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Analysis of different genetic traits and their association with biofilm formation in Staphylococcus epidermidis isolates from central venous catheter infections.

The aim of the present study was to characterize clinical isolates of Staphylococcus epidermidis, one of the bacterial species most often implicated in foreign-body-associated infections, for their ability to form biofilms and for the presence of mecA and IS256 element. Sixty-seven Staphylococcus epidermidis clinical isolates, obtained from implantable medical devices, were investigated. Overall, 70% of the strains were positive for ica operon genes, 85% possessed atlE, and 46% contained aap. In 89% of the population, the Congo red agar test confirmed the correlation between the presence of ica genes and slime expression. Almost all of the strains could be classified as biofilm producers by both the crystal violet assay and microscopy. The bacterial population studied showed a very high frequency of strains positive for mecA as well as for the IS256 element. Although well-structured biofilms have been previously observed only in those strains possessing genes belonging to the ica operon, this study demonstrates that strains lacking specific biofilm-formation determinants can be isolated from catheters and can form a biofilm in vitro. Hence, different and yet-to-be identified factors may work together in the formation and organization of complex staphylococcal microbial communities and sustain infections associated with implanted medical devices.

Anti-Bacterial Agents↗

Staphylococcus epidermidis ventriculitis treated with vancomycin and rifampin.

Two cases of ventriculitis with Staphylococcus epidermidis that failed on therapy with an antistaphylococcal penicillin are presented. Both infections responded to a combination of intravenous and intraventricular vancomycin and rifampin. Vancomycin and rifampin represent an important antibiotic regimen for the management of resistant infections of the central nervous system, especially with those due to S. epidermidis or methicillin-resistant Staphylococcus aureus.

Anti-Bacterial Agents↗

Purification and properties of a bacteriocin of Staphylococcus epidermidis isolated from dental plaque.

An extracellular bacteriocin of Staphylococcus epidermidis isolated from dental plaque was purified and characterized. Its molecular mass was 3500 Da and pI was 10.5. This bacteriocin inhibited the growth of Staphylococcus aureus, Streptococcus salivarius, and Streptococcus mitis, but Streptococcus mutans, Streptococcus sanguis and other oral indigenous bacterial species examined were not inhibited. The mode of inhibition was found to be bacteriostatic.

Bacterial Proteins↗

Experimental treatment of vascular graft infection due to Staphylococcus epidermidis by in situ replacement with a rifampin-bonded polyester graft.

In situ prosthetic graft replacement (ISPGR) of an infected prosthesis raises the risk of recurrent infection in the new graft, especially in cases involving drug-resistant microorganisms. The purpose of this animal study was to evaluate in situ replacement of a vascular graft infected by a highly rifampin-resistant strain of Staphylococcus epidermidis with the use of a rifampin-bonded polyester graft. Antibiotic bonding was obtained by soaking grafts in a high dose of rifampin solution (60 mg/mL). The infrarenal abdominal aorta of 20 dogs was replaced using a polyester prosthesis infected with a highly rifampin-resistant strain of Staphylococcus epidermidis. One week later, the 18 surviving animals were randomized into three groups. Group I (n = 6) did not undergo reoperation. Group II (n = 6) underwent ISPGR using a rifampin-bonded prosthesis. Group III (n = 6) underwent ISPGR using an untreated prosthesis. All surviving animals were killed 28 days after the first procedure. Infectious signs were noted and bacteriological study was carried out on explanted prostheses and various tissue samples. The findings of this experimental study show that soaking a polyester prosthesis in a high-dose rifampin solution can prevent reinfection after in situ replacement of a prosthesis infected by a highly rifampin-resistant Staphylococcus epidermidis.

Anastomosis, Surgical↗

Imipenem therapy of experimental Staphylococcus epidermidis endocarditis.

Imipenem was evaluated for its activity against Staphylococcus epidermidis in vitro and in a rabbit model of endocarditis. The MBC for imipenem of 55 methicillin-resistant S. epidermidis isolates from patients with prosthetic valve endocarditis increased by eightfold or greater with increasing inoculum size; there was no inoculum-associated increase in the imipenem MBC for 20 methicillin-susceptible S. epidermidis isolates. Endocarditis was produced in rabbits with either a methicillin-susceptible or a methicillin-resistant S. epidermidis isolate to investigate the correlation in vivo of the in vitro inoculum effect for imipenem. Six days of imipenem treatment eradicated methicillin-susceptible S. epidermidis from vegetations of infected rabbits significantly better than no therapy but was less effective against methicillin-resistant S. epidermidis in this regard. Among methicillin-resistant S. epidermidis-infected rabbits, 6 days of imipenem therapy (i) was not significantly better than that of the control and was significantly worse than that of vancomycin in eradicating bacteria from infected vegetations and (ii) increased the frequency of imipenem-resistant subpopulations in infected vegetations. Resistant subpopulations were not seen in vegetations from untreated or imipenem-treated, methicillin-susceptible S. epidermidis-infected rabbits. Imipenem may not be effective therapy for serious human methicillin-resistant S. epidermidis infections.

