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PubMed1993

Effect of growth conditions on expression and antigenicity of Staphylococcus epidermidis RP62A cell envelope proteins.

Staphylococcus epidermidis RP62A (ATCC 35984) was grown in tryptic soy broth (TSB), iron-depleted TSB (TSB-Fe), iron-reconstituted TSB-Fe (TSB+Fe), a chemically defined medium, and fetal calf serum (FCS) and on silastic disks in chambers that were sutured to the pig peritoneal wall. Bacterial cell wall proteins were extracted by digestion with recombinant lysostaphin, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and detected by silver staining. Cell wall proteins from TSB-, chemically defined medium-, or FCS-grown cells had a complex profile of greater than 25 protein bands spanning the full molecular mass range. By contrast, a digest obtained from in vivo-grown cells had only five major proteins of 40 kDa or greater. Proteins of 130 and 106 kDa were present in the cell envelopes of TSB-Fe- and in vivo-grown cells but not in those grown in TSB or TSB+Fe. A 43-kDa protein expressed by in vitro-grown cells and 52- and 96-kDa proteins expressed by in vivo-grown cells reacted with antisera from pigs with the chamber implants and from catheterized, paracatheter-inoculated pigs but not with hyperimmune sera from pigs immunized with TSB-grown cells. The data indicate that S. epidermidis, growing under in vivo conditions, expresses antigens distinct from those that are grown in vitro.

Animals↗

PubMed1991

Susceptibility to desferrioxamine: a new test for the identification of Staphylococcus epidermidis.

The ability to identify Staphylococcus epidermidis quickly and accurately has become increasingly important in clinical microbiology. Susceptibility to desferrioxamine, an iron-chelating agent, was investigated as a new test for the identification of S. epidermidis. All strains of S. epidermidis and S. hominis tested were susceptible to a 1000-micrograms disk of desferrioxamine when grown on brain heart infusion agar. All other strains of coagulase-negative staphylococci, S. aureus and micrococci were resistant. As a single test, susceptibility to desferrioxamine was 96.4% efficient in identifying S. epidermidis; when combined with additional tests such as alkaline phosphatase production and fermentation of trehalose, the efficiency improved to 100%. Desferrioxamine disks were easy to prepare, stable and inexpensive. The test was simple to perform and interpret and should readily find application in clinical microbiology laboratories.

Culture Media↗

PubMed1993

Vertebral osteomyelitis due to Staphylococcus epidermidis.

Vertebral osteomyelitis due to Staphylococcus epidermidis is extremely rare and usually occurs in the context of immunosuppression in association with an adequate portal of entry for infection. This paper reports a case of vertebral osteomyelitis due to S. epidermidis in a man with no evidence of immunosuppression or obvious portal of entry. The patient presented with severe back pain but there were no clinical signs of infection. Radiographs and computerized tomographs showed destruction of thoracic vertebral bodies and an adjacent soft tissue mass. S. epidermidis was grown from multiple blood cultures. All isolates had identical antibiograms and biochemical profiles. There was evidence of healing of the vertebral bodies and resolution of the soft tissue mass after appropriate antistaphylococcal treatment.

Aged↗

PubMed1988

Colonization of n-butyl-2-cyanoacrylate tissue adhesive by Staphylococcus epidermidis.

In vitro adhesion of Staphylococcus epidermidis to cyanoacrylate tissue adhesive surfaces was investigated by employing a modified Robbins device. Bacterial colonization of n-butyl-2-cyanoacrylate tissue adhesive polymer surfaces was demonstrated by employing scanning electron microscopy, epifluorescence technique, and aerobic plate counts. The S. epidermidis was found to rapidly colonize the n-butyl-2-cyanoacrylate polymer surface producing a biofilm composed of embedded bacteria in an extensive amorphous matrix that totally occludes the material surface. The results indicate that the bacteriocidal properties of the polymer to this bacteria are weak or absent.

Enbucrilate↗

PubMed1986

Interference with granulocyte function by Staphylococcus epidermidis slime.

