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Genomewide analysis of gene expression in Staphylococcus epidermidis biofilms: insights into the pathophysiology of S. epidermidis biofilms and the role of phenol-soluble modulins in formation of biofilms.

Many bacterial pathogens form cellular agglomerations known as biofilms, which considerably limit the success of both antibiotic treatment and the human immune defense. To gain insight into the pathophysiology of the leading nosocomial pathogen, Staphylococcus epidermidis, we analyzed the genome of biofilm-forming S. epidermidis, constructed a microarray representing its entire transcriptome, and performed expression profiling of an S. epidermidis biofilm. Gene-regulated processes in the biofilm led to a nonaggressive and protected form of bacterial growth with low metabolic activity, which is optimally suited to guarantee long-term survival during chronic infection. A class of peptides known as phenol-soluble modulins, which combine proinflammatory activity with a putative role in detachment of biofilms, evolved as potential key determinants controlling the switch between aggressive and quiescent modes of infection. Our data suggest that S. epidermidis adjusts its lifestyle to varying requirements during colonization and infection by means of an expansive change of gene expression. The observed physiological characteristics of the biofilm mode of growth--in particular, the contribution of surfactant-like peptides--might serve as a model for a variety of biofilm-forming pathogens.

Bacterial Toxins↗

Gene expression in Bacillus subtilis surface biofilms with and without sporulation and the importance of yveR for biofilm maintenance.

Five independent DNA microarray experiments were used to study the gene expression profile of a 5-day Bacillus subtilis air-liquid interface biofilm relative to planktonic cells. Both wild-type B. subtilis and its sporulation mutant (DeltaspoIIGB::erm) were investigated to discern the important biofilm genes (in the presence and absence of sporulation). The microarray results indicated that suspension cells were encountering anaerobic conditions, and the air-liquid interface biofilm was metabolically active. For the statistically significant differential expression (P < 0.05), there were 342 genes induced and 248 genes repressed in the wild-type biofilm, whereas 371 genes were induced and 128 genes were repressed in the sporulation mutant biofilm. The microarray results were confirmed with RNA dot blotting. A small portion of cells (1.5%) in the wild-type biofilm formed spores and sporulation genes were highly expressed. In the biofilm formed by the sporulation mutant, competence genes (comGA, srfAA, srfAB, srfAD, and comS) were induced which indicate a role for quorum sensing (bacterial gene expression controlled by sensing their population) in biofilms. There were 53 genes consistently induced in the biofilms of both the wild-type strain and its spoIIGB mutant-those genes have functions for transport, metabolism, antibiotic production-and 26 genes with unknown functions. Besides the large number of genes with known functions induced in the biofilm (121 genes in the wild-type biofilm and 185 genes in the sporulation mutant biofilm), some genes with unknown functions were also induced (221 genes in the wild-type biofilm and 186 genes in the sporulation mutant biofilm), such as the yve operon which appears to be involved in polysaccharide synthesis and the ybc operon which inhibits the growth of competitors for nutrients. A knockout mutant of yveR was constructed, and the mutant showed major defects in biofilm maintenance. Both the wild-type strain and its sporulation mutant formed normal biofilms, suggesting complete sporulation is not necessary for biofilm formation. The expression profiles of these two strains share more repressed genes than induced genes, suggesting that the biofilm cells repress similar pathways in response to starvation and high cell density.

Bacillus subtilis↗

Reproducibility of biofilm processes and the meaning of steady state in biofilm reactors.

