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Staphylococcus epidermidis infections.

Staphylococcus epidermidis, an organism routinely found on the skin and in the hospital environment, has become a primary pathogen in infections associated with prosthetic devices. Because these infections are indolent and often clinically silent, diagnosis and therapy are often difficult. Pathogens are often misidentified as contaminants. Their variable, often resistant antibiotic susceptibility pattern and the uncertain correlation of in-vitro beta-lactam sensitivity testing with therapeutic efficacy make selection of an effective antibiotic regimen difficult. Vancomycin combined with rifampin, gentamicin, or both, is recommended for empiric therapy of these infections. Usually, removal of the prosthetic device is also necessary and may contribute equally to a successful therapeutic outcome.

Anti-Bacterial Agents↗

First complete genome sequence of two Staphylococcus epidermidis bacteriophages.

Staphylococcus epidermidis is an important opportunistic pathogen causing nosocomial infections and is often associated with infections in patients with implanted prosthetic devices. A number of virulence determinants have been identified in S. epidermidis, which are typically acquired through horizontal gene transfer. Due to the high recombination potential, bacteriophages play an important role in these transfer events. Knowledge of phage genome sequences provides insights into phage-host biology and evolution. We present the complete genome sequence and a molecular characterization of two S. epidermidis phages, phiPH15 (PH15) and phiCNPH82 (CNPH82). Both phages belonged to the Siphoviridae family and produced stable lysogens. The PH15 and CNPH82 genomes displayed high sequence homology; however, our analyses also revealed important functional differences. The PH15 genome contained two introns, and in vivo splicing of phage mRNAs was demonstrated for both introns. Secondary structures for both introns were also predicted and showed high similarity to those of Streptococcus thermophilus phage 2972 introns. An additional finding was differential superinfection inhibition between the two phages that corresponded with differences in nucleotide sequence and overall gene content within the lysogeny module. We conducted phylogenetic analyses on all known Siphoviridae, which showed PH15 and CNPH82 clustering with Staphylococcus aureus, creating a novel clade within the S. aureus group and providing a higher overall resolution of the siphophage branch of the phage proteomic tree than previous studies. Until now, no S. epidermidis phage genome sequences have been reported in the literature, and thus this study represents the first complete genomic and molecular description of two S. epidermidis phages.

Base Sequence↗

Nosocomial septicemia due to multiply antibiotic-resistant Staphylococcus epidermidis.

Thirteen episodes of Staphylococcus epidermidis sepsis occurred over a 20-month period in 11 patients receiving general surgical and medical care. These episodes were characterized by fever, toxicity, multiple positive blood cultures, and uniformly colonized intravascular catheters. An additional 16 patients had possible sepsis. Four associated deaths occurred; all three patients autopsied had multiple pulmonary abscesses in which gram-positive cocci were profusely present. In individual patients, prolonged episodes of septicemia were confirmed by multiple blood culture isolates of S. epidermidis, identical in antibiotic resistance pattern, phage type, and biotype. A prominent feature of the S. epidermidis isolates was resistance to many commonly used antimicrobial agents. Case-control studies and review of laboratory records indicated a significant association between multiply resistant S. epidermidis blood isolates and prolonged hospitalization and parenteral hyperalimentation. Most of these patients were hospitalized in the intensive care unit; nose and hand cultures taken from the personnel showed frequent carriage of multiply resistant S. epidermidis Staphylococcus epidermidis associated with intravascular devices may produce life-threatening bloodstream infections.

Adult↗

Transduction of tetracycline resistance in Staphylococcus epidermidis.

Fourteen Staphylococcus epidermidis strains were biotyped according to the scheme of Baird-Parker and tested for their sensitivities to S. epidermidis typing phages. Tetracycline-susceptible strains were tested for the capacity to be transduced to resistance. Five strains, all of biotype 1, were lysed by one or more phages. Five of eight biotype 1 strains were transduced to tetracycline resistance with rates of 10(-8) to 10(-5). Three strains of other biotypes were not transduced, nor was PS 73, an unusual coagulase-negative strain. The transducing phage phi367 was isolated from the lysogenic tetracycline-resistant donor strain. The phage morphologically resembles serological group B S. aureus phages. Restriction and modification were indicated from the efficiency of plating on phage-sensitive strains.

