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PubMed2003

Involvement of adherence and adhesion Staphylococcus epidermidis genes in pacemaker lead-associated infections.

We explored three genes of attachment (fbe and atlE) and adhesion (ica) in 27 and 10 Staphylococcus epidermidis strains involved in pacemaker-related infections (PMI) and intravascular-catheter-related infections (IVCI), respectively, and in 25 saprophytic strains. The detection rates of fbe and atlE were identical in PMI and IVCI strains, but ica detection rates were identical in PMI and saprophytic strains.

Bacterial Adhesion↗

PubMed1993

Central line perforation associated with Staphylococcus epidermidis infection.

We present detailed case reviews of four very low birth weight (VLBW) infants in whom central venous silastic catheter perforation was associated with Staphylococcus epidermidis sepsis. The diagnostic and therapeutic dilemma presented by the intracavitary fluid collections occurring in all four of these cases proved to be of clinical interest. Additionally, we propose a model that may account for the etiology of catheter displacement--localized phlebitis as a result of S epidermidis infection with resultant extralumenal migration of the central venous catheter.

Algorithms↗

PubMed1990

Transformation of Staphylococcus epidermidis and other staphylococcal species with plasmid DNA by electroporation.

In this paper, the influence of various parameters on plasmid transformation by electroporation of Staphylococcus epidermidis Tü3298 was investigated. Cell growth conditions, various concentrations and forms of plasmid DNA, field strength, pulse duration and media for electroporation and regeneration were tested. In order to obtain optimal transformation efficiency, the cells were incubated for 30 min with DNA before pulsing. With the optimized procedure, other staphylococcal species such as S. aureus, S. staphylolyticus and S. carnosus were transformed with an efficiency up to 3 X 10(5) transformants per micrograms pC194 plasmid DNA.

DNA, Bacterial↗

PubMed1995

Variable degree of slime production is linked to different levels of beta-lactam susceptibility in Staphylococcus epidermidis phase variants.

Recently we demonstrated that lack of mecA transcription was responsible for the decreased methicillin resistance phenotype of strongly slime-negative Staphylococcus epidermidis phase variants [Mempel M, Feucht H, Ziehbuhr W, Endres M, Laufs R, Grüter L (1994) Antimicrob Agents Chemother 38: 1251-1255]. In the present study we compared the beta-lactam susceptibility and the slime production capacity of 60 phenotypic variants of S. epidermidis parent strain RP62A identified by their colony morphology on congo red agar. We could show that the variable degree of slime production is linked to different levels of beta-lactam susceptibility in intermediate-stage phase variants. The increased deltahemolysin production of slime-negative phase variants may indicate an accessory gene regulator-like control.

Anti-Bacterial Agents↗

PubMed2005

Prevalence of Staphylococcus epidermidis strains with biofilm-forming ability in isolates from conjunctiva and facial skin.

PURPOSE: To compare the prevalence of biofilm-forming strains of Staphylococcus epidermidis in the conjunctival and facial skin microflora. DESIGN: Experimental study. METHODS: The prevalence of biofilm-forming ability of 10 S epidermidis strains obtained from the conjunctival sac of healthy volunteers was compared with 40 strains obtained from the facial skin of healthy volunteers. The ability to form biofilm was determined by the presence of the icaA gene, and the production of biofilm was examined by qualitative (Congo red agar [CRA]) and quantitative (microtiter plate) assays. Additionally, the prevalence of 36 S epidermidis strains obtained from the conjunctival sac of precataract patients to form biofilm was investigated. RESULTS: The icaA gene was detected in 60% of the isolates from the conjunctival sac of volunteers and 15% of those from the facial skin. Fifty percent of the isolates from the conjunctiva of volunteers and 5% from the facial skin were CRA positive. Biofilm production was significantly greater in isolates from the conjunctiva of volunteers. Of the nine pairs of isolates found in the same volunteers, six conjunctival sac isolates were positive for the icaA gene with biofilm-forming ability except one strain, whereas only one of the facial skin isolates was positive for the icaA gene and none exhibited biofilm-forming phenotype. Sixty-nine percent and 44% of the isolates from the conjunctival sac of precataract patients were positive for icaA gene and CRA test, respectively. CONCLUSIONS: The prevalence of biofilm-forming S epidermidis isolates is higher in the conjunctival sac than the facial skin.

Aged↗

PubMed1988

The activity of antistaphylococcal drugs on nosocomial Staphylococcus epidermidis.