Animals↗

Effects of Staphylococcus epidermidis on cellular immunity to infection with Listeria monocytogenes.

With the present study, the effects of intravenous applications of Staphylococcus epidermidis (SE) on the course of experimental infections of mice with Listeria monocytogenes were evaluated. SE treatment 24 h prior to Listeria infection led to a reduced growth of Listeria organisms in both livers and spleens and to an increased resistance of infected animals against a lethal Listeria challenge. SE treatment 24 h after Listeria infection resulted in an enhanced growth of and retarded elimination of Listeria organisms from animal organs as well as in a reduction of delayed-type hypersensitivity to soluble Listeria antigen. Adoptive immunotherapy accomplished by transferring immune peritoneal exudate T-lymphocyte-enriched cells (PETLEs) to Listeria-infected recipients 24 h before SE treatment did not prevent the delay in clearance of Listeria organisms. When Listeria-infected recipients compromised in their immune response by SE treatment were infused with immune PETLEs either immediately or 24 h after the application of SE, the immunosuppression induced by SE proved to be reversible. It is concluded that, in analogy to other bacterial immunomodulators, Staphylococcus epidermidis is able to either nonspecifically activate macrophages or interfere with T-lymphocyte functions.

Animals↗

In-vitro selection of resistance to vancomycin and teicoplanin in Enterococcus faecium and Enterococcus faecalis compared with Staphylococcus epidermidis.

Clinical isolates of Enterococcus faecium, Enterococcus faecalis and Staphylococcus epidermidis were studied for their ability to develop resistance by two selection methods, one in broth and one on agar. The MICs for enterococci after exposure to vancomycin ranged from 0.5 mg/L to 4 mg/L, and those after exposure to teicoplanin ranged from 0.25 mg/L to 1 mg/L. No significant increases occurred for E. faecalis, whilst two isolates of E. faecium showed increases of up to eight-fold (from 0.125 mg/L to 1 mg/L). Vancomycin MICs ranged from 1 mg/L to 4 mg/L and teicoplanin MICs reached 32 mg/L for S. epidermidis. Pulsed-field gel electrophoresis analysis of SmaI-digested whole chromosomal DNA was performed to compare the genomic DNA of glycopeptide-exposed and wild-type strains. In-vitro exposure to vancomycin did not alter MICs significantly except in one S. epidermidis isolate for which the MIC reached 4 mg/L, whereas the teicoplanin-exposed cultures of S. epidermidis showed increases of up to 64 times the original MIC. Comparable results were achieved with the two selection methods.

Anti-Bacterial Agents↗

Role of granulocytes and monocytes in the prevention and therapy of experimental Staphylococcus epidermidis endocarditis in rabbits.

Rabbits with endocarditis caused by Staphylococcus epidermidis were studied to determine the parts played by granulocytes and monocytes in the prevention or outcome of therapy with cloxacillin. Both monocytes and granulocytes influenced prophylaxis with cloxacillin. The amount of cloxacillin needed to prevent infection in 50% of the rabbits was significantly less in control rabbits than in those selectively depleted of monocytes, as it was also in rabbits selectively depleted of monocytes compared with those that had both granulocytopenia and monocytopenia. Granulocytes strongly potentiated the effect of cloxacillin during prophylaxis, whereas the contribution of monocytes was merely additive. Monocytes also contributed to the effect of therapy with cloxacillin, partially via a cloxacillin-independent mechanism and partially by potentiation of the effect of cloxacillin. Granulocytes did not appear to affect cloxacillin therapy. Results of this study suggest that currently used regimens for prophylaxis and treatment of S. epidermidis endocarditis may need to be adjusted for neutropenic patients.

Animals↗

Key role of poly-gamma-DL-glutamic acid in immune evasion and virulence of Staphylococcus epidermidis.

Coagulase-negative staphylococci, with the leading species Staphylococcus epidermidis, are the predominant cause of hospital-acquired infections. Treatment is especially difficult owing to biofilm formation and frequent antibiotic resistance. However, virulence mechanisms of these important opportunistic pathogens have remained poorly characterized. Here we demonstrate that S. epidermidis secretes poly-gamma-DL-glutamic acid (PGA) to facilitate growth and survival in the human host. Importantly, PGA efficiently sheltered S. epidermidis from key components of innate host defense, namely antimicrobial peptides and neutrophil phagocytosis, and was indispensable for persistence during device-related infection. Furthermore, PGA protected S. epidermidis from high salt concentration, a key feature of its natural environment, the human skin. Notably, PGA was synthesized by all tested strains of S. epidermidis and a series of closely related coagulase-negative staphylococci, most of which are opportunistic pathogens. Our study presents important novel biological functions for PGA and indicates that PGA represents an excellent target for therapeutic maneuvers aimed at treating disease caused by S. epidermidis and related staphylococci.