The need for reproducing biofilm processes is undisputable - the quality of biofilm research depends on this reproducibility. However, as many biofilm researchers know, long-term biofilm processes are notoriously difficult to reproduce. To avoid problems related to biofilm reproducibility two strategies are used: (1) to study very young biofilms that have accumulated for a few hours to a few days only, and (2) to run biofilm experiments only once. The first approach trades reproducibility for relevance because natural biofilms are usually older, often much older than a few days. This approach can be applied to answer questions relevant to initial events of biofilm formation but not questions relevant to long-term biofilm accumulation. The second approach conceals the problem of biofilm reproducibility. To assure reproducibility of biofilm processes, we methodically followed a procedure for growing biofilms in terms of microbial makeup, media composition, temperature, surface preparation, etc. Despite all this effort the reproducibility of our results for long term growth is unimpressive. Consequently, the question had to be asked: Are biofilm processes reproducible? The experiments described in this paper address this question. Biofilms grown in two identical and identically operated biofilm reactors had comparable structure only until the first sloughing event. After that, biofilms had different patterns of accumulation.

Biofilms↗

Enhanced biofilm formation and increased resistance to antimicrobial agents and bacterial invasion are caused by synergistic interactions in multispecies biofilms.

Most biofilms in their natural environments are likely to consist of consortia of species that influence each other in synergistic and antagonistic manners. However, few reports specifically address interactions within multispecies biofilms. In this study, 17 epiphytic bacterial strains, isolated from the surface of the marine alga Ulva australis, were screened for synergistic interactions within biofilms when present together in different combinations. Four isolates, Microbacterium phyllosphaerae, Shewanella japonica, Dokdonia donghaensis, and Acinetobacter lwoffii, were found to interact synergistically in biofilms formed in 96-well microtiter plates: biofilm biomass was observed to increase by >167% in biofilms formed by the four strains compared to biofilms composed of single strains. When exposed to the antibacterial agent hydrogen peroxide or tetracycline, the relative activity (exposed versus nonexposed biofilms) of the four-species biofilm was markedly higher than that in any of the single-species biofilms. Moreover, in biofilms established on glass surfaces in flow cells and subjected to invasion by the antibacterial protein-producing Pseudoalteromonas tunicata, the four-species biofilms resisted invasion to a greater extent than did the biofilms formed by the single species. Replacement of each strain by its cell-free culture supernatant suggested that synergy was dependent both on species-specific physical interactions between cells and on extracellular secreted factors or less specific interactions. In summary, our data strongly indicate that synergistic effects promote biofilm biomass and resistance of the biofilm to antimicrobial agents and bacterial invasion in multispecies biofilms.

Acinetobacter↗

Biofilm morphology and nitrification activities: recovery of nitrifying biofilm particles covered with heterotrophic outgrowth.

Biofilm processes are commonly used for nitrification. Operationally, a whitish heterotrophic biofilm layer tends to develop onto nitrifying biofilm when it has been exposed to organic carbon-containing wastewater for a prolonged period. The development of a heterotrophic biofilm layer could lead to deterioration in nitrification activities and biofilm morphology. The recovery characteristics, in terms of biofilm morphology and nitrification activities, of deteriorated biofilm particles (i.e. nitrifying biofilm particles covered with heterotrophic outgrowth) were investigated by transferring the deteriorated biofilm particles (from an Ultra-Compact Biofilm Reactor (UCBR) which was part of a packed bed-UCBR system used for treating organic carbon and ammonia-containing wastewater) to a UCBR fed solely with ammonia wastewater (referred to as 'Recovery-UCBR'). At a hydraulic retention time (HRT) of 8.7 h and a sand-carrier concentration of 4.0% (v/v), density of the outer heterotrophic biofilm layer reduced progressively which led to subsequent detachment of this layer from the nitrifying biofilm particles. As a result, morphology of the nitrifying biofilm improved gradually in the Recovery-UCBR. A stable nitrification rate of up to 1.74 kg NH4+-N/m3 d was obtained in the Recovery-UCBR.

Ammonia↗

Interspecies variation in Candida biofilm formation studied using the Calgary biofilm device.

