Subsequent Biofilm Development Research Articles

An investigation into the effects of prosthesis relevant surfaces on the formation of Staphylococcus aureus biofilms

The aim of this study was to investigate the effects prosthesis relevant surfaces and finishes have on the formation of Staphylococcus aureus (S. aureus) biofilms, the leading causative pathogenic bacteria of periprosthetic joint infection (PJI). Microbiological biofilm analysis was conducted to quantify S. aureus biofilm growth on prosthesis relevant surface coatings and finishes. Through the use of a CDC Biofilm Reactor (CDC-BR), biofilms were grown under constant shear conditions on three different titanium surface finishes, including Plasma Spray (PS), Grit Blasted (GB) and an As-fabricated (AF) surface used for comparative and control purposes. Advanced metrological techniques were applied to characterise each surface. This advanced approach to surface characterisation, including functional volume parameters has been carried out to provide a detailed quantifiable description of the surface and one that better relates to the nature of growth upon a surface. Results show that the PS surface exhibits a significant increase in biofilm growth in comparison to the GB and AF surfaces, with the AF surface showing the lowest amount of biofilm growth. Additionally, the morphology of the features of the PS coating allows for the biofilm accumulation to flourish in the re-entrant features across its topography. These findings highlight the difficulties of biofilm eradication and further complicate the design process of prosthetics, where features implemented to promote osseointegration simultaneously offer favourable locations for bacterial cell attachment and subsequent biofilm development, leading potentially, to PJI. There is a general consensus throughout literature regarding an apparent trend between an increase in Sa and an increase in biofilm formation (Zheng et al 2021 Frontiers in Bioengineering and Biotechnology 9 643722; Bridgens et al 2015 Surface and Coatings Technology 284; James et al 2019 Aesthetic Plast. Surg. 43 490–497). This study has shown that whilst this may generally be the case, the location of this surface area increase within the topography may be a more important observation.

Manganese Acts as an Environmental Inhibitor of Pseudomonas aeruginosa Biofilm Development by Inducing Dispersion and Modulating c-di-GMP and Exopolysaccharide Production via RbdA.

The opportunistic human pathogen Pseudomonas aeruginosa causes chronic infections that involve multicellular aggregates called biofilms. Biofilm formation is modulated by the host environment and the presence of cues and/or signals, likely affecting the pool of the bacterial second messenger cyclic diguanylate monophosphate (c-di-GMP). The manganese ion Mn2+ is a divalent metal cation that is essential for pathogenic bacterial survival and replication during the infection in a host organism. In this study, we investigated how Mn2+ alters P. aeruginosa biofilm formation via the regulation of c-di-GMP levels. Exposure to Mn2+ was found to temporally enhance attachment but impair subsequent biofilm development, apparent by reduced biofilm biomass accumulation and lack of microcolony formation due to the induction of dispersion. Moreover, exposure to Mn2+ coincided with reduced production of the exopolysaccharides Psl and Pel, decreased transcriptional abundance of pel and psl, and decreased levels of c-di-GMP. To determine whether the effect of Mn2+ was linked to the activation of phosphodiesterases (PDEs), we screened several PDE mutants for Mn2+-dependent phenotypes (attachment and polysaccharide production) as well as PDE activity. The screen revealed that the PDE RbdA is activated by Mn2+ and is responsible for Mn2+-dependent attachment, inhibition of Psl production, and dispersion. Taken together, our findings suggest Mn2+ is an environmental inhibitor of P. aeruginosa biofilm development that acts through the PDE RbdA to modulate c-di-GMP levels, thereby impeding polysaccharide production and biofilm formation but enhancing dispersion. IMPORTANCE While diverse environmental conditions such as the availability of metal ions have been shown to affect biofilm development, little is known about the mechanism. Here, we demonstrate that Mn2+ affects Pseudomonas aeruginosa biofilm development by stimulating phosphodiesterase RbdA activity to reduce the signaling molecule c-di-GMP levels, thereby hindering polysaccharide production and biofilm formation but enhancing dispersion. Our findings demonstrate that Mn2+ acts as an environmental inhibitor of P. aeruginosa biofilms, further suggesting manganese to be a promising new antibiofilm factor.

