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PubMedDate not supplied

Reproducibility of Staphylococcus epidermidis plasmid profiles.

The plasmid profiles of six isolates of Staphylococcus epidermidis were repetitively evaluated over an 8-month period. Each isolate was subcultured and stored at three different temperatures (-70 degrees C, -20 degrees C, and room temperature) and plasmid DNA was prepared from each subculture at 0, 1, 4, and 8 months by two different methods of plasmid extraction [using mixed alkyltrimethylammonium bromide (ATAB) or Brij 58 and deoxycholate (modified Parisi)]. Plasmid DNA bands were lost from two isolates when subcultures were kept at room temperature. This plasmid loss was confirmed by repetitive extractions and electrophoresis, as well as by restriction endonuclease analysis of the ATAB preparations. Profiles were otherwise highly related to one another, with occasional exceptions being extra or missing plasmid DNA bands of high molecular size. The latter findings were not reproducible. Plasmid DNA extracted by the modified Parisi method was not reliably digested with restriction endonuclease enzymes. We conclude that the plasmid profiles of Staphylococcus epidermidis isolates are highly reproducible as long as isolates are stored at less than or equal to -20 degrees C. Minor discrepancies in the number of plasmid DNA bands of large molecular size may occur. These are resolvable by repetitive testing or restriction endonuclease analysis of ATAB-extracted plasmid DNA preparations.

Cold Temperature↗

PubMed1968

The glycerol teichoic acid from walls of Staphylococcus epidermidis I2.

1. Walls of Staphylococcus epidermidis I2 contain 30% (w/w) of a glycerol teichoic acid containing phosphate, d-alanine and d-glucose in the molecular proportions 1:0.25:0.50. 2. The teichoic acid was isolated by extraction with trichloroacetic acid and with dilute aqueous NN-dimethylhydrazine at pH7, and was shown to be a (1-->3)-linked poly(glycerol phosphate) containing beta-d-glucopyranosyl and d-alanyl ester substituents. 3. 2-O-beta-d-Glucopyranosylglycerol was isolated and characterized as its crystalline hexa-O-acetate. 4. Unlike that of certain other bacteria, the peptidoglycan component of the wall is not solubilized by NN-dimethylhydrazine. 5. The membrane teichoic acid is also a (1-->3)-linked poly(glycerol phosphate) but contains a smaller proportion of glucosyl substituents.

Alanine↗

PubMed1972

Production and purification of a Staphylococcus epidermidis bacteriocin.

Liquid cultures of Staphylococcus epidermidis 1580 contained rather small amounts of a bacteriocin, staphylococcin 1580, which was found both in the supernatant fluid and in the cell pellet. It could be extracted from the cells with 5% NaCl solution. The staphylococcin production could not be induced by ultraviolet irradiation or treatment with mitomycin C. Bacteria grown on semisolid medium produced a much larger amount of the compound with a high specific activity. The staphylococcin was purified by ammonium sulfate precipitation, ultracentrifugation, and chromatography on Sephadex columns. The purified material was homogeneous on polyacrylamide gel electrophoresis. The molecular weight was between 150,000 and 400,000. The bacteriocin was composed of protein, carbohydrate, and lipid and consisted of subunits exhibiting a molecular weight of about 20,000.

Ammonium Sulfate↗

PubMed1994

Protamine sulfate and vancomycin are synergistic against Staphylococcus epidermidis prosthesis infection in vivo.

