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The acetogen Clostridium ljungdahlii is capable of syngas fermentation and microbial electrosynthesis. Biofilm formation could benefit both these applications, but was not yet reported for C. ljungdahlii. Biofilm formation does not occur under standard growth conditions, but attachment or aggregation could be induced by different stresses. The strongest biofilm formation was observed with the addition of sodium chloride. After 3 days of incubation, the biomass volume attached to a plastic surface was 20 times higher with than without the addition of 200 mM NaCl to the medium. The addition of NaCl also resulted in biofilm formation on glass, graphite and glassy carbon, the latter two being often used electrode materials for microbial electrosynthesis. Biofilms were composed of extracellular proteins, polysaccharides, as well as DNA, while pilus-like appendages were observed with, but not without, the addition of NaCl. A transcriptome analysis comparing planktonic (no NaCl) and biofilm (NaCl addition) cells showed that C. ljungdahlii coped with the salt stress by the upregulation of the general stress response, Na+ export and osmoprotectant accumulation. A potential role for poly-N-acetylglucosamines and D-alanine in biofilm formation was found. Flagellar motility was downregulated, while putative type IV pili biosynthesis genes were not expressed. Moreover, the gene expression analysis suggested the involvement of the transcriptional regulators LexA, Spo0A and CcpA in stress response and biofilm formation. This study showed that NaCl addition might be a valuable strategy to induce biofilm formation by C. ljungdahlii, which can improve the efficacy of syngas fermentation and microbial electrosynthesis applications.

The frog skin-derived peptide Temporin 1Tb (TB) has gained increasing attention as novel antimicrobial agent for the treatment of antibiotic-resistant and/or biofilm-mediated infections. Nevertheless, such a peptide possesses a preferential spectrum of action against Gram-positive bacteria. In order to improve the therapeutic potential of TB, the present study evaluated the antibacterial and antibiofilm activities of two TB analogs against medically relevant bacterial species. Of the two analogs, TB_KKG6A has been previously described in the literature, while TB_L1FK is a new analog designed by us through statistical-based computational strategies. Both TB analogs displayed a faster and stronger bactericidal activity than the parental peptide, especially against Gram-negative bacteria in planktonic form. Differently from the parental peptide, TB_KKG6A and TB_L1FK were able to inhibit the formation of Staphylococcus aureus biofilms by more than 50% at 12 μM, while only TB_KKG6A prevented the formation of Pseudomonas aeruginosa biofilms at 24 μM. A marked antibiofilm activity against preformed biofilms of both bacterial species was observed for the two TB analogs when used in combination with EDTA. Analysis of synergism at the cellular level suggested that the antibiofilm activity exerted by the peptide-EDTA combinations against mature biofilms might be due mainly to a disaggregating effect on the extracellular matrix in the case of S. aureus, and to a direct activity on biofilm-embedded cells in the case of P. aeruginosa. Both analogs displayed a low hemolytic effect at the active concentrations and, overall, TB_L1FK resulted less cytotoxic toward mammalian cells. Collectively, the results obtained demonstrated that subtle changes in the primary sequence of TB may provide TB analogs that, used alone or in combination with adjuvant molecules such as EDTA, exhibit promising features against both planktonic and biofilm cells of medically relevant bacteria.

The presence of bacteria in human milk has been acknowledged since the seventies. For a long time, microbiological analysis of human milk was only performed in case of infections and therefore the presence of non-pathogenic bacteria was yet unknown. During the last decades, the use of more sophisticated culture-dependent and -independent techniques, and the steady development of the -omic approaches are opening up the new concept of the ‘milk microbiome’, a complex ecosystem with a greater diversity than previously anticipated. In this review, possible mechanisms by which bacteria can reach the mammary gland (contamination versus active migration) are discussed. In addition, the potential roles of human milk for both infant and maternal health are summarised. A better understanding of the link between the milk microbiome and health benefit, the potential factors influencing this relationship and whether or not it can be influenced by nutrition is required to open new avenues in the field of pregnancy and lactation.

