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PubMed1989

The effect of slime production on vancomycin efficacy in the treatment of experimental endocarditis due to Staphylococcus epidermidis.

We compared the efficacy of vancomycin in the treatment of experimental endocarditis caused by either a parent strain of Staphylococcus epidermidis that produced slime or a daughter strain that produced less slime. After subcutaneously administered vancomycin, we noted similar cure rates and similar microbiological characteristics of endocardial infections in rats challenged with these strains, regardless of slime produced.

Adhesiveness↗

PubMed1996

In vitro activities of two novel oxazolidinones (U100592 and U100766), a new fluoroquinolone (trovafloxacin), and dalfopristin-quinupristin against Staphylococcus aureus and Staphylococcus epidermidis.

Two oxazolidinones (U100592 and U100766), trovafloxacin, and a streptogramin combination (dalfopristin-quinupristin) were highly active in vitro against Staphylococcus aureus and Staphylococcus epidermidis, including methicillin-resistant strains. Trovafloxacin was more active than ciprofloxacin. Time-kill synergy studies demonstrated indifference for the oxazolidinones combined with vancomycin and rifampin against methicillin-resistant staphylococci. Spontaneous resistance was observed with all agents.

Acetamides↗

PubMed1993

Construction and overexpression in Escherichia coli of genetically engineered derivatives of penicillin-binding protein 2' of Staphylococcus epidermidis.

Removal of the putative amino-terminal membrane spanning region of penicillin-binding protein 2' (PBP-2') of Staphylococcus epidermidis WT55 was carried out by truncating the amino terminus-coding end of the mecA gene. PCR and site directed mutagenesis were used to introduce unique restriction sites at position 68 (HindIII) and at position 80 (NcoI) of the mecA gene, respectively. The coupling of the shortened coding regions to the trc promoter and gene fusion to the lacZ gene, aimed to facilitate subsequent protein purifications, resulted in strong expression in the cytoplasm of Escherichia coli and partial sequestration into insoluble protein granules. The truncated PBP-2' retained its penicillin-binding ability and also bound the monoclonal antibody directed against PBP-2' of Staphylococcus aureus.

Bacterial Proteins↗

PubMed1997

Identification of atypical strains of Staphylococcus epidermidis by use of molecular tools.

Four atypical coagulase-negative staphylococcal (CNS) isolates from clinical sources were compared with Staphylococcus epidermidis strains by ribotyping. The ribotypes of the four strains shared close rDNA restriction profiles with those of the S. epidermidis strains used. The DNA sequence encoding 16S rRNA demonstrated 99.9% homology with S. epidermidis. S1 nuclease experiments showed that these atypical strains formed a homogeneous genomic group. DNA-DNA homologies between the S. epidermidis type strain CCM 2124 and the four CNS isolates ranged from 70 to 89%. The guanine-plus-cytosine content of the deoxyribonucleic acid of the four strains ranged from 31 to 32 mol%.

Amino Acids↗

PubMed1986

Action of lombazole, and inhibitor of fungal ergosterol biosynthesis, on Staphylococcus epidermidis.

Lombazole had no effect on respiration at any tested concentration and had little effect on the K+ permeability of Staphylococcus epidermidis. Of the major metabolic processes investigated in this bacterium, only de novo synthesis of the cell envelope was inhibited by lombazole well in advance of an effect on growth. The time course of inhibition indicated that lombazole exerted its primary effect via inhibition of lipid synthesis; other induced changes, such as reduced synthesis of lipoteichoic acid and cell wall components, were considered to be secondary effects. Although the precise site of action in S. epidermidis has to be established, the absence of alterations in lipid patterns after treatment with lombazole suggests the toxicant may affect an essential step in lipid biosynthesis. In Candida albicans, lombazole inhibited the sterol C-14 demethylation step in the ergosterol biosynthesis pathway.

Anti-Bacterial Agents↗

PubMed1996

Identification and analysis of a gene encoding a Fur-like protein of Staphylococcus epidermidis.

