Biofilm Colonization Research Articles

Comparative Proteomics in Caries Active and Caries Free Associated S. Mutans Strains for Metabolic Remodeling Favouring Colonization

Introduction: A multifactorial condition, dental caries is primarily brought on by cariogenic bacteria that are frequently present in the mouth. Caries infections are commonly associated with the acid-producing bacteria Streptococcus mutans. If left untreated, dental caries can result in tooth decalcification, cavities, hypersensitivity, and even tooth loss. Comparative proteomics by Mass spectrometry was carried out to reveal caries-associated metabolic remodeling in S mutans isolates favouring biofilm formation and colonization. Methods: We conducted a differential proteomic analysis to determine the differences in protein expression between S.mutans strains linked to dental caries and those present in a healthy oral microbiome. High-performance mass spectrometry (MS) using Orbitrap, has led to the development of proteomics in larger-scale protein analysis. Five clinical specimens from individuals who had caries disease (Caries active) were compared with two isolates from the healthy oral dental microbiota (Caries free). Protein samples were digested, and a peptide mixture investigation was done. Results and Discussion: 3276 proteins that were expressed at comparable amounts in both groups of bacteria and were found by proteomic analysis. Only 39 of these proteins were unique and distinct to group 1 (those without caries), whereas 444 proteins were specific to group 2. (S.mutans from caries patients). Significant differences in the grouping of the control (Caries free) and caries (Caries active) samples were seen through PCA analysis. 23 Significantly regulated proteins in S. mutans bacteria were discovered (p < 0.05, t-test). Among these, 23 differentially expressed proteins, 10 were upregulated, and 13 downregulated. By Identifying differential peptides and proteins, their quantification, and appropriate bioinformatic analysis, there are insights into formation of biofilm by S. mutans in dental caries and potential targets for intervention using various antagonists/ nanoparticle approaches as alternatives to conventional antibiotics.

Antibiotic susceptibility and biofilm-forming ability of Veillonella strains

IntroductionVeillonella, known as early colonizers in oral biofilms, take part in some infections in human. Biofilm refers to complex, sessile communities of microbes, which function as strong barriers for bacteria to survive. Biofilm matrixes surrounding bacteria enable them to withstand harsh conditions, protect against immune cells, etc., and also make them resistant to antimicrobial treatments. Thus, the knowledge of antibiotic susceptibility and biofilm formation of Veillonella will shed light on their resistance mechanism. Materials and methodTheir morphology was observed by scanning electron microscopy (SEM). According to the performance standards for antibiotic susceptibility testing of the Clinical & Laboratory Standards Institute, the Agar dilution method was used to study the susceptibility of Veillonella strains to eight antibiotics (ampicillin, piperacillin-tazobactam, cefoxitin, tetracycline, moxifloxacin, clindamycin, metronidazole, and vancomycin). In addition, we applied the crystal violet staining method to reveal the processes of biofilm formation of these Veillonella strains. ResultsV. rogosae, V. nakazawae, and V. parvula were isolated from oral cavities of healthy adults and V. ratti was isolated from dairy goat droppings. Observations by scanning electron microscopy showed that Veillonella were spherical and arranged in single or short chains. The diameter of a single cell was about 0.3–0.5 μm. The Minimum Inhibitory Concentrations (MICs) of the antibiotics were determined and the results showed that these four strains were all sensitive to cefoxitin, tetracycline, moxifloxacin, clindamycin and metronidazole. Among the four strains, V. ratti was resistant to piperacillin-tazobactam, and V. rogosae and V. nakazawae were resistant to ampicillin. The vancomycin susceptibility of the four Veillonella strains varied greatly. The MICs of vancomycin against V. rogosae and V. ratti were greater than 256 μg/mL but the MICs of vancomycin against V. nakazawae and V. parvula were less than 2 μg/mL. V. parvula had significantly higher biofilm-forming ability than the other three strains (p < 0.05) and V. nakazawae had the weakest biofilm-forming ability. ConclusionIn this study, V. rogosae, V. nakazawae, V. parvula and V. ratti were isolated and identified. The four strains of Veillonella showed differences in MIC values for different antibiotics and biofilm-forming ability.

The role of biofilm matrix composition in controlling colony expansion and morphology.

