Step In Biofilm Formation Research Articles

Initial Deposition and Pioneering Colonization on Polymeric Membranes of Anaerobes Isolated from an Anaerobic Membrane Bioreactor (AnMBR).

Membrane biofouling constitutes a great challenge in anaerobic membrane bioreactor (AnMBR). Here, we studied the initial deposition of anaerobes, the first step in biofilm formation, with a consortium isolated from an AnMBR on membranes with different surface properties and under two shear rate conditions without filtration. We found that the cell transfer coefficient, calculated from the initial deposition experiments, was similar under the two shear rates for the hydrophobic membranes, but much higher under low shear rate and much lower under high shear rate, for the hydrophilic membrane. The cell transfer coefficient measured under filtration mode and at a higher shear rate showed a similar trend. The pioneer bacteria and archaea (without filtration) were identified by next-generation sequencing. The results showed that the selective force for the dissimilarity of the pioneer bacterial and archaeal diversity was the shear rate and the membrane surface properties, respectively. However, statistical analyses revealed minor changes in the pioneer bacteria (class level) and archaea (order level) populations under the various conditions. These results shed light on the first step of biofilm formation on the membranes in AnMBRs and emphasize the importance of hydrodynamic shear and membrane surface properties on the initially deposited anaerobes.

Targeting Pathogenic Biofilms: Newly Developed Superhydrophobic Coating Favors a Host-Compatible Microbial Profile on the Titanium Surface.

Polymicrobial infections are one of the most common reasons for inflammation of surrounding tissues and failure of implanted biomaterials. Because microorganism adhesion is the first step for biofilm formation, physical-chemical modifications of biomaterials have been proposed to reduce the initial microbial attachment. Thus, the use of superhydrophobic coatings has emerged because of their anti-biofilm properties. However, these coatings on the titanium (Ti) surface have been developed mainly by dual-step surface modification techniques and have not been tested using polymicrobial biofilms. Therefore, we developed a one-step superhydrophobic coating on the Ti surface by using a low-pressure plasma technology to create a biocompatible coating that reduces polymicrobial biofilm adhesion and formation. The superhydrophobic coating on Ti was created by the glow discharge plasma using Ar, O2, and hexamethyldisiloxane gases, and after full physical, chemical, and biological characterizations, we evaluated its properties regarding oral biofilm inhibition. The newly developed coating presented an increased surface roughness and, consequently, superhydrophobicity (contact angle over 150°) and enhanced corrosion resistance (p < 0.05) of the Ti surface. Furthermore, proteomic analysis showed a unique pattern of protein adsorption on the superhydrophobic coating without drastically changing the biologic processes mediated by proteins. Additionally, superhydrophobic treatment did not present a cytotoxic effect on fibroblasts or reduction of proliferation; however, it significantly reduced (≈8-fold change) polymicrobial adhesion (bacterial and fungal) and biofilm formation in vitro. Interestingly, superhydrophobic coating shifted the microbiological profile of biofilms formed in situ in the oral cavity, reducing by up to ≈7 fold pathogens associated with the peri-implant disease. Thus, this new superhydrophobic coating developed by a one-step glow discharge plasma technique is a promising biocompatible strategy to drastically reduce microbial adhesion and biofilm formation on Ti-based biomedical implants.

Spatio-temporal formation of biofilms and extracellular matrix analysis in Azospirillum brasilense.

