Maximum Biofilm Thickness Research Articles

Response of antibiotic resistance genes expression and distribution on extracellular polymeric substances and microbial community in membrane biofilm during greywater treatment

This study evaluated how organic loading affects antibiotic resistance genes (ARGs) expression and distribution in the membrane biofilm. Organic surface loading rate of 4.65 g chemical oxygen demand (COD)/m2·d achieved the maximum biofilm thickness, concentration and linear alkylbenzene sulfonate (LAS) removal ratio of 136.9 ± 4.7 μm, 5.4 ± 0.1 g VSS/m2 and 99.4 %, respectively. Extracellular polymeric substances (EPS), EPS-attached LAS, and ARGs gradually increased in the membrane air inlet, middle and air outlet. AGRs and Intl1 were abundant in biofilm. LAS promoted EPS secretion, biofilm growth and ARGs proliferation. EPS, protein and carbohydrate were significantly correlated with most of biofilm ARGs, but not corrected with liquid-based ARGs. Microbial community structure impacted ARGs proliferation and transfer in the system. The findings indicated that EPS and microbial community play a crucial role in ARGs proliferation, spread and distribution, which lay the foundation for front-end control of ARGs during biofilm-based wastewater treatment.

Cell surface hydrophobicity and petroleum hydrocarbon degradation by biofilm-forming marine bacterium Pseudomonas furukawaii PPS-19 under different physicochemical stressors

Biofilm-forming marine bacterium Pseudomonas furukawaii PPS-19 showed strong hydrophobicity under different physicochemical stressors, such as pH and salinity. Strong aggregation of P. furukawaii PPS-19 was observed at hydrophobic interfaces of n-dodecane and crude oil, while uptake of pyrene resulted in blue fluorescence of the bacterium. Changes in biofilm microcolonies were observed under different physicochemical stressors with maximum biofilm thickness of 15.15 µm and 15.77 µm at pH 7% and 1% salinity, respectively. Relative expression analysis of alkB2 gene revealed the maximum expression in n-dodecane (10.5 fold) at pH 7 (1 fold) and 1% salinity (8.3 fold). During the degradation process, a significant drop in surface tension resulted in increased emulsification activity. P. furukawaii PPS-19 showed the respective n-dodecane and pyrene degradation of 94.3% and 81.5% at pH 7% and 94.5% and 83% at 1% salinity. A significant positive correlation was obtained between cell surface hydrophobicity (CSH), biofilm formation, and PHs degradation (P < 0.05) under all the physicochemical stressors, with the highest value at pH 7% and 1% salinity. Analysis of metabolites indicated that mono-terminal oxidation and multiple pathways were followed for n-dodecane and pyrene biodegradation, respectively. Thus, P. furukawaii PPS-19 is an efficient hydrocarbonoclastic bacterium that may be exploited for large-scale oil pollution abatement.

Biofilm formation by sulphate-reducing bacteria on different metals and their prospective role in titanium corrosion

ABSTRACT This study describes the biofilm formation by sulphate-reducing bacteria (SRB) on different materials, which has implications for the biomedical, pharmaceutical, food and chemical process industries. SRB was chosen as a model organism being an anaerobic bacterium. Biofilm formation on different materials and corrosion of titanium by SRB were monitored with time using confocal laser scanning microscopy and fluorescent FISH probes were used to authenticate the SRB strain. The thickness of the mono-culture SRB biofilm has ranged from 4 to 24 µm during thed 12–84 hr; however, the maximum biofilm thickness (24 µm) was recorded after 60 hr of growth. Planktonic growth of the SRB strain showed a log phase up to 48 hr and the sulphide production ranged from 2 to 14 mg l−1. For a comparative account, the SRB biofilm formation on copper was chosen as a positive control. Finally, the putative role of extracellular electron transfer by SRB in the biocorrosion process and the plausible mechanism of pitting corrosion of titanium is described in detail.

Biofilm thickness controls the relative importance of stochastic and deterministic processes in microbial community assembly in moving bed biofilm reactors.