In reporting on the activity of cephalothin, cefamandole, FCE 22101, gentamicin, netilmicin, amikacin, rifampicin, clindamycin, josamycin, ofloxacin, ciprofloxacin, vancomycin and teicoplanin on 72 Staphylococcus epidermidis strains clinically isolated in the hospital, the Authors observed a high percentage of methicillin-resistance (68.05%) as well as resistance to other important drugs such as gentamicin (72.22%), rifampicin (27.7%), clindamycin (36.1%), and josamycin (40.27%). They also recorded good inhibitory activity of the studied beta-lactam drugs. However, this activity was not confirmed against methicillin-resistant strains when the test was performed under particular technical conditions (hypertonic medium, incubation at 30 degrees C, inoculum = 10(6)). The Authors also emphasize the poor bactericidal activity against these strains. The activity of quinolones was good; the activity of vancomycin and teicoplanin was very good on all strains studied.

Anti-Bacterial Agents↗

PubMed1997

Staphylococcus epidermidis produces a cell-associated proteinaceous fraction which causes bone resorption by a prostanoid-independent mechanism: relevance to the treatment of infected orthopaedic implants.

Staphylococcus epidermidis is the most commonly isolated coagulase-negative staphylococcus from the skin, gut and upper respiratory tract. Although it is less virulent than Staphylococcus aureus, it has emerged in recent years as an important causative agent of infections associated with metal implants, prosthetic devices and i.v. lines. We have previously demonstrated that a saline wash of S. aureus contained proteins which had potent bone-resorbing activity in vitro. The purpose of this study was to determine whether gently washing S. epidermidis in saline also released osteolytically active proteins. The so-called surface-associated material (SAM) eluted from S. epidermidis in saline was able to induce osteolysis, and activity was heat and trypsin sensitive, suggesting that the active component was proteinaceous. Fractionation studies have suggested that activity is due to a small number of cationic proteins. This SAM-induced bone resorption was not inhibited by the cyclo-oxygenase inhibitor, indomethacin, or the 5-lipoxygenase inhibitors BWA70C and MK886. However, it was partially inhibited by high concentrations of interleukin-1 receptor antagonist and was completely blocked by a neutralizing antibody to tumour necrosis factor-alpha. Thus, the SAM from this organism is a potent osteolytic agent which differs from that of S. aureus SAM in not being inhibited by cyclo-oxygenase inhibitors. As adjunctive therapy is becoming necessary in infectious diseases, either as a result of the side-effects of antibiotics or their lack of efficacy, consideration should be given to the future treatment of bone infections with staphylococci in the light of the different mechanisms of action of the surface proteins produced by these bacteria.

Animals↗

PubMed1998

Identification of multiresistant Staphylococcus epidermidis in neonates of a secondary care hospital using pulsed field gel electrophoresis and quantitative antibiogram typing.

AIMS: To determine the diversity of types of Staphylococcus epidermidis in a neonatal care unit of a secondary care hospital in the Netherlands. METHODS: In a prospective study, specimens from nose, ear, axilla, umbilicus, and groin were taken from patients twice a week during a period of up to two weeks. All isolates were typed by both pulsed field gel electrophoresis (PFGE) and antibiogram analysis. RESULTS: Fifty three S epidermidis isolates from 15 of 24 patients were obtained in one to four surveys. Fourteen isolates from six patients had a common PFGE pattern and were of one multiresistant antibiogram type. The remaining 39 isolates were allocated to 24 sporadic PFGE types and were more susceptible to antibiotics. Colonisation with the multiresistant strain correlated with a long period of stay and with the use of specific antibiotics. The multiresistant isolates were related closely to isolates of S epidermidis found in a recent study in a teaching hospital in the vicinity of the secondary care hospital. CONCLUSION: Repeated sampling and the use of two typing methods allowed the identification of two closely related multiresistant S epidermidis strains in two hospitals in the same area.

Bacterial Typing Techniques↗

PubMed2001

Pharmacodynamics of moxifloxacin and levofloxacin against Staphylococcus aureus and Staphylococcus epidermidis in an in vitro pharmacodynamic model.

An in vitro pharmacokinetic model was used to compare the pharmacodynamics of moxifloxacin and levofloxacin against 3 Staphylococcus aureus and 3 Staphylococcus epidermidis strains. Logarithmic-phase cultures were inoculated into the peripheral compartment of hollow-fiber cartridges and exposed to the peak serum concentrations achieved in humans with oral doses of moxifloxacin (400 mg) and levofloxacin (500 mg). Drugs were added at 0 and 24 h, elimination kinetics were simulated, and changes in viable bacterial counts were evaluated over the course of 36 h. Moxifloxacin was bactericidal against all 6 staphylococci (times to 99.9% kill, 1-3 h). Against most strains, bacterial killing continued through 36 h, with total kills exceeding 5.5 logs. Levofloxacin was bactericidal against 5 of the strains, with similar times to 99.9% kill. In contrast to moxifloxacin, however, resistant subpopulations emerged in 4 strains during therapy with levofloxacin, and this could have important implications for treatment of staphylococcal infections. These in vitro observations warrant the clinical evaluation of moxifloxacin in the treatment of staphylococcal infections.

