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An inflammatory polypeptide complex from Staphylococcus epidermidis: isolation and characterization.

Staphylococcus epidermidis releases factors that activate the HIV-1 long terminal repeat, induce cytokine release, and activate nuclear factor B in cells of macrophage lineage. The active material had a mass of 34,500 daltons, was inactivated by proteases and partitioned into the phenol layer on hot aqueous phenol extraction, and thus was termed phenol-soluble modulin (PSM). High performance liquid chromatography (HPLC) of crude PSM yielded two peaks of activity designated PSM peak 1 and peak 2. MALDI-TOF (matrix-assisted laser desorption ionization-time of flight) mass spectroscopy indicated the presence of two components in peak 1, which were designated PSM and PSM. Peak 2 contained a single component, designated PSM. Separation of PSM and PSM in peak 1 could be achieved by a second HPLC procedure. The structure of each component was determined by amino acid sequence analysis and identification and sequencing of their genes. PSM, PSM, and PSM were 22-, 44-, and 25-amino acid, respectively, strongly hydrophobic polypeptides. PSM was identified as Staphylococcus epidermidis delta toxin, whereas PSM and PSM exhibited more distant homology to previously described staphylococcal toxins. They appeared to exist as a complex or aggregate with activity greater than the component parts. The properties of the S. epidermidis PSMs suggest that they may contribute to the systemic manifestations of Gram-positive sepsis.

Amino Acid Sequence↗

Mycotic aneurysm of the abdominal aorta in a patient undergoing hemodialysis: an unusual complication of Staphylococcus aureus bacteremia.

When Staphylococcus aureus is repeatedly positive in blood cultures even under effective antibiotics therapy (vancomycin, teicoplanin, or rifampin), computed tomography scan and sonography should be performed early to exclude mycotic aneurysm of the deeply seated arteries, especially in patients with abdominal aortic calcification. Before 1990, the most common causative organism of suprarenal aortic mycotic aneurysm was Salmonella; since 1990, it has been gram-positive cocci (i.e., Streptococcus and Staphylococcus) rather than gram-negative bacilli (i.e., Salmonella), possibly because of the more invasive procedures performed in clinical settings, but this hypothesis needs further investigation.

Aged↗

Characterization of the relationship between polysaccharide intercellular adhesin and hemagglutination in Staphylococcus epidermidis.

To determine whether a relationship exists between biofilm formation and hemagglutination in Staphylococcus epidermidis, 20 skin isolates and 19 prosthetic valve endocarditis isolates were characterized for biofilm formation, hemagglutination, and the presence of a 357-bp polymerase chain reaction product within icaA. A strong association existed between biofilm formation, which has been linked to strains that produce polysaccharide intercellular adhesin (PIA), and hemagglutination. Strains that produced biofilm were significantly (P<.001) more likely to mediate hemagglutination (16 biofilm-positive/hemagglutination-positive strains and 19 biofilm-negative/hemagglutination-negative strains) within the 39 clinical strains tested. In addition, Staphylococcus carnosus TM300, a biofilm-negative, hemagglutination-negative strain, carrying the ica operon-containing plasmid pCN27, produced significant biofilm on glass and mediated hemagglutination (>/=1/128). It was concluded that production of PIA and hemagglutination are strongly associated and that PIA, at least in part, mediates hemagglutination in S. epidermidis.

Adhesins, Bacterial↗

Staphylococcus epidermidis with intermediate resistance to vancomycin: elusive phenotype or laboratory artifact?

The recent and troubling isolation of Staphylococcus aureus and coagulase-negative staphylococci that have increased resistance to glycopeptide antibiotics has prompted the use of aggressive surveillance measures in the clinical microbiology laboratory to aid in the recognition of these strains. Despite increasing awareness, the confirmation of glycopeptide resistance among staphylococci can be problematic; we present a case of catheter-associated peritonitis caused by Staphylococcus epidermidis to illustrate the dilemma.

Anti-Bacterial Agents↗

Increasing prevalence of methicillin-resistant Staphylococcus aureus infection in California jails.