Biofilms are biological viscoelastic gels composed of bacterial cells embedded in a self-secreted polymeric extracellular matrix (ECM). In environmental settings, such as in the rhizosphere and phyllosphere, biofilm colonization occurs at the solid-air interface. The biofilms' ability to colonize and expand over these surfaces depends on the formation of osmotic gradients and ECM viscoelastic properties. In this work, we study the influence of biofilm ECM components on its viscoelasticity and expansion, using the model organism Bacillus subtilis and deletion mutants of its three major ECM components, TasA, EPS and BslA. Using a multi-scale approach, we quantified macro-scale viscoelasticity and expansion dynamics. Furthermore, we used a microsphere assay to visualize the micro-scale expansion patterns. We find that the viscoelastic phase angle is likely the best viscoelastic parameter correlating to biofilm expansion dynamics. Moreover, we quantify the sensitivity of the biofilm to changes in substrate water potential as a function of ECM composition. Finally, we find that the deletion of ECM components significantly increases the coherence of micro-scale colony expansion patterns. These results demonstrate the influence of ECM viscoelasticity and substrate water potential on the expansion of biofilm colonies on wet surfaces at the air-solid interface, commonly found in natural environments.

Carbon nanotubes encapsulating FeS2 micropolyhedrons as an anode electrocatalyst for improving the power generation of microbial fuel cells

The low power density originating from poor electroactive bacteria (EAB) adhesion and sluggish extracellular electron transfer (EET) at the anode interface, is a major impediment preventing the practical implementation of microbial fuel cells (MFCs). Tailoring the surface properties of anodes is an effective and powerful strategy for addressing this issue. In this study, we successfully fabricated an efficient anode electrocatalyst, consisting of carbon nanotubes encapsulating iron disulfide (FeS2@CNT) micropolyhedrons, using simple hydrothermal and freeze-drying methods, which not only strengthened the anode interaction with EAB but also promoted the EET process at the anode interface. As expected, the MFCs with a FeS2@CNT anode yielded an outstanding power density of 1914 mWm−2 at a current density of 4350 mA m−2, which significantly exceeded those of pure CNT (1096.2mW m−2, 2703.3 mA m−2) and carbon cloth (426.8mWm−2, 965.6 mA m−2) anodes. The high-power output can be attributed to the synergistic effect between FeS2 and CNTs, endowing the anode with biocompatibility for biofilm adhesion and colonization, nutrient diffusion, and the presence of abundant Fe and S active sites for EET mediation. Owing to the low cost, facile fabrication process, and excellent electrocatalytic performance toward the redox reactions in biofilms, the synthesized FeS2@CNT electrocatalyst is a promising material for high-performance and cost-effective MFCs with commercial applications.

Effect of UV Light and Sodium Hypochlorite on Formation and Destruction of Pseudomonas fluorescens Biofilm In Vitro

Pseudomonas fluorescens is one of the first colonizers of bacterial biofilm in water systems and a member of opportunistic premise plumbing pathogens (OPPPs). The aim of this study was to examine the effect of UV light and sodium hypochlorite on the formation and destruction of mature P. fluorescens biofilm on ceramic tiles. Planktonic bacteria or bacteria in mature biofilm were exposed to UV light (254 nm) for 5, 20 s. and to 0.4 mg/L sodium hypochlorite for 1 min. Mature biofilm was also exposed to increased concentration of sodium hypochlorite of 2 mg/L for 0.5, 1 and 2 h and combined with UV. Prolonged action of sodium hypochlorite and an increase in its concentration in combination with UV gave the best results in the inhibition of biofilm formation after the pre-treatment and destruction of mature biofilm. The effect of hyperchlorination in combination with UV radiation shows better results after a long exposure time, although even after 120 min there was no completely destroyed biofilm. Furthermore, the mechanism of the effect of combined methods should be explored as well as the importance of mechanical cleaning that is crucial in combating bacterial biofilm in swimming pools.

Convergence of biofilm successional trajectories initiated during contrasting seasons.