In it together: Candida-bacterial oral biofilms and therapeutic strategies.

Under natural environmental settings or in the human body, the majority of microorganisms exist in complex polymicrobial biofilms adhered to abiotic and biotic surfaces. These microorganisms exhibit symbiotic, mutualistic, synergistic, or antagonistic relationships with other species during biofilm colonization and development. These polymicrobial interactions are heterogeneous, complex and hard to control, thereby often yielding worse outcomes than monospecies infections. Concerning fungi, Candida spp., in particular, Candida albicans is often detected with various bacterial species in oral biofilms. These Candida-bacterial interactions may induce the transition of C. albicans from commensal to pathobiont or dysbiotic organism. Consequently, Candida-bacterial interactions are largely associated with various oral diseases, including dental caries, denture stomatitis, periodontitis, peri-implantitis, and oral cancer. Given the severity of oral diseases caused by cross-kingdom consortia that develop hard-to-remove and highly drug-resistant biofilms, fundamental research is warranted to strategically develop cost-effective and safe therapies to prevent and treat cross-kingdom interactions and subsequent biofilm development. While studies have shed some light, targeting fungal-involved polymicrobial biofilms has been limited. This mini-review outlines the key features of Candida-bacterial interactions and their impact on various oral diseases. In addition, current knowledge on therapeutic strategies to target Candida-bacterial polymicrobial biofilms is discussed.

Nutrient starvation intensifies chlorine disinfection-stressed biofilm formation

Bacterial surface attachment and subsequent biofilm expansion represent an essential adaptation to environmental signals and stresses, which are of great concern for many natural and engineered ecosystems. Yet the underlying mechanisms and driving forces of biofilm formation in a chlorinated and nutrient-restricted system remain sketchy. In this study, we coupled an experimental investigation and modeling simulation to understand how chlorination and nutrient limitation conspire to form biofilm using Pseudomonas aeruginosa as a model bacterium. Experimental results showed that moderate chlorination at 1.0 mg/L led to biofilm development amplified to 2.6 times of those without chlorine, while additional nutrient limitation (of 1/50-diluted or 0.4 g/L LB broth culture) achieved 4.6 times increment as compared to those of undiluted scenarios (of 20 g/L LB broth culture) with absence of chlorination after 24 h exposure. Meanwhile, intermediate chlorination stimulated instant flagellar motility and subsequently extracellular polymeric substances (EPS) secretion, particularly under limited nutrient condition (of 1/50-diluted or 0.4 g/L LB broth culture) that retarded chlorine consumption and provoked bacterial nutrient-limitation response. From our simulations, chlorine and resource levels along with associated spatio-temporal variations collectively drove bacterial cell movement and EPS excretion. Our results demonstrated that restraining nutrient intensified chlorination-excited cell movement and EPS production that reinforced biological and cell-surface interactions, thereby encouraging bacterial surface attachment and subsequent biofilm development. The findings provide the insights into the linkage of disinfectant and nutrient-regulated bacterial functional responses with consequent micro-habitats and biofilm dynamics.

A Selection of Platforms to Evaluate Surface Adhesion and Biofilm Formation in Controlled Hydrodynamic Conditions.

The early colonization of surfaces and subsequent biofilm development have severe impacts in environmental, industrial, and biomedical settings since they entail high costs and health risks. To develop more effective biofilm control strategies, there is a need to obtain laboratory biofilms that resemble those found in natural or man-made settings. Since microbial adhesion and biofilm formation are strongly affected by hydrodynamics, the knowledge of flow characteristics in different marine, food processing, and medical device locations is essential. Once the hydrodynamic conditions are known, platforms for cell adhesion and biofilm formation should be selected and operated, in order to obtain reproducible biofilms that mimic those found in target scenarios. This review focuses on the most widely used platforms that enable the study of initial microbial adhesion and biofilm formation under controlled hydrodynamic conditions—modified Robbins devices, flow chambers, rotating biofilm devices, microplates, and microfluidic devices—and where numerical simulations have been used to define relevant flow characteristics, namely the shear stress and shear rate.

