Medical Device-related Infections Research Articles

Synthesis of conjugated PIA-rSesC and immunological evaluation against biofilm-forming Staphylococcus epidermidis.

Staphylococcus epidermidis is an opportunistic pathogen and a leading cause of morbidity and premature mortality in patients with medical device-related infections, which is a concern in hospitalized patients. Developing a strategy to raise opsonic antibodies against polysaccharide intracellular adhesin (PIA) could be promising for the elimination of colonizing and biofilm-forming S. epidermidis. Following the purification of truncated rSesC protein and PIA, for the first time, PIA was conjugated to rSesC as a carrier to increase the immunogenicity of PIA and its efficacy in mice was evaluated. The structure of the conjugate was analysed using the Fourier transform infrared spectroscopy (FTIR) and proton nuclear magnetic resonance spectroscopy (H1- NMR) methods. Afterwards, the immune response was evaluated by measuring the total IgG, IgG2a and IgG2b titres. The immunization of mice with the PIA-rSesC conjugate raised the levels of opsonic antibodies, and the vaccinated mice were protected when challenged intravenously by wild-type S. epidermidis strain 1457. Further studies indicated that the conjugated vaccine was able to eliminate S. epidermidis biofilm formation in in vitro or in vivo assays. This study confirms the proposal that the immunization of mice with PIA-rSesC conjugate vaccine could protect against S. epidermidis infection.

Cold plasma effect on the proteome of Pseudomonas aeruginosa - Role for bacterioferritin.

Cold atmospheric-pressure plasma (CAP) is a relatively new method used for bacterial inactivation. CAP is ionized gas that can be generated by applying an electric current to air or a feeding gas. It contains reactive species and emits UV radiation, which have antibacterial activity. Previous data suggests that CAP is effective in microbial inactivation and can decontaminate and sterilize surfaces, but its exact mode of action is still under debate. This study demonstrates the effect of CAP on the whole proteome of Pseudomonas aeruginosa PAO1 biofilms, which is a dominant pathogen in cystic fibrosis and medical device-related infections. Liquid chromatography-mass spectrometry (LC-MS) was used to identify differentially regulated proteins of whole cell P. aeruginosa extracts. A total of 16 proteins were identified to be affected by plasma treatment compared to the control. Eight of the identified proteins have functions in transcription and translation and their expression changes are likely to be part of a general physiological response instead of a CAP-specific adaptation. However, CAP also affected bacterioferritin (Bfr), Isocitrate dehydrogenase (Idh), Trigger factor (Tig) and a chemotaxis protein, which may be involved in P. aeruginosa’s specific response to CAP. We confirm that bacterioferritin B plays a role in the bacterial response to CAP because ΔbfrB mutants of both PAO1 and PA14 are more susceptible to plasma-induced cell-death than their corresponding wild-type strains. To our knowledge, this is the first study showing the effect of plasma on the whole proteome of a pathogenic microorganism. It will help our understanding of the mode of action of CAP-mediated bacterial inactivation and thus support a safe and effective routine use of CAP in clinical and industrial settings.

Endotracheal Tube Biofilm and its Impact on the Pathogenesis of Ventilator-Associated Pneumonia.

Ventilator-associated pneumonia (VAP) is a common and serious nosocomial infection in mechanically ventilated patients and results in high mortality, prolonged intensive care unit- (ICU) and hospital-length of stay and increased costs. In order to reduce its incidence, it is imperative to better understand the involved mechanisms and to identify the source of infection. The role of the endotracheal tube (ET) in VAP pathogenesis became more prominent over the last decades, along with extensive research dedicated to medical device-related infections and biofilms. ET biofilm formation is an early and constant process in intubated patients. New data regarding its temporal dynamics, composition, germ identification and consequences enhance knowledge about VAP occurrence, microbiology, treatment response and recurrence.This paper presents a structured analysis of the medical literature to date, in order to outline the role of ET biofilm in VAP pathogenesis and to review recommended methods to identify ET biofilm microorganisms and to prevent or decrease VAP incidence.

