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PubMed2004

The delivery of benzyl penicillin to Staphylococcus aureus biofilms by use of liposomes.

The delivery of benzyl penicillin [penicillin G (pen-G)] encapsulated in cationic liposomes to a pen-G-sensitive strain of Staphylococcus aureus immobilized in biofilms has been investigated. The cationic liposomes prepared by extrusion (VETs, diameter approximately 140 nm) were composed of dipalmitoylphosphatidylcholine (DPPC), cholesterol, and dimethylammonium ethane carbamoyl cholesterol (DC-chol) at a molar ratio of 1.0:0.49:0.43. This composition containing 22 mole% of the cationic lipid DC-chol has been found previously (Kim et al. Colloids Surfaces A 1999, 149, 561-570) to be optimum for adsorption of the liposomes on S. aureus biofilms. The effectiveness of the liposomes to deliver pen-G to the biofilms immobilized on microtitre plates was assessed from the rate of growth of the cells after exposure to the liposomal drug carrier relative to free pen-G at the same concentration. The time to reach maximum growth rate from biofilms was investigated as a function of overall drug concentration in a range 2.9 x 10- 3 mM to 1.09 mM and as a function of time of exposure to liposomal drug in a range 1.5 s to 2 h. Liposomal drug delivery was most effective relative to free drug at low overall drug concentrations and short times of exposure. The time to reach maximum growth rate from S. aureus biofilms could be extended by a factor of approximately 4 relative to free drug by the use of liposomally encapsulated pen-G. The results were supported by direct measurements of the distribution of pen-G between biofilm and supernatant which showed enhanced values relative to free drug and a transient preferential uptake of drug induced by the liposomes. The study demonstrates that for low drug concentrations and short exposure times liposomal drug delivery greatly enhances the effectiveness of pen-G for inhibiting the growth of bacterial biofilms of the potentially pathogenic bacterium Staphylococcus aureus.

Adsorption↗
PubMed1998

Response of single species biofilms and microcosm dental plaques to pulsing with chlorhexidine.

The aim of this study was to determine the effect of pulsing chlorhexidine gluconate, at concentrations commonly used in mouthwashes, on Streptococcus sanguis biofilms and microcosm dental plaques in vitro. Biofilms were grown on bovine enamel and nutrients were supplied in the form of artificial saliva. Pulsing experiments were carried out on steady-state biofilms using 0.05 or 0.2% chlorhexidine solutions delivered twice daily for 1 min. In a separate study, the enamel discs on which the biofilms were formed were pre-treated with chlorhexidine and pulsed directly after inoculation and then at regular intervals. With both concentrations of chlorhexidine used, a c.2 log10 reduction in the viable counts of S. sanguis was achieved with the initial pulse, but as pulsing continued, the bacterial population recovered, albeit not to the previous level. A c.1 log10 reduction in the total viable counts of the microcosm plaques was seen after the first pulse with 0.2% chlorhexidine. The total count then recovered rapidly and, after the fifth pulse, the total viable counts were not significantly different from those before pulsing. The total counts then remained at a similar level throughout the course of the experimental runs. Pre-treatment of the enamel discs with 0.2% chlorhexidine before inoculation produced viable counts of c.10(5) cfu/mm2, a 1 log10 reduction compared with untreated discs. After pulsing with 0.2% chlorhexidine at 8 h, a 3 log10 reduction was seen in the total aerobic and anaerobic counts, but again the viable counts subsequently increased despite twice-daily chlorhexidine pulsing. Regardless of the nature of the biofilm, pulsing initially achieved substantial kills, but the viability of the biofilms subsequently increased despite continued pulsing. Chlorhexidine was effective at reducing the viability of microcosm plaques when it was applied to the substratum before exposure to bacteria and subsequently pulsed on to the biofilms.

Animals↗
PubMed2000

An evaluation of the potential of the multiple antibiotic resistance operon (mar) and the multidrug efflux pump acrAB to moderate resistance towards ciprofloxacin in Escherichia coli biofilms.