Staphylococcus aureus clinical isolates obtained from patients who were inmates of the San Francisco County jail system showed an increase in the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) from 29%, in 1997, to 74%, in 2002; 91% of the MRSA isolates carried staphylococcal chromosomal cassette mec (SCCmec) type IV. Pulsed field gel electrophoresis and multilocus sequence typing demonstrated 2 major clonal groups. One of these clonal groups is genetically indistinguishable from the strain responsible for an outbreak of MRSA in the Los Angeles County jail system in 2002.

Anti-Bacterial Agents↗

Two cases of fatal shock after transfusion of platelets contaminated by Staphylococcus aureus: role of superantigenic toxins.

We detected Staphylococcus aureus superantigenic toxins in the platelet infusion bags that had been used for 2 patients who subsequently developed transfusion-associated Staphylococcus aureus infection. Both patients, who were immunodeficient, developed manifestations of toxic shock syndrome and septic shock, and they died soon after the onset of infection.

Aged↗

Risk factors for hematogenous complications of intravascular catheter-associated Staphylococcus aureus bacteremia.

BACKGROUND: The role of both host and pathogen characteristics in hematogenous seeding following Staphylococcus aureus bacteremia is incompletely understood. METHODS: Consecutive patients with intravascular catheter-associated Staphylococcus aureus bacteremia were prospectively recruited over a 91-month period. The corresponding bloodstream isolates were examined for the presence of 35 putative virulence determinants. Patient and bacterial characteristics associated with the development of hematogenous complications (HCs) (i.e., septic arthritis, vertebral osteomyelitis, or endocarditis) were defined. RESULTS: HC occurred in 42 (13%) of 324 patients. Patient characteristics at diagnosis that were associated with HC included community onset (relative risk [RR], 2.25; 95% confidence interval [CI], 1.24-4.07; P=.007), increased symptom duration (odds ratio for each day, 1.14; 95% CI, 1.06-1.2; P<.001), presence of a long-term intravascular catheter or noncatheter prosthesis (RR, 4.02; 95% CI, 1.74-9.27; P<.001), hemodialysis dependence (RR, 3.84; 95% CI, 2.08-7.10; P<.001), and higher APACHE II score (P=.02). Bacterial characteristics included sea (RR, 2.03; 95% CI, 1.16-3.55; P=.011) and methicillin-resistant S. aureus (MRSA) (RR, 2.09; 95% CI, 1.19-3.67; P=.015). Subsequent failure to remove a catheter was also associated with HC (RR, 2.28; 95% CI, 1.22-4.27; P=.011). On multivariable analysis, symptom duration, hemodialysis dependence, presence of a long-term intravascular catheter or a noncatheter device, and infection with MRSA remained significantly associated with HC. CONCLUSIONS: This investigation identifies 4 host- and pathogen-related risk factors for hematogenous bacterial seeding and reaffirms the importance of prompt catheter removal.

Adult↗

Lack of wall teichoic acids in Staphylococcus aureus leads to reduced interactions with endothelial cells and to attenuated virulence in a rabbit model of endocarditis.

Wall teichoic acids (WTAs) are major surface components of gram-positive bacteria that have recently been shown to play a key role in nasal colonization by Staphylococcus aureus. In the present study, we assessed the impact that WTAs have on endovascular infections by using a WTA-deficient S. aureus mutant ( Delta tagO). There were no significant differences detected between the isogenic parental strain (SA113) and the Delta tagO mutant in polymorphonuclear leukocyte-mediated opsonophagocytosis; killing by a prototypic platelet microbicidal protein; or binding to platelets, fibronectin, or fibrinogen. However, compared with the parental strain, the Delta tagO mutant adhered considerably less well to human endothelial cells, especially under flow conditions (70.3% reduction; P<.05). Beads coated with WTA bound to endothelium in a dose-dependent manner, suggesting that WTA contributes specifically to this interaction. These in vitro data closely paralleled those from a rabbit model of infective endocarditis in which the Delta tagO mutant was compared with the parental strain. Clearances of staphylococcus from the bloodstream were equivalent, but the Delta tagO mutant showed a significantly reduced capacity to both colonize sterile cardiac vegetations (P<.05) and proliferate within these vegetations, the kidneys, and the spleen (P<.001). We conclude that WTA is an important factor in the induction and progression of endovascular S. aureus infection, likely through a specific interaction with endothelial cells.