Elucidation of biofilm structure formation in the plant growth-promoting rhizobacterium Azospirillum brasilense is necessary to gain a better understanding of the growth of cells within the extracellular matrix and its role in the colonization of plants of agronomic importance. We used immunofluorescence microscopy and confocal laser scanning microscopy to study spatio-temporal biofilm formation on an abiotic surface. Observations facilitated by fluorescence microscopy revealed the presence of polar flagellin, exopolysaccharides, outer major membrane protein (OmaA) and extracellular DNA in the Azospirillum biofilm matrix. In static culture conditions, the polar flagellum disaggregated after 3 days of biofilm growth, but exopolysaccharides were increasing. These findings suggest that the first step in biofilm formation may be attachment, in which the bacterium first makes contact with a surface through its polar flagellum. After attaching to the surface, the long flagella and OmaA intertwine the cells to form a network. These bacterial aggregates initiate biofilm development. The underlying mechanisms dictating how the biofilm matrix components of A. brasilense direct the overall morphology of the biofilm are not well known. The methods developed here might be useful in further studies that analyze the differential spatial regulation of genes encoding matrix components that drive biofilm construction.

How Functionalized Surfaces Can Inhibit Bacterial Adhesion and Viability.

Device-associated infections (DAI) remain a serious concern in modern healthcare. Bacterial attachment to a surface is the first step in biofilm formation, which is one of the main causes of DAIs. The development of materials capable of preventing or inhibiting bacterial attachment constitutes a promising approach to deal with this problem. The multifactorial nature of biofilm maturation and antibiotic resistance directs the research for multitargeted or combinatorial therapeutic approaches. One attractive strategy is the modification or the engineering of surfaces in order to provide antiadhesive and/or antimicrobial properties. Currently, several different approaches that involve physical and chemical surface modification deliver some possible alternatives to achieve this goal. The engineered surfaces can be coated with molecules capable of inhibiting the bacterial adhesion or with active agents that kill microorganisms. In addition, surfaces can also be modified in order to be stimuli-responsive, responding to a particular trigger and then delivering the consequent antimicrobial outcome. Here, we review the prevailing strategies to modify surfaces in order to create an antimicrobial surface and discuss how different surface functionalization can affect bacterial adhesion and/or viability.

Flagellar Stators Stimulate c-di-GMP Production by Pseudomonas aeruginosa.

Flagellar motility is critical for surface attachment and biofilm formation in many bacteria. A key regulator of flagellar motility in Pseudomonas aeruginosa and other microbes is cyclic diguanylate (c-di-GMP). High levels of this second messenger repress motility and stimulate biofilm formation. c-di-GMP levels regulate motility in P. aeruginosa in part by influencing the localization of its two flagellar stator sets, MotAB and MotCD. Here, we show that while c-di-GMP can influence stator localization, stators can in turn impact c-di-GMP levels. We demonstrate that the swarming motility-driving stator MotC physically interacts with the transmembrane region of the diguanylate cyclase SadC. Furthermore, we demonstrate that this interaction is capable of stimulating SadC activity. We propose a model by which the MotCD stator set interacts with SadC to stimulate c-di-GMP production under conditions not permissive to motility. This regulation implies a positive-feedback loop in which c-di-GMP signaling events cause MotCD stators to disengage from the motor; then disengaged stators stimulate c-di-GMP production to reinforce a biofilm mode of growth. Our studies help to define the bidirectional interactions between c-di-GMP and the flagellar machinery.IMPORTANCE The ability of bacterial cells to control motility during early steps in biofilm formation is critical for the transition to a nonmotile, biofilm lifestyle. Recent studies have clearly demonstrated the ability of c-di-GMP to control motility via a number of mechanisms, including through controlling transcription of motility-related genes and modulating motor function. Here, we provide evidence that motor components can in turn impact c-di-GMP levels. We propose that communication between motor components and the c-di-GMP synthesis machinery allows the cell to have a robust and sensitive switching mechanism to control motility during early events in biofilm formation.

Visualizing Biomaterial Degradation by Candida albicans Using Embedded Luminescent Molecules To Report on Substrate Digestion and Cellular Uptake of Hydrolysate.