Deterministic and stochastic processes are believed to play a combined role in microbial community assembly, though little is known about the factors determining their relative importance. We investigated the effect of biofilm thickness on community assembly in nitrifying moving bed biofilm reactors using biofilm carriers where maximum biofilm thickness is controlled. We examined the contribution of stochastic and deterministic processes to biofilm assembly in a steady state system using neutral community modelling and community diversity analysis with a null-modelling approach. Our results indicate that the formation of biofilms results in habitat filtration, causing selection for phylogenetically closely related community members, resulting in a substantial enrichment of Nitrospira spp. in the biofilm communities. Stochastic assembly processes were more prevalent in biofilms of 200 µm and thicker, while stronger selection in thinner (50 µm) biofilms could be driven by hydrodynamic and shear forces at the biofilm surface. Thicker biofilms exhibited greater phylogenetic beta-diversity, which may be driven by a variable selection regime caused by variation in environmental conditions between replicate carrier communities, or by drift combined with low migration rates resulting in stochastic historical contingency during community establishment. Our results indicate that assembly processes vary with biofilm thickness, contributing to our understanding of biofilm ecology and potentially paving the way towards strategies for microbial community management in biofilm systems.

Impact of the restraint of biofilm volume and thickness on the performance and microbial composition in anaerobic moving-bed biofilm reactors (AnMBBRs)

The effect of limiting biofilm volume using carriers with maximum thickness control on the performance and microbial community composition was investigated in anaerobic moving-bed biofilm reactors (AnMBBRs). Three parallel, continuous AnMBBRs (4 L) were operated (288 d, 24/36 °C) to treat the soluble fraction of a wastewater from a food-manufacturing facility. Two different biofilm carriers controlling maximum biofilm thickness at 200 and 1000 µm (AnoxK™Z-200 and Z-1000) were used in the three reactors, targeting the same projected surface area for biofilm growth (410 m2/reactor): R200 (Z-200), R1000 (Z-1000) and RMix (both Z-200 and Z-1000). The composition of the bacterial (16S rRNA V1-V3) and methanogenic (mcrA gene) communities was analysed using amplicon sequencing. Under relatively constant organic loads (3.6–4.2 gSCOD/Ld, 34–39 gSCOD/m2d, hydraulic retention time=7.7–8 h), R200 presented lower soluble chemical oxygen demand (SCOD) removals (51–60%) than the other reactors (R100 = 91–93%, RMix = 86–91%), likely due to a limited amount of active protected biofilm volume. Lower SCOD removals were associated with higher concentrations of volatile fatty acids in the effluent (R200 =400–800 mgCOD/L; R1000 and RMix<200 mgCOD/L) and lower methane production (R200 =0.66; R1000 =0.72; RMix=0.73 gCH4-COD/gSCODremoved). The Z-200 carriers contained a different fermentative/acidogenic bacterial community abundant in representatives of the families Christensenellaceae, Anaerovoracaceae and Synergistaceae, and led to less amount of biofilm biomass growth, albeit more active than that of carriers allowing for thicker biofilms (Z-1000). In contrast, methanogenic populations were less sensitive to biofilm thickness restraint by these carriers. This study shows for the first time that limiting biofilm volume in MBBR carriers can impact the performance and bacterial community in AnMBBR biofilms.

R-based method for quantitative analysis of biofilm thickness by using confocal laser scanning microscopy.

Microscopy is mostly the method of choice to analyse biofilms. Due to the high local heterogeneity of biofilms, single and punctual analyses only give an incomplete insight into the local distribution of biofilms. In order to retrieve statistically significant results a quantitative method for biofilm thickness measurements was developed based on confocal laser scanning microscopy and the programming language R. The R‐script allows the analysis of large image volumes with little hands‐on work and outputs statistical information on homogeneity of surface coverage and overall biofilm thickness. The applicability of the script was shown in microbial fuel cell experiments. It was found that Geobacter sulfurreducens responds differently to poised anodes of different material so that the optimum potential for MFC on poised ITO anodes had to be identified with respect to maximum current density, biofilm thickness and MFC start‐up time. Thereby, a positive correlation between current density and biofilm thickness was found, but with no direct link to the applied potential. The optimum potential turned out to be +0.1 V versus SHE. The script proved to be a valuable stand‐alone tool to quantify biofilm thickness in a statistically valid manner, which is required in many studies.