The chromosomal multiple antibiotic resistance operon, mar, is widely represented amongst Gram-negative bacteria and has been implicated in resistance towards oxidative stress agents, organic solvents and a large number of structurally unrelated antimicrobial agents. The major mechanism associated with such increased resistance is an upregulation of the efflux pump acrAB. Growth as a biofilm is often associated with similar generalized reductions in susceptibility to inimical agents. Escherichia coli K12 (AG100), an isogenic mutant of AG100 constitutive for mar expression (AG102) and an isolate deleted of the mar locus (MCH164) were grown as biofilms in cellulose-fibre depth filters and perfused with a simple salts, minimal medium (CDM) over 120 h. Biofilms were exposed to various concentrations of ciprofloxacin (0.004, 0.015 and 0.1 mg/L) for 42 h. The numbers of viable cells within the perfusate and within the biofilm were estimated throughout. Whereas no differences were seen between the wild-type and mar-deleted isolates, that constitutive for mar displayed reduced susceptibility to ciprofloxacin at concentrations of 0.004 mg/L (MIC for AG100 was 0.0052 mg/L). Similar antibiotic perfusion experiments were conducted using isolates in which the efflux pump acrAB was either deleted (AG100-A) or constitutively expressed (AG100-B). Exposure of AG100-A biofilms to ciprofloxacin at 0.004 and 0.1 mg/L showed similar susceptibilities to those seen in the wild-type (AG100) and mar-deleted (MCH164) isolates and suggested that acrAB was not induced within the attached population. On the other hand, constitutive expression of acrAB (AG100-B) protected biofilms against the lower concentration of ciprofloxacin used (0.004 mg/L). This protection was again lost at concentrations of 0.1 mg/L. Overall, these results show that ciprofloxacin resistance in biofilms is not mediated by the upregulation of the mar or acrAB operons.

Ampicillin↗
PubMed2005

Telavancin: in vitro activity against staphylococci in a biofilm model.

OBJECTIVES: To assess the in vitro activity of the novel lipoglycopeptide telavancin against staphylococcal biofilms using an in vitro pharmacokinetic model. METHODS: Using the Sorbarod model, biofilms were established. The strains tested included methicillin-susceptible and -resistant strains of Staphylococcus aureus and coagulase-negative staphylococci, as well as glycopeptide-intermediate S. aureus (GISA). The biofilms were exposed to exponentially decreasing concentrations of telavancin and four comparator antibiotics, vancomycin, teicoplanin, linezolid and moxifloxacin and the bactericidal activity of the antibiotics was assessed. The concentrations of the antibiotics used in these experiments corresponded to peak serum levels achievable in humans and the rates at which drug concentrations were decreased corresponded to their elimination half-lives. RESULTS: All of the drugs tested produced a reduction in the number of bacteria eluted from the biofilms. Telavancin was more effective than the commercially available glycopeptides, vancomycin and teicoplanin, and of the three, was the most active agent against both the non-GISA and GISA strains. Of all the antibiotics tested, moxifloxacin produced the greatest reduction in biofilm cells, but only against the non-GISA strains. CONCLUSIONS: Telavancin exhibited substantial antimicrobial activity against staphylococcal biofilms, including GISA strains. This study supports the case for the evaluation of telavancin in the treatment of staphylococcal biofilm-associated infections.

Aminoglycosides↗
PubMed2005

Comparative assessment of antibiotic susceptibility of coagulase-negative staphylococci in biofilm versus planktonic culture as assessed by bacterial enumeration or rapid XTT colorimetry.