Microbial pathogens use hydrolases as a virulence strategy to spread disease through tissues and colonize medical device surfaces; however, visualizing this process is a technically challenging problem. To better understand the role of secreted fungal hydrolases and their role in Candida albicans virulence, we developed an in situ model system using luminescent Re(I) and Ir(III) containing probe molecules embedded in a biodegradable (poly(lactic-co-glycolic acid), PLGA) polymer and tracked their uptake using epifluorescent imaging. We found that secretion of esterases can explain how physically embedded probes are acquired by fungal cells through the degradation of PLGA since embedded probes could not be liberated from nonbiodegradable polystyrene (PS). It was important to verify that epifluorescent imaging captured the fate of probe molecules rather than naturally occurring fungal autofluorescence. For this, we exploited the intense luminescent signals and long spectral relaxation times of the Re and Ir containing probe molecules, resolved in time using a gated imaging system. Results provide a visual demonstration of a key virulence trait of C. albicans: the use of hydrolases as a means to degrade materials and acquire hydrolysis products during fungal growth and hyphal development. These results help to explain the role of nonspecific hydrolases using a degradable material that is relevant to the study of fungal pathogenesis on biotic (tissues) surfaces. Additionally, understanding how fungal pathogens condition surfaces by using nonspecific hydrolases is important to the study of fungal attachment on abiotic surfaces, the first step in biofilm formation on medical devices.

Rapid Lab Grown Biofilm As a Tool for Engineering Sensor Networks of Biofouling-Resistant Electrochemical Sensors

Robust and rapid biofilm formation is not only a major interest in the development of microbial fuel cell, but it is also be of relevance to the development of indwelling environmental sensor networks. Sensor networks offer real-time, online and long-term monitoring which is an attractive monitoring strategy for water industry to prepare for increasing demand for clean water due to increased urbanisation and the pressures of climate change. Sensor networks support the development of robust and managed drinking water supply networks that are essential to human health. Fouling of sensors due to biofilm formation is very common in indwelling situations and it is a major drawback in the development of sensor networks. Sensors will fail once it is fouled and it can happen in just a couple days or weeks. Engineering robust sensor networks begins with designing a biofouling resistant sensors. Electrochemical sensors are inexpensive and simple to construct and operate, and it is widely used in water industry. Understanding how biofilms affect electrochemical sensor performance is important to enable detection of malfunction in situ, and can inform the development of methodology to restore the performance in situ. Networks can consist of hundreds of sensors and automated restoration of function in biofouled sensors is thus essential. Lab-based models play a vital role. However, it is not always possible to recreate the real conditions for biofilm formation in the lab and it can take too long for it to form. In a situation in which it is not possible to build a permanent testing setup, rapid lab grown biofilm is desirable. Adhesion of pioneering bacterial cells to the surface is the most crucial step in biofilm formation. Adhesion of cells to surfaces can be accelerated by altering the surface chemistry of the surfaces. One way to do this is by plasma treatment of the surface. Formation of a lawn of biofilm is desirable to model early biofouling event. Polysaccharide production that forms the extrapolymeric substance (EPS) of biofilm can be induced by common biocidal substance such as triclosan. Electrochemically generated local concentration of hydrogen peroxide has similar consequences. In this work, we show that it is possible to form some coverage of biofilm on electrode surface in just two hours, which represents early biofouling event. It has a potential for the development of rapid lab grown biofilm that can be useful in developing fail-proof sensor networks.

The trimeric autotransporter adhesin BadA is required for in vitro biofilm formation by Bartonella henselae