An in vitro assay to study multiple Candida biofilms, in parallel, has been carried out using the Calgary biofilm device (CBD). We here report: i) standardization of the CBD for Candida albicans biofilm formation, ii) kinetics of C. albicans biofilm formation, iii) biofilm formation by five Candida species, and iv) effect of dietary carbohydrates on biofilm formation. The biofilm metabolic activity on all CBD pegs was similar (p=0.6693) and C. albicans biofilm formation revealed slow growth up to 36 h and significantly higher growth up to 48 h (p<0.001). Significant differences in total biofilm metabolic activity were seen for glucose, fructose and lactose grown C. albicans compared with sucrose and maltose grown yeasts. Candida krusei developed the largest biofilm mass (p<0.05) relative to C. albicans, C. glabrata, C. dubliniensis and C. tropicalis. Scanning electron microscopy revealed that C. krusei produced a thick multilayered biofilm of pseudohyphal forms embedded within the polymer matrix, whereas C. albicans, C. dubliniensis and C. tropicalis biofilms consisted of clusters or chains of cells with sparse extracellular matrix material. We conclude that CBD is a useful, simple, low cost miniature device for parallel study of Candida biofilms and factors modulating this phenomenon.

Biofilms↗

Biofilm development in a membrane-aerated biofilm reactor: effect of flow velocity on performance.

The effect of liquid flow velocity on biofilm development in a membrane-aerated biofilm reactor was investigated both by mathematical modeling and by experiment, using Vibrio natriegens as a test organism and acetate as carbon substrate. It was shown that velocity influenced mass transfer in the diffusion boundary layer, the biomass detachment rate from the biofilm, and the maximum biofilm thickness attained. Values of the overall mass transfer coefficient of a tracer through the diffusion boundary layer, the biofilm, and the membrane were shown to be identical during different experiments at the maximum biofilm thickness. Comparison of the results with published values of this parameter in membrane attached biofilms showed a similar trend. Therefore, it was postulated that this result might indicate the mechanism that determines the maximum biofilm thickness in membrane attached biofilms. In a series of experiments, where conditions were set so that the active layer of the membrane attached biofilm was located close to the membrane biofilm interface, it was shown that the most critical effect on process performance was the effect of velocity on biofilm structure. Biofilm thickness and effective diffusivity influenced reaction and diffusion in a complex manner such that the yield of biomass on acetate was highly variable. Consideration of endogenous respiration in the mathematical model was validated by direct experimental measurements of yield coefficients. Good agreement between experimental measurements of acetate and oxygen uptake rates and their prediction by the mathematical model was achieved.

Biofilms↗

Overcoming oxygen limitations in membrane-attached biofilms--investigation of flux and diffusivity in an anoxic biofilm.

The possibility of overcoming oxygen limitations in membrane-attached biofilms has been investigated by using nitrate as an electron acceptor instead of oxygen in an extractive membrane bioreactor (EMB) degrading toluene. The effect of nitrate concentration on toluene flux, the effective diffusivity in the biofilm and the biofilm activity has been investigated. A counter-diffusion-reaction model is also presented, describing the pollutant flux versus biofilm thickness. The toluene flux decreased with increasing biofilm thickness under excess nitrate concentrations, similar to the experiment with low nitrate. Mathematical modelling indicated that this was either due to decreasing activity, and/or different diffusivities in the biofilm. The effective diffusivity was investigated by using an inert tracer molecule. It remained constant for biofilm thicknesses up to 1.8mm, with a value twice that in water. The biofilm activity was investigated by inactivating a mature biofilm using sodium azide. The toluene flux remained the same before and after the addition of sodium azide, suggesting that the activity in the biofilm is very low. We conclude that the decreasing toluene flux with increasing biofilm thickness is due to the diffusional resistance of the inactive biofilm.

Biofilms↗

Biofilm growth and characteristics in an alternating pumped sequencing batch biofilm reactor (APSBBR).