Anti-Infective Agents↗

PubMed1991

Characterization of clinically significant isolates of Staphylococcus epidermidis from patients with cerebrospinal fluid shunt infections.

Biotyping, slime production, antibiograms, extrachromosomal DNA banding and total DNA restriction analysis were used to characterize Staphylococcus epidermidis strains causing cerebrospinal fluid shunt infections in 11 patients. Infections considered to be community acquired and those acquired in the first 2 weeks of hospital admission were due to oxacillin-susceptible isolates. Multiply resistant strains were isolated from patients who were in hospital for more than 1 month before tube implantation. Slime was detected in staphylococci for 54% of cases, but its expression varied. Strains from different patients could be differentiated from one another by the extrachromosomal DNA bandings and total DNA restriction patterns, but isolates from the same patient were usually similar. During the period of external drainage, epidemiological markers were useful in differentiating persistence of infection from contamination or re-infection by a new strain.

Adult↗

PubMed1991

Selection of colony, plasmid, and virulence variants of Staphylococcus epidermidis NRC853 during growth in continuous cultures exposed to erythromycin.

A continuous-culture system was developed to study changes in the structure of Staphylococcus epidermidis populations exposed to subminimum inhibitory concentrations of erythromycin. Continuous-culture experiments were carried out in a dextrose-free, tryptic soy broth medium supplemented with lactic acid and sodium lactate (MTSB-D). The multiresistant (penicillin-, tetracycline-, and erythromycin-resistant) S. epidermidis strain NRC853 was subjected to a series of experiments: (i) growth individually in continuous culture in the absence and presence of erythromycin and (ii) growth in mixed culture with the erythromycin-susceptible S. epidermidis strain NRC852 in the absence and presence of erythromycin. Strain NRC853 produced colony morphology variants during continuous culture in the presence of 0.05 and 0.1 microgram of erythromycin per ml. Variants (A, B, and C) were different from their wild-type parent on the basis of colony size, sector pattern, and/or the ability to transmit light. A variants rapidly lost a 2.7-MDa tetracycline resistance plasmid. B and C variants formed an ermC plasmid multimer series from unit size to a 16-mer and exhibited an approximately twofold increase in erythromycin MIC over that of the wild-type parent. They slowly lost the tetracycline resistance plasmid. The small-colony B variant demonstrated an increased virulence in the neonatal mouse weight gain test and an increase in fibronectin binding compared with the wild-type parent. The presence of a competing strain drastically increased the frequency of all variants.

Animals↗

PubMed1989

Activities of daptomycin and teicoplanin against Staphylococcus haemolyticus and Staphylococcus epidermidis, including evaluation of susceptibility testing recommendations.

The in vitro activities of daptomycin, teicoplanin, and three other antimicrobial agents were determined against 105 strains of Staphylococcus haemolyticus and 92 strains of Staphylococcus epidermidis. The MICs for 90% of strains tested (MIC90s) of fusidic acid and rifampin were less than or equal to 0.25 microgram/ml. The MIC90s of daptomycin and vancomycin were less than or equal to 4 micrograms/ml. Teicoplanin had a comparable MIC90 of less than or equal to 4 micrograms/ml for isolates of S. epidermidis. However, MIC90s were 8 and 16 micrograms/ml for oxacillin-susceptible and oxacillin-resistant S. haemolyticus, respectively. Disk diffusion tests were evaluated for daptomycin and teicoplanin. Disks with 30 micrograms of teicoplanin performed satisfactorily when S. epidermidis was tested, but when S. haemolyticus was tested, there was a very major error rate of 10% and a minor error rate of 38%.

Anti-Bacterial Agents↗

PubMed1993

Isolation and characterization of transposon mutants of Staphylococcus epidermidis deficient in capsular polysaccharide/adhesin and slime.

