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Biofilm Structure (biofilm + structure)
Selected AbstractsInfluences of milk components on biofilm formation of Cronobacter spp. (Enterobacter sakazakii)LETTERS IN APPLIED MICROBIOLOGY, Issue 6 2009G.I. Dancer Abstract Aim:, To determine the critical component(s) of skim milk for biofilm formation of Cronobacter species. Methods and Results:, Biofilm forming ability of 72 Cronobacter strains in skim milk preparation was assayed by crystal violet staining. The results revealed that whey protein and casein are more important determinants of skim milk for biofilm formation than lactose, although there was a wide variation in biofilm forming ability. Biofilm structure and capsular material of six strains exhibiting different biofilm forming ability was investigated via electron microscopes. Scanning electron microscopy showed visually that while the strong biofilm formers (E27B, FSM 30 and 2·82) resulted in almost complete coagulation of skim milk, the weak biofilm formers (55, FSM 290 and 2·84) caused less coagulation. No capsule was clearly delineated in transmission electron micrographs of either strong or weak biofilm formers. Conclusion:, These results indicate that, for biofilm formation of Cronobacter species in skim milk, nitrogen source is probably a more important determinant than carbohydrate, and that strong biofilm formers are responsible for substantial coagulation of skim milk. Significance and Impact of the Study:, This study provides information for better understanding of the underlying mechanisms by which Cronobacter species form biofilm in infant formula milk. [source] Acid microenvironments in microbial biofilms of antarctic endolithic microecosystemsENVIRONMENTAL MICROBIOLOGY, Issue 4 2003Asunción Los Ríos Summary Antarctic endolithic microecosystems harbour distinct biofilms. The lithic substrate and the microorganisms comprising these films are intimately linked, leading to complex mineral,microbe interactions. Hence, the microhabitats and microenvironments of these microecosystems are not only determined by the physicochemical features of the lithic substrate, but are also conditioned by the biological components of these biofilms. The Antarctic biofilms analysed in this study are characterized by the presence of extracellular polymer substances and acid microenvironments in the proximity of the cells; cyanobacteria appearing as key components. On ultrastructural analysis, these endolithic cyanobacteria showed differences in sheath organization, probably related to their spatial position in the lithic substrate. It is proposed that in this type of ecosystem, biofilm structure could favour the formation of microsites with specific physicochemical conditions appropriate for the survival of microbial communities in this extreme environment. [source] Microbial interactions and differential protein expression in Staphylococcus aureus ,Candida albicans dual-species biofilmsFEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 3 2010Brian M. Peters Abstract The fungal species Candida albicans and the bacterial species Staphylococcus aureus are responsible for a majority of hospital-acquired infections and often coinfect critically ill patients as complicating polymicrobial biofilms. To investigate biofilm structure during polymicrobial growth, dual-species biofilms were imaged with confocal scanning laser microscopy. Analyses revealed a unique biofilm architecture where S. aureus commonly associated with the hyphal elements of C. albicans. This physical interaction may provide staphylococci with an invasion strategy because candidal hyphae can penetrate through epithelial layers. To further understand the molecular mechanisms possibly responsible for previously demonstrated amplified virulence during coinfection, protein expression studies were undertaken. Differential in-gel electrophoresis identified a total of 27 proteins to be significantly differentially produced by these organisms during coculture biofilm growth. Among the upregulated staphylococcal proteins was l -lactate dehydrogenase 1, which confers resistance to host-derived oxidative stressors. Among the downregulated proteins was the global transcriptional repressor of virulence factors, CodY. These findings demonstrate that the hyphae-mediated enhanced pathogenesis of S. aureus may not only be due to physical interactions but can also be attributed to the differential regulation of specific virulence factors induced during polymicrobial growth. Further characterization of the intricate interaction between these pathogens at the molecular level is warranted, as it may aid in the design of novel therapeutic strategies aimed at combating fungal,bacterial polymicrobial infection. [source] Temporal Dynamics of River Biofilm in Constant Flows: A Case Study in a Riverside Laboratory FlumeINTERNATIONAL REVIEW OF HYDROBIOLOGY, Issue 2 2010Stéphanie Boulętreau Abstract A 15-week experiment was performed in a riverside laboratory flume (with diverted river water) to check variations of river biofilm structure (biomass, algal and bacterial compositions) and function (community gross primary production GPP and respiration) under constant flow while water quality went through natural temporal variations. One major suspended matter pulse coinciding with one river flood was recorded after 10 weeks of experiment. Epilithic biofilm first exhibited a 10-week typical pattern of biomass accrual reaching 33 g ash-free dry matter (AFDM) m,2 and 487 mg chlorophyll- a m,2 and then, experienced a shift to dominance of loss processes (loss of 60% AFDM and 80% chlorophyll- a) coinciding with the main suspended matter pulse. Algal diversity remained low and constant during the experiment: Fragilaria capucina and Encyonema minutum always contribute over 80% of cell counts. DGGE banding