Real-time monitoring of biofilm thickness allows for determination of acetate limitations in bio-anodes

Several studies have reported that current produced by electro-active bacteria (EAB) is dependent on anode potential and substrate concentration. However, information about the relation between biofilm growth and current density is scarce. In this study, biofilm thickness was monitored in-situ and this relation explored at three anode potentials and three acetate concentrations. The highest current densities of 3.7 A·m−2 were obtained for biofilms thinner than 40 μm, even though thicknesses up to 88 μm were measured. Fick's law was used to estimate the acetate penetration depth in the biofilm, acetate diffusion rates in the biofilm, and specific acetate utilization rates. A maximum biofilm thickness of a non-acetate limited biofilm of 55 μm and an acetate diffusion rate of 2.68 × 10−10 m2·s−1 were estimated at −0.2 V vs Ag/AgCl. The results provide information on the target biofilm thickness for which no acetate limitations occur and provide data for modeling works with bio-anodes.

Biofilm carrier type affects biogenic sulfur-driven denitrification performance and microbial community dynamics in moving-bed biofilm reactors

Autotrophic denitrification with biosulfur (ADBIOS) provides a sustainable technological solution for biological nitrogen removal from wastewater driven by biogenic S0, derived from biogas desulfurization. In this study, the effect of different biofilm carriers (conventional AnoxK™ 1 and Z-200 with a pre-defined maximum biofilm thickness) on ADBIOS performance and microbiomics was investigated in duplicate moving bed-biofilm reactors (MBBRs). The MBBRs were operated parallelly in continuous mode for 309 days, whilst gradually decreasing the hydraulic retention time (HRT) from 72 to 21 h, and biosulfur was either pumped in suspension (days 92–223) or supplied in powder form. Highest nitrate removal rates were approximately 225 (±11) mg/L·d and 180 (±7) mg NO3−-N/L·d in the MBBRs operated with K1 and Z-200 carriers, respectively. Despite having the same protected surface area for biofilm development in each MBBR, the biomass attached onto the K1 carrier was 4.8-fold more than that on the Z-200 carrier, with part of the biogenic S0 kept in the biofilm. The microbial communities of K1 and Z-200 biofilms could also be considered similar at cDNA level in terms of abundance (R = 0.953 with p = 0.042). A relatively stable microbial community was formed on K1 carriers, while the active portion of the microbial community varied significantly over time in the MBBRs using Z-200 carriers.

Microscopic techniques to evaluate the biofilm formation ability of a marine bacterium Pseudomonas aeruginosa PFL-P1 on different substrata.

Biofilm formation in bacteria is strongly affected by the nature of substrata. Different substrata such as glass, polystyrene, steel, ceramic, and rubber were used to assess the biofilm forming ability of a marine bacterium Pseudomonas aeruginosa PFL-P1 using a scanning electron microscope (SEM), atomic force microscope (AFM), and confocal laser scanning microscope (CLSM). The bacterium formed dense biofilms with varied aggregation on different substrata. SEM study revealed small rod-shaped cells with diverse arrangements within the biofilms on all the substrata under study. The AFM study revealed the highest roughness of 545 nm on the ceramic substratum. The biofilms formed on ceramic substratum were characterized with maximum roughness (742 nm), maximum peak height (1,480 nm), and maximum arithmetic mean height (611 nm), significantly higher than all the other substrata (p < .05). AFM studies confirmed that P. aeruginosa PFL-P1 exhibited biofilm heterogeneity on all the substrata. The CLSM study indicated a higher fraction of nucleic acids to α-polysaccharides ratio in the biofilms. COMSTAT analysis revealed the highest biofilm biomass of ~18 μm3 /μm2 on the ceramic substratum. The maximum biofilm thickness of ~50 μm in the native state on the ceramic substratum was significantly higher than glass (p=.0015), polystyrene (p=.0001), steel (p=.0035), and rubber substrata (p=.0001). The higher surface roughness of ceramic substratum is accountable for more area for colonization, as evident from higher biomass and thickness of the biofilm. This study provides insight into the substratum properties, which modulate the biofilm forming ability in bacteria.