The interaction of Staphylococcus epidermidis slime with human neutrophils (PMN) was examined by using isolated slime and allowing bacteria to elaborate slime and other extracellular products in situ. S. epidermidis slime was found to contain a chemoattractant. Incubation of PMN with 50 micrograms or more of slime per ml inhibited subsequent chemotaxis of the PMN to n-formyl-methionyl-leucyl-phenylalanine by 27% and to zymosan-activated serum by 44 to 67% with increasing slime concentrations. S. epidermidis slime stimulated little degranulation of untreated PMN. After pretreatment of PMN with 5 micrograms of cytochalasin b per ml, slime predominantly induced release of specific granule contents (33.8% lactoferrin release by 250 micrograms of slime per ml versus 10% myeloperoxidase release by 250 micrograms of slime per ml). By a surface phagocytosis assay, PMN uptake of radiolabeled S. epidermidis which were incubated for 18 h on a plastic surface for slime expression was less than that for S. epidermidis adhered to the plastic for 2 h or grown in unsupplemented nutrient broth. These results suggest that S. epidermidis slime interaction with PMN may be potentially detrimental to host defense and may contribute to the ability of this organism to persist on surfaces of foreign bodies in the vascular or central nervous system.

Chemotaxis, Leukocyte↗

PubMedDate not supplied

[In vitro adherence of Staphylococcus epidermidis and Pseudomonas alcaligenes to intraocular lenses].

PURPOSE: To quantify and compare the in vitro adherence of Staphylococcus epidermidis and Pseudomonas alcaligenes to different intraocular lenses (IOLs). METHODS: Fourteen intraocular lenses were used in the experiment. Four of polymethylmethacrylate (PMMA), four of silicone, four of hydrogel and two of acrylic. Eight intraocular lenses were placed in eight test tubes containing 4 ml of Pseudomonas alcaligenes suspension, and six intraocular lenses were placed in six test tubes containing 4 ml of Staphylococcus epidermidis suspension. The bacterial suspension used for adherence tests was 10(8) colony-forming units per milliliter (CFU/mL) which corresponds to 0.5 in the scale of McFarland. The lenses were incubated at 37 degrees for two hours. After this, intraocular lenses were removed from the test tubes and dried twice with the use of distilled and sterile water. The material was spread on blood-agar for cultivation at 35-37 degrees C and was evaluated every 24 hours up to 72 hours. In the samples where there was bacterial growth, the colonies were counted using the conventional laboratory methods. All assays were performed in duplicate. RESULTS: Adherence of Staphylococcus epidermidis to PMMA intraocular lenses was lower than to hydrogel and silicone intraocular lenses. Adherence of Pseudomonas alcaligenes to hydrogel intraocular lenses was lower than to PMMA, acrylic and silicone intraocular lenses. CONCLUSIONS: Results suggest that in vitro adherence of Staphylococcus epidermidis and Pseudomonas alcaligenes to intraocular lenses is influenced by type of material of the lens and by microorganism species. Bacterial adherence may play a role in the pathogenesis of some forms of endophthalmitis after cataract surgery. More in vitro and clinical studies are necessary to elucidate the mechanisms by which Staphylococcus epidermidis and Pseudomonas alcaligenes cause endophthalmitis.

Bacterial Adhesion↗

PubMed1987

Staphylococcus epidermidis: a significant nosocomial pathogen.

Staphylococcus epidermidis is an organism formerly believed to be nonpathogenic. It is now recognized as a pathogen, causing infections on implanted devices and among immunosuppressed patients. Further, it has been involved in the development of resistance to a number of antibiotics. The epidemiology of this organism, its pathogenesis, and its treatment are important to infection control practitioners.

Anti-Bacterial Agents↗

PubMed1992

Frequency and transferability of trimethoprim and sulfonamide resistance in methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis.