Bacteriolysis↗

Rifampin in the management of early prosthetic staphylococcus epidermidis endocarditis.

Staphylococcus epidermidis endocarditis occurred four days following aortic valve replacement with a Björk-Shiley prosthesis. Antimicrobial therapy, shown to be effective by in vitro and in vivo studies, failed to eradicate the infection. When rifampin was added to the existing antibiotic regimen, peak serum bactericidal activity increased, the patient defervesced, and blood cultures became negative.

Adult↗

Biotypes of Staphylococcus epidermidis and Micrococcus organisms, isolated from intramammary infections, reclassified into species of the genus Staphylococcus (epidermidis, hyicus, xylosus, and sciuri).

In a previous report (5), strains of Staphylococcus epidermidis and Micrococcus isolated from bovine intramammary infections were classified by the Baird-Parker (B-P) system and by serologic typing of the proteolytic enzymes. Since then, newer methods for distinguishing the genera Staphylococcus and Micrococcus and for defining new species have been reported. By utilizing these methods, the organisms from the original study were reclassified as follows: S. epidermidis B-P subgroup II, proteinase group F strains as S. epidermidis; S. epidermidis B-P subgroup III, proteinase group B strains as Staphylococcus hyicus subsp. chromogenes (pigmented) or subsp. hyicus (nonpigmented) and proteinase groups H and BH strains that were coagulase positive and nonpigmented as subsp. hyicus; B-P Micrococcus subgroup 6, proteinase group NR strains as Staphylococcus xylosus and subgroup 6, proteinase group G strains as Staphylococcus sciuri and S. xylosus. The reclassification of the strains increased the percentage of intramammary infections attributed to staphylococci from 62%, as first reported, to at least 86%.

Animals↗

Comparison of the SmaI-digested chromosomes of Staphylococcus epidermidis and the closely related species Staphylococcus capitis and Staphylococcus caprae.

Pulsed-field gel electrophoresis was used to examine the chromosomal polymorphisms existing within and between four closely related members of the Staphylococcus epidermidis species group, S. epidermidis, Staphylococcus caprae, Staphylococcus capitis subsp. capitis, and S. capitis subsp. ureolyticus. SmaI was chosen as the restriction endonuclease for this study because it generated only a few well-separated chromosomal fragments. Each of the species and subspecies showed distinct SmaI digest patterns. The strains examined in this study were collected over a 20-year period from various geographical locations. The results indicate that DNA fragment patterns are unique to each species and subspecies and represent a reasonably stable component in the chromosome structure. S. caprae and S. capitis demonstrated considerable conservation in chromosome structure as indicated by the large numbers of conserved SmaI digest fragments. The polymorphisms found within each species appear to be linked to the species' character variability. The genome size of each Staphylococcus strain was extrapolated from the SmaI digest fragment pattern obtained by pulsed-field gel electrophoresis. The average genome size for S. epidermidis is 2,364 +/- 119 kb; for S. caprae strains from humans it is 2,600 +/- 157 kb and for S. caprae strains from goats it is 2,493 +/- 15 kb; for S. capitis subsp. capitis it is 2,456 +/- 71 kb; and for S. capitis subsp. ureolyticus it is 2,276 +/- 90 kb.

Animals↗

Catheter sepsis due to Staphylococcus epidermidis during parenteral nutrition.

Staphylococcus epidermidis is a pathogenic organism with increasing importance in total parenteral nutrition therapy. Strict asepsis during catheter insertion prolongs the interval free from Staphylococcus epidermidis infection. Staphylococcus epidermidis colonizes the catheter after migrating from the skin. For protection, we advise a long subcutaneous tunnel for all catheters that are to be indwelling for longer than three weeks. Prompt recatheterization of a patient with Staphylococcus epidermidis sepsis can result in hematogenous seeding of the new catheter and persistence of the infection. Catheter related Staphylococcus epidermidis sepsis has subsided after catheter withdrawal, and there is no need for antibiotic therapy provided that other prosthetic materials are not placed in the vascular tree. Immunologic status of the patients is not related to the frequency or severity of Staphylococcus epidermidis infections, or both.