Pan-genome analysis of the genus Finegoldia identifies two distinct clades, strain-specific heterogeneity, and putative virulence factors

Finegoldia magna, a Gram-positive anaerobic coccus, is an opportunistic pathogen, associated with medical device-related infections. F. magna is the only described species of the genus Finegoldia. We report the analysis of 17 genomes of Finegoldia isolates. Phylogenomic analyses showed that the Finegoldia population can be divided into two distinct clades, with an average nucleotide identity of 90.7%. One clade contains strains of F. magna, whereas the other clade includes more heterogeneous strains, hereafter tentatively named “Finegoldia nericia”. The latter species appears to be more abundant in the human microbiome. Surface structure differences between strains of F. magna and “F. nericia” were detected by microscopy. Strain-specific heterogeneity is high and previously identified host-interacting factors are present only in subsets of “F. nericia” and F. magna strains. However, all genomes encode multiple host factor-binding proteins such as albumin-, collagen-, and immunoglobulin-binding proteins, and two to four copies of CAMP (Christie-Atkins-Munch-Petersen) factors; in accordance, most strains show a positive CAMP reaction for co-hemolysis. Our work sheds new light of the genus Finegoldia and its ability to bind host components. Future research should explore if the genomic differences identified here affect the potential of different Finegoldia species and strains to cause opportunistic infections.

Self-sterilizing ormosils surfaces based on photo-synzthesized silver nanoparticles.

Medical device-related infections represent a major healthcare complication, resulting in potential risks for the patient. Antimicrobial materials comprise an attractive strategy against bacterial colonization and biofilm proliferation. However, in most cases these materials are only bacteriostatic or bactericidal, and consequently they must be used in combination with other antimicrobials in order to reach the eradication condition (no viable microorganisms). In this study, a straightforward and robust antibacterial coating based on Phosphotungstate Ormosil doped with core-shell (SiO2@TiO2) was developed using sol-gel process, chemical tempering, and Ag nanoparticle photoassisted synthesis (POrs-CS-Ag). The coating was characterized by X-ray Fluorescence Spectroscopy (XRF), Field Emission Scanning Electron Microscopy (FE-SEM), Atomic Force Microscopy (AFM) and X-ray Photoelectron Microscopy (XPS). The silver free coating displays low antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa, in opposition to the silver loaded ones, which are able to completely eradicate these strains. Moreover, the antimicrobial activity of these substrates remains high until three reutilization cycles, which make them a promising strategy to develop self-sterilizing materials, such as POrs-CS-Ag-impregnated fabric, POrs-CS-Ag coated indwelling metals and polymers, among other materials.

Rifampicin resistance in Staphylococcus epidermidis: molecular characterisation and fitness cost of rpoB mutations

The molecular mechanisms and characteristics of rifampicin (RIF) resistance in Staphylococcus epidermidis are poorly characterised, even though S. epidermidis is one of the most common nosocomial pathogens associated with indwelling medical device-related infections. The aim of this study was to investigate the evolution of RIF resistance and to characterise the associated molecular mechanisms in S. epidermidis. RIF-resistant mutants from two RIF-susceptible S. epidermidis strains (RP62A and IDRL-8883) were selected through in vitro and in vivo exposure to RIF. A total of 16 colonies with an RP62A background and 63 colonies with an IDRL-8883 background were analysed for rpoB mutations. The fitness of RIF-susceptible and isogenic RIF-resistant strains was assessed using a paired competition assay and by comparing generation times. All mutations detected were in cluster I of rpoB. The following five amino acid substitutions were selected in vitro: Asp471→Asn; Asp471→Gly; Asp471→Val; Ser486→Tyr; and His481→Tyr. The following three amino acid substitutions were selected in vivo: His481→Tyr; Gln468→Lys; and Ser486→Phe. Asp471→Asn and Asp471→Gly changes were associated with susceptible minimal inhibitory concentrations (MICs). In vitro competition assays revealed that all RIF-resistant mutants other than Ser486→Tyr and Ser486→Phe had a relative fitness of <1.0. His481→Tyr mutations had their own specific fitness costs and effects on growth rate, irrespective of strain background. In conclusion, the current study presents molecular characterisations and fitness costs of several rpoB mutations in S. epidermidis.