Biofilm communities play a major role in explaining the temporal variation of biogeochemical conditions in freshwater ecosystems, and yet we know little about how these complex microbial communities change over time (aka succession), and from different initial conditions, in comparison to other stream communities. This has resulted in limited knowledge on how biofilm community structure and microbial colonization vary over relevant time scales to become mature biofilms capable of significant alteration of the freshwater environment in which they live. Here, we monitored successional trajectories of biofilm communities from summer and winter in a headwater stream and evaluated their structural state over time by DNA high-throughput sequencing. Significant differences in biofilm composition were observed when microbial colonization started in the summer vs. winter seasons, with higher percentage of algae (Bacillariophyta) and Bacteroidetes in winter-initiated samples but higher abundance of Proteobacteria (e.g., Rhizobiales, Rhodobacterales, Sphingomonadales, and Burkholderiales), Actinobacteria, and Chloroflexi in summer-initiated samples. Interestingly, results showed that despite seasonal effects on early biofilm succession, biofilm community structures converged after 70 days, suggesting the existence of a stable, mature community in the stream that is independent of the environmental conditions during biofilm colonization. Overall, our results show that algae are important in the early development of biofilm communities during winter, while heterotrophic bacteria play a more critical role during summer colonization and development of biofilms.

Control of Line Complications with KiteLock (CLiCK) in the critical care unit: study protocol for a multi-center, cluster-randomized, double-blinded, crossover trial investigating the effect of a novel locking fluid on central line complications in the critical care population

BackgroundInsertion of a central venous access device (CVAD) allows clinicians to easily access the circulation of a patient to administer life-saving interventions. Due to their invasive nature, CVADs are prone to complications such as bacterial biofilm production and colonization, catheter-related bloodstream infection, occlusion, and catheter-related venous thrombosis. A CVAD is among the most common interventions for patients in the intensive care unit (ICU), exposing this vulnerable population to the risk of nosocomial infection and catheter occlusion. The current standard of care involves the use of normal saline as a catheter locking solution for central venous catheters (CVCs) and peripherally inserted central catheter (PICC) lines, and a citrate lock for hemodialysis catheters. Saline offers little prophylactic measures against catheter complications. Four percent of tetrasodium ethylenediaminetetraacetic acid (EDTA) fluid (marketed as KiteLock Sterile Locking Solution™) is non-antibiotic, possesses antimicrobial, anti-biofilm, and anti-coagulant properties, and is approved by Health Canada as a catheter locking solution. As such, it may be a superior CVAD locking solution than the present standard of care lock in the ICU patient population.MethodsOur team proposes to fill this knowledge gap by performing a multi-center, cluster-randomized, crossover trial evaluating the impact of 4% tetrasodium EDTA on a primary composite outcome of the incidence rate of central line-associated bloodstream infection (CLABSI), catheter occlusion leading to removal, and use of alteplase to resolve catheter occlusion compared to the standard of care. The study will be performed at five critical care units.DiscussionIf successful, the results of this study can serve as evidence for a shift of standard of care practices to include EDTA locking fluid in routine CVAD locking procedures. Completion of this study has the potential to improve CVAD standard of care to become safer for patients, as well as provides an opportunity to decrease strain on healthcare budgets related to treating preventable CVAD complications. Success and subsequent implementation of this intervention in the ICU may also be extrapolated to other patient populations with heavy CVAD use including hemodialysis, oncology, parenteral nutrition, and pediatric patient populations. On a global scale, eradicating biofilm produced by antibiotic-resistant bacteria may serve to lessen the threat of “superbugs” and contribute to international initiatives supporting the termination of antibiotic overuse.Trial registrationClinicalTrials.gov NCT04548713, registered on September 9th, 2020.

Filamentous morphology of bacterial pathogens: regulatory factors and control strategies.