Adult↗

Nucleotide sequence of the constitutive macrolide-lincosamide-streptogramin B resistance plasmid pNE131 from Staphylococcus epidermidis and homologies with Staphylococcus aureus plasmids pE194 and pSN2.

The complete nucleotide sequence of the Staphylococcus epidermidis plasmid pNE131 is presented. The plasmid is 2,355 base pairs long and contains two major open reading frames. A comparison of the pNE131 DNA sequence with the published DNA sequences of five Staphylococcus aureus plasmids revealed strong regional homologies with two of them, pE194 and pSN2. The region of pNE131 containing the reading frame which encodes the constitutive ermM gene is almost identical to the inducible ermC gene region of pE194, except for a 107-base-pair deletion which removes the mRNA leader sequence required for inducible expression. A second region of pNE131 contains an open reading frame with homology to the small cryptic plasmid pSN2 and potentially encodes a 162-amino-acid protein.

Amino Acid Sequence↗

Detection and preliminary characterization of extrachromosomal DNA in clinical isolates of Staphylococcus aureus and Staphylococcus epidermidis.

Isolates of Staphylococcus aureus and Staphylococcus epidermidis frequently harboured extrachromosomal DNA. From the 20 Staphylococcus aureus strains examined 85% contained extrachromosomal DNA and 65% carried more than one plasmid DNA species. The results obtained with the 19 Staphylococcus epidermidis strains analyzed were similar, 84% of the strains had extrachromosomal DNA and 68% contained several DNA species. The agarose gel electrophoresis of the DNA obtained from cleared lysates of the staphylococci proved to be efficient for the detection of plasmids and the determination of their molecular weights. The gel method appeared therefore to be suitable for the establishment of the role of plasmids in the phage typability of staphylococci. Using a Staphylococcus epidermidis strain as an example it was shown by electron microscopy that the extrachromosomal DNA demonstrated by the agarose gel electrophoresis represented covalently closed circular plasmid DNA. This plasmid DNA could be sufficiently labeled with 3H-thymidine to undertake studies on the relationship and molecular characterization of staphylococcal plasmids.

DNA, Bacterial↗

Genome-based analysis of virulence genes in a non-biofilm-forming Staphylococcus epidermidis strain (ATCC 12228).

Staphylococcus epidermidis strains are diverse in their pathogenicity; some are invasive and cause serious nosocomial infections, whereas others are non-pathogenic commensal organisms. To analyse the implications of different virulence factors in Staphylococcus epidermidis infections, the complete genome of Staphylococcus epidermidis strain ATCC 12228, a non-biofilm forming, non-infection associated strain used for detection of residual antibiotics in food products, was sequenced. This strain showed low virulence by mouse and rat experimental infections. The genome consists of a single 2499 279 bp chromosome and six plasmids. The chromosomal G + C content is 32.1% and 2419 protein coding sequences (CDS) are predicted, among which 230 are putative novel genes. Compared to the virulence factors in Staphylococcus aureus, aside from delta-haemolysin and beta-haemolysin, other toxin genes were not found. In contrast, the majority of adhesin genes are intact in ATCC 12228. Most strikingly, the ica operon coding for the enzymes synthesizing interbacterial cellular polysaccharide is missing in ATCC 12228 and rearrangements of adjacent genes are shown. No mec genes, IS256, IS257, were found in ATCC 12228. It is suggested that the absence of the ica operon is a genetic marker in commensal Staphylococcus epidermidis strains which are less likely to become invasive.

Adhesins, Bacterial↗

Cloning and expression of Staphylococcus epidermidis urease gene sequences in Staphylococcus carnosus.

The urease gene sequences of Staphylococcus epidermidis CNS23 were cloned and expressed in Staphylococcus carnosus TM300. In vitro translation of the cloned sequences revealed four polypeptides (60, 17, 11 and 7.5 kDa) that were associated with enzyme activity. Southern hybridisation experiments showed high homologies with the urease genes of Staphylococcus saprophyticus.

Blotting, Southern↗

In vivo transfer of high-level mupirocin resistance from Staphylococcus epidermidis to methicillin-resistant Staphylococcus aureus associated with failure of mupirocin prophylaxis.