Sterilization of hydrogels for biomedical applications: A review

Despite the beneficial properties and outstanding potential of hydrogels for biomedical applications, several unmet challenges must be overcome, especially regarding to their known sensitivity to conventional sterilization methods. It is crucial for any biomaterial to withstand an efficient sterilization to obtain approval from regulatory organizations and to safely proceed to clinical trials. Sterility assurance minimizes the incidence of medical device-related infections, which still constitute a major concern in health care. In this review, we provide a detailed and comprehensive description of the published work from the past decade regarding the effects of sterilization on different types of hydrogels for biomedical applications. Advances in hydrogel production methods with simultaneous sterilization are also reported. Terminal sterilization methods can induce negative or positive effects on several material properties (e.g., aspect, size, color, chemical structure, mechanical integrity, and biocompatibility). Due to the complexity of factors involved (e.g., material properties, drug stability, sterilization conditions, and parameters), it is important to note the virtual impossibility of predicting the outcome of sterilization methods to determine a set of universal rules. Each system requires case-by-case testing to select the most suitable, effective method that allows for the main properties to remain unaltered. The impact of sterilization methods on the intrinsic properties of these systems is understudied, and further research is needed. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2472-2492, 2018.

Prevalence and antibiogram of methicillin resistant Staphylococcus aureus isolated from medical device-related infections; a retrospective study in Lahore, Pakistan.

With the advancement of medicine and surgery, various types of medical devices have become part of treatment strategies. Identification and antimicrobial sensitivity testing were done according to CLSI guidelines following standard microbiological practices. Urinary catheter infections (31%) were most frequent followed by central venous catheter (18%) and orthopedic implants (15%). Methicillin resistant Staphylococcus aureus (MRSA) was a major cause of device-related infection after Escherichia coli (21%); other pathogens were Klebsiella pneumoniae (14%), Pseudomonas spp. (10%), Acinetobacter spp. (8%) and Candida species (7%). None of MRSA was resistant to vancomycin (MIC ≥16µg/mL). Resistance rates were 98% and 97% for ofloxacin and ciprofloxacin, respectively. Escherichia coli and MRSA are major pathogens of medical device-related infections.

Frequency of bap and cpaA virulence genes in drug resistant clinical isolates of Acinetobacter baumannii and their role in biofilm formation.

Objective(s):Acinetobacter baumannii has a high propensity to form biofilm and frequently causes medical device-related infections with multiple-drug-resistance in hospitals. The aim of this work is to study antimicrobial resistance and the role of bap and cpaA genes in biofilm formation by A. baumannii to understand how this pathogen persists in the hospital environment.Materials and Methods:The antibiotic resistance profile and in vitro biofilm-forming ability of one hundred clinical isolates of A. baumannii was evaluated by disc diffusion and crystal-violet staining methods, respectively. Isolates were tested for the presence of bap and cpaA genes.Results:The isolates were highly resistant to cefepime, third-generation cephalosporins, ciprofloxacin, cotrimoxazole, aminoglycosides and carbapenems. Moreover, four isolates were resistant to colistin. Quantification of biofilm showed that 43% of the isolates were strong biofilm-producer. Furthermore, 32% of the isolates exhibited moderate biofilm-formation and showed initial binding activity. Frequency of bap and cpaA were determined 92% and 36%, respectively.Conclusion:There was strong association between the presence of bap gene and biofilm formation by A. baumannii isolates (P=0.003). In addition, multidrug resistant isolates produced stronger biofilm than other isolates (P=0.0001). These results indicate importance of biofilm in resistance of isolates and effect of presence of bap gene in biofilm formation by A. baumannii strains.