Several studies have demonstrated that when exposed to physical, chemical, and biological stresses in the environment, many bacteria (Gram-positive and Gram-negative) change their morphology from a normal cell to a filamentous shape. The formation of filamentous morphology is one of the survival strategies against environmental stress and protection against phagocytosis or protist predators. Numerous pathogenic bacteria have shown filamentous morphologies when examined in vivo or in vitro. During infection, certain pathogenic bacteria adopt a filamentous shape inside the cell to avoid phagocytosis by immune cells. Filamentous morphology has also been seen in biofilms formed on biotic or abiotic surfaces by certain bacteria. As a result, in addition to protecting against phagocytosis by immune cells or predators, the filamentous shape aids in biofilm adhesion or colonization to biotic or abiotic surfaces. Furthermore, these filamentous morphologies of bacterial pathogens lead to antimicrobial drug resistance. Clinically, filamentous morphology has become one of the most serious challenges in treating bacterial infection. The current review went into great detail about the various factors involved in the change of filamentous morphology and the underlying mechanisms. In addition, the review discussed a control strategy for suppressing filamentous morphology in order to combat bacterial infections. Understanding the mechanism underlying the filamentous morphology induced by various environmental conditions will aid in drug development and lessen the virulence of bacterial pathogens. KEY POINTS: • The bacterial filamentation morphology is one of the survival mechanisms against several environmental stress conditions and protection from phagocytosis by host cells and protist predators. • The filamentous morphologies in bacterial pathogens contribute to enhanced biofilm formation, which develops resistance properties against antimicrobial drugs. • Filamentous morphology has become one of the major hurdles in treating bacterial infection, hence controlling strategies employed for inhibiting the filamentation morphology from combating bacterial infections.

Elucidating biofouling over thermal and spatial gradients in seawater membrane distillation in hot climatic conditions

Biofouling is a hurdle of seawater desalination that increases water costs and energy consumption. In membrane distillation (MD), biofouling development is complicated due to the temperature effect that adversely affects microbial growth. Given the high relevance of MD to regions with abundant warm seawater, it is essential to explore the biofouling propensity of microbial communities with higher tolerance to elevated temperature conditions. This study presents a comprehensive analysis of the spatial and temporal biofilm distribution and associated membrane fouling during direct contact MD (DCMD) of the Red Sea water. We found that structure and composition of the biofilm layer played a significant role in the extent of permeate flux decline, and biofilms that built up at 45°C had lower bacterial concentration but higher extracellular polymeric substances (EPS) content as compared to biofilms that formed at 55 °C and 65°C. Pore wetting and bacterial passage to the permeate side were initially observed but slowed down as operating time increased. Intact cells in biofilms dominated over the damaged cells at any tested condition emphasizing the high adaptivity of the Red Sea microbial communities to elevated feed temperatures. A comparison of microbial abundance revealed a difference in bacterial distribution between the feed and biofilm samples. A shift in the biofilm microbial community and colonization of the membrane surface with thermophilic bacteria with the feed temperature increase was observed. The results of this study improve our understanding of biofouling propensity in MD that utilizes temperature-resilient feed waters.

Bioaccessibility of Microplastic-Associated Antibiotics in Freshwater Organisms: Highlighting the Impacts of Biofilm Colonization via an In Vitro Protocol.

Microplastics in the environment can be colonized by microbes capable of forming biofilms, which may act as reactive coatings to affect the bioaccessibility of pollutants in organisms. This study investigated the dynamic evolution of biofilm colonization on microplastics and its impacts and mechanisms on the bioaccessibility of microplastic-associated sulfamethazine (SMT) via microcosm incubation in surface water and sediment. After 60 days of incubation, the microbial communities formed in microplastics were distinct and more diverse than those untethered in surroundings, and photoaging treatment decreased the affinity of biofilms on microplastics due to decreased hydrophobicity. Biofilm formation further enhanced the desorption and bioaccessibility of microplastic-sorbed SMT in organisms. In vitro experiments indicated that the critical effects were mainly related to the stronger interaction of gastrointestinal components (i.e., pepsin, bovine serum albumin (BSA), and NaT) with biofilm components (e.g., extracellular polymer substances) than with the pure surface of microplastics, which competed for binding sites in microplastics for SMT more significantly. Photoaging decreased the enhancing effects of biofilms due to their lower accumulation in aged microplastics. This study is the first attempt to reveal the role of biofilms in the bioaccessibility of microplastics with associated antibiotics and provide insights into the combined risk of microplastics in the environment.

