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  • 1
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Quorum sensing (QS) governs the production of virulence factors and the architecture and sodium dodecyl sulphate (SDS) resistance of biofilm-grown Pseudomonas aeruginosa. P. aeruginosa QS requires two transcriptional activator proteins known as LasR and RhlR and their cognate autoinducers PAI-1 (N-(3-oxododecanoyl)-l-homoserine lactone) and PAI-2 (N-butyryl-l-homoserine lactone) respectively. This study provides evidence of QS control of genes essential for relieving oxidative stress. Mutants devoid of one or both autoinducers were more sensitive to hydrogen peroxide and phenazine methosulphate, and some PAI mutant strains also demonstrated decreased expression of two superoxide dismutases (SODs), Mn-SOD and Fe-SOD, and the major catalase, KatA. The expression of sodA (encoding Mn-SOD) was particularly dependent on PAI-1, whereas the influence of autoinducers on Fe-SOD and KatA levels was also apparent but not to the degree observed with Mn-SOD. β-Galactosidase reporter fusion results were in agreement with these findings. Also, the addition of both PAIs to suspensions of the PAI-1/2-deficient double mutant partially restored KatA activity, while the addition of PAI-1 only was sufficient for full restoration of Mn-SOD activity. In biofilm studies, catalase activity in wild-type bacteria was significantly reduced relative to planktonic bacteria; catalase activity in the PAI mutants was reduced even further and consistent with relative differences observed between each strain grown planktonically. While wild-type and mutant biofilms contained less catalase activity, they were more resistant to hydrogen peroxide treatment than their respective planktonic counterparts. Also, while catalase was implicated as an important factor in biofilm resistance to hydrogen peroxide insult, other unknown factors seemed potentially important, as PAI mutant biofilm sensitivity appeared not to be incrementally correlated to catalase levels.
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  • 2
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature biotechnology 23 (2005), S. 1378-1379 
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] Excessive use of antibiotics has increased resistance of many microbes to these drugs. In a recent issue of Nature, Hoffman et al. show that too little antibiotic can also be detrimental. They demonstrate that subinhibitory levels of the aminoglycoside-class antibiotic tobramycin increase biofilm ...
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  • 3
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Bacteria growing in biofilms experience gradients of environmental conditions, including varying levels of nutrients and oxygen. Therefore, bacteria within biofilms may enter distinct physiological states, depending on the surrounding conditions. In this study, rpoS expression and RpoS levels were measured as indicators of stationary phase growth within thick continuously-fed Pseudomonas aeruginosa biofilms. The level of rpoS expression in a 3-day-old biofilm was found to be three-fold higher than the average expression in stationary phase planktonic culture. RpoS levels in biofilms, indicated by immunoblot analysis, were similar to levels in stationary phase planktonic cultures. In planktonic cultures, oxygen limitation did not lead to increased levels of RpoS, suggesting that oxygen limitation was not the environmental signal causing increased expression of rpoS. These results suggest that bacteria within P. aeruginosa biofilms may exhibit stationary phase characteristics even when cultured in flow conditions that continually replenish nutrients.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Applied microbiology and biotechnology 30 (1989), S. 34-40 
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Stresses exerted by a growing entrapped colony of Escherichia coli up to 3 atmospheres were measured by incorporating a pressure transducer into a specially designed immobilized cell reactor. This stress is comparable in magnitude to the turgor pressure generated by Gram negative bacteria. In complementary experiments, cell densities as high as 850 grams dry weight per liter were measured in aggregates of starved E. coli subjected to controlled applied stresses up to 9 atmospheres. Cell volume reduction was quantitatively described by a model which incorporated the fundamental osmotic properties of the cell. compression of entrapped cells was qualitatively corroborated by electron microscopic examination. These results suggest that entrapped growing bacteria can exert a substantial stress on their surroundings and that dewatering of the starved cell population in an entrapped system may occur. Both of these consequences of entrapped cell growth can be understood in terms of the osmotic behavior of the cells.
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  • 5
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Biofilms are surface-attached microbial communities with characteristic architecture and phenotypic and biochemical properties distinct from their free-swimming, planktonic counterparts. One of the best-known of these biofilm-specific properties is the development of antibiotic resistance that ...