Revisiting the Microscopic Processes of Biofilm Formation on Organic Carriers: A Study under Variational Shear Stresses.

The microscopic process of biofilm development on carriers is critical for interfacial regulation of biofilms in attached-growth wastewater treatment. However, the process under shear stress has not been well understood. The study purposed to revisit the processes of biofilm formation on organic carriers under different shear stresses with special highlights on bacterial reversible adhesion and pioneers in the microbial community. Biofilm formation on high-density polyethylene, polyamide, acrylonitrile butadiene styrene plastic, polyvinyl chloride, and polycarbonate carriers under shear stresses ranging from 1.0 to 2.5 Pa was investigated using Couette-Taylor reactors. Employing extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) theory, the bacterial reversible adhesion regions ranging from 3.74 ± 0.20 to 5.51 ± 0.24 nm on an organic carrier were quantified for the first time, elucidating significant differences among different carriers (p < 0.01). The colonization of pioneers in the microbial community was significantly altered by shear stress rather than carrier properties (p < 0.01). In particular, the diversity of the biofilm microbial community was pronouncedly enhanced by a higher shear stress (p < 0.01). XDLVO analysis suggested that extracellular polymeric substances had a negative feedback on subsequent microbial adhesion and biofilm development, especially the transition from reversible to irreversible bacterial adhesion. This study contributed to a better understanding of the biofilm formation process at the microscopic scale and shed light on micro-interfacial manipulation for biofilm accumulation or renewal.

A curcumin-sophorolipid nanocomplex inhibits Candida albicans filamentation and biofilm development

Candida albicans is an opportunistic fungal pathogen that is highly resistant to contemporary antifungals, due to their biofilm lifestyle. The ability of C. albicans to invade human tissues is due to its filamentation. Therefore, inhibition of biofilms and filamentation of the yeast are high value targets to develop the next-generation antifungals. Curcumin (CU) is a natural polyphenol with excellent pharmacological attributes, but limitations such as poor solubility, acid, and enzyme tolerance have impeded its practical utility. Sophorolipids (SL) are biologically-derived surfactants that serve as efficient carriers of hydrophobic molecules such as curcumin into biofilms. Here, we synthesised a curcumin-sophorolipid nanocomplex (CUSL), and comprehensively evaluated its effects on C. albicans biofilms and filamentation. Our results demonstrated that sub-inhibitory concentration of CUSL (9.37 μg/mL) significantly inhibited fungal adhesion to substrates, and subsequent biofilm development, maturation, and filamentation. This effect was associated with significant downregulation of a select group of biofilm, adhesins, and hyphal regulatory genes. In conclusion, the curcumin-sophorolipid nanocomplex is a potent inhibitor of the two major virulence attributes of C. albicans, biofilm formation and filamentation, thus highlighting its promise as a putative anti-fungal agent with biofilm penetrative potential.

Bacillus subtilis extracellular polymeric substances conditioning layers inhibit Escherichia coli adhesion to silicon surfaces: A potential candidate for interfacial antifouling additives.

Biofouling on material surfaces is a ubiquitous problem in a variety of fields. In aqueous environments, the process of biofouling initiates with the formation of a layer of macromolecules called the conditioning layer on the solid-liquid interface, followed by the adhesion and colonization of planktonic bacteria and the subsequent biofilm development and maturation. In this study, the extracellular polymeric substances (EPS) secreted by Bacillus subtilis were collected and used to prepare conditioning layers on inert surfaces. The morphologies and antifouling performances of the EPS conditioning layers were investigated. It was found that the initial adhesion of Escherichia coli was inhibited on the surfaces precoated with EPS conditioning layers. To further explore the underlying antifouling mechanisms of the EPS conditioning layers, the respective roles of two constituents of B. subtilis EPS (γ-polyglutamic acid and surfactin) were investigated. This study has provided the possibility of developing a novel interfacial antifouling additive with the advantages of easy preparation, nontoxicity, and environmental friendliness.