OBJECTIVES: To quantitatively compare the antibiotic susceptibility of biofilms formed by the coagulase-negative staphylococci (CoNS) Staphylococcus epidermidis and Staphylococcus haemolyticus with the susceptibility of planktonic cultures. METHODS: Several CoNS strains were grown planktonically or as biofilms to determine the effect of the mode of growth on the level of susceptibility to antibiotics with different mechanisms of action. The utility of a new, rapid colorimetric method that is based on the reduction of a tetrazolium salt (XTT) to measure cell viability was tested by comparison with standard bacterial enumeration techniques. A 6 h kinetic study was performed using dicloxacillin, cefazolin, vancomycin, tetracycline and rifampicin at the peak serum concentration of each antibiotic. RESULTS: In planktonic cells, inhibitors of cell wall synthesis were highly effective over a 3 h period. Biofilms were much less susceptible than planktonic cultures to all antibiotics tested, particularly inhibitors of cell wall synthesis. The susceptibility to inhibitors of protein and RNA synthesis was affected by the biofilm phenotype to a lesser degree. Standard bacterial enumeration techniques and the XTT method produced equivalent results both in biofilms and planktonic assays. CONCLUSIONS: This study provides a more accurate comparison between the antibiotic susceptibilities of planktonic versus biofilm populations, because the cell densities in the two populations were similar and because we measured the concentration required to inhibit bacterial metabolism rather than to eradicate the entire bacterial population. While the biofilm phenotype is highly resistant to antibiotics that target cell wall synthesis, it is fairly susceptible to antibiotics that target RNA and protein synthesis.

Anti-Bacterial Agents↗
PubMed2005

Susceptibility of Streptococcus mutans biofilms to photodynamic therapy: an in vitro study.

OBJECTIVES: The purpose of this study was to evaluate the antimicrobial effect of toluidine blue O (TBO), in combination with either a helium/neon (HeNe) laser or a light-emitting diode (LED), on the viability and architecture of Streptococcus mutans biofilms. METHODS: Biofilms were grown on hydroxyapatite discs in a constant depth film fermentor fed with artificial saliva that was supplemented with 2% sucrose four times a day, thus producing a typical 'Stephan pH curve'. Photodynamic therapy was subsequently carried out on biofilms of various ages with light from either the HeNe laser or LED using energy densities of between 49 and 294 J/cm(2). RESULTS: Significant decreases in the viability of S. mutans biofilms were only observed when biofilms were exposed to both TBO and light, when reductions in viability of up to 99.99% were observed with both light sources. Overall, the results showed that the bactericidal effect was light dose-dependent and that older biofilms were less susceptible to photodynamic therapy. Confocal laser scanning microscopy images suggested that lethal photosensitization occurred predominantly in the outermost layers of the biofilms. CONCLUSIONS: Photodynamic therapy may be a useful approach in the treatment of dental plaque-related diseases.

Biofilms↗
PubMed2003

Effects of biofilm formation on haemodialysis monitor disinfection.

BACKGROUND: Biofilms are composed of communities of micro-organisms adhering to essentially any surface. We evaluated whether biofilm formation in the hydraulic circuit of a purposely contaminated haemodialysis monitor would modify the efficacy of different disinfection modalities against bacteria and endotoxin concentrations. METHODS: A water-borne Pseudomonas aeruginosa (109) suspension was recirculated for 1 h and was left standing for 72 h (stationary phase) in the hydraulic circuit of the monitor. The monitor was then washed and disinfected by different physical (heat, 85 degrees C) or chemical (hypochlorite or peracetic acid) disinfection modalities (protocol A). In protocol B, the bacterial suspension was also recirculated for 1 h, but the monitor was then immediately washed and disinfected by different chemical disinfection modalities (hypochlorite or peracetic acid). RESULTS: Biofilm formation was revealed by scanning and confocal laser electron microscopy after the stationary phase (protocol A), but was absent when the monitor was immediately washed and disinfected (protocol B). In the presence of biofilm (protocol A), heat in association with citric acid was the most effective modality for reducing both colony forming units and endotoxin concentrations, whereas heat by itself was the least effective method of disinfection. Dwelling (60 h) with diluted peracetic acid completely prevented the formation of biofilm. In the absence of biofilm (protocol B), chemical disinfection proved to be effective against both colony forming units and endotoxin concentrations. CONCLUSIONS: We found that biofilm formation may markedly reduce the efficacy of presently available disinfection modalities. Therefore, different disinfection modalities and the combined action of descaling (by citric acid) and disinfection (physical/chemical agents) should be used periodically in haemodialysis monitors. In addition, dwelling with diluted peracetic acid should be adopted whenever monitors are not in use.

