONMENTAL MICROBIOLOGY, Issue 11 2006Susanne Ude Summary The ability to form biofilms is seen as an increasingly important colonization strategy among both pathogenic and environmental bacteria. A survey of 185 plant-associated, phytopathogenic, soil and river Pseudomonas isolates resulted in 76% producing biofilms at the air,liquid (A,L) interface after selection in static microcosms. Considerable variation in biofilm phenotype was observed, including waxy aggregations, viscous and floccular masses, and physically cohesive biofilms with continuously varying strengths over 1500-fold. Calcofluor epifluorescent microscopy identified cellulose as the matrix component in biofilms produced by Pseudomonas asplenii, Pseudomonas corrugata, Pseudomonas fluorescens, Pseudomonas marginalis, Pseudomonas putida, Pseudomonas savastanoi and Pseudomonas syringae isolates. Cellulose expression and biofilm formation could be induced by the constitutively active WspR19 mutant of the cyclic-di-GMP-associated, GGDEF domain-containing response regulator involved in the P. fluorescens SBW25 wrinkly spreader phenotype and cellular aggregation in Pseudomonas aeruginosa PA01. WspR19 could also induce P. putida KT2440, which otherwise did not produce a biofilm or express cellulose, as well as Escherichia coli K12 and Salmonella typhimurium LT2, both of which express cellulose yet lack WspR homologues. Statistical analysis of biofilm parameters suggest that biofilm development is a more complex process than that simply described by the production of attachment and matrix components and bacterial growth. This complexity was also seen in multivariate analysis as a species-ecological habitat effect, underscoring the fact that in vitro biofilms are abstractions of those surface and volume colonization processes used by bacteria in their natural environments. [source] Biofilms in chronic bacterial prostatitis (NIH-II) and in prostatic calcificationsFEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 3 2010Sandra Mazzoli Abstract The prevalence of inflammatory conditions of the prostate gland is increasing. In Italy, there is a high incidence of prostatitis (13.3%), also accompanied by prostatic calcifications. Cat NIH-II chronic bacterial prostatitis (CBPs) are the most frequent. Their aetiology theoretically involves the whole range of bacterial species that are able to form biofilms and infect prostate cells. The aim of our study was to isolate potential biofilm-producing bacteria from CBP patients, to evaluate their ability to produce in vitro biofilms, and to characterize intraprostatic bacteria and prostatic calcifications using scanning electron microscopy. The 150 clinical bacterial strains isolated from chronic prostatitis NIH-II patients were: 50 Enterococcus faecalis; 50 Staphylococcus spp.; 30 Escherichia coli; 20 gram-negative miscellanea. Quantitative assay of biofilm production and adhesion was performed according to the classic Christensen microwell assay. Isolates were classified as nonproducers, weak, moderate or strong producers. The majority of E. coli, gram-negative bacteria, Staphylococci and Enterococci strains were strong or medium producers: 63,30%, 75,15%, 46,36%, and 58,14%, respectively. Prostatic calcifications consisted of bacteria-like forms similar to the species isolated from biological materials and calcifications of patients. Our study proves, for the first time, that bacterial strains able to produce biofilms consistently are present in CBP. Additionally, prostatic calcifications are biofilm-related. [source] Determining the spatial distribution of viable and nonviable bacteria in hydrated microcosm dental plaques by viability profilingJOURNAL OF APPLIED MICROBIOLOGY, Issue 3 2002C.K. Hope Aims: The aim of this study was to use confocal laser scanning microscopy (CLSM) to examine the spatial distribution of both viable and nonviable bacteria within microcosm dental plaques grown in vitro. Previous in vivo studies have reported upon the distribution of viable bacteria only. Methods and Results: Oral biofilms were grown on hydroxyapatite (HA) discs in a constant-depth film fermenter (CDFF) from a saliva inoculum. The biofilms were stained with the BacLightTM LIVE/DEAD system and examined by CLSM. Fluorescence intensity profiles through the depth of the biofilm showed an offset between the maximum viable intensity and the maximum nonviable intensity. Topographical differences between the surface properties of the viable and nonviable biofilm virtual surfaces were also measured. Conclusions: The profile of fluorescence intensity from viable and nonviable staining suggested that the upper layers of the biofilm contain proportionally more viable bacteria than the lower regions of the biofilm. Significance and Impact of Study: Viability profiling records the transition from predominantly viable to nonviable bacteria through biofilms suggesting that this technique may be of use for quantifying the effects of antimicrobial compounds upon biofilms. The distribution of viable bacteria was similar to that found in dental plaque in vivo suggesting that the CDFF produces in vitro biofilms which are comparable to their in vivo counterparts in terms of the spatial distribution of viable bacteria. [source] | ||||||||||