Plastisphere in lake waters: Microbial diversity, biofilm structure, and potential implications for freshwater ecosystems

Once dispersed in water, microplastic (MP) particles are rapidly colonised by aquatic microbes, which can adhere and grow onto solid surfaces in the form of biofilms. This study provides new insights on microbial diversity and biofilm structure of plastisphere in lake waters. By combining Fourier Confocal Laser Scanning Microscopy (CLSM), Transform Infrared Spectroscopy (FT-IR) and high-throughput DNA sequencing, we investigated the microbial colonization patterns on floating MPs and, for the first time, the occurrence of eukaryotic core members and their possible relations with biofilm-forming bacterial taxa within the plastisphere of four different lakes. Through PCR-based methods (qPCR, LAMP-PCR), we also evaluated the role of lake plastisphere as long-term dispersal vectors of potentially harmful organisms (including pathogens) and antibiotic resistance genes (ARGs) in freshwater ecosystems. Consistent variation patterns of the microbial community composition occurred between water and among the plastisphere samples of the different lakes. The eukaryotic core microbiome was mainly composed by typical freshwater biofilm colonizers, such as diatoms (Pennales, Bacillariophyceaea) and green algae (Chlorophyceae), which interact with eukaryotic and prokaryotic microbes of different trophic levels. Results also showed that MPs are suitable vectors of biofilm-forming opportunistic pathogens and a hotspot for horizontal gene transfer, likely facilitating antibiotic resistance spread in the environments.

Bacterial biofilm colonization and succession in tropical marine waters are similar across different types of stone materials used in seawall construction.

Seawalls are important in protecting coastlines from currents, erosion, sea-level rise, and flooding. They are, however, associated with reduced biodiversity, due to their steep orientation, lack of microhabitats, and the materials used in their construction. Hence, there is considerable interest in modifying seawalls to enhance the settlement and diversity of marine organisms, as microbial biofilms play a critical role facilitating algal and invertebrate colonization. We assessed how different stone materials, ranging from aluminosilicates to limestone and concrete, affect biofilm formation. Metagenomic assessment of marine microbial communities indicated no significant impact of material on microbial diversity, irrespective of the diverse surface chemistry and topography. Based on KEGG pathway analysis, surface properties appeared to influence the community composition and function during the initial stages of biofilm development, but this effect disappeared by Day 31. We conclude that marine biofilms converged over time to a generic marine biofilm, rather than the underlying stone substrata type playing a significant role in driving community composition.

Antibiofilm activity of secondary metabolites from bacterial endophytes of Red Sea soft corals

Endophytic marine bacteria are a significant unexplored resource for the search of new compounds with antifouling activity. In this study, endophytic marine bacteria isolated from Red Sea soft corals were subjected to secondary metabolites extraction and evaluation of the extracts for the antibiofilm activity to reveal compounds that can prevent biofilm bacterial colonization on surfaces. Six bacteria were isolated from two species of soft corals: Sarcophyton glaucum and Heteroxenia fuscescens. Extracts obtained from each strain were screened for antibiofilm activity against three biofilm-forming bacteria and evaluated to detect compounds with antibiofilm properties by Gas chromatography-mass spectrometry (GC-MS). The isolated strains are closely related and belong to the family Bacillaceae and all except one belong to the Bacillus genus. The extracts from the endophytes show significant antibacterial activity against the tested organisms. Biofilm formation inhibition activity of the extracts ranges from 3.16 to 98.41%. The extracts also showed significant activity against biofilm formation across all the tested strains. Compounds detected in the extracts include known compounds with antibiofilm properties such as 2,4-dimethyl-2-nitro-, 2,4-bis(1,1-dimethylethyl)-phenol, Pyrrolo[1,2-a] pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl)-; Bis(tridecyl) phthalate, Heinecosane, Pentane, and Undecane.This indicates the ability of the bacteria to produce compounds relevant in preventing biofilm colonization on surfaces which could serve as a novel source of antifouling compounds.

Vegetation Formation in Staphylococcus Aureus Endocarditis Inversely Correlates With RNAIII and sarA Expression in Invasive Clonal Complex 5 Isolates.