Fish Bone Derived Bi-Phasic Calcium Phosphate Coatings Fabricated by Pulsed Laser Deposition for Biomedical Applications.

We report on new biomaterials with promising bone and cartilage regeneration potential, from sustainable, cheap resources of fish origin. Thin films were fabricated from fish bone-derived bi-phasic calcium phosphate targets via pulsed laser deposition with a KrF * excimer laser source (λ = 248 nm, τFWHM ≤ 25 ns). Targets and deposited nanostructures were characterized by SEM and XRD, as well as by Energy Dispersive X-ray (EDX) and FTIR spectroscopy. Films were next assessed in vitro by dedicated cytocompatibility and antimicrobial assays. Films were Ca-deficient and contained a significant fraction of β-tricalcium phosphate apart from hydroxyapatite, which could contribute to an increased solubility and an improved biocompatibility for bone regeneration applications. The deposited structures were biocompatible as confirmed by the lack of cytotoxicity on human gingival fibroblast cells, making them promising for fast osseointegration implants. Pulsed laser deposition (PLD) coatings inhibited the microbial adhesion and/or the subsequent biofilm development. A persistent protection against bacterial colonization (Escherichia coli) was demonstrated for at least 72 h, probably due to the release of the native trace elements (i.e., Na, Mg, Si, and/or S) from fish bones. Progress is therefore expected in the realm of multifunctional thin film biomaterials, combining antimicrobial, anti-inflammatory, and regenerative properties for advanced implant coatings and nosocomial infections prevention applications.

Plant components affect bacterial biofilms development by altering their cell surface physicochemical properties: a predictability study using Actinomyces naeslundii.

We hypothesized that the initial events leading to biofilm formation by bacteria, in general, are predominantly mediated by cell surface physicochemical interactions, and that natural products can impact the process by altering cell surface physicochemical properties. We exemplified this phenomenon using Actinomyces naeslundii as the model organism, and using tea products to modify its cell surface physicochemical properties. To test the hypothesis, a non-linear multiple regression model incorporating a normal distribution curve was constructed to explain the impact of tea extracts on the physiochemical processes of biofilm formation by A. naeslundii. The model utilized tea extract-induced changes in cell surface physicochemical properties as independent variables, and the corresponding biofilm formation as a dependent variable. Five different tea extracts were used to treat A. naeslundii, and their impact on the cell surface hydrophobicity, charge, auto-aggregation, attachment and biofilm formation on four different hard surfaces were measured and the data were used to construct the model. The established model was then tested in independent experiments involving other plant extracts and purified phytochemicals. Experimental results showed that the tea extracts significantly reduced cell surface hydrophobicity (by up to 21.3%), increased cell surface charge and auto-aggregation (by up to 4.5mV and 14.9%, respectively), inhibited attachment (by 0.6-2.5 log CFU cm-2) and affected biofilm formation (by up to 0.6 log CFU cm-2). The model indicated that both cell surface hydrophobicity and charge played an important role in bacterial auto-aggregation and attachment, and that the latter two phenomena significantly correlated with subsequent biofilm development. The accuracy of the model construct was approximately 64%. This modelling approach can be employed for other microbial colonization systems to predict biofilm formation, and to study the impact of cell surface physicochemical properties in biofilm development.

Intravital Multiphoton Examination of Implant-Associated Staphylococcus aureus Biofilm Infection.