Animals↗

Further characterization of Staphylococcus epidermidis transposon mutants deficient in primary attachment or intercellular adhesion.

Biofilm formation is suggested to be the result of primary attachment of Staphylococcus epidermidis cells to a polymer surface followed by accumulation in multilayered cell clusters. Here we describe the further characterization of transposon (Tn917) mutants of Staphylococcus epidermidis O-47 having been biofilm-negative in a polystyrene microtiter plate adhesion assay. Among 5000 Tn917 insertion strains, we isolated four biofilm-negative mutants, each carrying one copy of Tn917. The mutants could be divided into two phenotypic classes: class A (mut1 and mut1a) and class B (mut2 and mut2a). Mutants of phenotype class A lacked four cell surface proteins and were affected in the primary attachment to polystyrene, but remained able to form multilayered cell clusters and to produce PIA. Mutants of phenotype class B were able to attach to polystyrene, but did not form multilayered cell clusters nor produce PIA. The cell surface protein pattern relative to the wild type was unchanged in class B mutants. On Congo red agar, the wild type and class A mutants formed black colonies (positive reaction on Congo red agar) while class B mutant colonies were red (negative reaction). The initial binding of cells to polystyrene and the ability to form multilayered cell clusters were found to be phenotypically and genetically distinct traits.

Adhesins, Bacterial↗

Staphylococcus epidermidis: emerging resistance and need for alternative agents.

Previously, Staphylococcus epidermidis and other coagulase-negative staphylococci isolated from the blood of hospitalized patients were often considered contaminants. Now, coagulase-negative staphylococci are among the leading causes of nosocomial blood infections. Multidrug resistance could predict a true nosocomial infection rather than a blood culture contaminant. Recent studies indicated the emergence of resistance to the quinolones, particularly to ciprofloxacin. Tolerance and occasional resistance to vancomycin have been reported recently. In addition, several reports indicated that vancomycin and other glycopeptide antibiotics lose their effectiveness against S. epidermidis organisms embedded in the biofilm environment on the surface of medical devices. Alternative agents have been proposed in the prevention and treatment of device-related and glycopeptide-tolerant S. epidermidis infections. These agents include minocycline, rifampin, and, more recently, quinupristin/dalfopristin and the oxazolidinones.

4-Quinolones↗

Site-specific adhesion of Staphylococcus epidermidis (RP12) in Ti-Al-V metal systems.

Staphylococcus epidermidis (RP12) adhesion patterns were studied on the following titanium (Ti)-aluminium (Al)-vanadium (V) metal systems: (i) microfabricated samples consisting of Ti, Al and V islands deposited onto Ti or V substrata, (ii) pure Ti, Al and V metals, and (iii) medical grade Ti6Al4-V alloy. All of these surfaces were covered with their respective oxides formed upon exposure of the metals to air. Quantitative analysis of the number of cells bound per unit area indicates that S. epidermidis (RP12) exhibits greatest adhesion to pure V surfaces. When exposed to surfaces having controlled spatial variations in chemical composition on the 10 microns scale (microfabricated samples), the bacteria preferentially populate V islands versus Ti or Al substrata. In the case of the biphasic Ti6Al4V alloy, the bacteria tend to adhere to V-rich, mixed phase regions and phase boundaries. These findings demonstrate that enhanced and preferential adhesion of S. epidermidis (RP12) occurs on V surfaces in Ti-Al-V metal systems and suggest that bacterial interactions are influenced by surface oxide composition.

Alloys↗

Nosocomial infections by Staphylococcus epidermidis: how a commensal bacterium turns into a pathogen.

Staphylococcus epidermidis is a commensal bacterium of the human skin. However, S. epidermidis and other coagulase-negative staphylococci (CNS) emerge also as common nosocomial pathogens infecting immunocompromized patients carrying medical devices. Antibiotic resistance and the ability of many nosocomial S. epidermidis isolates to form biofilms on inert surfaces make these infections hard to treat. Epidemiological analyses using multilocus sequence typing (MLST) and genetic studies suggest that S. epidermidis isolates in the hospital environment differ from those obtained outside of medical facilities with respect to biofilm formation, antibiotic resistance, and the presence of mobile DNA elements. Since S. epidermidis isolates exhibit high genome flexibility, they are now regarded as reservoirs for the evolution and spread of resistance traits within nosocomial bacterial communities.