Bartonellahenselae (Bh) is a Gram-negative rod transmitted to humans by a scratch from the common house cat. Infection of humans with Bh can result in a range of clinical diseases including lymphadenopathy observed in cat-scratch disease and more serious disease from persistent bacteremia. It is a common cause of blood-culture negative endocarditis as the bacterium is capable of growing as aggregates, and forming biofilms on infected native and prosthetic heart valves. The aggregative growth requires a trimeric autotransporter adhesin (TAA) called Bartonella adhesin A (BadA). TAAs are found in all Bartonella species and many other Gram-negative bacteria. Using Bh Houston-1, Bh Houston-1 ∆badA and Bh Houston-1 ∆badA/pNS2PTrcbadA (a partial complement of badA coding for a truncated protein of 741 amino acid residues), we analyze the role of BadA in adhesion and biofilm formation. We also investigate the role of environmental factors such as temperature on badA expression and biofilm formation. Real-time cell adhesion monitoring and electron microscopy show that Bh Houston-1 adheres and forms biofilm more efficiently than the Bh Houston-1 ∆badA. Deletion of the badA gene significantly decreases adhesion, the first step in biofilm formation in vitro, which is partially restored in Bh Houston-1 ∆badA/pNS2PTrcbadA. The biofilm formed by Bh Houston-1 includes polysaccharides, proteins, and DNA components and is susceptible to enzymatic degradation of these components. Furthermore, both pH and temperature influence both badA expression and biofilm formation. We conclude that BadA is required for optimal adhesion, agglutination and biofilm formation.

Evaluation of microbial proliferation on cementitious materials exposed to biogas systems

ABSTRACT Understanding the interactions between biofilm and cementitious materials in biogas production systems is an essential step toward the development of durable concrete for this expanding sector. Although the action of the liquid phase medium on the material has been the subject of several research studies, the possible impact of the material’s properties on biofilm formation and composition has been little investigated, if at all. The aim of this paper is to evaluate the characteristics of the biofilm according to the surface properties of the materials. Four cementitious materials with different chemical and mineralogical compositions, and various topological surface characteristics (pastes of CEM I, CEM III/C and CAC, and CEM I paste treated with oxalic acid) were exposed to the liquid phase of a fermenting biowaste for 10 weeks. The steps of biofilm formation were observed using SEM. Even though all the cementitious material surfaces were intensely colonized at the end of the experiments, the establishment of the biofilm seems to have been delayed on the oxalate-treated CEM I and on CAC coupons. Roughness and surface pH effects were not of prime importance for the biofilm development. The analysis of bacterial population diversity using 16S rDNA sequencing showed a less diversified microbial flora in the biofilm than in the reaction medium.

Physical Determinants of Amyloid Assembly in Biofilm Formation.

A wide range of bacterial pathogens have been shown to form biofilms, which significantly increase their resistance to environmental stresses, such as antibiotics, and are thus of central importance in the context of bacterial diseases. One of the major structural components of these bacterial biofilms are amyloid fibrils, yet the mechanism of fibril assembly and its importance for biofilm formation are currently not fully understood. By studying fibril formation in vitro, in a model system of two common but unrelated biofilm-forming proteins, FapC from Pseudomonas fluorescens and CsgA from Escherichia coli, we found that the two proteins have a common aggregation mechanism. In both systems, fibril formation proceeds via nucleated growth of linear fibrils exhibiting similar measured rates of elongation, with negligible fibril self-replication. These similarities between two unrelated systems suggest that convergent evolution plays a key role in tuning the assembly kinetics of functional amyloid fibrils and indicates that only a narrow window of mechanisms and assembly rates allows for successful biofilm formation. Thus, the amyloid assembly reaction is likely to represent a means for controlling biofilm formation, both by the organism and by possible inhibitory drugs.IMPORTANCE Biofilms are generated by bacteria, embedded in the formed extracellular matrix. The biofilm's function is to improve the survival of a bacterial colony through, for example, increased resistance to antibiotics or other environmental stresses. Proteins secreted by the bacteria act as a major structural component of this extracellular matrix, as they self-assemble into highly stable amyloid fibrils, making the biofilm very difficult to degrade by physical and chemical means once formed. By studying the self-assembly mechanism of the fibrils from their monomeric precursors in two unrelated bacteria, our experimental and theoretical approaches shed light on the mechanism of functional amyloid assembly in the context of biofilm formation. Our results suggest that fibril formation may be a rate-limiting step in biofilm formation, which in turn has implications on the protein self-assembly reaction as a target for potential antibiotic drugs.