We used transposon (Tn) mutagenesis to study the role of capsular polysaccharide/adhesin (PS/A) and slime in adherence of Staphylococcus epidermidis to catheters. pLTV1, containing Tn917-LTV1, was transformed into S. epidermidis M187 by protoplast fusion with S. aureus RN4220(pLTV1), creating M187(pLTV1). Tn mutants were isolated following growth at 42 degrees C; mutants deficient in PS/A and slime production were selected. PS/A- and slime-deficient Tn mutants had a 10-fold decrease in vitro in the initial phase of adherence to catheters, comparable to levels of strains that do not produce PS/A. Introduction of Tn917-LTV1-interrupted DNA from PS/A-deficient mutant M187sn3 into the parental strain via transformation of protoplasts yielded recipients with inserts identical to those of the Tn mutant that were PS/A and slime deficient. Chromosomal DNA flanking the Tn in mutant M187sn3 was cloned into Escherichia coli. The cloned DNA was found to hybridize to approximately 5-kb EcoRI fragments from the parental strain and from control Tn mutants that express parental levels of PS/A and to either approximately 9- or approximately 14-kb EcoRI fragments from other highly adherent, PS/A-producing strains. Mapping studies demonstrated that in the eight PS/A-deficient mutants that have been isolated, the Tn insertions all occur within a region of approximately 11.6 kb that is defined by three EcoRI sites. These results support previous findings indicating that in S. epidermidis PS/A is involved with in vitro adherence to plastic biomaterials and elaboration of PS/A is closely associated with slime production.

Bacterial Adhesion↗

PubMed2000

Antibiotics in the irrigating solutions reduce Staphylococcus epidermidis adherence to intraocular lenses.

PURPOSE: To investigate the effect of antibiotics in the irrigating solutions on hydrophobicity, slime production and the adherence of Staphylococcus epidermidis to intraocular lenses (IOLs). METHODS: A standard culture of S. epidermidis was incubated with a control phosphate-buffered saline (PBS) or PBS containing vancomycin (20 micrograms/ml) or gentamicin (8 micrograms/ml) or a combination of gentamicin and vancomycin (8 and 20 micrograms/ml, respectively) for 30, 60 and 120 min at 35 degrees C. The bacteria were harvested by centrifugation, and washed with PBS before incubation with IOLs for 1 h. Adhesion of bacterial cells to IOLs was determined by counting the viable cells attached to the lenses. Slime production on IOLs was measured using safranin staining. Hydrophobicity of the control cultures and cultures treated with antibiotics was assayed on the basis of the hexadecane droplet method. RESULTS: Bacterial exposure to antibiotics produced a time-dependent significant decrease in bacterial hydrophobicity and adherence to IOLs compared with the untreated control cells (p < 0.001). Hydrophobicity showed a significant correlation with adherence (r = 0.89, p < 0.001). Gentamicin was significantly more effective than vancomycin, and the synergistic combination of gentamicin and vancomycin was the most effective in reducing bacterial adherence to IOLs, hydrophobicity and slime production. CONCLUSIONS: The use of antibiotics in the irrigating solutions during cataract surgery may be useful in reducing bacterial adherence to IOLs. Further studies are needed to determine the clinical implications of these findings in reducing the incidence of post-operative endophthalmitis associated with IOL implantation.

Anti-Bacterial Agents↗

PubMed1994

Comparative analysis of a biofilm-forming Staphylococcus epidermidis strain and its adhesion-positive, accumulation-negative mutant M7.

We have isolated a stable slime-negative mutant, M7, from the wild-type Staphylococcus epidermidis RP62A by mitomycin mutagenesis. Besides its inability to produce slime in the test tube this mutant differed also in two other properties from its parent strain: it lacked the ability to accumulate on a surface, and it did not produce a 115 kDa and a 18 kDa extracellular protein. In all other tested properties such as initial adherence, growth rate, cell-wall composition, surface characteristics, DNA restriction profile, the presence of a 29 kb antibiotic resistance plasmid, and antimicrobial susceptibility profile, M7 was indistinguishable from its wild-type. The mutant is an important basis for further study of the pathogenesis of polymer-associated S. epidermidis infections.

Bacterial Adhesion↗

PubMed2006

Pheno- and genotyping of Staphylococcus epidermidis isolated from bovine milk and human skin.

The purpose of this study was to improve our knowledge concerning the epidemiology and strain diversity of Staphylococcus epidermidis isolated from bovine milk in commercial dairy herds. A total of 341 S. epidermidis isolates obtained from cows' milk (317), farmers (17) and patients (7) were characterized. Of these 105 isolates were from cows' milk in two farms, where also 17 isolates were sampled from farmers. The remaining 212 isolates from cows' milk were from 170 farms. All isolates were examined by antimicrobial susceptibility, whereas 202 were examined by pulsed-field gel electrophoresis (PFGE) and 122 by ribotyping. PFGE showed single patterns in the human strains with one exception; one strain was categorised as the same clone as four of the milk strains. PFGE divided 73 of the milk strains into 62 different patterns. The PFGE method had high discriminatory power and shows that many different S. epidermidis types exist in milk samples. Antibiotic resistance patterns matched the SmaI profiles closely in the two herds, but poorly in the routinely collected milk samples. Isolates from herd 1 showed one to five patterns, depending on the typing method used. Isolates from the milker's skin showed one pattern, which was identical to the most common pattern found in the milk isolates. Isolates from herd 2 showed three to four patterns, two of these being identical to skin isolates from the milker. As dairy cows are not a natural host for S. epidermidis the results suggest a human source of these udder infections.