We have previously demonstrated that the quaternary amine, protamine sulfate (PS), is bactericidal against Staphylococcus epidermidis. In an attempt to decrease genitourinary prosthesis infection rates, we examined the ability of PS as a wound irrigant to inhibit Staphylococcus epidermidis viability. Eighty-seven Sprague-Dawley rats were studied by implanting a sterile silicone pellet in their dorsum. The pellet was inoculated with Staphylococcus epidermidis and the rats were divided into four groups based on the wound irrigant employed after inoculation: (1) control (sterile water) (2) vancomycin; (3) PS; (4) vancomycin + PS. All rats received perioperative and daily intramuscular vancomycin, and the pellets were explanted on postoperative day 28 and cultured. The infection rates were: (1) control 77%, (2) vancomycin 50%, (3) protamine sulfate 67%, and (4) protamine sulfate and vancomycin 19%. The differences between (2) vancomycin versus (4) vancomycin + PS and (3) PS versus (4) vancomycin + PS were significant (p = 0.05 and p < 0.005). The data suggest that PS potentiates vancomycin as a wound irrigant in prosthesis implantation.

Animals↗

PubMed1987

Action of clindamycin-phosphate in foreign body infections due to Staphylococcus epidermidis in mice.

The virulence of Staphylococcus epidermidis strain slime producer was examined in an experimental model of foreign body infection in mice. In the course of this experimental infection the mice were injected with two antibiotics (clindamycin and cefazolin) active in vitro toward the Staphylococcus strain used. The results obtained after a week of antibiotic therapy show that clindamycin alone has a therapeutic action against the infection caused by S. epidermidis. Cefazolin showed a very poor therapeutic effect. The results are discussed on the basis of inflammatory reaction elicited from the foreign body and the characteristics of clindamycin in connection with the host's defense mechanisms.

Animals↗

PubMed1999

Some characteristics of platelet concentrates contaminated with Staphylococcus epidermidis.

The study investigates platelet concentrates (PCs) contaminated with Staphylococcus epidermidis. The work was conducted using 12 pairs of PCs. Light transmission through the PCs was observed over 7 days. Immediately after preparation, one PC was inoculated with Staphylococcus epidermidis (17-43 colony forming units (CFU)/ ml) and the other served as a non-contaminated control. In the contaminated PCs the CFU/ml and swirling were determined on days 1, 3 and 7. In all PCs determination of pH and the extent of shape change (ESC) was carried out on days 3 and 7. All contaminated PCs had visible swirling on day 3 but they contained 2.0 X 10 4 to 9.0 X 10 8 CFU/ml. pH (P=0.05) and ESC (P<0.05) then were slightly lower. On day 7, eight of 12 of the contaminated PCs failed to display transmission changes despite deranged analytical results and impaired swirling. During early storage the contaminated units had unimpaired swirling and only slightly impaired in vitro quality. They appear to be difficult to identify in the routine.

Journal Article↗

PubMed1999

Detection of Staphylococcus aureus and Staphylococcus epidermidis in clinical samples by 16S rRNA-directed in situ hybridization.

Staphylococcus epidermidis and Staphylococcus aureus are the most common causes of medical device-associated infections, including septicemic loosenings of orthopedic implants. Frequently, the microbiological diagnosis of these infections remains ambiguous, since at least some staphylococci have the capacity to reduce their growth rate considerably. These strains exhibit a small-colony phenotype, and often they are not detectable by conventional microbiological techniques. Moreover, clinical isolates of S. aureus and S. epidermidis adhere to polymer and metal surfaces by the generation of thick, multilayered biofilms consisting of bacteria and extracellular polysaccharides. This study reports improved detection and identification of S. aureus and S. epidermidis by an in situ hybridization method with fluorescence-labeled oligonucleotide probes specific for staphylococcal 16S rRNA. The technique has proven to be suitable for the in situ detection of staphylococci, which is illustrated by the identification of S. epidermidis in a connective tissue sample obtained from a patient with septicemic loosening of a hip arthroplasty. We also show that this technique allows the detection of intracellularly persisting bacteria, including small-colony variants of S. aureus, and the differentiation of S. epidermidis from other clinically relevant staphylococci even when they are embedded in biofilms. These results suggest that the 16S rRNA in situ hybridization technique could represent a powerful diagnostic tool for the detection and differentiation of many other fastidious microorganisms.

Genes, rRNA↗

PubMed1963

Identity of Staphylococcus epidermidis.