A novel biofilm reactor-alternating pumped sequencing batch biofilm reactor (APSBBR)-was developed to treat synthetic dairy wastewater at a volumetric chemical oxygen demand (COD) loading rate of 487 g COD m(-3) d(-1) and an areal loading rate of 5.4 g COD m(-2) d(-1). This biofilm reactor comprised two tanks, Tanks 1 and 2, with two identical plastic biofilm modules in each tank. The maximum volume of bulk fluid in the two-tank reactor was the volume of one tank. The APSBBR was operated as a sequencing batch biofilm reactor with five operational phases-fill (25 min), anoxic (9 h), aerobic (9 h), settle (6 h) and draw (5 min). The fill, anoxic, settle and draw phases occurred in Tank 1. In the aerobic phase, the wastewater was circulated between the two tanks with centrifugal pumps and aeration was mainly achieved through oxygen absorption by micro-organisms in the biofilms when they were exposed to the air. In this paper, the biofilm growth and characteristics in the APSBBR were studied in a 98-day laboratory-scale experiment. During the course of the study, it was found that the biofilm thickness (delta) in Tank 1 ranged from 1.2 to 7.2 mm and that in Tank 2 from 0.5 to 2.2 mm; the biofilm growth against time (t) can be simulated as delta=0.07t0.99 (R2 = 0.97, P = 0.002) in Tank 1 and delta = 0.08t0.66 (R2 = 0.81, P = 0.04) in Tank 2. The biomass yield coefficient, Y, was 0.18 g volatile solids (VS) g(-1) COD removal. The biofilm density in both tanks, X, decreased as the biofilm thickness increased and can be correlated to the biofilm thickness, delta .

Absorption↗

Persister cells, the biofilm matrix and tolerance to metal cations in biofilm and planktonic Pseudomonas aeruginosa.

In this study, we examined Pseudomonas aeruginosa ATCC 27853 biofilm and planktonic cell susceptibility to metal cations. The minimum inhibitory concentration (MIC), the minimum bactericidal concentration (MBC) required to eradicate 100% of the planktonic population (MBC 100), and the minimum biofilm eradication concentration (MBEC) were determined using the MBEC trade mark-high throughput assay. Six metals - Co(2+), Ni(2+), Cu(2+), Zn(2+), Al(3+) and Pb(2+)- were each tested at 2, 4, 6, 8, 10 and 27 h of exposure to biofilm and planktonic cultures grown in rich or minimal media. With 2 or 4 h of exposure, biofilms were approximately 2-25 times more tolerant to killing by metal cations than the corresponding planktonic cultures. However, by 27 h of exposure, biofilm and planktonic bacteria were eradicated at approximately the same concentration in every instance. Viable cell counts evaluated at 2 and 27 h of exposure revealed that at high concentrations, most of the metals assayed had killed greater than 99.9% of biofilm and planktonic cell populations. The surviving cells were propogated in vitro and gave rise to biofilm and planktonic cultures with normal sensitivity to metals. Further, retention of copper by the biofilm matrix was investigated using the chelator sodium diethlydithiocarbamate. Formation of visible brown metal-chelates in biofilms treated with Cu(2+) suggests that the biofilm matrix may coordinate and sequester metal cations from the aqueous surroundings. Overall, our data suggest that both metal sequestration in the biofilm matrix and the presence of a small population of 'persister' cells may be contributing factors in the time-dependent tolerance of both planktonic cells and biofilms to high concentrations of metal cations.

Animals↗

Streptococcus gordonii biofilm formation: identification of genes that code for biofilm phenotypes.

Viridans streptococci, which include Streptococcus gordonii, are pioneer oral bacteria that initiate dental plaque formation. Sessile bacteria in a biofilm exhibit a mode of growth that is distinct from that of planktonic bacteria. Biofilm formation of S. gordonii Challis was characterized using an in vitro biofilm formation assay on polystyrene surfaces. The same assay was used as a nonbiased method to screen isogenic mutants generated by Tn916 transposon mutagenesis for defective biofilm formation. Biofilms formed optimally when bacteria were grown in a minimal medium under anaerobic conditions. Biofilm formation was affected by changes in pH, osmolarity, and carbohydrate content of the growth media. Eighteen biofilm-defective mutants of S. gordonii Challis were identified based on Southern hybridization with a Tn916-based probe and DNA sequences of the Tn916-flanking regions. Molecular analyses of these mutants showed that some of the genes required for biofilm formation are involved in signal transduction, peptidoglycan biosynthesis, and adhesion. These characteristics are associated with quorum sensing, osmoadaptation, and adhesion functions in oral streptococci. Only nine of the biofilm-defective mutants had defects in genes of known function, suggesting that novel aspects of bacterial physiology may play a part in biofilm formation. Further identification and characterization of biofilm-associated genes will provide insight into the molecular mechanisms of biofilm formation of oral streptococci.