Biofilms are detrimental in many industrial and biomedical applications and prevention of biofilm formation has been a prime challenge for decades. Biofilms consist of communities of adhering bacteria, supported and protected by extracellular-polymeric-substances (EPS), the so-called house of biofilm organisms. EPS consists of water, proteins, polysaccharides and extracellular-DNA (eDNA). eDNA, being the longest molecule in EPS, connects the different EPS components and therewith holds an adhering biofilm together. eDNA is associated with bacterial cell surfaces by specific and non-specific mechanisms, mediating binding of other biopolymers in EPS. eDNA therewith assists in facilitating adhesion, aggregation and maintenance of biofilm structure. Here, a new method is described to prevent biofilm formation on surfaces by applying a DNase I enzyme coating to polymethylmethacrylate, using dopamine as an intermediate. The intermediate coupling layer and final DNase I coating are characterized by water-contact-angle measurements and X-ray photoelectron-spectroscopy. The DNase I coating strongly reduces adhesion of Staphylococcus aureus (95%) and Pseudomonas aeruginosa (99%) and prevents biofilm formation up to 14 h, without affecting mammalian cell adhesion and proliferation. Also agarose-gel-electrophoresis indicates loss of enzyme activity between 8 and 24 h. This duration however, is similar to many local antibiotic-delivery devices, which makes it an ideal coating for biomaterial implants and devices, known to fail due to biofilm formation with disastrous consequences for patients and high costs to the healthcare system. With threatening increases in antibiotic resistance, the DNase I coating may provide a timely, potent new approach to biofilm prevention on biomaterial implants and devices.

Biofilms are widespread in nature and constitute an important strategy implemented by microorganisms to survive in sometimes harsh environmental conditions. They can be beneficial or have a negative impact particularly when formed in industrial settings or on medical devices. As such, research into the formation and elimination of biofilms is important for many disciplines. Several new methodologies have been recently developed for, or adapted to, biofilm studies that have contributed to deeper knowledge on biofilm physiology, structure and composition. In this review, traditional and cutting-edge methods to study biofilm biomass, viability, structure, composition and physiology are addressed. Moreover, as there is a lack of consensus among the diversity of techniques used to grow and study biofilms. This review intends to remedy this, by giving a critical perspective, highlighting the advantages and limitations of several methods. Accordingly, this review aims at helping scientists in finding the most appropriate and up-to-date methods to study their biofilms., The authors would like to acknowledge the support from the EU COST Action BacFoodNet FA1202.

BackgroundProsthetic meshes are increasingly popular in abdominal wall reconstructive surgery owing to a reduction in hernia recurrence rate. Individual meshes have been evaluated with respect to the formation of infectious biofilms, but no comprehensive comparison yet exists. The aim of this study was to evaluate the effect of the material and morphology of surgical meshes on biofilm growth.MethodsBiofilms of a coagulase-negative staphylococcus, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa were grown in vitro on different meshes. Intact biofilms were stained fluorescently and evaluated with confocal laser scanning microscopy to distinguish dead from live bacteria and slime.ResultsNumbers of adhering bacteria and the amount of slime in the biofilm were dependent on the morphology of the mesh. Hydrophobicity and the presence of niches in multifilament meshes contributed most to increased biofilm growth.ConclusionThe morphological properties of surgical meshes as well as their hydrophobicity influence bacterial growth and slime production. Differences in slime production may explain why the effectiveness of antibiotic treatment varies for different types of mesh.

Item does not contain fulltext The fate of secondary biomaterial implants was determined by bio-optical imaging and plate counting, after antibiotic treatment of biomaterials-associated-infection (BAI) and surgical removal of an experimentally infected, primary implant. All primary implants and tissue samples from control mice showed bioluminescence and were culture-positive. In an antibiotic treated group, no bioluminescence was detected and only 20% of all primary implants and no tissue samples were culture-positive. After revision surgery, bioluminescence was detected in all control mice. All the implants and 80% of all tissue samples were culture-positive. In contrast, in the antibiotic treated group, 17% of all secondary implants and 33% of all tissue samples were culture-positive, despite antibiotic treatment. The study illustrates that due to the BAI of a primary implant, the infection risk of biomaterial implants is higher in revision surgery than in primary surgery, emphasizing the need for full clearance of the infection, as well as from surrounding tissues prior to implantation of a secondary implant. 01 oktober 2010