Bacterial Infections↗
PubMed2005

Biofilm theory can guide the treatment of device-related orthopaedic infections.

Direct observations of the surfaces of orthopaedic prostheses that have failed and of bone affected by osteomyelitis with and without the presence of a prosthesis have shown that the bacteria that cause these infections live in well-developed biofilms. The cells within these matrix-enclosed surface-associated communities are protected from host defenses and antibiotics, and clinical experience has shown that they must be removed physically before the infection can be resolved. The biofilm etiology of these diseases demands new diagnostic methods because biofilm cells typically do not grow on agar plates when recovered by scraping or swabbing. I will recommend new molecular and immunologic diagnostic methods that have been useful in other biofilm infections. These diseases progress through quiescent periods that alternate with acute exacerbations, and clinicians must realize that antibiotic therapy can control the acute phases but cannot resolve the basic biofilm nidus of the infection. Now that it has been realized that these orthopaedic infections are caused by relatively common biofilm-forming bacterial pathogens, new technologies that deliver very high concentrations of antibiotics locally and "on demand" and novel molecular "mimics" that block the signals that control biofilm formation need to be examined.

Anti-Bacterial Agents↗
PubMed2005

Molecular and imaging techniques for bacterial biofilms in joint arthroplasty infections.

Biofilm formation on surfaces is an ancient and integral strategy for bacterial survival. Billions of years of adaptation provide microbes with the ability to colonize any surface, including those used in orthopaedic surgery. Although remarkable progress has been made in the treatment of orthopaedic diseases with implanted prostheses, infection rates remain between 1% and 2%, and are higher for revision surgeries. The chronic nature of implant infections, their nonresponsiveness to antibiotics, and their frequent culture negativity can be explained by the biofilm paradigm of infectious disease. However, the role of biofilms in orthopaedic implant infections and aseptic loosening is controversial. To address these issues, we developed molecular diagnostic and confocal imaging techniques to identify and characterize biofilms associated with infected implants. We designed PCR and reverse transcription (RT)-PCR-based assays that can be used to detect bacterial infections associated with culture-negative joint effusions that distinguish between physiologically active Staphylococcus aureus and Staphylococcus epidermidis. Using clinical isolates of Pseudomonas aeruginosa, we constructed a series of reporter strains expressing colored fluorescent proteins to observe biofilms growing on 316L stainless steel and titanium orthopaedic screws. Three-dimensional structures of Pseudomonas aeruginosa and staphylococci biofilms growing on the screws were documented using confocal microscopy. The application of these tools for clinical diagnosis and biofilm research in animal and in vitro models is discussed.

Arthroplasty↗
PubMed2000

Effect of probiotic bacteria on prevalence of yeasts in oropharyngeal biofilms on silicone rubber voice prostheses in vitro.

The proliferation of yeasts in the mixed bacterial and fungal biofilms colonising silicone rubber voice prostheses in laryngectomised patients is the main cause of malfunctioning of the valve mechanism on the oesophageal side of the prostheses. Indwelling voice prostheses usually have to be replaced every 3-4 months. The consumption of probiotic bacteria is largely motivated by health claims related to the urogenital and lower digestive tract, but not to the upper digestive tract. The present study examined the influence of probiotic bacteria on the prevalence of yeasts in oropharyngeal biofilms on silicone rubber voice prostheses, as formed in a modified Robbins device. Exposure of oropharyngeal biofilms on voice prostheses to suspensions of Bifidobacterium infantis 420 or Enterococcus faecium 603 did not significantly reduce the number of yeasts in the biofilm. However, suspensions of Lactobacillus fermentum B54, L. rhamnosus 744 or L. lactis cremoris SK11 led to a reduction in the number of yeasts harvested from the voice prostheses. Suspensions of L. casei Shirota and Streptococcus thermophilus B significantly reduced the number of yeasts in the biofilm to 39% and 33%, respectively. The reduction brought about in yeast prevalence in the mixed biofilm was greatest by exposure to a suspension of L. lactis 53, with yeast prevalence only 4% of the control. In conclusion, the study demonstrated that the prevalence of yeasts in oropharyngeal biofilms on silicone rubber voice prostheses might be controlled by consumption of probiotic bacteria.