Bacterial infections associated with implanted medical devices represents a healthcare crisis due to their persistence, antibiotic tolerance, and immune avoidance. Indwelling devices are rapidly coated with host plasma and extracellular matrix proteins which can then be exploited by bacterial pathogens for adherence and subsequent biofilm development. Our understanding of the host-pathogen interface that determines the fate of biofilm-mediated infections is limited to the experimental models employed by laboratories studying these organisms. Current in vivo models of biofilm-mediated infection, while certainly useful, are typically limited to end-point analyses of bacterial burden enumeration, immune cell profiling, and cytokine/chemokine analysis. Thus, with these models, the complex, real-time assessment of biofilm development and innate immune cell activity remains imperceptible. Here, we describe a novel murine biofilm infection model employing time-lapse intravital multiphoton microscopy which permits concurrent and real-time visualization of Staphylococcus aureus biofilm formation and immune cell activity. Using cell tracking, we found that S. aureus biofilms impede neutrophil chemotaxis, redirecting their migration patterns to prevent biofilm invasion. This approach is the first to directly examine device-associated biofilm development and host-pathogen interactions and will serve to both further our understanding of infection development and help reveal the effects of future antibiofilm treatment strategies.

Analyzing the inhibitory effect of metabolic uncoupler on bacterial initial attachment and biofilm development and the underlying mechanism

Metabolic uncouplers inhibit biofilm and biofouling formation in membrane bioreactor (MBR) systems, which have been considered as a potential biofouling control alternative. To better understand the inhibitory mechanism of uncoupler on biofouling, this study investigated the impact of the uncoupler 3, 3′, 4′, 5-tetrachlorosalicylanilide (TCS) on biofilm formation of B. subtilis in different development stages. Significant reductions in both the initial bacterial attachment stage and the subsequent biofilm development stage were caused by TCS at 100 μg/L. The motility of B. subtilis in semisolid medium was inhibited by TCS, which explicitly explained the reduction in initial bacterial attachment. Meanwhile, a reduction of extracellular polymeric substance (EPS) secretion owing to TCS suggested why biofilm development was suppressed. In addition, the fluorescent materials in tight-bound EPS (TB-EPS) and loose-bound EPS (LB-EPS) of Bacillus subtilis cultured in different TCS concentrations were distinguished and quantified by three-dimensional excitation-emission matrix (EEM) fluorescence spectroscopy coupled with parallel factor analysis (PARAFAC). The results of this study suggested that the biofilm inhibitory mechanism of the uncoupler was both a inhibition in bacterial motor ability and a reduction in EPS secretion.

Nanoporous anodic alumina reduces Staphylococcus biofilm formation.

Staphylococcus epidermidis and Staphylococcus aureus, two bacterial strains commonly associated with biofilm-related medical infections and food poisoning, can rapidly colonize biotic and abiotic surfaces. The present study investigates the ability of anodic alumina surfaces with nanoporous surface topography to minimize the attachment and biofilm formation mediated by these pathogenic bacterial strains. Early attachment and subsequent biofilm development were retarded on surfaces with nanopores of 15-25nm in diameter compared to surfaces with 50-100nm pore diameter and nanosmooth surfaces. After 30min of incubation in nutritive media, the biomass accumulation per unit surface area was 2·93±1·72µm3 µm-2 for the 15nm, 3·49±1·97µm3 µm-2 for the 25nm, as compared to 14·04±6·39µm3 µm-2 for the nanosmooth, 11·88±9·72µm3 µm-2 for the 50nm and 12·09±11·84µm3 µm-2 for the 100nm surfaces respectively. These findings suggest that anodic alumina with small size nanoscale pores could reduce the incidence of staphylococcal biofilms and infections, and shows promise as a material for a variety of medical applications and food contact surfaces. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper reports on a simple, robust and scientifically sound method to reduce attachment and biofilm formation by Staphylococcus aureus and Staphylococcus epidermidis to abiotic surfaces using a carefully designed nanoscale topography. This approach can help to reduce the incidence of staphylococcal biofilms and infections without imposing selective stresses on bacteria, thus preventing the creation of resistant strains.

Penetrating the air-liquid interface is the key to colonization and wrinkly spreader fitness.