Quorum Sensing - Dependent Virulence Factors and Biofilm Formation of Pseudomonas Aeruginosa Isolates from Retrieved Orthopedic Implants

Background: Quorum sensing (QS) plays a crucial role in medical device-related infections. However, epidemiological analysis of QS- dependent virulence factors among Pseudomonas aeruginosa (P. aeruginosa) isolates from orthopedic implants-related infections has rarely been reported. Objective: to assess some QS-dependent virulence factors (as pyocyanin, elastase, protease, rhamnolipid, and biofilm production) in P. aeruginosa isolates from retrieved different orthopedic implants. Methodology: Twenty-four P. aeruginosa isolates obtained from 300 retrieved orthopedic implants with proven diagnosis of osteoarticular infection directly related to orthopedic implants were investigated for production of QS-dependent virulence factors. The pyocyanin production was assayed using King A media, rhamnolipid production was checked utilizing cetyltrimethylammonium bromide (CTAB) agar test. Elastin lytic activity was assayed using elastin Congo red, total protease was estimated using the modified skimmed milk assay technique, and the biofilm forming ability was assayed by microtiter plate assay. N-acyl homoserine lactones (3-oxo-C 12 -HSL, C 4 -HSL) were detected by Cross-feeding bioassay. Polymerase chain reaction (PCR) was used to assess the presence of the QS genes lasI, lasR, rhlI, and rhlR in P. aeruginosa isolates. Results: Seventeen isolates (70.8%) displayed the QS-dependent phenotypes and seven isolates were found to be lacking all tested virulence factors. The lack of production of pyocyanin, Rhamnolipid, las B elastase, total protease, and biofilm formation by the seven isolates proposed that they might have a defective QS system. Among the Seven isolates exhibiting impaired QS-dependent phenotypes, AHLs autoinducers were present in three isolates, though the residual four isolates were lacking both autoinducers and therefore are considered QS-deficient. PCR analysis declared that two isolates contained lasI, lasR, rhlI and rhlR genes while one isolate lacked lasR gene, and one isolate was lacking the four tested genes. Conclusion: Although QS seems to be a crucial regulator of virulence factors and biofilm formation in P. aeruginosa, it is equipped to produce implant infection in humans despite an impaired QS system. These results do not challenge the concept that QS plays a foremost role in P. aeruginosa pathogenicity, but highlight that along with known virulence factors, perhaps there are other virulence factors, that may not be strictly under QS control.

Silicone rubber films functionalized with poly(acrylic acid) nanobrushes for immobilization of gold nanoparticles and photothermal therapy

Prevention and treatment of medical device-related infections demands novel strategies for hindering bacteria adhesion and proliferation. The aim of this work was to immobilize gold nanoparticles onto silicone rubber (SR) and then exploit the plasmon resonance properties for the photothermal killing of adhered bacteria. To obtain a good rate of gold deposition, SR films were grafted with PAAc nanobrushes (SR-g-AAc) using a 60Co gamma source. Effects of absorbed dose, monomer concentration, temperature and reaction time on the grafting percentage, swelling and mechanical properties of the films were first investigated. Gold nanoparticles were immobilized on SR and SR-g-AAc following different strategies. Immersion in gold ions (HAuCl4) solution and subsequent reduction made the films to be uniformly coated by 10–20 nm gold nanoparticles as shown by SEM-EDX and XRD. Photothermal features were evidenced as an increase in 6–7 °C in the first 5 min of irradiation with a 514 nm laser at a power density of 0.5 W/cm2. Finally, SR-g-AAc films were challenged against Staphylococcus aureus and effects of irradiation at 514 nm on bacteria survival evaluated. Overall, the gold-immobilized films were shown to be stable against autoclave sterilization and ultrasounds and exhibited outstanding ability to damage adhered bacteria after laser irradiation.

Microbiological diagnosis of device-related biofilm infections.