CFD study of the hydrodynamics and biofilm growth effect of an anaerobic inverse fluidized bed reactor operating in the laminar regime

The unsteady laminar flow of an inverse fluidized bed reactor was addressed in this study using computational fluid dynamics (CFD). Three-dimensional (3D) and two-dimensional (2D) simulations were carried out using the Eulerian-Eulerian approach with the Syamlal O'Brien and Gidaspow models for modeling the drag between the solid and liquid phases. Reported data of solids expansion and bed porosity were used to validate the simulations. Results show that 2D simulations can satisfactorily reproduce experimental data of fluidized bed reactors, which are essentially equal to those obtained on 3D simulations. The model of Syamlal O'Brien was found to be in better agreement with experiments than that of Gidaspow. It is shown that commonly used correlations for predicting bed porosity fail to provide a reasonable overall description of the evaluated operating conditions. The flow patterns for both phases in the bed section exhibit chaotic recirculation zones with high values of shear stress, and after this, the flow quickly becomes fully developed. The effect of biofilm thickness increase on bed porosity, under similar apparent densities of colonized particles reported for this system, is analyzed. Biofilm growth generates a decrease in the Archimedes number (i.e., decreases the buoyancy of particles) and an increase in the particle Reynolds numbers (Rep), affecting significantly the final porosity of the bed. However, this effect vanishes for Rep < 0.25. Results from this work can be used to develop scale‐up criteria and to establish safe operating conditions (e.g., maximum liquid velocity and biofilm thickness) to avoid loss of support particles.

Microbiota of spent nuclear fuel pool water with emphasis on their biofilm forming ability on stainless steel (SS-304L)

Spent nuclear fuel (SNF) pool is an essential unit of a nuclear power plant infrastructure, where radioactive fuel rods are kept for cooling and shielding, before reprocessing. This study explored the presence of bacteria in SNF pool water with emphasis on their capability to form biofilms on pool wall cladding material stainless steel (SS-304L). Bacteria were isolated from SNF pool water and were characterized using 16S rRNA gene sequencing. The six bacterial isolates (Bacillus subtilis, Staphylococcus sps., S. arlettae, S. epidermidis, S. auricularis and Chryseobacterium gleum) can grow and form biofilms at very low nutrient condition as well as in chronic radioactivity. The bacterial isolates formed biofilm on SS-304L and glass. However, the biofilm parameters assessed by CLSM microscopy showed that the strains preferred SS-304L surface for biofilm formation. On SS-304L, the maximum biomass (0.45 l μm3/l μm2) was formed by S. arlettae when compared to maximum biomass (0.054 l μm3/l μm2) by Staphylococcus sp., on glass. Maximum biofilm thickness on SS- 304L was observed by Staphylococcus sp. (8.81 l μm) when compared to that of S. epidermidis (4.16 l μm) on the glass surface. The biofilm formation on SS-304L surface suggests the possible risk of microbial-induced corrosion of SNF pool cladding material. This study highlights the need for mandatory monitoring of microbial biofilm formation in an extreme environment such as SNF pool.

Un-steady state modeling for free cyanide removal and biofilm growth in a RBC batch process

Biofilm growth and free cyanide biological removal from gold mine wastewater were modeled and simulated using a bench-scale rotating biological contactor (RBC). Eight batch cultures were run in three independent compartments (1.7 L, each) of the RBC. The system worked under the following conditions: [CNi-] = 0.3 g/L, pH = 10.5 ± 0.5, T = 20 ± 5 °C, ω =5 rpm, and 40.5 % of disc submersion. During each culture, biofilm thickness, biomass, and free cyanide concentration in the liquid were quantified. Subsequently, μmax, KCN-,Y'X/CN-, qmax, b', Df, k, and JCN- were determined using experimental data to later model and simulate the biofilm thickness and free cyanide biological removal with Wolfram Mathematica software. After the experiments, free cyanide biological removal was 96.33 % after three days, and maximum biofilm thickness was 0.0292 cm in the 16th day. Moreover, biofilm growth and free cyanide consumption models were adjusted to the experimental data with r2 = 0.90 and r2 = 0.99. Also, there was an equivalent error of 7.89 and 7.38 and a standard deviation of 10.89 % and 10.17 %, between the models and their experimental data, respectively. Finally, the proposed models will allow improvement of reactor operation and its design.

Metabolic modeling of a chronic wound biofilm consortium predicts spatial partitioning of bacterial species.