Evaluation of Surface Roughness and &amp;lt;i&amp;gt;Streptococcus mutans&amp;lt;/i&amp;gt; Adhesion to Bulk-Fill Resin Composites Polished with Different Systems

Purpose: Bacterial adhesion represents the initial step in biofilm formation, dental caries and decay. This study aimed to evaluate and compare surface roughness and bacterial adhesion to bulk fill resin composites polished with different systems. Methods: Filtek Z350 XT (Incremental-fill resin composite), Filtek Bulk-fill Posterior (Bulk-fill resin composite), and Tetric N Ceram (Bulk-fill resin composite) were used as resin composites. The polishing systems used in this study were Sof-Lex multi-step, PoGo one step, and Mylar strip. Scanning electron microscope (SEM) was used to examine the surface roughness and adhesion of Streptococcus mutans ATCC 25175 standard strain to bulk-fill resin composites. Results: The type of restorative materials did not affect the surface roughness or bacterial adhesion (p > 0.05) but the polishing systems were significant (p < 0.05) influencing factors. Furthermore, Pearson correlation revealed a statistically significant (p < 0.001) association (R = 0.943) between surface roughness and bacterial adhesion to the tested surfaces. Conclusion: Regardless of the restorative material, Mylar polishing system revealed the smoothest surface and the lowest adhesion of S. mutans as compared to Pogo one step and Sof-Lex multi-step polishing systems.

Nanozymes in Nanofibrous Mats with Haloperoxidase-like Activity To Combat Biofouling.

Electrospun polymer mats are widely used in tissue engineering, wearable electronics, and water purification. However, in many environments, the polymer nanofibers prepared by electrospinning suffer from biofouling during long-term usage, resulting in persistent infections and device damage. Herein, we describe the fabrication of polymer mats with CeO2- x nanorods that can prevent biofouling in an aqueous environment. The embedded CeO2- x nanorods are functional mimics of natural haloperoxidases that catalyze the oxidative bromination of Br- and H2O2 to HOBr. The generated HOBr, a natural signaling molecule, disrupted the bacterial quorum sensing, a critical step in biofilm formation. The polymer fibers provide porous structures with high water wettability, and the embedded cerium oxide nanozymes act as a catalyst that can efficiently trigger oxidative bromination, as shown by a haloperoxidase assay. Additionally, the embedded nanozymes enhance the mechanical property of polymer mats, as shown by a single-fiber bending test using atomic force microscopy. We envision that the fabricated polymer mats with CeO2- x nanorods may be used to provide mechanically robust coatings with antibiofouling properties.

Surface Modification by Combination of Dip-Pen Nanolithography and Soft Lithography for Reduction of Bacterial Adhesion

Dip-pen nanolithography (DPN) and soft lithography are techniques suitable to modify the surface of biomaterials. Modified surfaces might play a role in modulating cells and reducing bacterial adhesion and biofilm formation. The main objective of this study was threefold: first, to create patterns at microscale on model surfaces using DPN; second, to duplicate and transfer these patterns to a real biomaterial surface using a microstamping technique; and finally, to assess bacterial adhesion to these developed patterned surfaces using the cariogenic species Streptococcus mutans. DPN was used with a polymeric adhesive to create dot patterns on model surfaces. Elastomeric polydimethylsiloxane was used to duplicate the patterns and silica sol to transfer them to the medical grade stainless steel 316L surface by microstamping. Optical microscopy and atomic force microscopy (AFM) were used to characterize the patterns. S. mutans adhesion was assessed by colony-forming units (CFUs), MTT viability assay, and scanning electron microscopy (SEM). DPN allowed creating microarrays from 1 to 5 µm in diameter on model surfaces that were successfully transferred to the stainless steel 316L surface via microstamping. A significant reduction up to one order of magnitude in bacterial adhesion to micropatterned surfaces was observed. The presented experimental approach may be used to create patterns at microscale on a surface and transfer them to other surfaces of interest. A reduction in bacterial adhesion to patterned surfaces might have a major impact since adhesion is a key step in biofilm formation and development of biomaterial-related infections.