Bifidobacterium↗
PubMed2001

Assessment of lectin-binding analysis for in situ detection of glycoconjugates in biofilm systems.

An assessment of lectin-binding analysis for the characterization of extracellular glycoconjugates as part of the extracellular polymeric substances in environmental microbial communities was performed using fully hydrated river biofilms. The applicability of the method was evaluated for single, dual and triple staining with a panel of fluor-conjugated lectins. It was shown that lectin-binding analysis was able to stain glycoconjugates within biofilm communities. Lectin staining also demonstrated spatial heterogeneity within the biofilm matrix. Furthermore, the application of two or even three lectins was possible if suitable combinations were selected. The lectin-binding analysis can be combined with general nucleic acid stains to collect both nucleic acid and glycoconjugate signals. The effects of incubation time, lectin concentration, fluor labelling, carbohydrate inhibition, order of addition and lectin interactions were studied. An incubation time of 20 min was found to be sufficient for completion of lectin binding. It was not possible to ascertain saturating concentration for individual lectins, therefore a standard concentration was used for the assay. Carbohydrate inhibition tests indicated that fluorescein isothiocyanate (FITC)-conjugated lectins had more specific binding characteristics than tetramethyl rhodamine isothiocyanate (TRITC)- or cyanine dye (CY5)-labelled lectins. The order of addition and the nature of the fluor conjugate were also found to influence the binding pattern of the lectins. Therefore the selection of a panel of lectins for investigating the EPS matrix must be based on a full evaluation of their behaviour in the biofilm system to be studied. Despite this necessity, lectin-binding analysis represents a valuable tool to examine the glycoconjugate distribution in fully hydrated biofilms. Thereby, chemical heterogeneities within extracellular biofilm locations can be identified in order to examine the role (e.g. sorption properties, microenvironments, cell-extracellular polymeric substance interactions) of the extracellular polymeric substances in environmental biofilm systems.

Bacteria↗
PubMed2001

The cep quorum-sensing system of Burkholderia cepacia H111 controls biofilm formation and swarming motility.

Burkholderia cepacia and Pseudomonas aeruginosa often co-exist as mixed biofilms in the lungs of patients suffering from cystic fibrosis (CF). Here, the isolation of random mini-Tn5 insertion mutants of B. cepacia H111 defective in biofilm formation on an abiotic surface is reported. It is demonstrated that one of these mutants no longer produces N-acylhomoserine lactones (AHLs) due to an inactivation of the cepR gene. cepR and the cepI AHL synthase gene together constitute the cep quorum-sensing system of B. cepacia. By using a gene replacement method, two defined mutants, H111-I and H111-R, were constructed in which cepI and cepR, respectively, had been inactivated. These mutants were used to demonstrate that biofilm formation by B. cepacia H111 requires a functional cep quorum-sensing system. A detailed quantitative analysis of the biofilm structures formed by wild-type and mutant strains suggested that the quorum-sensing system is not involved in the regulation of initial cell attachment, but rather controls the maturation of the biofilm. Furthermore, it is shown that B. cepacia is capable of swarming motility, a form of surface translocation utilized by various bacteria to rapidly colonize appropriate substrata. Evidence is provided that swarming motility of B. cepacia is quorum-sensing-regulated, possibly through the control of biosurfactant production. Complementation of the cepR mutant H111-R with different biosurfactants restored swarming motility while biofilm formation was not significantly increased. This result suggests that swarming motility per se is not essential for biofilm formation on abiotic surfaces.

Bacterial Adhesion↗
PubMed1996

Susceptibility of oral bacterial biofilms to antimicrobial agents.