Biofilms↗

Increased colonization of indwelling medical devices by quorum-sensing mutants of Staphylococcus epidermidis in vivo.

Infections with the leading nosocomial pathogen Staphylococcus epidermidis are characterized by biofilm development on indwelling medical devices. We demonstrate that the quorum-sensing regulator agr affects the biofilm development of S. epidermidis in an unexpected fashion and is likely involved in promoting biofilm detachment. An isogenic agr mutant showed increased biofilm development and colonization in a rabbit model. In addition, nonfunctional agr occurred more frequently among strains isolated from infections of joint prostheses. Lack of functionality was based on mutations, including insertion of an IS256 element. Relative to other bacterial pathogens, quorum sensing in S. epidermidis thus has a different role during biofilm development and biofilm-associated infection. Our results indicate that disabling agr likely enhances the success of S. epidermidis during infection of indwelling medical devices. The permanent elimination of quorum-sensing regulation used by S. epidermidis represents a surprising and unusual means to adapt to a certain environment and type of infection.

Animals↗

Anaerobic conditions induce expression of polysaccharide intercellular adhesin in Staphylococcus aureus and Staphylococcus epidermidis.

Products of the intercellular adhesion (ica) operon in Staphylococcus aureus and Staphylococcus epidermidis synthesize a linear beta-1,6-linked glucosaminylglycan. This extracellular polysaccharide mediates bacterial cell-cell adhesion and is required for biofilm formation, which is thought to increase the virulence of both pathogens in association with prosthetic biomedical implants. The environmental signal(s) that triggers ica gene product and polysaccharide expression is unknown. Here we demonstrate that anaerobic in vitro growth conditions lead to increased polysaccharide expression in both S. aureus and S. epidermidis, although the regulation is less stringent in S. epidermidis. Anaerobiosis also dramatically stimulates ica-specific mRNA expression in ica- and polysaccharide-positive strains of both S. aureus and S. epidermidis. These data suggest a mechanism whereby ica gene expression and polysaccharide production may act as a virulence factor in an anaerobic environment in vivo.

Aerobiosis↗

Analysis of two Staphylococcus epidermidis plasmids coding for resistance to streptogramin A.

The two Staphylococcus epidermidis plasmids pIP1629 (7.5 kb) and pIP1630 (14.4 kb) contain the vga gene conferring resistance to streptogramin A. All the sequences of pIP1629, except two of the four 22-nt iterons preceding the replication gene, were found in pIP1630. The additional 6.9-kb fragment of pIP1630 is similar to the mobilizable S. epidermidis plasmid pSK639, carrying the dfrA-thyE-orf140 operon and thought to replicate by an iteron controlled theta-type replication mechanism. The replication-mobilization elements of pIP1629 and pSK639 are very similar despite having been isolated in France and in Australia, respectively, showing that they are geographically widely dispersed in S. epidermidis. The gene thyE encoding thymidylate synthetase carried by pSK639 is not present in pIP1630. pIP1630 probably arose by the recombination of two homologous plasmids carrying distinct resistance determinants.

Australia↗

Treatment of Staphylococcus epidermidis endophthalmitis with intravitreal moxifloxacin in a rabbit model.

Staphylococcus epidermidis is one of the most common causes of postoperative infectious endophthalmitis, which is a serious complication of ocular surgery and penetrating trauma. Moxifloxacin is a newly developed fluoroquinolone with a potent antimicrobial activity. Corticosteroids are used in endophthalmitis to suppress devastating intraocular inflammatory response. This study was designed to assess the efficacy of intravitreal moxifloxacin alone and in combination with intravitreal dexamethasone. To the best of our knowledge, there is no published report demonstrating the effect of intravitreal moxifloxacin on bacterial endophthalmitis. One eye of each rabbit (n=24) was infected by inoculation of 10(5) colony-forming units (CFU) of S. epidermidis into the vitreus cavity. Rabbits received intravitreal injection of moxifloxacin (50 microg) or a combination of moxifloxacin (50 microg) and dexamethasone (400 microg). No treatment was given to control group. Clinical and histopathological examination scores and microbiological analysis of vitreus aspirates were compared. In the treatment groups, the clinical and histopathological scores and mean CFU were significantly lower than those in the control group (p <0.05) but showed no significant difference between the treatment groups. These results suggest that intravitreal injection of moxifloxacin is effective against S. epidermidis in this experimental rabbit model. Moxifloxacin may be a promising agent in the treatment of bacterial endophthalmitis.

Adrenal Cortex Hormones↗