Comparative study of virulence factors among methicillin resistant Staphylococcus aureus clinical isolates

BackgroundMethicillin resistant Staphylococcus aureus (MRSA) is recognized worldwide as a leading cause of hospital and community infections. Biofilm formation by MRSA is an extremely important virulence factor to be understood. Our aim was to establish phenotypic and genotypic characterization of virulence factors among 43 MRSA clinical isolates in a Tunisian hospital.MethodsWe investigated enzymatic profiles, biofilm production and prevalences of genes encoding intracellular adhesion molecules (icaA and icaD), Microbial Surface Components Recognizing Adhesive Matrix Molecules genes (fnbA, fnbB and cna) and exoenzymes genes (geh, sspA and sspB).ResultsOur findings revealed that caseinase, gelatinase, lipase and lecithinase activities were detected in 100%, 100%, 76.6% and 93.3% of cases respectively. This study showed that 23 strains (76.7%) were slime producers on Congo red medium. Furthermore, 46.5% and 53.5% of isolates were respectively highly and moderately biofilm-forming on polystyrene. Significant association was found between both biofilm tests. PCR detection showed that 74.4%, 18.6%, 69.8%, 65.1% and 74.4% of isolates harbored fnbA, fnbB, icaA, icaD and cna genes respectively. In addition, 34.9%, 18.6% and 30.2% of MRSA strains were found positive for sspA, sspB and geh genes respectively. Further, statistical data showed that the presence of the fnbA and fnbB genes was significantly associated with a high biofilm production on polystyrene. However, no statistical association was observed for the icaA, icaD and cna genes.ConclusionsThis study indicates that the detection of fnbA and fnbB contributing to the first step of biofilm formation has been predictable of high biofilm production. As studied factors contribute to MRSA virulence, this research could be of value in orienting towards the development of new preventive and therapeutic measures.

Zirconium Nitride Coating Reduced Staphylococcus epidermidis Biofilm Formation on Orthopaedic Implant Surfaces: An In Vitro Study.

One of the most commonly identified pathogens responsible for orthopaedic implant infection is Staphylococcus epidermidis, which can form biofilms on surfaces. Currently, orthopaedic implants made of various surface materials are available, each with features influencing osseointegration, biocompatibility, and adherence of bacteria to the surface, which is the first step in biofilm formation. The aim of this experimental study was to investigate the effect of a high tribologic-resistant 2.5-µm zirconium nitride top coat on an antiallergic multilayer ceramic-covered cobalt-chromium-molybdenum surface on the formation of S. epidermidis biofilm compared with other commonly used smooth and rough orthopaedic implant surface materials. (1) When evaluating the surfaces of a cobalt-chromium-molybdenum (CoCrMo) alloy with a zirconium (Zr) nitride coating, a CoCrMo alloy without a coating, titanium alloy, a titanium alloy with a corundum-blasted rough surface, and stainless steel with a corundum-blasted rough surface, does a Zr coating reduce the number of colony-forming units of S. epidermidis in an in vitro setting? (2) Is there quantitatively less biofilm surface area on Zr-coated surfaces than on the other surfaces tested in this in vitro model? To determine bacterial adhesion, five different experimental implant surface discs were incubated separately with one of 31 different S. epidermidis strains each and subsequently sonicated. Twenty test strains were obtained from orthopaedic patients undergoing emergency hip prosthesis surgeries or revision of implant infection and 10 further strains were obtained from the skin of healthy individuals. Additionally, one reference strain, S. epidermidis DSM 3269, was tested. After serial dilutions, the number of bacteria was counted and expressed as colony-forming units (CFUs)/mL. For biofilm detection, discs were stained with 0.1% Safranin-O for 15 minutes, photographed, and analyzed with computer imaging software. The lowest bacterial count was found in the CoCrMo + Zr surface disc (6.6 x 10 CFU/mL ± 4.6 x 10 SD) followed by the CoCrMo surface (1.1 x 10 CFU/mL ± 1.9 x 10 SD), the titanium surface (1.36 x 10 CFU/mL ± 1.8 x 10 SD), the rough stainless steel surface (2.65 x 10 CFU/mL ± 3.8 x 10 SD), and the rough titanium surface (2.1 x 10 CFU/mL ± 3.0 x 10 SD). The mean CFU count was lower for CoCrMo + Zr discs compared with the rough stainless steel surface (mean difference: 2.0 x 10, p = 0.021), the rough titanium alloy surface (mean difference: 1.4 x 10, p = 0.002), and the smooth titanium surface (mean difference: 7.0 x 10, p = 0.016). The results of biofilm formation quantification show that the mean covered area of the surface of the CoCrMo + Zr discs was 19% (± 16 SD), which was lower than CoCrMo surfaces (35% ± 23 SD), titanium alloy surface (46% ± 20 SD), rough titanium alloy surface (66% ± 23 SD), and rough stainless steel surface (58% ± 18 SD). These results demonstrate that a multilayer, ceramic-covered, CoCrMo surface with a 2.5-µm zirconium nitride top coat showed less S. epidermidis biofilm formation compared with other surface materials used for orthopaedic implants. CoCrMo with a 2.5-µm zirconium nitride top coat seems to be a promising surface modification technology able to reduce bacterial attachment on the surface of an implant and, hence, may further prevent implant infection with S. epidermidis biofilm formation.