Medical device-related infections cause undue patient distress, increased morbidity and mortality and pose a huge financial burden on healthcare services. The pathogens are frequently distributed heterogeneously in biofilms, which can persist without being effectively cleared by host immune defenses and antibiotic therapy. At present, there is no 'gold standard' available to reveal the presence of device-related biofilm infections. However, adequate sample collection and logistics, standardised diagnostic methods, and interpretation of results by experienced personnel are important steps in efficient diagnosis and treatment of these infections. The focus of this mini review is on prosthethic joint and cardiovascular implantable device infections, which exemplify permanent devices that are placed in a sterile body site. These device-related infections represent some of the most challenging in terms of both diagnosis and treatment.

Osteopontin adsorption to Gram-positive cells reduces adhesion forces and attachment to surfaces under flow

ABSTRACTThe bovine milk protein osteopontin (OPN) may be an efficient means to prevent bacterial adhesion to dental tissues and control biofilm formation. This study sought to determine to what extent OPN impacts adhesion forces and surface attachment of different bacterial strains involved in dental caries or medical device–related infections. It further investigated if OPN’s effect on adhesion is caused by blocking the accessibility of glycoconjugates on bacterial surfaces. Bacterial adhesion was determined in a shear-controlled flow cell system in the presence of different concentrations of OPN, and interaction forces of single bacteria were quantified using single-cell force spectroscopy before and after OPN exposure. Moreover, the study investigated OPN’s effect on the accessibility of cell surface glycoconjugates through fluorescence lectin-binding analysis. OPN strongly affected bacterial adhesion in a dose-dependent manner for all investigated species (Actinomyces naeslundii, Actinomyces viscosus, Lactobacillus paracasei subsp. paracasei, Staphylococcus epidermidis, Streptococcus mitis, and Streptococcus oralis). Likewise, adhesion forces decreased after OPN treatment. No effect of OPN on the lectin-accessibility to glycoconjugates was found. OPN reduces the adhesion and adhesion force/energy of a variety of bacteria and has a potential therapeutic use for biofilm control. OPN acts upon bacterial adhesion without blocking cell surface glycoconjugates.

Evaluation of the ability of Acinetobacter baumannii to form biofilms on six different biomedical relevant surfaces.

The human opportunistic pathogen, Acinetobacter baumannii, has the propensity to form biofilms and frequently cause medical device-related infections in hospitals. However, the physio-chemical properties of medical surfaces, in addition to bacterial surface properties, will affect colonization and biofilm development. The objective of this study was to compare the ability of A.baumannii to form biofilms on six different materials common to the hospital environment: glass, porcelain, stainless steel, rubber, polycarbonate plastic and polypropylene plastic. Biofilms were developed on material coupons in a CDC biofilm reactor. Biofilms were visualized and quantified using fluorescent staining and imaged using confocal laser scanning microscopy (CLSM) and by direct viable cell counts. Image analysis of CLSM stacks indicated that the mean biomass values for biofilms grown on glass, rubber, porcelain, polypropylene, stainless steel and polycarbonate were 0·04, 0·26, 0·62, 1·00, 2·08 and 2·70μm(3) /μm(2) respectively. Polycarbonate developed statistically more biofilm mass than glass, rubber, porcelain and polypropylene. Viable cell counts data were in agreement with the CLSM-derived data. In conclusion, polycarbonate was the most accommodating surface for A.baumannii ATCC 17978 to form biofilms while glass was least favourable. Alternatives to polycarbonate for use in medical and dental devices may need to be considered. In the hospital environment, Acinetobacter baumannii is one of the most persistent and difficult to control opportunistic pathogens. The persistence of A.baumannii is due, in part, to its ability to colonize surfaces and form biofilms. This study demonstrates that A.baumannii can form biofilms on a variety of different surfaces and develops substantial biofilms on polycarbonate - a thermoplastic material that is often used in the construction of medical devices. The findings highlight the need to further study the invitro compatibility of medical materials that could be colonized by A.baumannii and allow it to persist in hospital settings.