A gene (fur) for a Fur-like protein was identified on a 1.1 kb chromosomal DNA fragment of Staphylococcus epidermidis BN 280; the fur gene is followed by an open reading frame coding for the N-terminus of a putative superoxide dismutase. Within the -35 promoter region of both genes, as sequence motif was detected with low similarity to Fur-binding regulatory DNA segments, the so-called Fur boxes. Fur titration in Escherichia coli strain H1717 demonstrated that the E. coli Fur protein binds to the Fur box of the promoter region of the S. epidermidis fur gene. The S. epidermidis Fur protein was expressed in E. coli as indicated by the formation of inactive dimers with the chimeric repressor CI(N)-Fur(C) (Stojiljkovic, I. and Hantke. K. (1995) Mol. Gen. Genet. 247, 199-205), but was not able to complement the Fur mutation in E. coli H1681.

Amino Acid Sequence↗

PubMed1997

Activity of quinupristin/dalfopristin against Staphylococcus epidermidis in biofilms: a comparison with ciprofloxacin.

Bactericidal effects of quinupristin/dalfopristin and ciprofloxacin have been studied against six strains of Staphylococcus epidermidis, with varying erythromycin resistance phenotypes and varying abilities to form slime and adhere, growing as biofilms. Ciprofloxacin and quinupristin/dalfopristin killed (2-3 log10 cfu/mL) planktonic and sessile cells slowly, but progressively, over the course of 48 h. Rates of killing by quinupristin/dalfopristin were independent of concentration over the range 5-30 mg/L, and were somewhat slower than those seen with ciprofloxacin. These findings suggest that quinupristin/dalfopristin may be useful in the treatment of line-associated infections.

Anti-Bacterial Agents↗

PubMed1988

Cell surface antigen of encapsulated Staphylococcus epidermidis SE-360 protects mice from homologous infection.

Cell surface antigen was mechanically extracted from encapsulated strain SE-360 of Staphylococcus epidermidis and purified by DEAE-Sephadex A 25 (Cl- form) ion exchange chromatography. This antigen manifested type-specific activity and major sugar constituents were galactose, glucose and N-acetyl-glucosamine at the molar ratio 1.00:9.05:1.65. alpha-D-glucosyl- and N-acetyl-glucosaminyl-residues were closely correlated to the antigenic determinant. In mice, protection against homologous microorganisms could be achieved by active immunization with thus purified antigen. Type-specific opsonin in rabbit anti-SE-360 serum could also be absorbed.

Animals↗

PubMed1987

Protective antibodies in human sera against encapsulated strains of Staphylococcus epidermidis.

Passive protective antibodies in 100 samples of normal human sera against challenge with three representative capsular type strains of Staphylococcus epidermidis in mice were examined. Six of them passively protected mice against capsular type I; 17 protected against capsular type II; one against capsular type III; and one against both capsular types I and II. The activities were sensitive to 2-mercaptoethanol and were absorbed out either with rabbit anti-human IgG serum, rabbit anti-human IgA serum or rabbit anti-human IgM serum. Also, the sera activities absorbed out with cell surface polysaccharide extracted from three representative capsular type strains. These results indicate that the protective activities were specifically related to three major immunoglobulins against the above cell surface polysaccharides.

Animals↗

PubMed1984

In-vitro synergy testing of triple antibiotic combinations against Staphylococcus epidermidis isolates from patients with endocarditis.

In-vitro synergy testing was performed against ten blood or valve isolates of Staphylococcus epidermidis taken from patients with endocarditis. A three-dimensional microtitre checkerboard method was used for evaluation of vancomycin-rifampicin-gentamicin. The triple combination of vancomycin plus rifampicin plus gentamicin was found to be synergistic in 70% of the isolates. Vancomycin plus rifampicin was not synergistic. Oral agents including dicloxacillin, rifampicin, and fusidic acid were also evaluated. Four methicillin-resistant isolates were relatively resistant to dicloxacillin (MIC greater than 0.79 mg/l) but all four isolates were susceptible to fusidic acid alone and rifampicin alone. The triple combination of dicloxacillin plus fusidic acid plus rifampicin was found to be synergistic in 50% of the isolates and generally superior to any two-drug combination raising the possibility of an effective oral combination of antibiotics.

Anti-Bacterial Agents↗

PubMed1991

Multiply antibiotic-resistant Staphylococcus epidermidis in patients, staff and environment--a one-week survey in a bone marrow transplant unit.