There is great interest in the use of antimicrobial agents for the prevention and treatment of plaque-related oral diseases and many publications have reported the results of studies in which the minimum inhibitory concentrations of agents for cariogenic and periodontopathogenic bacteria have been determined. However, such data are relevant only to situations where the organisms of interest are in aqueous suspension, whereas in caries and the inflammatory periodontal diseases the target organisms are in the form of biofilms. On the basis of studies with medically important bacteria, it has been established that bacteria in biofilms are invariably less susceptible to antimicrobial agents than their planktonic counterparts. Therefore, in the laboratory assessment of agents which may be suitable for treating plaque-related diseases, the target organisms should be in the form of biofilms. While laboratory evaluation of chemical agents for the prevention of plaque formation has usually employed biofilm-based models, the search for antimicrobial agents effective in the treatment of plaque-related diseases has not. Therefore, there are few data available regarding those characteristics of antimicrobial agents (e.g., their biofilm eliminating concentrations or biofilm killing concentrations) that could be used to judge their suitability for treating plaque-related diseases. In this review the limited information available concerning the antimicrobial susceptibility of oral bacteria in biofilms is presented.

Anti-Bacterial Agents↗
PubMed2007

Formation and properties of in vitro biofilms of ica-negative Staphylococcus epidermidis clinical isolates.

Coagulase-negative Staphylococcus epidermidis has become the leading cause of foreign-body infections due to its biofilm formation on all kinds of medical-device surfaces. The biofilm development of S. epidermidis includes two steps: the initial attachment phase and the accumulative phase. In the accumulative phase, the polysaccharide intercellular adhesin (PIA), encoded by the icaADBC locus, is the major component mediating intercellular adhesion. However, recent studies have revealed the emergence of biofilm-positive/ica-negative staphylococcal clinical isolates. In this report, two ica-negative S. epidermidis clinical strains, SE1 and SE4, exhibited their heterogeneity in biofilm architecture under static and flow conditions, compared with the biofilm-positive/ica-positive RP62A strain. Strains with this type of absence of PIA from biofilms also displayed intermediate resistance to vancomycin. More importantly, the cells of both SE1 and SE4 strains were more tolerant than those of RP62A to exposure to lysostaphin and vancomycin. Based on the results, it is suggested that the biofilm-positive/ica-negative strain represents a newly emergent subpopulation of S. epidermidis clinical strains, arising from selection by antibiotics in the nosocomial milieu, which displays a survival advantage in its host environment. Recent epidemiological data support this suggestion, by showing a tendency towards an increasing proportion of this subpopulation in staphylococci-associated infections.

Anti-Bacterial Agents↗
PubMed2003

Persistence of biofilm-associated Escherichia coli and Pseudomonas aeruginosa in groundwater and treated effluent in a laboratory model system.

This study was based on the hypothesis that groundwater-derived biofilms may provide a reservoir for coliform or pathogenic bacteria as has been observed in drinking water distribution systems. Escherichia coli, labelled with green fluorescent protein, was found to colonize all layers of mixed-population biofilms developed in association with indigenous groundwater micro-organisms in a laboratory-scale reactor. Biofilm-associated E. coli was removed at a slower rate from the reactor flasks than planktonic E. coli under a continuous flow regime. During flow-through of groundwater, planktonic E. coli removal was slower in flasks containing coverslips for enhanced biofilm development compared to a control flask without coverslips. Conversely, during flow-through of treated effluent, planktonic E. coli removal was faster in flasks with coverslips compared to without. Removal of attached E. coli was also fastest in the coverslip-containing flasks with effluent flow-through. This suggests that an increase in available nutrients may reduce E. coli survival potential due to either enhanced competition for nutrients or enhanced antagonism by the indigenous microbial population. Under identical conditions, GFP-labelled Pseudomonas aeruginosa was found to persist in the biofilms for longer than E. coli, most notably when exposed to flow-through of treated effluent. However, prolonged persistence of P. aeruginosa in the effluent could not be attributed to an association with the biofilms. This study has shown that under certain conditions the presence of mixed-population biofilms may limit the survival potential of enteric bacterial pathogens introduced into groundwater.