Co-opting the Lap System of Pseudomonas fluorescens To Reversibly Customize Bacterial Cell Surfaces.

Initial attachment to a surface is a key and highly regulated step in biofilm formation. In this study, we present a platform for reversibly functionalizing bacterial cell surfaces with an emphasis on designing biofilms. We engineered the Lap system of Pseudomonas fluorescens Pf0-1, which is normally used to regulate initial cell surface attachment, to display various protein cargo at the bacterial cell surface and control extracellular release of the cargo in response to changing levels of the second messenger c-di-GMP. To accomplish this goal, we fused the protein cargo between the N-terminal retention module and C-terminal secretion signal of LapA and controlled surface localization of the cargo with natural signals known to stimulate or deplete c-di-GMP levels in P. fluorescens Pf0-1. We show this system can tolerate large cargo in excess of 500 amino acids, direct P. fluorescens Pf0-1 to surfaces it does not typically colonize, and program this microbe to sequester the toxic medal cadmium.

Evidence-based strategy for dental biofilms: Current evidence of mouthwashes on dental biofilm and gingivitis.

SummaryTherapeutic mouthwash (MW) is an adjunctive tool along with a regular oral hygiene routine of daily tooth brushing and daily flossing. Previous systematic reviews have demonstrated that it is effective against dental biofilm and gingival inflammation, for prevention of dental caries, and for managing one’s bad breath condition according to the active ingredients. MWs prevent the microorganisms from bacterial adhesion that corresponds to the initial step in biofilm formation.This review summarized the current state of evidence such as anti-biofilm, anti-gingivitis and cariostatic properties of MWs by evaluating systematic reviews from the past six years. The anti-biofilm property has been proven to be effective, with strong evidence of three main clinical efficacies.The most commonly studied active agent was chlorhexidine gluconate (CHX), followed by essential oil (EO) and cetylpyridinium chloride. All the systematic reviews are in complete agreement that CHX and EO provide statistically significant improvements in terms of plaque and gingival indices. These effects have held up over the years as the number of studies has increased. While the use of fluoride MW is proven to be effective in improving the oral health of both children and adults, the quality of evidence is still regarded as low.

New Thiazole Nortopsentin Analogues Inhibit Bacterial Biofilm Formation.