Oral biofilm can never be fully removed by oral hygiene measures. Biofilm left behind after brushing is often left behind on the same sites and exposed multiple times to antimicrobials from toothpastes and mouthrinses, after which removal becomes increasingly difficult. On the basis of this observation, we hypothesize that oral bacteria adhering to salivary conditioning films become more difficult to remove after adsorption of antimicrobials due to stiffening of their adhesive bond. To verify this hypothesis, bacteria adhering to bare and saliva-coated glass were exposed to 3 different mouthrinses, containing chlorhexidine-digluconate, cetylpyridinium-chloride, or amine-fluoride, after which bacterial vibration spectroscopy was carried out or a liquid-air interface was passed over the adhering bacteria to stimulate their detachment. Brownian motion–induced nanoscopic vibration amplitudes of 4 oral streptococcal strains, reflecting their bond stiffness, decreased after exposure to mouthrinses. Concurrently, the percentage detachment of adhering bacteria upon the passage of a liquid-air interface decreased after exposure to mouthrinses. A buffer control left both vibration amplitudes and detachment percentages unaffected. Exposure to either of the selected mouthrinses yielded more positively charged bacteria by particulate microelectrophoresis, suggesting antimicrobial adsorption to bacterial cell surface components. To rule out that exposure of adhering bacteria to the mouthrinses stimulated polysaccharide production with an impact on their detachment, Fourier transform infrared spectroscopy was carried out on bacteria adhering to an internal reflection element, prior to and after exposure to the mouthrinses. Infrared absorption band areas indicated no significant change in amount of polysaccharides after exposure of adhering bacteria to mouthrinses, but wave number shifts demonstrated stiffening of polysaccharides in the bond, as a result of antimicrobial adsorption to the bacterial cell surface and in line with changes in surface charge. Clinically, these findings suggest that accumulation of oral biofilm exposed to antimicrobials should be prevented (interdental cleaning aids, floss use), as removal becomes progressively more difficult upon multiple exposures.

Background: Staphylococcal species are the most common organisms causing prosthetic mesh infections, however, infections due to rapidly growing mycobacteria are increasing. This study evaluates the resistance of biomaterial for abdominal wall prostheses against the development of postoperative infection in a rat model. Material and methods: In 75 rats, we intramuscularly implanted three different types of prostheses: (1) low-density polypropylene monofilament mesh (PMM), (2) high-density PMM, and (3) a composite prosthesis composed of low-density PMM and a nonporous hydrophilic film. Meshes were inoculated with a suspension containing 108 colony-forming units of Staphylococcus aureus, Staphylococcus epidermidis, Mycobacterium fortuitum, or Mycobacterium abscessus before wound closure. Animals were sacrificed on the eighth day postoperatively for clinical evaluation, and the implants were removed for bacteriologic analyses. Results: Prostheses infected with S aureus showed a higher bacterial viability, worse integration, and clinical outcome compared with infection by other bacteria. Composite prostheses showed a higher number of viable colonies of both M fortuitum and Staphylococcus spp., with poorer integration in host tissue. However, when the composite prosthesis was infected with M abscessus, a lower number of viable bacteria were isolated and a better integration was observed compared with infection by other bacteria. Conclusions: Considering M abscessus, a smaller collagen-free contact surface shows better resistance to infection, however, depending on the type of bacteria, prostheses with a large surface, and covered with collagen shows reduced resistance to infection, worse integration, and worse clinical outcome. (C) 2016 Elsevier Inc. All rights reserved.

Biofilm has been recognized as a well-protected form of living for bacteria, contributing to bacterial pathogenicity, particularly for opportunistic species. Biofilm-associated infections are marked by their persistence. Extensive research has been devoted to the formation and composition of biofilms. The immune response against biofilms remains rather unexplored, but there is the notion that bacteria within a biofilm are protected from host defences. Here we glance at the mechanisms by which neutrophils recognize and face biofilms in implant infections and discuss the implications of this interplay, as well as speculate on its significance.