A total of 374 Staphylococcus aureus and 126 Staphylococcus epidermidis strains from 14 countries were studied for their resistance to methicillin, trimethoprim (Tp) and sulfonamides (Su), alone and combined (TpSu). The frequency of resistance to Tp, Su and TpSu was much higher in methicillin-resistant S. epidermidis (MRSE) than in methicillin-resistant S. aureus (MRSA). Considerable differences, however, existed in isolates from different countries. Resistance to Tp, Su or TpSu in MRSA was low or absent in isolates from Switzerland, Spain, Japan, Mexico, Argentina and Chile, but high in isolates from Germany and Brazil. High level Tp resistance mostly resided on large plasmids. It could be transferred in 17 out of 97 strains. Su resistance was never cotransferred. Strains cured of their large Tp resistance plasmids remained Su-resistant, which suggests a chromosomal location of Su resistance.

Anti-Bacterial Agents↗

PubMed1984

Pathogenesis and management of Staphylococcus epidermidis 'plastic' foreign body infections.

Staphylococcus epidermidis infections on foreign bodies made of plastic are caused by special and complex mechanisms. The staphylococcal cells are able to adhere to and grow on polymer surfaces in vivo and in vitro. In the course of colonization they produce an extracellular substance ('slime') which eventually covers them. It is thought that the staphylococcal slime has several biological functions, including promoting adhesion and protection against both antibiotics and host defence mechanisms. In patients, the removal of a colonized device should be accompanied by the parenteral administration of highly effective antistaphylococcal drugs, such as vancomycin.

Adhesiveness↗

PubMed2004

Specificity grouping of the accessory gene regulator quorum-sensing system of Staphylococcus epidermidis is linked to infection.

Staphylococcus epidermidis represents the most frequent pathogen involved in nosocomial infections and infections of indwelling medical devices. The strain-to-strain variation of the gene encoding the quorum-sensing pheromone of S. epidermidis as well as the correlation between specificity groups and origin from infection were determined. The pro-pheromone gene was highly conserved and showed infrequent, non-synonymous, single-nucleotide polymorphisms that led to conservative amino acid exchanges only. Importantly, one specificity group was significantly more frequent among strains isolated from infection. The finding that quorum-sensing specificity groups are linked to infection demonstrates the relevance of quorum-sensing for virulence in this critical human pathogen and contributes to the scientific basis needed for the development of quorum-sensing-targeting drugs.

Amino Acid Sequence↗

PubMed1990

Defining Staphylococcus epidermidis cell wall proteins.

Three Staphylococcus epidermidis isolates of differing bacteriophage types were studied to define proteins confined to the cell wall, which were surface exposed and thus available to interact with the host. Three major proteins of 37, 41, and 51 kDa were identified in all whole-cell lysates and cell wall extracts by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. Two additional proteins of 18 and 25 kDa became evident by using 125I labeling to delineate surface-exposed proteins. A classification scheme using P1 to P5 to delineate the 51-, 41-, 37-, 25- and 18-kDa proteins is proposed. Additionally, murine immune sera were used to identify two immunodominant proteins of 51 and 25 kDa (P1 and P4, respectively).

Bacterial Proteins↗

PubMed1991

Phage typing and phage induction in carrier and invasive Staphylococcus epidermidis isolates.

Fifty-nine Staphylococcus epidermidis isolates (30 carrier and 29 invasive) were phage typed using the phage set developed by Pulverer. Poor results were obtained as only 24% of the invasive and 27% of the carrier isolates were typable, with no differences between these percentages (P greater than 0.05). Isolation of lysogenic phages present in the isolates was attempted by three different methods, induction being satisfactory only by overnight incubation with 0.1 mg l-1 mitomycin C and detection with 2,3,5-triphenyl-tetrazolium chloride. All isolates produced phages, and the lytic activity of these phages was expressed on more than 50% of the isolates in all cases (except one phage which lysed 10% of the carrier isolates and 13.8% of the invasive isolates). The percentages of isolates lysed in the two groups (carrier and invasive) were statistically different (P less than 0.05) only for phages derived from four isolates.

Bacteriophage Typing↗

PubMed2004

Biofilm detection and the clinical significance of Staphylococcus epidermidis isolates.