New thiazole nortopsentin analogues were conveniently synthesized and evaluated for their activity as inhibitors of biofilm formation of relevant Gram-positive and Gram-negative pathogens. All compounds were able to interfere with the first step of biofilm formation in a dose-dependent manner, showing a selectivity against the staphylococcal strains. The most active derivatives elicited IC50 values against Staphylococcus aureus ATCC 25923, ranging from 0.40–2.03 µM. The new compounds showed a typical anti-virulence profile, being able to inhibit the biofilm formation without affecting the microbial growth in the planktonic form.

Differential Production of Psl in Planktonic Cells Leads to Two Distinctive Attachment Phenotypes in Pseudomonas aeruginosa.

Exponentially growing bacteria in a well-mixed planktonic culture are generally assumed to be physiologically and phenotypically uniform and distinct from their genetically identical counterparts living in biofilms. Using a combination of high spatiotemporal microscopy and a bacterial tracking algorithm, in this study, we showed that planktonic cells of Pseudomonas aeruginosa differently attached to surfaces even when they remained in the exponential phase. We consistently observed that fast- and slow-attaching phenotypes coexist in planktonic cells, regardless of their growth phase. Furthermore, we found that (i) the distinct attaching phenotypes of planktonic cells resulted from the differential production of Psl and (ii) the RsmYZ/RsmA signaling pathway mainly regulated the differential production of Psl. Our results indicate that the differential production of Psl in P. aeruginosa plays a significant role in biofilm development and formation.IMPORTANCE The attachment of planktonic cells to surfaces is the first and most crucial step in biofilm formation. In this paper, we show that planktonic cells of Pseudomonas aeruginosa differently attach to surfaces. Typically, in the later exponential phase, approximately 80% of the cells can quickly attach to surfaces within 15 min, whereas approximately 20% of the cells slowly attach to surfaces, which greatly affects the initial stage of biofilm formation in the presence of flows. This is because fast-attaching cells are more likely to attach on surfaces to form microcolonies, whereas slow-attaching cells are more likely to remain in the mobile phase. This scenario is different from the previous understanding of biofilm formation in the initial stage, in which planktonic cells were thought to uniformly attach to surfaces. Most notably, the results of this study show that the different attachment manner of planktonic cells to surfaces affects the subsequent stages of biofilm formation. This research highlights that the phenotypic variations in planktonic cells plays significant roles in various stages of biofilm formation.

Antibacterial and antibiofouling clay nanotube-silicone composite.

IntroductionInvasive medical devices are used in treating millions of patients each day. Bacterial adherence to their surface is an early step in biofilm formation that may lead to infection, health complications, longer hospital stays, and death. Prevention of bacterial adherence and biofilm development continues to be a major healthcare challenge. Accordingly, there is a pressing need to improve the anti-microbial properties of medical devices.Materials and MethodsPolydimethylsiloxane (PDMS) was doped with halloysite nanotubes (HNTs), and the PDMS-HNT composite surfaces were coated with PDMS-b-polyethylene oxide (PEO) and antibacterials. The composite material properties were examined using SEM, energy dispersive spectroscopy, water contact angle measurements, tensile testing, UV-Vis spectroscopy, and thermal gravimetric analysis. The antibacterial potential of the PDMS-HNT composites was compared to commercial urinary catheters using cultures of E. coli and S. aureus. Fibrinogen adsorption studies were also performed on the PDMS-HNT-PEO composites.ResultsHNT addition increased drug load during solvent swelling without reducing material strength. The hydrophilic properties provided by PEO were maintained after HNT addition, and the composites displayed protein-repelling properties. Additionally, composites showed superiority over commercial catheters at inhibiting bacterial growth.ConclusionPDMS-HNT composites showed superiority regarding their efficacy at inhibiting bacterial growth, in comparison to commercial antibacterial catheters. Our data suggest that PDMS-HNT composites have potential as a coating material for anti-bacterial invasive devices and in the prevention of institutional-acquired infections.