Objectives The purpose of this study was to assess the efficacy of different cleansing agents in killing mixed species biofilms on silicone facial prostheses. Materials and methods Two bacterial and three yeast strains, isolated from silicone facial prostheses, were selected for the mixed species biofilms. A variety of agents used to clean facial prostheses were employed, viz., antibacterial soap, essential-oil-containing mouth rinse, ethanol 27 %, chlorhexidine mouth rinse, and buttermilk. Colony forming units (CFUs) and live/dead staining were analyzed to assess the efficacy of these cleansing agents against 24-h and 2-week biofilms and regrown biofilms on silicone samples. Results Chlorhexidine was the most effective cleansing agent. Chlorhexidine killed 8 log unit CFUs (>99.99 % killing) in a 24-h biofilm and 5 log unit CFUs (>99.99 % killing) in 2-week biofilms. Also, after regrowth and repeated treatment of the biofilm, chlorhexidine was the most effective cleansing agent showing no detectable CFUs. The essential-oil-containing mouth rinse (containing 26.9 % ethanol) showed a similar efficacy as ethanol (27 %) alone. Antibacterial soap and buttermilk were the least effective agents tested. Conclusions Chlorhexidine showed the highest reduction in CFUs in 24-h, 2-week, and regrown mixed species biofilm of microorganisms isolated from silicone facial prostheses. Clinical relevance Chlorhexidine mouth rinse (easy obtainable and relatively cheap) is very effective in killing bacteria and yeast present in biofilms on silicone facial prostheses. When applied on a regular basis, cleansing a facial prosthesis with chlorhexidine will presumably increase its lifetime and reduce skin irritations.

Bile acid resistance by Lactococcus lactis depends on the ABC-type multidrug transporter LmrCD. Upon deletion of the lmrCD genes, cells can reacquire bile acid resistance upon prolonged exposure to cholate, yielding the Δ lmrCD r strain. The resistance mechanism in this strain is non-transporter based. Instead, cells show a high tendency to flocculate, suggesting cell surface alterations. Contact angle measurements demonstrate that the Δ lmrCD r cells are equipped with an increased cell surface hydrophilicity compared to those of the parental and wild-type strains, while the surface hydrophilicity is reduced in the presence of cholate. Δ lmrCD r cells are poor in biofilm formation on a hydrophobic polystyrene surface, but in the presence of subinhibitory concentrations of cholate, biofilm formation is strongly stimulated. Biofilm cells show an enhanced extracellular polymeric substance production and are highly resistant to bile acids. These data suggest that non-transporter-based cholate resistance in L. lactis is due to alterations in the cell surface that stimulate cells to form resistant biofilms.

This review presents the current state of Bioluminescence and Fluorescent Imaging technologies (BLI and FLI) as applied to Biomaterial-Associated Infections (BAI). BLI offers the opportunity to observe the in vivo course of BAI in small animals without the need to sacrifice animals at different time points after the onset of infection. BLI is highly dependent on the bacterial cell metabolism which makes BLI a strong reporter of viable bacterial presence. Fluorescent sources are generally more stable than bioluminescent ones and specifically targeted, which renders the combination of BLI and FLI a promising tool for imaging BAI. The sensitivity and spatial resolution of both imaging tools are, however, dependent on the imaging system used and the tissue characteristics, which makes the interpretation of images, in terms of the location and shape of the illuminating source, difficult. Tomographic reconstruction of the luminescent source is possible in the most modern instruments, enabling exact localization of a colonized implant material, spreading of infecting organisms in surrounding tissue and immunological tissue reactions. BLI studies on BAI have successfully distinguished between different biomaterials with respect to the development and clearance of BAI in vivo, simultaneously reducing animal use and experimental variation. It is anticipated that bio-optical imaging will become an indispensable technology for the in vivo evaluation of antimicrobial coatings. (C) 2009 Elsevier Ltd. All rights reserved.