Jones, Dorothy (American Meat Institute Foundation, Chicago, Ill.), R. H. Deibel, and C. F. Niven, Jr. Identity of Staphylococcus epidermidis. J. Bacteriol. 85:62-67. 1963.-The species Staphylococcus epidermidis is defined more precisely, and compared with other staphylococci. The physiological characteristics considered to be of primary import are: (i) ability to grow anaerobically in a defined complex medium with glucose or pyruvate as the energy source; (ii) inability to produce coagulase; (iii) ability to ferment serine as an energy source; (iv) the requirement for biotin and uracil (under anaerobic conditions) in a semisynthetic medium; and (v) inability to reduce nitrate beyond nitrite.

Coagulase↗

PubMed2001

Infection and/or colonization by methicillin-resistant Staphylococcus epidermidis (MRSE).

We analyzed the parameters predictive of identification of methicillin-resistant Staphylococcus epidermidis (MRSE) in sample performed in hospitalized patients. One hundred six Staphylococcus epidermidis strains (60 MRSE and 46 MSSE) were collected. Three variables were independently linked to MRSE isolation in multivariate analysis: hospitalization during the month preceding the current admission; on-going antimicrobial therapy before sampling, and on-going infection at the time of sampling. MRSE isolation was associated with a poor vital prognosis. The air and surfaces sampling in the rooms of two patients with nasal MRSE carriage yielded the same strains as those carried by the patient, and could play a role in the epidemiological chain of hospital-acquired MRSE infections.

Carrier State↗

PubMed1984

Comparison of adherence and urine growth rate properties of Staphylococcus saprophyticus and Staphylococcus epidermidis.

To compare the adherence properties of Staphylococcus saprophyticus and Staphylococcus epidermidis in vitro studies were conducted with clinical isolates and cell culture monolayers (Hela, Vero, MDCK). Staphylococcus saprophyticus exhibited greater cell adherence, but no significant difference in mean urine growth rate. Its cell-adherence properties seem to explain in part its virulence.

Adhesiveness↗

PubMed1995

Bacteria/blood/material interactions. I. Injected and preseeded slime-forming Staphylococcus epidermidis in flowing blood with biomaterials.

Blood-material interactions were studied using in vitro recirculation with human blood, slime-forming Staphylococcus epidermidis, and cardiovascular materials. Staphylococcus epidermidis, under preseeded or injected conditions, adhered to nonsmooth materials and elevated plasma levels of fibrinopeptide A (FpA) and C3a in the presence of all materials. Increased white blood cell (WBC) and platelet adhesion and thrombospondin and platelet factor 4 (PF4) release were noted for respective materials in the presence of injected bacteria. Materials that adhered significant quantities of injected S. epidermidis exhibited low levels of adsorbed proteins. Materials with high levels of preseeded S. epidermidis showed high levels of adsorbed proteins. Adhesion of preseeded bacteria and blood plasma elevations of C3a and FpA were lowest on semicrystalline polymer substrates, intermediate on halogenated substrates, and highest on amorphous substrates. In the presence of injected bacteria, WBCs and platelets adhered at earlier recirculation times to amorphous substrates than to semicrystalline substrates.

Bacterial Adhesion↗

PubMed2004

Reduced triclosan susceptibility in methicillin-resistant Staphylococcus epidermidis.

Triclosan MIC determination showed that recent Staphylococcus aureus clinical isolates (n = 100) were highly susceptible to triclosan, with a 50% minimal inhibitory concentration (MIC(50)) of 0.12 microg/ml and a MIC(90) of 0.25 microg/ml. Staphylococcus epidermidis isolates (n = 96) were less susceptible, with a MIC(50) of 0.12 microg/ml and a MIC(90) of 8 microg/ml. Decreased susceptibility to triclosan was more prevalent among methicillin-resistant S. epidermidis than among methicillin-sensitive S. epidermidis isolates.