BackgroundChronic wounds are often colonized by consortia comprised of different bacterial species growing as biofilms on a complex mixture of wound exudate. Bacteria growing in biofilms exhibit phenotypes distinct from planktonic growth, often rendering the application of antibacterial compounds ineffective. Computational modeling represents a complementary tool to experimentation for generating fundamental knowledge and developing more effective treatment strategies for chronic wound biofilm consortia.ResultsWe developed spatiotemporal models to investigate the multispecies metabolism of a biofilm consortium comprised of two common chronic wound isolates: the aerobe Pseudomonas aeruginosa and the facultative anaerobe Staphylococcus aureus. By combining genome-scale metabolic reconstructions with partial differential equations for metabolite diffusion, the models were able to provide both temporal and spatial predictions with genome-scale resolution. The models were used to analyze the metabolic differences between single species and two species biofilms and to demonstrate the tendency of the two bacteria to spatially partition in the multispecies biofilm as observed experimentally. Nutrient gradients imposed by supplying glucose at the bottom and oxygen at the top of the biofilm induced spatial partitioning of the two species, with S. aureus most concentrated in the anaerobic region and P. aeruginosa present only in the aerobic region. The two species system was predicted to support a maximum biofilm thickness much greater than P. aeruginosa alone but slightly less than S. aureus alone, suggesting an antagonistic metabolic effect of P. aeruginosa on S. aureus. When each species was allowed to enhance its growth through consumption of secreted metabolic byproducts assuming identical uptake kinetics, the competitiveness of P. aeruginosa was further reduced due primarily to the more efficient lactate metabolism of S. aureus. Lysis of S. aureus by a small molecule inhibitor secreted from P. aeruginosa and/or P. aeruginosa aerotaxis were predicted to substantially increase P. aeruginosa competitiveness in the aerobic region, consistent with in vitro experimental studies.ConclusionsOur biofilm modeling approach allows the prediction of individual species metabolism and interspecies interactions in both time and space with genome-scale resolution. This study yielded new insights into the multispecies metabolism of a chronic wound biofilm, in particular metabolic factors that may lead to spatial partitioning of the two bacterial species. We believe that P. aeruginosa lysis of S. aureus combined with nutrient competition is a particularly relevant scenario for which model predictions could be tested experimentally.Electronic supplementary materialThe online version of this article (doi:10.1186/s12918-016-0334-8) contains supplementary material, which is available to authorized users.

Individual or Combined Effects of Meropenem, Imipenem, Sulbactam, Colistin, and Tigecycline on Biofilm-Embedded Acinetobacter baumannii and Biofilm Architecture.

Acinetobacter baumannii biofilms are difficult to eradicate. We investigated the effects of meropenem (2 mg/liter), imipenem (2 mg/liter), sulbactam (4 mg/liter), colistin (2 mg/liter), and tigecycline (2 mg/liter), alone or in combination, on biofilm-embedded carbapenem-resistant and carbapenem-susceptible A. baumannii (CRAb and CSAb, respectively) cells, as well as on the architecture of the biofilms. A. baumannii ATCC 15151 (Ab15151) and its OXA-82-overproducing transformant, along with two clinical CSAb and two clinical CRAb isolates of differing clonalities, were used. The minimal bactericidal concentrations for biofilm-embedded cells of the six tested isolates were >50-fold those of their planktonic cells. When used individually, meropenem exhibited a higher killing effect than the other four antimicrobials on biofilm-embedded CSAb cells in the colony biofilm assay. For two clinical CRAb isolates, meropenem plus sulbactam or sulbactam plus tigecycline showed >100-fold the bactericidal effect exhibited by these agents used alone after 48 h of treatment. The effect of antimicrobials on the architecture of Ab15151 biofilm emitting green fluorescence was determined by confocal laser scanning microscopy using COMSTAT software. Significant decreases in the maximum biofilm thickness were observed after exposure to meropenem and imipenem. Meropenem plus sulbactam significantly decreased the biomass and mean thickness and increased the roughness coefficient of biofilms, but sulbactam plus tigecycline only decreased the maximum and mean biofilm thickness compared to any of these agents used alone. Meropenem was active against biofilm-embedded CSAb, whereas meropenem plus sulbactam exhibited synergism against biofilm-embedded CRAb and caused significantly more damage to the biofilm architecture than did any of the agents used alone.

Characterization of Electrical Current-Generation Capabilities from Thermophilic Bacterium Thermoanaerobacter pseudethanolicus Using Xylose, Glucose, Cellobiose, or Acetate with Fixed Anode Potentials.