Curing biomaterial-associated infection (BAI) frequently includes antibiotic treatment, implant removal and re-implantation. However, revision implants are at a greater risk of infection as they may attract bacteria from their infected surroundings. Polymer brush-coatings attract low numbers of bacteria, but the virtue of polymer brush-coatings in vivo has seldom been investigated. Here, we determine the possible benefits of polymer brush-coated versus pristine silicone rubber in revision surgery, using a murine model.BAI was induced in 26 mice by subcutaneous implantation of silicone rubber discs with a biofilm of Staphylococcus aureus Xen29. During the development of BAI, half of the mice received rifampicin/vancomycin treatment. After 5 days, the infected discs were removed from all mice, and either a polymer brush-coated or pristine silicone rubber disc was re-implanted. Revision discs were explanted after 5 days, and the number of cfu cultured from the discs and the surrounding tissue was determined.None of the polymer brush-coated discs after antibiotic treatment appeared colonized by staphylococci, whereas 83% of the pristine silicone rubber discs were re-infected. Polymer brush-coated discs also showed reduced colonization rates in the absence of antibiotic treatment when compared with pristine silicone rubber discs. Tissue surrounding the discs was culture-positive in all cases.Polymer brush-coatings are less prone to re-infection than pristine silicone rubber when used in revision surgery, i.e. when implanted in a subcutaneous pocket infected by a staphylococcal BAI. Antibiotic pre-treatment during the development of BAI hardly had any effect in preventing the colonization of pristine silicone rubber.

11 p.-6 fig., Streptococcus pneumoniae is a frequent member of the microbiota of the human nasopharynx. Colonization of the nasopharyngeal tract is a first and necessary step in the infectious process and often involves the formation of sessile microbial communities by this human pathogen. The ability to grow and persist as biofilms is an advantage for many microorganisms, because biofilm-grown bacteria show reduced susceptibility to antimicrobial agents and hinder recognition by the immune system. The extent of host protection against biofilm-related pneumococcal disease has not been determined yet. Using pneumococcal strains growing as planktonic cultures or as biofilms, we have investigated the recognition of S. pneumoniae by the complement system and its interactions with human neutrophils. Deposition of C3b, the key complement component, was impaired on S. pneumoniae biofilms. In addition, binding of C-reactive protein and the complement component C1q to the pneumococcal surface was reduced in biofilm bacteria, demonstrating that pneumococcal biofilms avoid the activation of the classical complement pathway. In addition, recruitment of factor H, the downregulator of the alternative pathway, was enhanced by S. pneumoniae growing as biofilms. Our results also show that biofilm formation diverts the alternative complement pathway activation by a PspC-mediated mechanism. Furthermore, phagocytosis of pneumococcal biofilms was also impaired. The present study confirms that biofilm formation in S. pneumoniae is an efficient means of evading both the classical and the PspC-dependent alternative complement pathways the host immune system., This work was supported by grants SAF2009-10824 and SAF2012-39444-C02-01/02 from Dirección General de Investigación Científica y Técnica and MINECO, respectively.

International audience; Aims The purpose of this study was to quantify the extracellular matrix of Listeria monocytogenes biofilm. A preliminary study was carried out to establish a relationship between phylogenetic lineage of 27 strains and their ability to form biofilm in various conditions. Methods and Results Biofilm formation on microtitre plates of 27 strains of L.monocytogenes belonging to lineages I or II was evaluated in different conditions [two temperatures (37 and 22 degrees C) and two media (tryptone soy broth yeast extract medium (TSBYE) and MCDB 202 defined medium)] using crystal violet assay. Lineage II strains produced significantly more biofilm than lineage I strains. In microtitre plates assay, biofilm quantities were greater in MCDB 202 vs TSBYE medium [confirmed by scanning electron microscopy (SEM) analysis] and at 37 vs 22 degrees C. Cultivable bacteria from biofilm population on Petri dishes were enumerated in greater quantities in TSBYE than in MCDB 202 medium. The SEM investigation established that L.monocytogenes biofilms produce extracellular matrix in both media at 37 degrees C. The amount of exopolymers in the extracellular matrix and the pH values were significantly higher in TSBYE than in MCDB 202 medium. The exception was the ScottA strain that presented similar pH values and exopolymer contents in both media. Proteins were the most abundant exopolymer components, followed by DNA and polysaccharides. Conclusions The interpretation of results of biofilm quantification was depending on the growth conditions, the viability of the bacteria and the analysis method. The quantities of proteins, DNA and polysaccharides were different according to the strains and the medium. Significance and Impact of the Study This study screened the potential of a wide panel of L.monocytogenes strains to synthesize exopolymers in biofilm growing condition. The characterization of L.monocytogenes biofilm composition may help to develop new strategies to prevent the formation of biofilms and to remove the biofilms.