patterns discriminated between two groups that corresponded to samples before and after biomass loss, indicating major changes in the bacterial community composition. GPP/R remained high during the experiment, suggesting that photoautotrophic metabolism prevailed and detachment was not autogenic (i.e., due to algal senescence or driven by heterotrophic processes within the biofilm). Observational results suggested that silt deposition into the biofilm matrix could have triggered biomass loss. (© 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Genetic analysis of functions involved in the late stages of biofilm development in Burkholderia cepacia H111MOLECULAR MICROBIOLOGY, Issue 2 2002Birgit Huber Summary Burkholderia cepacia and Pseudomonas aeruginosa often co-exist as mixed biofilms in the lungs of patients suffering from cystic fibrosis (CF). Here, we report the isolation of 13 random mini-Tn 5 insertion mutants of B. cepacia H111 that are defective in biofilm formation on a polystyrene surface. We show that the screening procedure used in this study is biased towards mutants defective in the late stages of biofilm development. A detailed quantitative analysis of the biofilm structures formed by wild-type and mutant strains revealed that the isolated mutants are impaired in their abilities to develop a typical three-dimensional biofilm structure. Molecular investigations showed that the genes required for biofilm maturation fall into several classes: (i) genes encoding for surface proteins; (ii) genes involved in the biogenesis and maintenance of an integral outer membrane; and (iii) genes encoding regulatory factors. It is shown that three of the regulatory mutants produce greatly reduced amounts of N -octanoylhomoserine lactone (C8-HSL). This compound serves as the major signal molecule of the cep quorum-sensing system. As this density-dependent regulatory system is involved in the regulation of biofilm maturation, we investigated the interplay between the three regulatory genes and the quorum-sensing cascade. The results of these investigations show that the identified genes encode for regulatory elements that are positioned upstream of the cep system, indicating that the quorum-sensing system of B. cepacia is a major checkpoint for biofilm formation. [source] Interaction of biofilm structure and bulk liquid flowBIOTECHNOLOGY & BIOENGINEERING, Issue 1 2010Article first published online: 16 JUL 2010 No abstract is available for this article. [source] Online assessment of biofilm development, sloughing and forced detachment in tube reactor by means of magnetic resonance microscopyBIOTECHNOLOGY & BIOENGINEERING, Issue 1 2010Michael Wagner Abstract Magnetic resonance microscopy (MRM) was successfully applied for non-invasive online monitoring of biofilm development, sloughing, and forced detachment. Biofilm cultivation was performed in a tube reactor directly placed in the MRM scanner. Based on the differences in relaxation time of free and bound protons, the distributed water signal was allocated to the bulk and the biofilm phase. The velocity of the flowing water in the tube reactor was measured in all three directions (x, y, and z) at spatial resolutions of 78,µm. From the velocity data, maps of flow gradients (shear rates) were derived. The experiments showed that a more compact biofilm structure is sloughed off in total with nearly no biomass left on the substratum. Continued biofilm cultivation resulted in filamentous biofilm structures, which did not show any sloughing. Experiments at higher Reynolds numbers were performed in order to force biofilm detachment. Continuous measuring of proton velocity and biomass was used to characterize the different stages of biofilm development. The measurements revealed that biofilms are able to resist extremely high local shear stress being raised up to factor of 20 compared to the mean local shear stress acting on the complete biofilm surface. The maximum local shear stress of single biofilm structures exposed to flow was found to be on average seven times higher compared to the mean local shear stress of the entire biofilm surface. MRM was able to visualize and quantify the development of biofilms and interaction of biofilms with the surrounding fluid at the meso-scale. It is suggested that detachment and sloughing depends on both internal and external structural parameters. Biotechnol. Bioeng. 2010;107: 172,181. © 2010 Wiley Periodicals, Inc. [source] Analyzing characteristic length scales in biofilm structuresBIOTECHNOLOGY & BIOENGINEERING, Issue 2 2009K. Milferstedt Abstract The quantification of biofilm structure based on image analysis requires a statistical measure like representative elemental areas (REA) to determine the necessary size of biofilm area to be imaged. In this study, REAs for biofilm structure were calculated for the descriptors Gray level and Correlation (COR) derived from a spatial gray level dependence matrix analysis (SGLDM). An important difference between these two descriptors is their response to structural features at different spatial scales. Gray level is a scale-independent descriptor, whereas COR is scale-dependent. For scale-independent descriptors, the size of the individual images is not relevant when determining REAs. This is in contrast to scale-dependent descriptors for which REAs can only be determined when the area of each image covers the range of structural variability of the biofilm. We used COR to analyze scale dependence of structural heterogeneity at different length scales. A characteristic length of 400 µm in biofilm images provides structural information relevant for mass transport phenomena in biofilms. Overall REAs for gray level and COR were on average 3.4 mm2. The scale-dependent descriptor COR could not in all cases accurately be determined from combining individual image analysis results,even when the combined area resulted in the REA. Microscope and camera specifications define the upper and lower limit of detectable characteristic length that can be extracted from images and should therefore be considered in the experimental design when choosing an imaging technique. Biotechnol. Bioeng. 