Thermoanaerobacter pseudethanolicus 39E (ATCC 33223), a thermophilic, Fe(III)-reducing, and fermentative bacterium, was evaluated for its ability to produce current from four electron donors-xylose, glucose, cellobiose, and acetate-with a fixed anode potential (+ 0.042 V vs SHE) in a microbial electrochemical cell (MXC). Under thermophilic conditions (60 °C), T. pseudethanolicus produced high current densities from xylose (5.8 ± 2.4 A m(-2)), glucose (4.3 ± 1.9 A m(-2)), and cellobiose (5.2 ± 1.6 A m(-2)). It produced insignificant current when grown with acetate, but consumed the acetate produced from sugar fermentation to produce electrical current. Low-scan cyclic voltammetry (LSCV) revealed a sigmoidal response with a midpoint potential of -0.17 V vs SHE. Coulombic efficiency (CE) varied by electron donor, with xylose at 34.8% ± 0.7%, glucose at 65.3% ± 1.0%, and cellobiose at 27.7% ± 1.5%. Anode respiration was sustained over a pH range of 5.4-8.3, with higher current densities observed at higher pH values. Scanning electron microscopy showed a well-developed biofilm of T. pseudethanolicus on the anode, and confocal laser scanning microscopy demonstrated a maximum biofilm thickness (Lf) greater than ~150 μm for the glucose-fed biofilm.

Distribution of biofilm thickness in porous media and implications for permeability models

The distribution of biofilm thickness on individual sand grains in a two-dimensional sand-filled cell maintained under anaerobic conditions was investigated. The cell was inoculated with a mixed microbial culture fed with methanol. Concentrations of biomass attached to the sand and suspended in the water in the cell were determined by protein analysis. The biofilm thickness on individual sand grains was investigated with confocal laser scanning microscopy (CLSM), and was found to follow a normal distribution with a mean range of 59–316 μ and standard deviations of 30–77 μ. The bulk average permeability reduction factor is 8. To investigate the implications of the variability of biofilm thicknesses, four models were used to calculate reductions in porous media permeability. Taylor’s model predicted a reduction by a factor ranging from 14 to 5,000 (from minimum to maximum mean biofilm thickness). Vandevivere’s model predicted a reduction in permeability by a factor ranging from 769 to 3,846 (from minimum to maximum mean biofilm thickness). Seki’s model did not give physically meaningful results in this study. Clement’s model predicted reduction by a factor ranging from 1 to 1.14 over the range of biomass levels observed in the cell. Data on the statistical and normal distributions of biofilm thickness in porous media, and assessment of their implications with respect to different permeability models, could lead to better understanding of the extent of bioclogging associated with field implementation of bioremediation of contaminants in aquifers.

Porphyromonas gingivalis and Treponema denticola Synergistic Polymicrobial Biofilm Development

Chronic periodontitis has a polymicrobial biofilm aetiology and interactions between key bacterial species are strongly implicated as contributing to disease progression. Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia have all been implicated as playing roles in disease progression. P. gingivalis cell-surface-located protease/adhesins, the gingipains, have been suggested to be involved in its interactions with several other bacterial species. The aims of this study were to determine polymicrobial biofilm formation by P. gingivalis, T. denticola and T. forsythia, as well as the role of P. gingivalis gingipains in biofilm formation by using a gingipain null triple mutant. To determine homotypic and polymicrobial biofilm formation a flow cell system was employed and the biofilms imaged and quantified by fluorescent in situ hybridization using DNA species-specific probes and confocal scanning laser microscopy imaging. Of the three species, only P. gingivalis and T. denticola formed mature, homotypic biofilms, and a strong synergy was observed between P. gingivalis and T. denticola in polymicrobial biofilm formation. This synergy was demonstrated by significant increases in biovolume, average biofilm thickness and maximum biofilm thickness of both species. In addition there was a morphological change of T. denticola in polymicrobial biofilms when compared with homotypic biofilms, suggesting reduced motility in homotypic biofilms. P. gingivalis gingipains were shown to play an essential role in synergistic polymicrobial biofilm formation with T. denticola.