Animals↗

Evaluation of an intervention designed to decrease the rate of nosocomial methicillin-resistant Staphylococcus aureus infection by encouraging decreased fluoroquinolone use.

OBJECTIVE: Society for Health Care Epidemiology guidelines recommend decreasing the use of fluoroquinolone antibiotics in institutions where methicillin-resistant Staphylococcus aureus (MRSA) is endemic. We evaluated whether an intervention to limit fluoroquinolone use was associated with a lower rate of nosocomial MRSA infection and summarized changes in antibiotic use, changes in other variables potentially correlated with a lower rate of MRSA infection, and rates of nosocomial infections due to other pathogens. DESIGN: Single-center quasi-experimental design. A time series of nosocomial MRSA infections was measured at monthly intervals from July 2001 through June of 2004; there were 80 MRSA infections recorded. Segmented regression analysis (ie, quasi-Poisson generalized linear models) was used to evaluate variables possibly associated with the nosocomial MRSA infection rate. SETTING: An 87-bed Veterans Affairs teaching hospital with an extended-care facility. INTERVENTION: A physician-directed computer-generated intervention designed to limit the use of fluoroquinolone antibiotics was initiated, and institutional changes in antibiotic use and nosocomial MRSA infection rates were tracked. RESULTS: After the intervention, fluoroquinolone use decreased by approximately 34%, and levofloxacin use decreased by approximately 50%. Decreased fluoroquinolone use was offset by increased cephalosporin, piperacillin-tazobactam, and trimethoprim-sulfamethoxazole use. The nosocomial MRSA infection rate decreased from 1.37 to 0.63 episodes per 1,000 patient-days after the study intervention (P=.02). Coagulase-negative Staphylococcus and Enterococcus infection rates also decreased. However, the rate of infection with gram-negative organisms increased. The rate of MRSA infection was positively correlated with levofloxacin use (P=.01) and azithromycin use (P=.08), whereas it was negatively correlated with summer season (P=.05). In a subsequent model, the rate of MRSA infection was negatively correlated with the study intervention (P=.04). CONCLUSION: Reduction in the institutional use of fluoroquinolones may be associated with a lower nosocomial MRSA infection rate.

Cross Infection↗

Community-associated methicillin-resistant Staphylococcus aureus causing chronic pneumonia.

A young woman presented with pneumonia of a 3-month duration with predominantly nodular pulmonary infiltrates. Methicillin-resistant Staphylococcus aureus was identified in multiple cultures of sputum specimens. According to findings of pulsed-field gel electrophoresis, the isolate was identical to USA 300 and carried a type IV Staphylococcus cassette chromosome mec type IV gene and the genes for Panton-Valentine leukocidin.

Adult↗

Evidence of delays in transferring patients with methicillin-resistant Staphylococcus aureus or vancomycin-resistant Enterococcus to long-term-care facilities.

This retrospective case-control study examined whether there was a difference in length of time awaiting long-term-care placement for patients identified as having methicillin-resistant Staphylococcus aureus or vancomycin-resistant Enterococcus compared to controls. Thirty-nine patients with methicillin-resistant Staphylococcus aureus or vancomycin-resistant Enterococcus waited for placement an average of 61 days longer than controls (P<.0002). The average number of requests for placement was 2.5 compared to 1.7 for controls (P=.015).

Aged↗

Healthcare-associated Staphylococcus aureus bacteremia and the risk for methicillin resistance: is the Centers for Disease Control and Prevention definition for community-acquired bacteremia still appropriate?