In radiating populations of Pseudomonas fluorescens SBW25, adaptive wrinkly spreader (WS) mutants are able to gain access to the air-liquid (A-L) interface of static liquid microcosms and achieve a significant competitive fitness advantage over other non-biofilm-forming competitors. Aerotaxis and flagella-based swimming allows SBW25 cells to move into the high-O2 region located at the top of the liquid column and maintain their position by countering the effects of random cell diffusion, convection and disturbance (i.e. physical displacement). However, wild-type cells showed significantly lower levels of enrichment in this region compared to the archetypal WS, indicating that WS cells employ an additional mechanism to transfer to the A-L interface where displacement is no longer an issue and a biofilm can develop at the top of the liquid column. Preliminary experiments suggest that this might be achieved through the expression of an as yet unidentified surface active agent that is weakly associated with WS cells and alters liquid surface tension, as determined by quantitative tensiometry. The effect of physical displacement on the colonization of the high-O2 region and A-L interface was reduced through the addition of agar or polyethylene glycol to increase liquid viscosity, and under these conditions the competitive fitness of the WS was significantly reduced. These observations suggest that the ability to transfer to the A-L interface from the high-O2 region and remain there without further expenditure of energy (through, for example, the deployment of flagella) is a key evolutionary innovation of the WS, as it allows subsequent biofilm development and significant population increase, thereby affording these adaptive mutants a competitive fitness advantage over non-biofilm-forming competitors located within the liquid column.

Unravelling the core microbiome of biofilms in cooling tower systems

In this study, next generation sequencing and catalyzed reporter deposition fluorescence in situ hybridization, combined with confocal microscopy, were used to provide insights into the biodiversity and structure of biofilms collected from four full-scale European cooling systems. Water samples were also analyzed to evaluate the impact of suspended microbes on biofilm formation. A common core microbiome, containing members of the families Sphingomonadaceae, Comamonadaceae and Hyphomicrobiaceae, was found in all four biofilms, despite the water of each coming from different sources (river and groundwater). This suggests that selection of the pioneer community was influenced by abiotic factors (temperature, pH) and tolerances to biocides. Members of the Sphingomonadaceae were assumed to play a key role in initial biofilm formation. Subsequent biofilm development was driven primarily by light availability, since biofilms were dominated by phototrophs in the two studied ‘open’ systems. Their interactions with other microbial populations then shaped the structure of the mature biofilm communities analyzed.

Bioactive ZnO Coatings Deposited by MAPLE-An Appropriate Strategy to Produce Efficient Anti-Biofilm Surfaces.

Deposition of bioactive coatings composed of zinc oxide, cyclodextrin and cefepime (ZnO/CD/Cfp) was performed by the Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique. The obtained nanostructures were characterized by X-ray diffraction, IR microscopy and scanning electron microscopy. The efficient release of cefepime was correlated with an increased anti-biofilm activity of ZnO/CD/Cfp composites. In vitro and in vivo tests have revealed a good biocompatibility of ZnO/CD/Cfp coatings, which recommend them as competitive candidates for the development of antimicrobial surfaces with biomedical applications. The release of the fourth generation cephalosporin Cfp in a biologically active form from the ZnO matrix could help preventing the bacterial adhesion and the subsequent colonization and biofilm development on various surfaces, and thus decreasing the risk of biofilm-related infections.

Biocompatible cephalosporin-hydroxyapatite-poly(lactic-co-glycolic acid)-coatings fabricated by MAPLE technique for the prevention of bone implant associated infections

In this study we aimed to obtain functionalized thin films based on hydroxyapatite/poly(lactic-co-glycolic acid) (HAp/PLGA) containing ceftriaxone/cefuroxime antibiotics (ATBs) deposited by Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique. The prepared thin films were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-Ray diffraction (XRD), selected area electron diffraction (SAED), and infra red (IR) analysis. HAp/PLGA/ATBs thin films sustained the growth of human osteoblasts, proving their good biocompatibility. The microscopic evaluation and the culture-based quantitative assay of the E. coli biofilm development showed that the thin films inhibited the initial step of microbial attachment as well as the subsequent colonization and biofilm development on the respective surfaces. This study demonstrates that MAPLE technique could represent an appealing technique for the fabrication of antibiotics-containing polymeric implant coatings. The bioevaluation results recommend this type of surfaces for the prevention of bone implant microbial contamination and for the enhanced stimulation of the implant osseointegration process.