Purpose Biomaterial-associated infections (BAI) remain a major concern in modern health care. BAI is difficult to treat and often results in implant replacement or removal. Pathogens can be introduced on implant surfaces during surgery and compete with host cells attempting to integrate the implant. Here we studied the influence of prophylactically given cephatholin in the competition between highly virulent Staphylococcus aureus and human osteoblast-like cells (U-2 OS, ATCC HTB-94) for a poly(methyl methacrylate) surface in vitro using a peri-operative contamination model. Method S. aureus was seeded on the acrylic surface in a parallel plate flow chamber prior to adhesion of U-2 OS cells. Next, S. aureus and U-2 OS cells were allowed to grow simultaneously under shear (0.14 1/s) in a modified culture medium containing cephatholin for 8 h, the time period this drug is supposed to be active in situ. Subsequently, the flow was continued with modified culture medium for another 64 h. Results In the absence of cephatholin, highly virulent S. aureus caused U-2 OS cell death within 18 h. In contrast, the presence of cephatholin for 8 h resulted in survival of U-2 OS cell up to 72 h during simultaneous growth of U-2 OS cells and bacteria. Not all adhering bacteria were killed however, but they showed a delayed growth. Conclusions These findings are in line with the recalcitrance of biofilms against antibiotic treatment observed clinically, and represent another support for the use of in vitro co-culture models in mimicking the clinical situation.

Bacteria adhere to virtually all natural and synthetic surfaces [1,2]. Although there are a number of different reasons as to why bacteria adhere to a surface, the summarizing answer is brief: ‘‘Adhesion to a surface is a survival mechanism for bacteria’’. Nutrients in aqueous environments have the tendency to accumulate at surfaces [1,3], giving adhering bacteria a benefit over free floating, so-called planktonic ones. This is why mountain creeks may contain crystal clear, drinkable water, while stepping stones underneath the water may be covered with a slippery film of adhering microbes. In the oral cavity, adhesion to dental hard and soft tissues is life-saving to the organisms, because microbes that do not manage to adhere and remain planktonic in saliva are swallowed with an almost certain death in the gastrointestinal tract. Bacterial adhesion is generally recognized as the first step in biofilm formation, and for the human host, the ability of a bacterium to adhere is a definite virulence factor, especially in immunocompromised patients and in the growing number of elderly patients relying on biomaterials implants and devices for the restoration of function after (oncological) intervention surgery, trauma, or wear [4]. Well-known examples of biomaterials implants are dental implants, vascular grafts, and prosthetic hips and knee joints. Bacterial adhesion is a virulence factor, because it stimulates the organism to produce extracellular polymeric substances (EPSs), such as polysaccharides, proteins, nucleic acids, and lipids [5], through which they embed themselves in a protective matrix. This protective matrix provides mechanical stability to a biofilm and constitutes the main difference between planktonic bacteria and bacteria adhering to a surface in a socalled biofilm mode of growth. Bacteria organized in biofilms are at least ten to 1,000 times more resistant to antibiotics [6] than bacteria in a planktonic state and can cope much better with unfavorable external conditions in the host immune system than their planktonic counterparts. Not surprisingly, the fate of an infection associated with a biomaterials implant is mostly removal and replacement of the implant [7], at high costs to the health care system and great inconvenience to the patient.

The aim of this study was to investigate the role of extracellular DNA (eDNA) on the adhesion strength of Streptococcus mutans LT11 on substrata with different hydrophobicities at high and low ionic strengths. AFM adhesion forces to a hydrophilic and hydrophobic substratum increased with increasing surface-delay times and ionic strength and were stronger on a hydrophobic than on a hydrophilic substratum. The presence of eDNA on the streptococcal cell surface enhanced its adhesion force to a hydrophobic substratum significantly more than to a hydrophilic substratum, especially after bond maturation. Bond maturation on a hydrophilic substratum was accompanied by an increasing number of minor adhesion peaks, indicating the involvement of acid base interactions, whereas on the hydrophobic substratum surface the number of minor adhesion peaks remained low. More minor adhesion peaks developed on the hydrophilic substratum at low ionic strength than at high ionic strength. The final rupture distance in retraction force distance curves was independent of ionic strength on a hydrophilic substratum and increased with increasing surface delay time. On the hydrophobic surface, the final rupture distance did not increase with surface delay time but was significantly smaller at low than at high ionic strength. Final rupture distances were different in presence and absence of eDNA, and the lower values of this difference coincided with the decrease in hydrodynamic radius of the streptococci upon increasing ionic strength, measured using dynamic light scattering. AFM also yielded higher values for the ionic strength induced difference in final rupture distance because in AFM rupture is forced, while in dynamic light scattering differences in radius are only induced by ionic strength differences.