2009;102: 368,379. © 2008 Wiley Periodicals, Inc. [source] Genetic analysis of functions involved in the late stages of biofilm development in Burkholderia cepacia H111MOLECULAR MICROBIOLOGY, Issue 2 2002Birgit Huber Summary Burkholderia cepacia and Pseudomonas aeruginosa often co-exist as mixed biofilms in the lungs of patients suffering from cystic fibrosis (CF). Here, we report the isolation of 13 random mini-Tn 5 insertion mutants of B. cepacia H111 that are defective in biofilm formation on a polystyrene surface. We show that the screening procedure used in this study is biased towards mutants defective in the late stages of biofilm development. A detailed quantitative analysis of the biofilm structures formed by wild-type and mutant strains revealed that the isolated mutants are impaired in their abilities to develop a typical three-dimensional biofilm structure. Molecular investigations showed that the genes required for biofilm maturation fall into several classes: (i) genes encoding for surface proteins; (ii) genes involved in the biogenesis and maintenance of an integral outer membrane; and (iii) genes encoding regulatory factors. It is shown that three of the regulatory mutants produce greatly reduced amounts of N -octanoylhomoserine lactone (C8-HSL). This compound serves as the major signal molecule of the cep quorum-sensing system. As this density-dependent regulatory system is involved in the regulation of biofilm maturation, we investigated the interplay between the three regulatory genes and the quorum-sensing cascade. The results of these investigations show that the identified genes encode for regulatory elements that are positioned upstream of the cep system, indicating that the quorum-sensing system of B. cepacia is a major checkpoint for biofilm formation. [source] Online assessment of biofilm development, sloughing and forced detachment in tube reactor by means of magnetic resonance microscopyBIOTECHNOLOGY & BIOENGINEERING, Issue 1 2010Michael Wagner Abstract Magnetic resonance microscopy (MRM) was successfully applied for non-invasive online monitoring of biofilm development, sloughing, and forced detachment. Biofilm cultivation was performed in a tube reactor directly placed in the MRM scanner. Based on the differences in relaxation time of free and bound protons, the distributed water signal was allocated to the bulk and the biofilm phase. The velocity of the flowing water in the tube reactor was measured in all three directions (x, y, and z) at spatial resolutions of 78,µm. From the velocity data, maps of flow gradients (shear rates) were derived. The experiments showed that a more compact biofilm structure is sloughed off in total with nearly no biomass left on the substratum. Continued biofilm cultivation resulted in filamentous biofilm structures, which did not show any sloughing. Experiments at higher Reynolds numbers were performed in order to force biofilm detachment. Continuous measuring of proton velocity and biomass was used to characterize the different stages of biofilm development. The measurements revealed that biofilms are able to resist extremely high local shear stress being raised up to factor of 20 compared to the mean local shear stress acting on the complete biofilm surface. The maximum local shear stress of single biofilm structures exposed to flow was found to be on average seven times higher compared to the mean local shear stress of the entire biofilm surface. MRM was able to visualize and quantify the development of biofilms and interaction of biofilms with the surrounding fluid at the meso-scale. It is suggested that detachment and sloughing depends on both internal and external structural parameters. Biotechnol. Bioeng. 2010;107: 172,181. © 2010 Wiley Periodicals, Inc. [source] Analyzing characteristic length scales in biofilm structuresBIOTECHNOLOGY & BIOENGINEERING, Issue 2 2009K. Milferstedt Abstract The quantification of biofilm structure based on image analysis requires a statistical measure like representative elemental areas (REA) to determine the necessary size of biofilm area to be imaged. In this study, REAs for biofilm structure were calculated for the descriptors Gray level and Correlation (COR) derived from a spatial gray level dependence matrix analysis (SGLDM). An important difference between these two descriptors is their response to structural features at different spatial scales. Gray level is a scale-independent descriptor, whereas COR is scale-dependent. For scale-independent descriptors, the size of the individual images is not relevant when determining REAs. This is in contrast to scale-dependent descriptors for which REAs can only be determined when the area of each image covers the range of structural variability of the biofilm. We used COR to analyze scale dependence of structural heterogeneity at different length scales. A characteristic length of 400 µm in biofilm images provides structural information relevant for mass transport phenomena in biofilms. Overall REAs for gray level and COR were on average 3.4 mm2. The scale-dependent descriptor COR could not in all cases accurately be determined from combining individual image analysis results,even when the combined area resulted in the REA. Microscope and camera specifications define the upper and lower limit of detectable characteristic length that can be extracted from images and should therefore be considered in the experimental design when choosing an imaging technique. Biotechnol. Bioeng. 2009;102: 368,379. © 2008 Wiley Periodicals, Inc. [source] | ||||||||||