Biofilms↗
PubMed2005

The pel genes of the Pseudomonas aeruginosa PAK strain are involved at early and late stages of biofilm formation.

Pseudomonas aeruginosa is a Gram-negative bacterium associated with nosocomial infections and cystic fibrosis. Chronic bacterial infections are increasingly associated with the biofilm lifestyle in which microcolonies are embedded in an extracellular matrix. Screening procedures for identifying biofilm-deficient strains have allowed the characterization of several key determinants involved in this process. Biofilm-deficient P. aeruginosa PAK strains affected in a seven-gene cluster called pel were characterized. The pel genes encode proteins with similarity to components involved in polysaccharide biogenesis, of which PelF is a putative glycosyltransferase. PelG was also identified as a putative component of the polysaccharide transporter (PST) family. The pel genes were previously identified in the P. aeruginosa PA14 strain as required for the production of a glucose-rich matrix material involved in the formation of a thick pellicle and resistant biofilm. However, in PA14, the pel mutants have no clear phenotype in the initiation phase of attachment. It was shown that pel mutations in the PAK strain had little influence on biofilm initiation but, as in PA14, appeared to generate the least robust and mature biofilms. Strikingly, by constructing pel mutants in a non-piliated P. aeruginosa PAK strain, an unexpected effect of the pel mutation in the early phase of biofilm formation was discovered, since it was observed that these mutants were severely defective in the attachment process on solid surfaces. The pel gene cluster is conserved in other Gram-negative bacteria, and mutation in a Ralstonia solanacearum pelG homologue, ragG, led to an adherence defect.

Bacterial Adhesion↗
PubMed2005

Bap-dependent biofilm formation by pathogenic species of Staphylococcus: evidence of horizontal gene transfer?

The biofilm-associated protein (Bap) is a surface protein implicated in biofilm formation by Staphylococcus aureus isolated from chronic mastitis infections. The bap gene is carried in a putative composite transposon inserted in SaPIbov2, a mobile staphylococcal pathogenicity island. In this study, bap orthologue genes from several staphylococcal species, including Staphylococcus epidermidis, Staphylococcus chromogenes, Staphylococcus xylosus, Staphylococcus simulans and Staphylococcus hyicus, were identified, cloned and sequenced. Sequence analysis comparison of the bap gene from these species revealed a very high sequence similarity, suggesting the horizontal gene transfer of SaPIbov2 amongst them. However, sequence analyses of the flanking region revealed that the bap gene of these species was not contained in the SaPIbov2 pathogenicity island. Although they did not contain the icaADBC operon, all the coagulase-negative staphylococcal isolates harbouring bap were strong biofilm producers. Disruption of the bap gene in S. epidermidis abolished its capacity to form a biofilm, whereas heterologous complementation of a biofilm-negative strain of S. aureus with the Bap protein from S. epidermidis bestowed the capacity to form a biofilm on a polystyrene surface. Altogether, these results demonstrate that Bap orthologues from coagulase-negative staphylococci induce an alternative mechanism of biofilm formation that is independent of the PIA/PNAG exopolysaccharide.

Adhesins, Bacterial↗
PubMed2006

Biofilm formation by Pseudomonas fluorescens WCS365: a role for LapD.

A role for the outer-membrane-associated LapA protein in early biofilm formation by Pseudomonas fluorescens WCS365 has previously been shown. This paper reports that lapD, a gene located adjacent to the lapA gene, also plays a role in biofilm formation. A mutation in lapD results in a conditional biofilm defect in a static assay - this biofilm phenotype is exacerbated when biofilm formation is assayed in a flow-cell system. Furthermore, a lapD mutation shows a partial defect in the transition from reversible to irreversible attachment, consistent with an early role for the lapD gene product in biofilm formation. LapD is shown to be localized to the inner membrane of P. fluorescens. The data show decreased LapA associated with the cell surface, but no apparent change in cytoplasmic levels of this protein or lapA transcription, in a lapD mutant. A model is proposed wherein the role of LapD in biofilm formation is modulating the secretion of the LapA adhesin.

Adhesins, Bacterial↗