Microbial Surface Colonization and Biofilm Development in Marine Environments.

Biotic and abiotic surfaces in marine waters are rapidly colonized by microorganisms. Surface colonization and subsequent biofilm formation and development provide numerous advantages to these organisms and support critical ecological and biogeochemical functions in the changing marine environment. Microbial surface association also contributes to deleterious effects such as biofouling, biocorrosion, and the persistence and transmission of harmful or pathogenic microorganisms and their genetic determinants. The processes and mechanisms of colonization as well as key players among the surface-associated microbiota have been studied for several decades. Accumulating evidence indicates that specific cell-surface, cell-cell, and interpopulation interactions shape the composition, structure, spatiotemporal dynamics, and functions of surface-associated microbial communities. Several key microbial processes and mechanisms, including (i) surface, population, and community sensing and signaling, (ii) intraspecies and interspecies communication and interaction, and (iii) the regulatory balance between cooperation and competition, have been identified as critical for the microbial surface association lifestyle. In this review, recent progress in the study of marine microbial surface colonization and biofilm development is synthesized and discussed. Major gaps in our knowledge remain. We pose questions for targeted investigation of surface-specific community-level microbial features, answers to which would advance our understanding of surface-associated microbial community ecology and the biogeochemical functions of these communities at levels from molecular mechanistic details through systems biological integration.

Relationship between surface chemistry, biofilm structure, and electron transfer in Shewanella anodes.

A better understanding of how anode surface properties affect growth, development, and activity of electrogenic biofilms has great potential to improve the performance of bioelectrochemical systems such as microbial fuel cells. The aim of this paper was to determine how anodes with specific exposed functional groups (-N(CH3)3 (+), -COOH, -OH, and -CH3), created using ω-substituted alkanethiolates self-assembled monolayers attached to gold, affect the surface properties and functional performance of electrogenic Shewanella oneidensis MR-1 biofilms. A combination of spectroscopic, microscopic, and electrochemical techniques was used to evaluate how electrode surface chemistry influences morphological, chemical, and functional properties of S. oneidensis MR-1 biofilms, in an effort to develop improved electrode materials and structures. Positively charged, highly functionalized, hydrophilic surfaces were beneficial for growth of uniform biofilms with the smallest cluster sizes and intercluster diffusion distances, and yielding the most efficient electron transfer. The authors derived these parameters based on 3D morphological features of biofilms that were directly linked to functional properties of the biofilm during growth and that, during polarization, were directly connected to the efficiency of electron transfer to the anode. Our results indicate that substratum chemistry affects not only primary attachment, but subsequent biofilm development and bacterial physiology.

Escherichia coli adhesion, biofilm development and antibiotic susceptibility on biomedical materials.

The aim of this work was to test materials typically used in the construction of medical devices regarding their influence in the initial adhesion, biofilm development and antibiotic susceptibility of Escherichia coli biofilms. Adhesion and biofilm development was monitored in 12-well microtiter plates containing coupons of different biomedical materials--silicone (SIL), stainless steel (SS) and polyvinyl chloride (PVC)--and glass (GLA) as control. The susceptibility of biofilms to ciprofloxacin and ampicillin was assessed, and the antibiotic effect in cell morphology was observed by scanning electron microscopy. The surface hydrophobicity of the bacterial strain and materials was also evaluated from contact angle measurements. Surface hydrophobicity was related with initial E. coli adhesion and subsequent biofilm development. Hydrophobic materials, such as SIL, SS, and PVC, showed higher bacterial colonization than the hydrophilic GLA. Silicone was the surface with the greatest number of adhered cells and the biofilms formed on this material were also less susceptible to both antibiotics. It was found that different antibiotics induced different levels of elongation on E. coli sessile cells. Results revealed that, by affecting the initial adhesion, the surface properties of a given material can modulate biofilm buildup and interfere with the outcome of antimicrobial therapy. These findings raise the possibility of fine-tuning surface properties as a strategy to reach higher therapeutic efficacy.