Pseudomonas aeruginosa Outer Membrane Vesicles Triggered by Human Mucosal Fluid and Lysozyme Can Prime Host Tissue Surfaces for Bacterial Adhesion

Pseudomonas aeruginosa is a leading cause of human morbidity and mortality that often targets epithelial surfaces. Host immunocompromise, or the presence of indwelling medical devices, including contact lenses, can predispose to infection. While medical devices are known to accumulate bacterial biofilms, it is not well understood why resistant epithelial surfaces become susceptible to P. aeruginosa. Many bacteria, including P. aeruginosa, release outer membrane vesicles (OMVs) in response to stress that can fuse with host cells to alter their function. Here, we tested the hypothesis that mucosal fluid can trigger OMV release to compromise an epithelial barrier. This was tested using tear fluid and corneal epithelial cells in vitro and in vivo. After 1 h both human tear fluid, and the tear component lysozyme, greatly enhanced OMV release from P. aeruginosa strain PAO1 compared to phosphate buffered saline (PBS) controls (∼100-fold). Transmission electron microscopy (TEM) and SDS-PAGE showed tear fluid and lysozyme-induced OMVs were similar in size and protein composition, but differed from biofilm-harvested OMVs, the latter smaller with fewer proteins. Lysozyme-induced OMVs were cytotoxic to human corneal epithelial cells in vitro and murine corneal epithelium in vivo. OMV exposure in vivo enhanced Ly6G/C expression at the corneal surface, suggesting myeloid cell recruitment, and primed the cornea for bacterial adhesion (∼4-fold, P < 0.01). Sonication disrupted OMVs retained cytotoxic activity, but did not promote adhesion, suggesting the latter required OMV-mediated events beyond cell killing. These data suggest that mucosal fluid induced P. aeruginosa OMVs could contribute to loss of epithelial barrier function during medical device-related infections.

Impact of Insertion Sequences and Recombination on the Population Structure of Staphylococcus haemolyticus.

Staphylococcus haemolyticus is one of the most common pathogens associated with medical-device related infections, but its molecular epidemiology is poorly explored. In the current study, we aimed to better understand the genetic mechanisms contributing to S. haemolyticus diversity in the hospital environment and their impact on the population structure and clinical relevant phenotypic traits. The analysis of a representative S. haemolyticus collection by multilocus sequence typing (MLST) has identified a single highly prevalent and diverse genetic lineage of nosocomial S. haemolyticus clonal complex (CC) 29 accounting for 91% of the collection of isolates disseminated worldwide. The examination of the sequence changes at MLST loci during clonal diversification showed that recombination had a higher impact than mutation in shaping the S. haemolyticus population. Also, we ascertained that another mechanism contributing significantly to clonal diversification and adaptation was mediated by insertion sequence (IS) elements. We found that all nosocomial S. haemolyticus, belonging to different STs, were rich in IS1272 copies, as determined by Southern hybridization of macrorestriction patterns. In particular, we observed that the chromosome of a S. haemolyticus strain within CC29 was highly unstable during serial growth in vitro which paralleled with IS1272 transposition events and changes in clinically relevant phenotypic traits namely, mannitol fermentation, susceptibility to beta-lactams, biofilm formation and hemolysis. Our results suggest that recombination and IS transposition might be a strategy of adaptation, evolution and pathogenicity of the major S. haemolyticus prevalent lineage in the hospital environment.

Nanomaterial-Based Approaches for Prevention of Biofilm-Associated Infections on Medical Devices and Implants.

Biofilm formation is a major problem in medical device-related infections leading to failure of implant-based therapies. Though various conventional approaches to counter biofilm formation like physical and/or mechanical removal, chemical removal, and the use of antimicrobials exist, they fail due to increased resistance of biofilms. This review discusses various nanomaterial-based approaches such as the use of metallic and metal oxide nanoparticles- and polymer-based nanocomposites, which are currently being developed for prevention and treatment of biofilms. Nanoparticles of transition metals and their oxides are toxic to microorganisms and exhibit their toxicity through the generation of reactive oxygen species at concentrations that are non-toxic to eukaryotic cells. Other approaches include the entrapment of bioactive agents in polymer/ceramic nanoparticles, for enhanced anti-biofilm activity due to the synergistic effect between them. These nanomaterial-based approaches could play an important role in control and eradication of biofilm related infections and complications associated with medical devices and implants.