A calibration protocol of a one-dimensional moving bed bioreactor (MBBR) dynamic model for nitrogen removal

This work suggests a procedure to correctly calibrate the parameters of a one-dimensional MBBR dynamic model in nitrification treatment. The study deals with the MBBR configuration with two reactors in series, one for carbon treatment and the other for nitrogen treatment. Because of the influence of the first reactor on the second one, the approach needs a specific calibration strategy. Firstly, a comparison between measured values and simulated ones obtained with default parameters has been carried out. Simulated values of filtered COD, NH(4)-N and dissolved oxygen are underestimated and nitrates are overestimated compared with observed data. Thus, nitrifying rate and oxygen transfer into the biofilm are overvalued. Secondly, a sensitivity analysis was carried out for parameters and for COD fractionation. It revealed three classes of sensitive parameters: physical, diffusional and kinetic. Then a calibration protocol of the MBBR dynamic model was proposed. It was successfully tested on data recorded at a pilot-scale plant and a calibrated set of values was obtained for four parameters: the maximum biofilm thickness, the detachment rate, the maximum autotrophic growth rate and the oxygen transfer rate.

A clinical assessment of the Mucus Shaver

: We evaluated a new device designed to clean the endotracheal tube in mechanically ventilated patients, the Mucus Shaver. : Prospective, randomized trial. : University hospital intensive care unit. : We enrolled 24 patients expected to remain ventilated for >72 hrs. : The Mucus Shaver is a concentric inflatable catheter for the removal of mucus and secretions from the interior surface of the endotracheal tube. The Mucus Shaver is advanced to the distal endotracheal tube tip, inflated, and subsequently withdrawn over a period of 3-5 secs. Patients were prospectively randomized within 2 hrs of intubation to receive standard endotracheal tube suctioning treatment or standard suctioning plus Mucus Shaver use until extubation. : During the study period, demographic data, recent medical history, adverse events, and staff evaluation of the Mucus Shaver were recorded. At extubation, each endotracheal tube was removed, cultured, and analyzed by scanning electron microscopy. Twelve patients were assigned to the study group and 12 were assigned to the control group. No adverse events related to the use of the Mucus Shaver were observed. At extubation, only one endotracheal tube from the Mucus Shaver group was colonized, whereas in the control group ten endotracheal tubes were colonized (8% vs. 83%; p < .001). Scanning electron microscopy showed little secretions on the endotracheal tubes from the study group, whereas thick bacterial deposits were present on all the endotracheal tubes from the control group (p < .001 by Fisher exact test, using a maximum biofilm thickness of 30 μm as cut-off). The nursing staff was satisfied by the overall safety, feasibility, and efficacy of the Mucus Shaver. : The Mucus Shaver is a safe, feasible, and efficient device for endotracheal tube cleaning in the clinical setting. The Mucus Shaver is helpful in preventing endotracheal tube colonization by potentially harmful microorganisms.

The impact of antimicrobial photodynamic therapy in an artificial biofilm model

The susceptibility of bacterial cultures in biofilm formations is important for a variety of clinical treatment procedures. Therefore, the aim of the study was to assess the impact of laser-induced antimicrobial photodynamic therapy on the viability of Streptococcus mutans cells employing an artificial biofilm model. Using sterile chambered coverglasses, a salivary pellicle layer was formed in 40 chambers. Streptococcus mutans cells were inoculated in a sterile culture medium. Employing a live/dead bacterial viability kit, bacteria with intact cell membranes stained fluorescent green. Each pellicle-coated test chamber was filled with 0.7ml of the bacterial suspension and analysed using a confocal laser scanning microscope within a layer of 10μm at intervals of 1μm from the pellicle layer. Phenothiazine chloride was used as a photosensitizer in all 40 test chambers. A diode laser (wavelength 660nm, output power 100mW) was used to irradiated 20 chambers for 2min. Fluorescence values in the test chambers after laser irradiation (median 2.1U, range 0.4-3.4U) were significantly lower than baseline values after adding the photosensitizer (median 3.6U, range 1.1-9.0; p < 0.05). The non-irradiated control chambers showed no change in fluorescence at the end of an additional photosensitizer residence time of 2min without laser irradiation (median 1.9U, range 0.7-3.6; median 1.9U, range 0.8-6.0, respectively; p > 0.05). The present study indicated that laser irradiation is an essential part of antimicrobial photodynamic therapy to reduce bacteria within a layer of 10μm. Further studies are needed to evaluate the maximum biofilm thickness that still allows a toxic effect on microorganisms.