Infective endocarditis (IE) is one of the most feared and lethal diseases caused by Staphylococcus aureus. Once established, the infection is fast-progressing and tissue destructive. S. aureus of the clonal complex 5 (CC5) commonly cause IE yet are severely understudied. IE results from bacterial colonization and formation of tissue biofilms (known as vegetations) on injured or inflamed cardiac endothelium. S. aureus IE is promoted by adhesins, coagulases, and superantigens, with the exotoxins and exoenzymes likely contributing to tissue destruction and dissemination. Expression of the large repertoire of virulence factors required for IE and sequelae is controlled by complex regulatory networks. We investigated the temporal expression of the global regulators agr (RNAIII), rot, sarS, sarA, sigB, and mgrA in 8 invasive CC5 isolates and established intrinsic expression patterns associated with IE outcomes. We show that vegetation formation, as tested in the rabbit model of IE, inversely correlates with RNAIII and sarA expression during growth in Todd-Hewitt broth (TH). Large vegetations with severe sequelae arise from strains with high-level expression of colonization factors but slower transition towards expression of the exotoxins. Overall, strains proficient in vegetation formation, a hallmark of IE, exhibit lower expression of RNAIII and sarA. Simultaneous high expression of RNAIII, sarA, sigB, and mgrA is the one phenotype assessed in this study that fails to promote IE. Thus, RNAIII and sarA expression that provides for rheostat control of colonization and virulence genes, rather than an on and off switch, promote both vegetation formation and lethal sepsis.

The Complex Mechanism of the Salmonella typhi Biofilm Formation That Facilitates Pathogenicity: A Review

Salmonella enterica serovar Typhi (S. typhi) is an intracellular pathogen belonging to the Enterobacteriaceae family, where biofilm (aggregation and colonization of cells) formation is one of their advantageous traits. Salmonella typhi is the causative agent of typhoid fever in the human body and is exceptionally host specific. It is transmitted through the fecal–oral route by consuming contaminated food or water. This subspecies is quite intelligent to evade the innate detection and immune response of the host body, leading to systemic dissemination. Consequently, during the period of illness, the gallbladder becomes a harbor and may develop antibiotic resistance. Afterwards, they start contributing to the continuous damage of epithelium cells and make the host asymptomatic and potential carriers of this pathogen for an extended period. Statistically, almost 5% of infected people with Salmonella typhi become chronic carriers and are ready to contribute to future transmission by biofilm formation. Biofilm development is already recognized to link with pathogenicity and plays a crucial role in persistency within the human body. This review seeks to discuss some of the crucial factors related to biofilm development and its mechanism of interaction causing pathogenicity. Understanding the connections between these things will open up a new avenue for finding therapeutic approaches to combat pathogenicity.

Revealing the effect of biofilm formation in partial nitritation-anammox systems: Start-up, performance stability, and recovery

Successful application of partial nitritation-anammox (PNA) processes is currently and primarily associated with biofilm systems. Biofilm characteristics significantly influence start-up, performance stability, and recovery. Here, two PNA systems with and without carriers were implemented simultaneously for treating wastewater containing 50 mg-NH4/L. The performance characteristics of these two PNA systems were compared. Stable nitrogen removal efficiencies of 76.3 ± 2.8% and 72.9 ± 1.6% were obtained for suspended sludge and biofilm systems, respectively. The slow process of biofilm colonization resulted in a long start-up time in the biofilm system. Biofilm enrichment and protection conferred stable performance when exposed to aeration shock. The suspended sludge system displayed good elasticity during performance recovery after shock compared to the slow recovery in the biofilm system. Moreover, suitable control of dissolved oxygen could improve the activity and abundance of the functional microbes. This study provides new insights into the operation and control of PNA systems for treating mainstream wastewater.

Non-typeable Haemophilus influenzae biofilm formation on primary human airway epithelia cultured at air-liquid interface

Introduction Primary ciliary dyskinesia (PCD) is an inherited heterogeneous disorder associated with defective motile cilia on airway epithelia, resulting in impaired mucociliary function and ultimately, bronchiectasis. Non-typeable Haemophilus influenzae (NTHi) is frequently isolated from PCD airways where it resides in biofilms. It is unclear if biological pathways in response to NTHi infection differ in PCD, contributing to increased susceptibility to biofilm-associated infection. Aim The development of a biologically representative co-culture model to investigate the interactions between PCD airway epithelia and NTHi biofilms. Methods Air-liquid interface (ALI)-cultured non-PCD primary nasal cells were grown in transwells. Barrier properties were assessed by trans-epithelial electrical resistance (TEER), FITC-dextran passage and tight junction staining. Ciliary function was assessed by high-speed video microscopy at 37°C. ALI cultures were infected with a PCD NTHi isolate at multiplicity of infections (MOIs) 10, 50 or 100 and co-cultured to form biofilms over 3 days. TEER was measured pre- and post-infection, NTHi recoverability assessed by conventional culture, and biofilm formation confirmed by scanning electron microscopy (SEM). Results Epithelial culture at ALI was successful, with tight barriers being formed and normal ciliary beat pattern and frequency recorded (mean 14Hz). Following co-culture, TEER increased relative to pre-infection at all MOIs but maximum NTHi recoverability was at MOI 50 (median colony forming units, 6.8x106per cm2). SEM confirmed NTHi biofilm formation at MOI 50. Conclusions A successful NTHi biofilm co-culture model on human primary epithelia was established. This will be used with PCD epithelia to investigate host-pathogen interactions during NTHi biofilm colonisation.