OBJECTIVES: We examined the molecular basis of the emergence of mupirocin resistance in a methicillin-resistant Staphylococcus aureus (MRSA) strain colonizing a nursing home resident undergoing mupirocin prophylaxis. PATIENT AND METHODS: A persistent carrier of mupirocin-susceptible MRSA participated in a trial of mupirocin for nasal decolonization among nursing home residents. During prophylaxis a high-level mupirocin-resistant MRSA emerged in the nasal isolates from this patient. S. aureus and coagulase-negative staphylococci were isolated prior to, during and after 14 days of mupirocin treatment. The staphylococcal isolates and their plasmids were examined by molecular genetic methods. RESULTS: All mupirocin-susceptible and -resistant MRSA isolates possessed the same genotype. The patient was also colonized by a single mupirocin-resistant Staphylococcus epidermidis strain. The mupirocin-resistant MRSA and S. epidermidis strains harboured identical plasmids that carried the mupA determinant and genes for conjugative DNA transfer in staphylococci. These plasmids could be transferred in vitro from both clinical isolates to S. aureus RN2677. CONCLUSIONS: The MRSA strain contained a conjugative plasmid expressing mupA that was identical with that found in the S. epidermidis strain which colonized the patient. These findings suggest that transfer of mupA from S. epidermidis to MRSA probably occurred during mupirocin prophylaxis.

Aged↗

Topical vancomycin for the treatment of Staphylococcus epidermidis and methicillin-resistant Staphylococcus aureus conjunctivitis.

Staphylococcus aureus and Staphylococcus epidermidis are organisms that frequently cause conjunctivitis or blepharoconjunctivitis. We describe a patient with methicillin-resistant S. aureus and S. epidermidis conjunctivitis who was treated successfully using an extemporaneously prepared topical ophthalmic solution of vancomycin hydrochloride 31 mg/mL. Studies describing the preparation, stability, and comfort of this solution, as well as reports pertaining to efficacy, are reviewed. Controlled clinical trials evaluating the safety and efficacy of vancomycin ophthalmic solution have not yet been performed.

Administration, Topical↗

Plasmid-pattern analysis for the differentiation of infecting from noninfecting Staphylococcus epidermidis.

Isolation of Staphylococcus epidermidis from cultures of blood was differentiated from culture contamination by the detection of identical isolates in two or more consecutive cultures from an infected patient. We used plasmid-pattern analysis as a tool for establishing the identity of individual isolates. In a control study of 15 patients with two or more cultures of blood contaminated with S. epidermidis, analysis revealed that none of the isolates had identical plasmid patterns. This reflected the variety of plasmid patterns among colonizing coagulase-negative staphylococci cultured from skin sites of uninfected patients. In contrast, plasmid-pattern identity was seen among sequential or paired S. epidermidis isolates from a given patient in 32 of 36 patients with documented S. epidermidis infection. The plasmid pattern of each set of isolates from patients was unique. Infections included prosthetic-valve endocarditis in 26 patients, cerebrospinal fluid-shunt or ventriculostomy infections in six patients, intravenous-catheter sepsis in two patients, urinary tract infection in one patient, and osteomyelitis in one patient. Plasmid-pattern analysis may therefore be useful in the diagnosis of S. epidermidis infections.

Anti-Bacterial Agents↗

[Diagnostic molecular microbiology--identification of Staphylococcus epidermidis].

The species Staphylococcus epidermidis is the predominant coagulase-negative staphylococci (CNS) isolated from clinical sources. S. epidermidis is now recognized as an important nosocomial pathogen. Identification of CNS is often performed using diagnostic kits based on biochemical or immunological reactions. However, these kits are often unreliable for the identification of CNS species including S. epidermidis. Currently, ribosomal RNA (rRNA) analyses are the most powerful methods for determining phylogenetic relationships among microorganisms and also for identification of species. Several aspects of construction of ribosomal probes for identification of CNS species are presented and discussed. Additionally, the application of restriction fragment length polymorphisms (RFLP) of rRNA genes for differentiation of clinical isolates of S. epidermidis is shown.

Cross Infection↗