Bacillus cereus infection: 57 case patients and a literature review

ObjectivesWe aimed to study the characteristics of patients presenting with a Bacillus cereus infection in a university hospital. MethodsWe performed a retrospective analysis of the clinical, biological, and treatment-related data of patients hospitalized in our university hospital between January 1st, 2008 and December 31st, 2012 and diagnosed with a B. cereus infection. We identified a subgroup of patients presenting with bacteremia and looked for risk factors for death within that group of patients. ResultsWe included 57 patients in our study; 31 (54.4%) were hospitalized in a medical ward. We identified 24 bacteremia case patients, including 17 patients presenting with bacteremia alone (29.8%). Other frequently observed infection sites were skin infections (16; 28.1%) and bone and joint infections (10; 17.5%). We recorded 9 deaths (11.8%); 2 patients, despite being on an appropriate antibiotic therapy, died from a medical device-related infection that had not been removed. The empirical administration of a beta-lactam antibiotic was significantly associated with death (P=0.022). Three patients presenting with recurrent bacteremia were identified. The patients only recovered once the infected device had been removed. ConclusionB. cereus infections may have various clinical presentations. Prospective data is needed to put forward a consensual treatment approach and guide physicians in choosing the appropriate antibiotic therapy and in removing the infected device.

Prevalence of fibronectin-binding protein (FnbA and FnbB) genes among clinical isolates of methicillin resistant Staphylococcus aureus

One of the most important stages in the occurrence of infection caused by Staphylococcus aureus is able to adhere of the bacterium to the cells and the extracellular matrix. Among adhesins, two fibronectinbinding proteins, (FnbA and FnbB) have been proved significantly to contribute to tissue colonization in various pathological conditions and indwelling medical device related infections. The aims of this study were to detect fnbA and fnbB genes and relation to the fibronectin binding ability in clinical isolates of methicillin resistant Staphylococcus aureus (MRSA). Of the 62 MRSA clinical isolates collected from selected hospitals in Tehran, Iran, fibronectin binding ability was determined by microtiter tissue culture coated by fibronectin plates. All MRSA isolates were examined for determination the fnbA and fnbB genes by using PCR method. The prevalence of fnbA and fnbB in MRSA strains was 82.2 and 46.7% respectively. In 26 (42%) MRSA strains both fnbA and fnbB genes were positive. All fnbA+ or fnbB+ strains were found to be positive also to in vitro fibronectin binding activity, while the fnb-negative strains were found negative to the in vitro binding assay. The finding that, fibronectin-adhesins are present in the most of the MRSA clinical isolates encourages the development of strategies to specifically block the interaction of this bacterium with fibronectin by antagonist molecules.

Two repetitive, biofilm-forming proteins from Staphylococci: from disorder to extension.

Staphylococcus aureus and Staphylococcus epidermidis are an important cause of medical device-related infections that are difficult to treat with antibiotics. Biofilms, in which bacteria are embedded in a bacterially-produced exopolymeric matrix, form on the surface of the implanted medical device. Our understanding of the molecular mechanisms underlying the initial surface attachment and subsequent intercellular interactions as the biofilm matures is improving. Biofilm accumulation can be mediated by a partially deacetylated form of poly-N-acetylglucosamine (PNAG) but, more recently, the role of bacterial surface proteins is being recognized. Here we describe the structure and function of two S. aureus cell surface proteins, FnBPA and SasG, implicated in host interactions and biofilm accumulation. These multifunctional proteins employ intrinsic disorder for distinct molecular outcomes. In the case of FnBPA, disorder generates adhesive arrays that bind fibronectin (Fn); in the case of SasG, disorder is, counterintuitively, used to maintain a strong extended fold.