Filling Materials Efficacy on Preventing Biofilm Formation Inside Screw Access Channels of Implant Abutments.

The choice of the material used to fill screw access channels in implant-supported prostheses depends, in most cases, on operator's preference, without considering the susceptibility of biofilm colonization. Therefore, the aim of this study was to determine and compare the total amount of biofilm formed on different materials used to fill screw access channels in implant abutments. For this propose, titanium implant analogs were attached on abutments and divided into 5 groups: positive control (no filling material); negative control (closed with resin); and filled with cotton, gutta-percha, or polytetrafluoroethylene (PTFE). The analogs with attached abutments were then immersed in a brain heart infusion medium containing Candida albicans (strain 10231 from American Type Culture Collection [ATCC]) and incubated aerobically at 37°C with gentle agitation. After 15 days, materials were removed, and total viable biofilm on each material was quantified by methyl tetrazolium reduction assay at 490 nm. All experiments were performed in triplicate. Data were processed by IBM SPSS Statistic software using 1-way analysis of variance and Bonferroni post hoc tests to analyze differences between groups, with an overall significance level of P < .001. A significant difference was observed between cotton and gutta-percha (P < .017) and between cotton and PTFE (P < .025). However, there was no statistical difference between gutta-percha and PTFE (P > .050). Thus, this in vitro experiment showed that gutta-percha and PTFE presented lower biofilm formation compared with cotton when used to fill screw access channels. These results can provide a basis for future clinical studies that can be a guide to decreasing the occurrence of gaps and bacterial growth inside the implant/abutment attachment site. In addition, controlled in vivo studies are necessary to confirm the clinical viability of findings of this study.

Individual based Modelling of Biofilm Colonisation

Bacterial biofilms on implants or host tissue can cause unwanted infections. One of the main factors for the pathogenic biofilm growth is the initial adhesion of the bacteria. In this project a framework for systematic studies of the bacterial adhesion is provided. Therefore, an Individual based Model (IbM) is set up in the IbM solver NUFEB, an extension of the molecular dynamics code LAMMPS, which integrates Newton's equations of motion for the cells that interact via various forces. Experimentally measured adhesion forces are implemented for the cell-wall and cell-cell interaction. Furthermore, the random movement of planktonic cells is included into the IbM software. A sensitivity analysis of important parameters and an exemplary simulation of the attachment are performed. Thus, the attachment of the bacterial cells and the initial growth of a biofilm can be qualitatively modelled.

Effect of biofilm colonization on Pb(II) adsorption onto poly(butylene succinate) microplastic during its biodegradation

Few studies have mentioned the enrichment of heavy metal pollutants on microplastics derived from degradable plastics. This study investigated the adsorption behavior of Pb(II) onto biodegradable poly(butylene succinate) (PBS) microplastics during its biodegradation. The results indicated that Pb(II) adsorbed by biofilm-colonized biodegraded-PBS microplastics (B-PBS) was about 10-folds higher than that of virgin PBS (647.09 μg·g−1 versus 64.13 μg·g−1) due to the biofilm colonization and the degradation of PBS. After removing the biofilm, the biodegraded PBS still had high Pb(II) adsorption capacity, which was attributed to the complexation of Pb(II) and the stably adhered extracellular polymeric substances (EPS). Pb(II) adsorption onto both virgin PBS and B-PBS was highly pH-dependent, its adsorption on virgin PBS was dominated by electrostatic interaction, while as for B-PBS, the adsorption mechanisms mainly involved the coordination/complexation of Pb(II) and the EPS components on the colonized biofilm, surface complexation, and electrostatic interaction. This study suggested that the enrichment of heavy metal pollutants onto the biodegradable microplastics may pose risks to the aquatic ecosystem.