The ability of Staphylococcus epidermidis to produce biofilm was compared in 147 clinically significant strains repeatedly isolated from blood cultures of patients with bloodstream infection and in 147 strains isolated from skin. The strains were examined for the presence of ica operone, for the ability to form biofilm by Christensen's test-tube method and for the production of slime by Congo Red agar method. The ica operone was found in 92 (62.6 %) blood isolates and in 44 (29.9) isolates from skin. Christensen's test-tube method was positive in 79 (53.7) and 33 (22.4), Congo Red agar method in 64 (43.5) and 31 (21.1) of blood and skin isolates, respectively. All three methods were more frequently positive in clinically significant isolates from blood than in strains isolated from skin. The detection of ica operone and the Christensen's test-tube method showed better correlation with the clinical significance than the Congo Red agar method.

Bacteremia↗

PubMed2006

Role of the luxS quorum-sensing system in biofilm formation and virulence of Staphylococcus epidermidis.

Nosocomial infections caused by Staphylococcus epidermidis are characterized by biofilm formation on implanted medical devices. Quorum-sensing regulation plays a major role in the biofilm development of many bacterial pathogens. Here, we describe luxS, a quorum-sensing system in staphylococci that has a significant impact on biofilm development and virulence. We constructed an isogenic DeltaluxS mutant strain of a biofilm-forming clinical isolate of S. epidermidis and demonstrated that luxS signaling is functional in S. epidermidis. The mutant strain showed increased biofilm formation in vitro and enhanced virulence in a rat model of biofilm-associated infection. Genetic complementation and addition of autoinducer 2-containing culture filtrate restored the wild-type phenotype, demonstrating that luxS repressed biofilm formation through a cell-cell signaling mechanism based on autoinducer 2 secretion. Enhanced production of the biofilm exopolysaccharide polysaccharide intercellular adhesin in the mutant strain is presumably the major cause of the observed phenotype. The agr quorum-sensing system has previously been shown to impact biofilm development and biofilm-associated infection in a way similar to that of luxS, although by regulation of different factors. Our study indicates a general scheme of quorum-sensing regulation of biofilm development in staphylococci, which contrasts with that observed in many other bacterial pathogens.

Animals↗

PubMed1991

Evaluation of three techniques for documenting Staphylococcus epidermidis vascular prosthetic graft infections.

Staphylococcus epidermidis (S. epidermidis) vascular prosthetic graft infections are notoriously hard to detect. Three different techniques of determining whether vascular prosthetic grafts were infected using a dog model were evaluated. Aortic angiograms were compared with nuclear magnetic resonance (NMR) imaging and systemic norepinephrine (NE) kinetics to determine if either newer technique would be more reliable than standard angiograms. Twelve dogs were randomized to control (n = 6) or infected groups (n = 6). All dogs had a 5 cm section of their infrarenal aorta replaced with knitted Dacron vascular prosthetic graft. The grafts in the infected group were contaminated by soaking them in a broth containing S. epidermidis. NE production and clearance rates were calculated for all animals after an infusion of 3H-NE using the steady-state radionuclide tracer methodology. One week following graft insertion, dogs were reanesthetized, and the 3H-NE infusion and measurements were repeated. Standard angiograms and NMR imaging were also performed. Once all tests were performed, the prosthetic grafts were removed for cultures. Comparisons between the initial and final norepinephrine measurements for each group were made using the nonparametric Wilcoxon two-sample test, while comparisons between the groups were made by chi square or the Student's t test. Angiogram results were similar for control and infected animals. Angiograms missed disruption of the proximal anastomosis found in three of the six infected dogs at graft removal. None of the six control animals, while five of the six infected animals, had localized areas of high signal intensity on NMR imaging (P less than 0.01) suggesting abscess formation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1987

Bacterial concentration correlations in experimental endocarditis caused by Staphylococcus epidermidis.