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 59 (1998), S. 261-272 
    ISSN: 0006-3592
    Keywords: effective diffusive permeability ; diffusion coefficient ; biofilm ; cell density ; review ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Experimental measurements of effective diffusive permeabilities and effective diffusion coefficients in biofilms are reviewed. Effective diffusive permeabilities, the parameter appropriate to the analysis of reaction-diffusion interactions, depend on solute type and biofilm density. Three categories of solute physical chemistry with distinct diffusive properties were distinguished by the present analysis. In order of descending mean relative effective diffusive permeability (De/Daq) these were inorganic anions or cations (0.56), nonpolar solutes with molecular weights of 44 or less (0.43), and organic solutes of molecular weight greater than 44 (0.29). Effective diffusive permeabilities decrease sharply with increasing biomass volume fraction suggesting a serial resistance model of diffusion in biofilms as proposed by Hinson and Kocher (1996). A conceptual model of biofilm structure is proposed in which each cell is surrounded by a restricted permeability envelope. Effective diffusion coefficients, which are appropriate to the analysis of transient penetration of nonreactive solutes, are generally similar to effective diffusive permeabilities in biofilms of similar composition. In three studies that examine diffusion of very large molecular weight solutes ( 〉 5000) in biofilms, the average ratio of the relative effective diffusion coefficient of the large solute to the relative effective diffusion coefficient of either sucrose or fluorescein was 0.64, 0.61, and 0.36. It is proposed that large solutes are effectively excluded from microbial cells, that small solutes partition into and diffuse within cells, and that ionic solutes are excluded from cells but exhibit increased diffusive permeability (but decreased effective diffusion coefficients) due to sorption to the biofilm matrix. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:261-272, 1998.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 45 (1995), S. 503-510 
    ISSN: 0006-3592
    Keywords: biofilm ; thickness ; heterogeneity ; roughness ; microscopy ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The thickness variability of biofilms of Pseudomonas aeruginosa, Klebsiella pneumoniae, and the binary population combination of these two species was quantified. The experimental method involved cryoembedding biofilms with a commercial tissue embedding agent, sectioning, and applying image analysis to construct thickness profiles along linear transects (up to 1 cm in length) across the substratum. Biofilms embedded and sectioned by this method were locally as thin as a single cell attached to the surface (〈5 μm) and as thick as 1000 μm. Week-old biofilms of three different species compositions displayed distinct structural features as indicated by their mean thicknesses and by a roughness coefficient. Monopopulation biofilms of P. aeruginosa (29 μm mean thickness) or K. pneumoniae (100 μm mean thickness) were thinner than the binary population biofilm (400 μm mean thickness). A roughness coefficient developed in this investigation corroborated the qualitative visual characterization of P. aeruginosa biofilms as relatively uniformly thick (mean roughness coefficient 0.15), K. pneumoniae biofilms as patchy (mean roughness coefficient 1.14), and the binary population biofilm as intermediate (mean roughness coefficient 0.26). Whereas P. aeruginosa and binary population biofilms covered the substratum completely, significant areas of essentially bare substratum were apparent in K. pneumoniae biofilms. The patchiness of K. pneumoniae biofilms may be due to the fact that this organism is nonmotile. A spatial correlation analysis of the thickness data indicated that thickness measurements were still correlated even when separated by distances that exceeded the mean biofilm thickness. Cell aggregates, some of them hundreds of microns in size, were observed in the effluent of K. pneumoniae and binary population biofilm reactors. Measurements of thickness variability and other observations reported in this article provide a quantitative basis for analysis of microscale structural heterogeneity of biofilms. © 1995 John Wiley & Sons, Inc.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 41 (1993), S. 111-117 
    ISSN: 0006-3592
    Keywords: biofilm ; detachment ; model ; physiology ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A general mathematical framework for modeling biofilm detachment is presented. The approach is founded on a material balance on biomass that equates the detachment rate to the product of a detachment frequency and a detaching particle mass. The model provides a theoretical basis for deriving many of the empirical detachment rate expressions in common use and can thus lend some insight into their physical and biological significance. By allowing for variation in the detachment frequency with depth in the biofilm, the model permits derivation of detachment expressions that reflect a dependence on chemical or physiological gradients in the biofilm. Analysis of literature data sets from two different biofilm systems suggests, in both cases, that detachment is a growth-associated phenomenon. © 1993 John Wiley & Sons, Inc.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 42 (1993), S. 111-117 
    ISSN: 0006-3592
    Keywords: biofilm ; particle ; Pseudomonas aeruginosa ; transport ; roughness ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Fluorescent latex microbeads added to a Pseudomonas aeruginosa biofilm as tracers of particle movement penetrated the biofilm and remained in it much longer than predicted by a model of advective displacement due to cell growth. Beads with a nominal diameter of 1 μm that were added in the bulk fluid became distributed throughout the biofilm depth. Some microbeads penetrated to the substratum within the 24-h bead addition period. The biofilms had a mean thickness of approximately 34 μm but have been previously shown to be quite rough. Measured rates of bead release from the biofilm corresponded to first order time coefficients of 0.01-0.03 h-1. These bead release rates were approximately an order of magnitude less than the predicted time scale of advective transport, which is just the experimentally measured specific cellular growth rate of 0.15 h-1. Computer simulations of bead transport using the biofilm model BIOSIM were compared with bead release rate data and with bead position distributions within the biofilm as determined by microscopic examination of thin cross sections of embedded biofilm. The model predicted much faster release of beads from the biofilm than actually occurred. It is hypothesized that both the ability of beads to penetrate the biofilm and the unexpectedly low advective displacement velocity of particles in the biofilm were due to the rough nature of the biofilm. © 1993 John Wiley & Sons, Inc.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 37 (1991), S. 824-833 
    ISSN: 0006-3592
    Keywords: immobilized cells ; growth rate ; autoradiography ; E. coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The growth of immobilized Escherichia coli was analyzed by pulse-chase radioisotope labeling of the cell mass with 35SO42- and subsequent liquid emulsion autoradiography of thin cross sections of the cell aggregate. Bacteria were retained in a planar aggregate on a microporous membrane and grown anaerobically on a phosphate-buffered medium with glucose as the sole carbon and energy source. A mathematical model of immobilized cell growth and convection was used to predict the distribution of label in the cell mass and permit information about both the magnitude and variation in the intrinsic growth rate to be extracted. Growing zone dimensions ranging from 4 to 48 μm and growth rates from 0.28 to 0.5 h-1 were found. Data collected at low glucose concentrations were consistent with a zero-order description of intrinsic growth kinetics. At high glucose concentrations, conditions under which the system was subject to significant pH inhibition, the data were best described by the prediction of a first-order kinetic model. When coupled with a suitable analytical framework, the combination of radioisotope labeling and autoradiography provides a general method for characterizing immobilized cell growth rates.
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