The distribution of Staphylococcus epidermidis resistant to ciprofloxacin and/or gentamicin was studied in an isolation unit for patients undergoing bone marrow transplantation. During 1 week all such strains colonizing patients or staff members or found on the clothes of staff or in the air were investigated. Antibiograms and plasmid profiles were used for epidemiological typing. Thirty-one different antibiograms were found. A few strains were widely spread and dominated quantitatively. Staff colonization was 23%, but contamination of their clothes was 82%. Two strains which colonized three of the patients were widely spread in the air. They were found in the corridor and in every patient room, even where the patient was not colonized. The main source of the environmental contamination seems to have been patients. Possible routes of infection were direct airborne transmission as well as passive transfer via staff.

Bone Marrow Transplantation↗

PubMedDate not supplied

Relationship of staphylococcal conjugative plasmids to large gentamicin-resistance nonconjugative plasmids in clinical isolates of Staphylococcus epidermidis.

Self-mobilization of large plasmids was not observed in 15 of 15 strains of gentamicin-resistant (Gmr) Staphylococcus epidermidis clinical isolates that were taken from bacteremic newborns in a neonatal intensive care unit. Alternatively, nine Gmr Staphylococcus aureus clinical isolates, which were isolated along with the S. epidermidis strains, were all shown previously to contain Gmr conjugative plasmids. All of the nonconjugative strains had plasmids that were similar in size to the S. aureus conjugative plasmids. Transfer of seven of these nonconjugative plasmids by protoplast transformation indicated that they carried Gmr determinants. The rationale of these studies was to detect the presence of genes for conjugation (tra) on the large Gmr nonconjugative plasmids. It was thought that these plasmids might contain either defective or deleted tra gene sequences. To gain insight into these possibilities, a 6.3-kb probe, which contained a major tra gene region, was hybridized with EcoRI and XbaI digests of nonconjugative plasmid DNA. Hybridization occurred with only one of eight plasmids. It was concluded that seven of the large S. epidermidis plasmids were not self-transmissible because they lacked tra genes. However, pMH6502 contained an excess of tra gene regions compared to prototype conjugative plasmids and was still not self-transmissible.

Conjugation, Genetic↗

PubMed1984

Drug resistance in Streptococcus faecalis, Streptococcus faecium and Staphylococcus epidermidis isolated from laboratory animals.

A total of 1,278 strains of Streptococcus faecalis, 688 of Str. faecium and 796 of Staphylococcus epidermidis were collected from laboratory animals of 20 colonies in 1980, and tested for their resistance to 20 drugs. Thirteen percent of Str. faecalis, 12% of Str. faecium and 25% of Staph. epidermidis were drug resistant. Multiple drug resistant organisms were frequently detected in laboratory animal colonies where antibiotic treatment had been undertaken. TC-resistant streptococci and EM-resistant Staph. epidermidis were distributed in about half of the colonies regardless of the history of antibiotic usage.

Animals↗

PubMed1989

Comparison of in vitro activity of daptomycin, vancomycin and fluoroquinolones in broth and serum against Staphylococcus epidermidis as determined by time-kill kinetics.

We compared the activity of daptomycin, vancomycin and fluoroquinolones against 46 blood isolates of Staphylococcus epidermidis. Daptomycin was more rapidly bactericidal than vancomycin and demonstrated less of an inoculum effect than the quinolones. Time-kill kinetic studies demonstrated no significant differences in survival in broth as compared to 50% human serum-supplemented broth. Daptomycin and the quinolones are suitable for study in clinical trials of efficacy as therapeutic alternatives to vancomycin for S. epidermidis infections.

Anti-Bacterial Agents↗

PubMed1998

Characterization of the N-acetylglucosaminyltransferase activity involved in the biosynthesis of the Staphylococcus epidermidis polysaccharide intercellular adhesin.

The polysaccharide intercellular adhesin (PIA) is an important factor in the colonization of medical devices by Staphylococcus epidermidis. The genes encoding PIA production are organized in the icaADBC (intercellular adhesion) operon. To study the function of the individual genes, we have established an in vitro assay with UDP-N-acetylglucosamine, the substrate for PIA biosynthesis, and analyzed the products by thin-layer chromatography and mass spectrometry. IcaA alone exhibited a low N-acetylglucosaminyltransferase activity and represents the catalytic enzyme. Coexpression of icaA with icaD led to a significant increase in activity. The newly identified icaD gene is located between icaA and icaB and overlaps both genes. N-Acetylglucosamine oligomers produced by IcaAD reached a maximal length of 20 residues. Only when icaA and icaD were expressed together with icaC were oligomer chains that react with PIA-specific antiserum synthesized. IcaA and IcaD are located in the cytoplasmic membrane, and IcaC also has all the structural features of an integral membrane protein. These results indicate a close interaction between IcaA, IcaD, and IcaC. Tunicamycin and bacitracin did not affect the in vitro synthesis of PIA intermediates or the complete PIA biosynthesis in vivo, suggesting that a undecaprenyl phosphate carrier is not involved. IcaAD represents a novel protein combination among beta-glycosyltransferases.