Animals↗

PubMed1982

Adherence of slime-producing strains of Staphylococcus epidermidis to smooth surfaces.

Slime production is not a generally recognized feature of Staphylococcus epidermidis. In a recent outbreak of S. epidermidis intravascular catheter-associated sepsis, we noted that 63% of clinically implicated strains grew as a slimy film coating the culture tube walls when propagated in tryptic soy broth. Only 37% of randomly collected blood culture contaminants and skin isolates demonstrated a similar phenomenon (p less than 0.05). Transmission electron micrographs of these coating bacteria showed them to be encased in an extracellular matrix that stained with alcian blue. Slime production was most evident in autoclaved media containing Casamino Acids and glucose supplementation (0.25% wt/vol). There were strain and media preparation variability of slime production in the presence of other carbohydrates. Some strains were not able to produce slime under any of the tested conditions. The production or nonproduction of slime did not influence growth rate. When grown in vitro, slime producers accumulated on the surface of intravascular catheters as macrocolonies, whereas non-slime, producers did not. Transmission and scanning electron micrographs showed slime producers to be encased in an adhesive layer on the catheter surface, whereas nonproducers were not encased. These results suggest that slime-mediated adherence may be a critical factor in the pathogenesis of S. epidermidis infections of medical devices.

Adhesiveness↗

PubMed2000

Adherence of ocular isolates of staphylococcus epidermidis to ACRYSOF intraocular lenses. A scanning electron microscopy and molecular biology study.

PURPOSE: To investigate the adherence of two clinically relevant ocular isolates of Staphylococcus epidermidis to ACRYSOF intraocular lenses (IOLs) (Alcon Surgical, Fort Worth, Texas) and to determine whether the strains under study carried the intercellular adhesion (ica) locus, which encodes production of S. epidermidis antigens mediating adherence to biomaterials. DESIGN: Experimental study. PARTICIPANTS: The authors examined the in vitro adherence of two clinically relevant ocular isolates of S. epidermidis (S. epidermidis 1 and S. epidermidis 2). Adherence was tested on six ACRYSOF IOLs. METHODS: Three IOLs were placed in three separate test tubes containing 5 ml of S. epidermidis 1 suspension, and three other IOLs were placed in three test tubes containing 5 ml of S. epidermidis 2 suspension. At different times (3, 30, and 90 minutes), the IOLs were removed from the test tubes and rinsed in sterile phosphate buffered solution. The lenses were then fixed in glutaraldehyde, postfixed in osmium tetroxide, and serially dehydrated in ethyl alcohol. After critical point drying, they were sputter-coated with gold and then examined with a scanning electron microscope. In addition, polymerase chain reaction amplification was used to investigate whether the isolates under study carried the ica locus. MAIN OUTCOME MEASURES: The number of adhered bacteria per area (square millimeters) of IOL optic was calculated. Statistical analysis included calculation of arithmetic means and 95% confidence intervals (t test). Polymerase chain reaction amplification products were visualized in ethidium bromide-stained agarose gel. RESULTS: Direct counting of adherent bacteria in scanning electron microscopy photographs revealed that the total amount of adhered bacteria per area of IOL optic after 3, 30, and 90 minutes of incubation in bacterial suspension was 1306/mm(2), 3389/mm(2), and 7195/mm(2) (S. epidermidis 1) and 778/mm(2), 1056/mm(2), and 3861/mm(2) (S. epidermidis 2). Differences at 30 and 90 minutes were statistically significant (P: = 0.01 and 0.02, respectively). Polymerase chain reaction amplification revealed that S. epidermidis 1 contained the ica locus, whereas S. epidermidis 2 was ica negative. CONCLUSIONS: Different ocular isolates of S. epidermidis may differ significantly with regard to adherence to ACRYSOF IOLs. Adherence appeared to be greater when the bacterial DNA contained the ica locus. Strains of S. epidermidis carrying the ica locus may play an important role in the pathogenesis of some forms of endophthalmitis occurring after cataract surgery.

Acrylates↗