Biofilms↗

Influence of biomass production and detachment forces on biofilm structures in a biofilm airlift suspension reactor

The influence of process conditions (substrate loading rate and detachment force) on the structure of biofilms grown on basalt particles in a Biofilm Airlift Suspension (BAS) reactor was studied. The structure of the biofilms (density, surface shape, and thickness) and microbial characteristics (biomass yield) were investigated at substrate loading rates of 5, 10, 15, and 20 kg COD/m3. day with basalt concentrations of 60 g/L, 150 g/L, and 250 g/L. The basalt concentration determines the number of biofilm particles in steady state, which is the main determining factor for the biofilm detachment in these systems. In total, 12 experimental runs were performed. A high biofilm density (up to 67 g/L) and a high biomass concentration was observed at high detachment forces. The higher biomass content is associated with a lower biomass substrate loading rate and therefore with a lower biomass yield (from 0.4 down to 0.12 gbiomass/gacetate). Contrary to general beliefs, the observed biomass detachment decreased with increasing detachment force. In addition, smoother (fewer protuberances), denser and thinner compact biofilms were obtained when the biomass surface production rate decreased and/or the detachment force increased. These observations confirmed a hypothesis, postulated earlier by Van Loosdrecht et al. (1995b), that the balance between biofilm substrate surface loading (proportional to biomass surface production rate, when biomass yield is constant) and detachment force determines the biofilm structure. When detachment forces are relatively high only a patchy biofilm will develop, whereas at low detachment forces, the biofilm becomes highly heterogeneous with many pores and protuberances. With the right balance, smooth, dense and stable biofilms can be obtained. Copyright 1998 John Wiley & Sons, Inc.

Journal Article↗

[Therapeutic study on biofilm of the urinary tract using a severely complicated bladder model (biofilm model of the urinary tract)--experimental study using an automatic simulator of urinary antimicrobial agent concentration, and clinical study].

For the purpose of conducting a therapeutic study on biofilm of the urinary tract, we devised a computer-controlled severely complicated bladder model (biofilm model of the urinary tract) enabling us to simulate the time-course of the concentration of antimicrobial agents in the urine. Using this model, we investigated clarithromycin (CAM), which has been reported to have anti-biofilm action, at concentrations close to its urinary levels at the time of clinical use in order to predict its effect on biofilm of the urinary tract. On the basis of those experimental results, we also conducted a clinical examination. The following results were obtained. 1. The action of ciprofloxacin (CPFX, MIC: 8 micrograms/ml) alone, which shows anti-P. aeruginosa activity, caused apparent elimination of P. aeruginosa from the model. However, regrowth of the microbes occurred when CPFX was removed from the bladder model. Moreover, the biofilm was not eliminated by the antimicrobial action of CPFX, and this was surmised to be the cause of the regrowth. 2. CAM (MIC: above 128 micrograms/ml), which has no anti-P. aeruginosa activity, was similarly tested as anti-biofilm agent when added alone to the biofilm model. The P. aeruginosa recovered to its initial concentration within 48 hours, but the biofilm disappeared due to the action of CAM. 3. The combined action of CPFX and CAM caused microbial elimination from the bladder model without microbial regrowth, even after these antimicrobial agents were removed from the bladder model. After the action of CPFX and CAM, the biofilm disappeared, and no microbial adherence was noted. 4. Measurement of time-course of the alginate content, which is the main component of P. aeruginosa biofilm, in the presence of CAM found that the alginate content decreased below the limit of detection after day 5. 5. The clinical study of complicated urinary tract infection revealed the microbial elimination rate and the efficacy rate to be higher in the combined CPFX-CAM administration group than in the CPFX-only administration group. 6. Based on the above results, we surmise that the combined use of an antimicrobial agent which is active against the causative microbe and anti-biofilm agent such as CAM will show some degree of efficacy in eliminating biofilm of the urinary tract.