Using 13 strains of Staphylococcus epidermidis to produce catheter-induced experimental endocarditis in rats, we found that bacterial concentrations in blood cultures obtained at the time of sacrifice correlated significantly with the number of organisms per gram of endocardial vegetation (P less than 0.001) and the total number of organisms per vegetation (P less than 0.001). Furthermore, blood culture concentrations correlated with vegetation weights (P less than 0.001) and sizes of infecting inocula (P less than 0.0001). Mean bacterial concentrations in vegetations more than doubled as bacterial concentrations in blood rose from less than 10 to greater than 100 CFU/ml. Mean values for vegetation weights, total organisms per vegetation, and sizes of infecting inocula were also reflected by the intensity of bacteremia. Moreover, intracardiac catheters were more likely colonized as bacterial concentrations in blood cultures increased, with all catheters culture positive in the 25 animals that exhibited high-grade bacteremia (greater than or equal to 100 CFU/ml). Slime production by the bacteria did not influence the above-mentioned correlations. These data indicate that the blood concentration of bacteria reflects the microbiologic status of infected vegetations in experimental infective endocarditis.

Animals↗

PubMed1990

Comparative treatment of experimental Staphylococcus epidermidis endophthalmitis.

We created experimental Staphylococcus epidermidis endophthalmitis of moderate severity in the aphakic rabbit eye by injecting 100,000 organisms of a standardized laboratory strain (ATCC 155) into the mid-vitreous cavity. This model of endophthalmitis self-sterilizes in about 4 days, but inflammatory signs continue to increase 5 to 7 days after the initial bacterial inoculum. Control eyes were compared with eyes treated with five different strategies 24 hours after bacterial inoculation: intravitreal cefazolin sodium, intravitreal cefazolin plus intramuscular corticosteroid, vitrectomy plus intravitreal antibiotics, vitrectomy plus intravitreal antibiotics and intramuscular corticosteroids, and vitrectomy plus intravitreal antibiotics and corticosteroids. Quantitative grading of inflammation and media clarity were compared at the end of weeks 1, 2, and 3 after treatment. At week 1, eyes treated with vitrectomy had significantly lower inflammatory scores; those treated with corticosteroids had significantly lower scores than those without. The two effects were independent. The best results were observed with treatment consisting of vitrectomy, intraocular antibiotics, and corticosteroids. This strategy also produced significantly more eyes with clear media at the end of week 3 than treatment with intravitreal antibiotics alone.

Adrenal Cortex Hormones↗

PubMed2001

Phenotypic and genotypic markers of Staphylococcus epidermidis virulence.

OBJECTIVES: To analyze Staphylococcus epidermidis strains, previously tested for their virulence in a mouse model of subcutaneous infection, for various phenotypic traits (biofilm density, extracellular polysaccharide, slime-associated antigen (SAA)) and for the presence of the ica gene cluster, to determine which of these phenotypic and genotypic methods best correlates with virulence in the mouse model. METHODS: The quantitative biofilm assay was performed on 10 strains of S. epidermidis, comprising (1) RP62A (ATCC 35984), (2) the strongest and weakest biofilm producers in our collection, (3) a pair of phenotypic variants, and (4) a strain whose biofilm density was enhanced in iron-limited media. Biofilm density was measured after growth at 37 degrees C and at ambient temperature, in trypticase soy broth (TSB) with and without glucose supplementation and using both chemical and heat fixation. Strains were assayed for SAA using a double immunodiffusion method. Extracellular polysaccharide was detected by transmission electron microscopy (TEM). A 546-base-pair segment of the ica gene cluster was amplified by PCR. RESULTS: Biofilm formation in TSB, glucose-enriched TSB, extracellular polysaccharide (observed by TEM), expression of SAA and presence of the ica gene predicted virulence of nine, nine, nine, eight and eight of 10 strains, respectively. The phenotypic expression of biofilm and related properties was medium and temperature dependent. We encountered one ica-positive strain that failed to express biofilm in standard TSB at 37 degrees C, but was virulent in a mouse model, and another strain that lacked ica, produced biofilm and was virulent in the model. CONCLUSIONS: Mouse virulence in our model can be predicted by any of the phenotypic or genotypic methods examined for > or = 80% of strains. Medium and incubation conditions affect the expression of phenotypic markers by some strains. For the remaining strains, possible reasons for inconsistencies between the presence of the ica gene, phenotypic markers and mouse virulence include (1) dependence of biofilm on genes other than ica, (2) sequence differences in ica, (3) dependence of biofilm expression in vivo on strain characteristics and media used to prepare inocula for in vivo studies.

Animals↗