Amino Acid Sequence↗

PubMed1988

Characterization of clinically significant isolates of Staphylococcus epidermidis from patients with endocarditis.

Biotyping, slime production, bacteriophage typing, serotyping, antibiograms, and plasmid profiles were used to characterize 19 Staphylococcus epidermidis strains isolated from 12 patients with prosthetic valve endocarditis and from 7 patients with native valve endocarditis. With the API Staph battery, 12 different biocodes with, at the most, three differences were obtained. Slime production was found for 10 strains (53%). Agglutinogens investigated by agglutination with two specific sera were found for 12 strains (63.1%). Three strains were phage typable (15.2%). Against a panel of nine antimicrobial agents, 15 different profiles were found. Multiply antibiotic-resistant strains were isolated from patients with prosthetic valve endocarditis when disease onset occurred less than 18 months after heart surgery and from patients with native valve endocarditis who received antibiotics immediately prior to their illness. All of the strains were available for plasmid analysis, and all the DNA profiles were distinct. On gels run in Tris-borate buffer, 73.7% of the strains had large plasmids of more than 30 megadaltons. A small plasmid of 2.8 megadaltons was found in multiply resistant strains and in strains resistant only to tetracyclines. None of the isolates appeared to be the same strain, and the bacteriological differences between the strains were confirmed mainly by the antibiotic susceptibility profile and the plasmid pattern analysis. These bacteriological results were in agreement with the clinical data.

Bacteriophage Typing↗

PubMed1995

Immunosuppression augments growth of graft-adherent Staphylococcus epidermidis.

OBJECTIVE: To determine if systemic suppression of host defenses during graft implantation alters the initial adherence and subsequent growth of Staphylococcus epidermidis on vascular prostheses. DESIGN: Dacron grafts 1 cm2 were implanted in the back subcutaneous tissue of Swiss-Webster mice (n = 247), followed by topical inoculation with 2 x 10(7), 2 x 10(5), 2 x 10(3), or 2 x 10(1) colony-forming units of S epidermidis. Half of the mice were immunosuppressed with cyclophosphamide (150 mg/kg intraperitoneally), to achieve a consistent, significant decrease in the white blood cell count and major histocompatibility complex class II (Ia) expression. Control mice received an equal volume of saline solution. Graft bacterial biofilm concentrations were determined at 1 day for adherence and within 2 weeks for bacterial growth, by using sonication and quantitative agar culture. RESULTS: Immunosuppression did not significantly alter the initial adherence of bacteria to vascular grafts. Immunosuppressed animals that were inoculated with 2 x 10(7) and 2 x 10(5) colony-forming units of S epidermidis had significantly higher bacterial biofilm concentrations as compared with those in control animals. Graft infection persisted at 14 days in all animals, with and without immunosuppression. CONCLUSIONS: Suppression of immune function during graft implantation augmented growth of adherent bacteria. The effect of short-term perioperative immunosuppression on late-appearing S epidermidis graft infection needs further study.

Animals↗

PubMed2001

Catecholamine inotropes as growth factors for Staphylococcus epidermidis and other coagulase-negative staphylococci.

Drugs commonly used in intensive care settings were assayed for their ability to affect the growth of Staphylococcus epidermidis in a minimal salts medium containing 30% serum. Of 28 compounds tested, the inotropic catecholamines adrenaline, dobutamine, dopamine, isoprenaline and noradrenaline significantly stimulated bacterial growth. These drugs, but not structurally similar compounds lacking a dihydroxybenzoyl moiety (such as tyramine, phenylephrine and salbutamol), were able to remove iron from iron-saturated transferrin and to supply transferrin-bound 55Fe to S. epidermidis cells. Similar results were observed with a range of coagulase-negative staphylococci associated with line infections, but not with Staphylococcus aureus (including MRSA).

Catecholamines↗