Ciprofloxacin↗

[Adaptation study for biofilm of the urinary tract via highly complicated bladder model (biofilm model of the urinary tract)--experimental study using automatic simulator of the urinary antimicrobial agent concentration].

With the view of making an adaptation study for biofilm of the urinary tract, we devised a computer-controlled highly complicated bladder model (biofilm model of the urinary tract) that allowed the simulation of time-course changes in the urinary concentration of antimicrobial agents. Clarithromycin (CAM), which is reported to have an anti-biofilm action, was examined at urinary levels approximating clinical concentrations and its effect on biofilm was determined. The following results were obtained. 1) Ofloxacin (OFLX, 200 mg x 2/day, MIC; 8 micrograms/ml), which is active against Pseudomonas aruginosa, caused apparent microbial elimination from the model at 42 hours, but bacterial regrowth occurred 4 hours after withdrawal of this agent. No disappearance of the biofilm was noted with OFLX suggesting that this was the cause of bacterial regrowth. 2) The combination of OFLX (200 mg x 2/day) and CAM (MIC; > 128 micrograms/ml, 200 mg x 2/day) on anti-biofilm agent, with no effect on P. aeruginosa, eliminated bacteria from the bladder model more rapidly and prolonged the regrowth time to 10 hours after withdrawal of the antimicrobial agents. Disappearance of most of the biofilm and only slight microbial adhesion was noted. 3) The combination of OFLX (200 mg x 2/day) and CAM (400 mg x 3/day) caused microbial elimination from the bladder model with no regrowth at 30 hours after withdrawal of the antimicrobial agents. The biofilm disappeared completely and no microbial adhesion was noted. 4) CAM alone (400 mg x 3/day) allowed microbial recovery to the initial level within 48 hours after withdrawal, but led to disappearance of the biofilm and the adhesion of microbes without a glycocalyx. 5) These results suggest that the anti-biofilm action of CAM is dose-dependent, and that combined use of an appropriate antimicrobial agent and anti-biofilm agent like CAM may be effective for biofilm infections of the urinary tract.

Anti-Bacterial Agents↗

Structural deformation of bacterial biofilms caused by short-term fluctuations in fluid shear: an in situ investigation of biofilm rheology.

The physical properties (rheology) of biofilms will determine the shape and mechanical stability of the biofilm structure and consequently affect both mass transfer and detachment processes. Biofilm viscoelasticity is also thought to increase fluid energy losses in pipelines. Yet there is very little information on the rheology of intact biofilms. This is due in part to the difficulty in using conventional testing techniques. The size and nature of biofilms makes them difficult to handle, while removal from a surface destroys the integrity of the sample. We have developed a method which allowed us to conduct simple stress-strain and creep experiments on mixed and pure culture biofilms in situ by observing the structural deformations caused by changes in hydrodynamic shear stress (tau(w)). The biofilms were grown under turbulent pipe flow (flow velocity (u) = 1 m/s, Reynolds number (Re) = 3600, tau(w) = 5. 09 N/m(2)) for between 12 and 23 days. The resulting biofilms were heterogeneous and consisted of filamentous streamers that were readily deformed by changes in tau(w). At tau(w) of 10.11 N/m(2) the streamers were flattened so that the thickness was reduced by 25%. We estimated that the shear modulus (G) of the mixed culture biofilm was 27 N/m(2) and the apparent elastic modulus (E(app)) of both biofilms was in the range of 17 to 40 N/m(2). The biofilms behaved like elastic and viscoelastic solids below the tau(w) at which they were grown but behaved like viscoelastic fluids at elevated tau(w). The implications of these results for fluid energy losses and the processes of mass transfer and detachment are discussed.