Biomaterials-associated infections (BAI) constitute a major clinical problem and often necessitate implant replacement. In this study, the race for the surface between Staphylococcus epidermidis ATCC 35983 and U2OS osteosarcoma cells is studied on biomaterials with different wettabilities and on a polymer-brush coating. S. epidermidis was deposited on the different surfaces in a parallel plate flow chamber and then U2OS cells were seeded. Subsequently, staphylococci and U2OS cells were allowed to grow simultaneously on the surfaces for 48 h under low flow conditions. The presence of staphylococci reduced cell growth on all surfaces, but adhering cells spread equally well in the absence and presence of staphylococci. A hydrophilic polymer-brush coating discouraged bacterial and cellular adhesion and growth. Thus, whereas the biomaterials evaluated support both biofilm formation and tissue integration, polymer-brush coatings support neither. Therewith, the outcome of the race for the surface on these surfaces remains uncertain, emphasizing the need for biofunctionalized surfaces that discourage biofilm formation and support tissue growth at the same time. (C) 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 94A: 533-538, 2010

International audience; This work describes the implementation of a new assay named the BioFilm Ring Test® to evaluate the ability of bacteria to form biofilms. This assay is based on the immobilisation (or not) of magnetic beads embedded by bacterial aggregates or mats (pattented concept). It is realised on modified polystyrene 96-wells microtiter plates with individual 8-wells slides. The kinetic of biofilm formation of four bacterial species, Listeria monocytogenes, Escherichia coli, Staphylococcus carnosus and Staphylococcus xylosus was evaluated with this new device by comparison with the standard crystal violet staining method of sessile cells. In parallel, the biofilm growth was visualized by Scanning Electron Microscopy (SEM) observations. The BioFilm Ring Test® gave similar but faster results than the crystal violet method. Moreover, the new assay was easier to implement, more reproducible and allowed high throughput screenings due to limited manipulations (no washing and staining steps) and rapid and accurate measurements of magnetic bead immobilisation by sessile bacterial cells.

Many studies have shown relationships of substratum hydrophobicity, charge or roughness with bacterial adhesion, although bacterial adhesion is governed by interplay of different physico-chemical properties and multiple regression analysis would be more suitable to reveal mechanisms of bacterial adhesion. The formation of a conditioning film of organic compounds adsorbed from seawater affects the properties of substratum surfaces prior to bacterial adhesion, which is a complicating factor in studying the mechanism of bacterial adhesion. In this paper, the impact of conditioning films adsorbed from natural seawater to four polyurethane coatings with different hydrophobicity, elasticity and roughness was studied for three different marine bacterial strains in a multiple linear regression analysis. The water contact angle on hydrophobic coatings decreased on average by 8 degrees and increased on average by the same amount on hydrophilic coatings. These changes were accompanied by increased concentrations of oxygen and nitrogen on the surface as determined by X-ray photoelectron spectroscopy, indicative of adsorption of proteinaceous material. Furthermore, the mean surface roughness increased on average by 4 nm after conditioning film formation. Multiple linear regression analysis revealed that changes in deposition due to conditioning film formation ofMarinobacter hydrocarbonoclasticus,Psychrobactersp. SW5H andHalomonas pacificain a stagnation-point flow chamber could be explained in a model comprising hydrophobicity and the prevalence of nitrogen-rich components on the surface for the most hydrophobic strain. For the two more hydrophilic strains, deposition was governed by a combination of surface roughness and hydrophobicity. Elasticity was not a factor in bacterial adhesion to conditioning films.