Anti-Infective Agents, Local↗

PubMed1986

Inhibition of human neutrophil bacteriocidal activity by extracellular substance from slime-producing Staphylococcus epidermidis.

The role of slime produced by Staphylococcus epidermidis as a virulence factor has been investigated by preincubating human neutrophils in fractions of extracellular material from cultures of slime-producing and non-slime-producing S. epidermidis. Extracellular product (molecular weight greater than 300,000) from slime-producing S. epidermidis did not affect neutrophil phagocytosis but did produce a generalized loss of bacteriocidal activity which was not found with non-slime-producing S. epidermidis, or broth.

Biological Products↗

PubMed2006

Self-reinforced ciprofloxacin-releasing polylactide-co-glycolide 80/20 inhibits attachment and biofilm formation by Staphylococcus epidermidis: an in vitro study.

We have observed the efficiency of antibiotic-releasing polylactide-co-glycolide (PLGA) 80/20 in preventing Staphylococcus epidermidis attachment and biofilm formation in vitro. The aim of the present study was to evaluate the effect of self-reinforced (SR) implants with enhanced antibiotic release on bacterial attachment and biofilm formation rates, and also on growth inhibition of Staphylococcus epidermidis. Cylindrical SR-PLGA+AB specimens (length 30 mm, diameter 3 mm) were examined by scanning electron microscopy (SEM) for attachment of S. epidermidis ATCC 35989 on biomaterial surface and formation of biofilm, after incubating with bacterial suspension of ca. 10 cfu/mL for 1, 3, 7, 14 and 21 days. SR-PLGA and SR-PLGA+AB implants were tested on agar plates by measuring the inhibition distance around implants. On the surface of SR-PLGA+AB, at days 1, 3, 7, 14 and 21, the percentage of areas with not a single bacteria attached, was 88.6%, 71.1%, 73.7%, 73.7%, and 68.4%, respectively. On the areas where bacteria were detected, the number of bacterial cells remained low during whole study period, and no significant increase by time was seen. There was no biofilm observed on 97-99% of the examined areas during the whole study period on SR-PLGA+AB. In agar plates, the SR-PLGA+AB showed inhibition of bacterial growth, with (mean) 53.2 mm diameter of inhibition area with peeled implants and 50.5 mm with non-peeled implants. There was no inhibition seen around implants without ciprofloxacin. Bioabsorbable ciprofloxacin-releasing self-reinforced PLGA (SR-PLGA+AB) was superior to plain SR-PLGA in preventing bacterial attachment, biofilm formation, and also the growth of Staphylococcus epidermidis.

Anti-Infective Agents↗

PubMed1996

A comparison of methods to determine whether clinical isolates of Staphylococcus epidermidis from the same patient are related.

Staphylococcus epidermidis is a major cause of hospital-acquired infections but also part of the normal skin flora. A common clinical question is whether repeated isolation of S. epidermidis from one patient represents the same strain; because if different strains are isolated, they are often thought to be contaminants. In this study, different typing methods were compared to answer this question. Twenty isolates of S. epidermidis from five different patients were investigated. The isolates from each patient had identical or very similar antibiograms, and were recovered on different occasions. Typing was performed by antibiogram, biotype, slime production, plasmid profile, and pulsed-field gel electrophoresis (PFGE) banding pattern of SmaI digests of chromosomal DNA. In addition, the level of resistance to methicillin was determined by growth curves in broth containing methicillin for a series of different inocula for each isolate. The results showed that the isolates from each patient belonged to the same clone, but examples of instabilities in their antibiograms, plasmid profiles, as well as their PFGE banding patterns were seen. A change in the level of methicilli, resistance was observed in one strain; otherwise this characteristic was found to be strain-specific and stable in vivo. It was concluded that in combination with biotyping and antibiotic resistance testing the level of resistance to methicillin could be used as an aid to distinguish between two or more clinical isolates of S. epidermidis from the same patient.