Bacteria↗

Biofilm accumulation model that predicts antibiotic resistance of Pseudomonas aeruginosa biofilms.

A computer model of biofilm dynamics was adapted to incorporate the activity of an antimicrobial agent on bacterial biofilm. The model was used to evaluate the plausibility of two mechanisms of biofilm antibiotic resistance by qualitative comparison with data from a well-characterized experimental system (H. Anwar, J. L. Strap, and J. W. Costerton, Antimicrob. Agents Chemother. 36:1208-1214, 1992). The two mechanisms involved either depletion of the antibiotic by reaction with biomass or physiological resistance due to reduced bacterial growth rates in the biofilm. Both mechanisms predicted the experimentally observed resistance of 7-day-old Pseudomonas aeruginosa biofilms compared with that of 2-day-old ones. A version of the model that incorporated growth rate-dependent killing predicted reduced susceptibility of thicker biofilms because oxygen was exhausted within these biofilms, leading to very slow growth in part of the biofilm. A version of the model that incorporated a destructive reaction of the antibiotic with biomass likewise accounted for the relative resistance of thicker biofilms. Resistance in this latter case was due to depletion of the antibiotic in the bulk fluid rather than development of a gradient in the antibiotic concentration within the biofilm. The modeling results predicted differences between the two cases, such as in the survival profiles within the biofilm, that could permit these resistance mechanisms to be experimentally distinguished.

Anti-Bacterial Agents↗

Measuring local flow velocities and biofilm structure in biofilm systems with magnetic resonance imaging (MRI).

The characterization of substrate transport in the bulk phase and in the biofilm matrix is one of the problems which has to be solved for the verification of biofilm models. Additionally, the surface structure of biofilms has to be described with appropriate parameters. Magnetic resonance imaging (MRI) is one of the promising methods for the investigation of transport phenomena and structure in biofilm systems. The MRI technique allows the noninvasive determination of flow velocities and biofilm structures with a high resolution on the sub-millimeter scale. The presented investigations were carried out for defined heterotrophic biofilms which were cultivated in a tube reactor at a Reynolds number of 2000 and 8000 and a substrate load of 6 and 4 g/m2d glucose. Magnetic resonance imaging provides both structure data of the biofilm surface and flow velocities in the bulk phase and at the bulk/biofilm interface. It is shown that the surface roughness of the biofilms can be determined in one experiment for the complete cross section of the test tubes both under flow and stagnant conditions. Furthermore, the local shear stress was calculated from the measured velocity profiles. In the investigated biofilm systems the local shear stress at the biofilm surface was up to 3 times higher compared to the mean wall shear stress calculated on the base of the mean flow velocity.

Algorithms↗

Plaque biofilms: the effect of chemical environment on natural human plaque biofilm architecture.

The architecture of microbial biofilms especially the outer regions have an important influence on the interaction between biofilm and local environment particularly on the flux of materials into and out of biofilm compartments and as a consequence, biofilm metabolic behaviour. In the case of dental plaque biofilms, architecture will determine access of nutrients including acidogenic substrates and therapeutic materials to the microbial biomass and to the underlying tooth surface. Manipulation of this architecture may offer a means of altering mass transfer into the whole biofilm and biomass and raises the possibility of improving access of therapeutics. Plaque biofilms formed in vivo on human enamel were subjected to a number of different chemical conditions while under observation by confocal laser scanning microscopy in reflection mode. In this way the outer 50-100 microm or so of the biofilms was examined. Density and distribution of biomass were recorded as degree of reflectance. The amount and density of biofilm biomass increased from the plaque saliva interface towards the interior. Plaque biofilms were robust and little affected by mechanical manipulation, high ionic strength or low pH (2.5). Detergent (SLS), however, often appeared to either remove biomass and/or dramatically reduce its density.

Biofilms↗