Bacterial Typing Techniques↗

PubMed1984

Effect of extracellular slime substance from Staphylococcus epidermidis on the human cellular immune response.

Staphylococcus epidermidis infection of plastic catheters is often associated with heavy deposits of slime. To test whether this slime affects the human cellular immune response, its effect on the lympho-proliferative response of mononuclear cells to polyclonal stimulators was measured. Slime drastically reduces this response. Its inhibitory action was not immediate but took place over a few days and resulted in destruction of affected cells. The effect is dose related. This inhibition of cellular response may contribute to S epidermidis infection of implanted prostheses.

Adhesiveness↗

PubMed1997

Detection of the intercellular adhesion gene cluster (ica) and phase variation in Staphylococcus epidermidis blood culture strains and mucosal isolates.

Staphylococcus epidermidis is a common cause of catheter-associated infections and septicemia in immunocompromised patients. To answer the question whether S. epidermidis skin isolates differ from isolates causing septicemic diseases, 51 strains obtained from blood cultures, 1 strain from shunt-associated meningitis, and 36 saprophytic isolates were characterized. The study demonstrates that most of the blood culture strains formed a multilayered biofilm on plastic material, whereas skin and mucosal isolates did not. Moreover, biofilm-producing strains were found to generate large bacterial autoaggregates in liquid culture. Autoaggregation and biofilm formation on polymer surfaces was associated with the presence of a DNA sequence encoding an intercellular adhesion gene cluster (ica) that mediates the production of a polysaccharide intercellular adhesin. The presence of the intercellular adhesion genes in blood culture isolates was also found to be correlated with the exhibition of black colonies on Congo red agar, whereas the adhesin-negative strains formed red colonies. Upon subcultivation on Congo red agar, the black colony forms of the blood culture strains exhibited red colony variants which were biofilm and autoaggregation negative and occurred at a frequency of 10(-5). The DNA analysis of these S. epidermidis variants by pulsed-field gel electrophoresis and Southern hybridization with an ica-specific gene probe revealed no detectable difference between the black and red colony types. Moreover, after repeated passage, the phenotype of the parent strain could be restored. Therefore, these colony forms were regarded as phase variants. This phenotypic change was observed exclusively in adhesin-positive clinical isolates and not in adhesin-negative saprophytic strains of S. epidermidis.

Bacteremia↗

PubMed1991

Sonication provides maximal recovery of staphylococcus epidermidis from slime-coated vascular prosthetics.

Staphylococcus epidermidis (S. epidermidis) prosthetic vascular graft infections are often difficult to detect. A reliable culture method is needed so that appropriate therapeutic decisions can be initiated in a timely fashion. Sonication of graft material has been proposed as a method of enhancing bacterial recovery. Theoretically, however, this could cause cell lysis and false-negative culture results. Several methods of obtaining bacterial cultures were compared to determine the best method of quantitative bacterial recovery from infected graft material. Polytetrafluoroetehylene (PTFE) and knitted Dacron graft segments (n = 192) were exposed to a slime-producing strain of S. epidermidis or Escherichia coli (E. coli) at four different bacterial concentrations. Recovery of bacteria from the segments was then compared using three different methods to obtain organisms. One-third of the segments were pressed directly onto the agar to transfer bacteria while the other two-thirds were placed in broth and then either sonicated or vortexed prior to plating onto agar. The direct technique was significantly less effective (P less than 0.01) at recovering bacteria than either sonicating or vortexing for both graft materials at bacterial concentrations of 10, 10(2), and 10(4) CFU/ml of S. epidermidis. Sonication was significantly better at recovery from either material at 10(2) and 10(4) CFU/ml of S. epidermidis than vortexing (P less than 0.05). E. coli graft adherence was poor, and significant recovery occurred only at 10(8) CFU/ml. Sonication is necessary to maximally recover S. epidermidis from infected prosthetic grafts.

Bacteriological Techniques↗