OBJECTIVE: To evaluate a new classification for bloodstream infections that differentiates hospital acquired, healthcare associated, and community acquired in patients with blood cultures positive for Staphylococcus aureus. DESIGN: Prospective, observational study. SETTING: Three tertiary-care, university-affiliated hospitals in Dublin, Ireland, and Strasbourg, France. PATIENTS: Two hundred thirty consecutive patients older than 18 years with blood cultures positive for S. aureus. METHODS: S. aureus bacteremia (SAB) was defined as hospital acquired if the first positive blood culture was performed more than 48 hours after admission. Other SABs were classified as healthcare associated or community acquired according to the definition proposed by Friedman et al. When available, strains of methicillin-resistant Staphylococcus aureus (MRSA) were analyzed by pulsed-field gel electrophoresis (PFGE). RESULTS: Eighty-two patients were considered as having community-acquired bacteremia according to the Centers for Disease Control and Prevention (CDC) classification. Of these 82 patients, 56% (46) had healthcare-associated SAB. MRSA prevalence was similar in patients with hospital-acquired and healthcare-associated SAB (41% vs 33%; P > .05), but significantly lower in the group with community-acquired SAB (11%; P < .03). PFGE of MRSA strains showed that most community-acquired and healthcare-associated MRSA strains were similar to hospital-acquired MRSA strains. On multivariate analysis, Friedman's classification was more effective than the CDC classification for predicting MRSA. CONCLUSION: These results support the call for a new classification for community-acquired bacteremia that would account for healthcare received outside the hospital by patients with SAB.

Aged↗

Impact of routine intensive care unit surveillance cultures and resultant barrier precautions on hospital-wide methicillin-resistant Staphylococcus aureus bacteremia.

BACKGROUND: Serial interventions are often used to reduce the risk of health care-associated methicillin-resistant Staphylococcus aureus (MRSA) infections. To our knowledge, the relative impact of these interventions has not previously been ascertained. METHODS: We conducted a retrospective study of 4 major infection control interventions using an interrupted time series design to evaluate their impact on MRSA bacteremia in an 800-bed hospital with 8 intensive care units (ICUs). Interventions were introduced 1 at a time during a 9-year period and involved the promotion of compliance with maximal sterile barrier precautions during central venous catheter placement, the institution of alcohol-based hand rubs for hand disinfection, the introduction of a hand hygiene campaign, and the institution of routine nares surveillance cultures for MRSA in all ICUs for patients on ICU admission and weekly thereafter while in the ICU. Positive cultures resulted in the initiation of contact isolation precautions. Using segmented regression analyses, we evaluated changes in monthly incidence and prevalence of MRSA bacteremia from their predicted values. Methicillin-susceptible Staphylococcus aureus bacteremia was monitored as a control. RESULTS: Routine surveillance cultures and subsequent contact isolation precautions resulted in substantial reductions in MRSA bacteremia in both ICUs and non-ICUs. In 16 months, the incidence density of MRSA bacteremia decreased by 75% in ICUs (P=.007) and by 40% in non-ICUs (P=.008), leading to a 67% hospital-wide reduction in the incidence density of MRSA bacteremia (P=.002). Methicillin-susceptible S. aureus bacteremia rates remained stable during this time. The other interventions were not associated with a statistically significant change in MRSA bacteremia. CONCLUSIONS: Routine surveillance for MRSA in ICUs allowed earlier initiation of contact isolation precautions and was associated with large and statistically significant reductions in the incidence of MRSA bacteremia in the ICUs and hospital wide. In contrast, no similar decrease was attributable to the other infection control interventions.

Bacteremia↗

Control of methicillin-resistant Staphylococcus aureus at a university hospital: one decade later.

OBJECTIVE: To investigate the cause of increasing rates of nosocomial methicillin-resistant Staphylococcus aureus (MRSA) infection at a university hospital. DESIGN: Review of data collected by prospective hospital wide surveillance regarding rates of nosocomial MRSA colonization and infection. SETTING: A 700-bed university hospital providing primary and tertiary care. PATIENTS: Patients admitted to the hospital between 1986 and 1993 who were found to be infected or colonized with MRSA. MAIN OUTCOME MEASUREMENT: Rates of MRSA infection and colonization. RESULTS: MRSA infection or colonization was identified in 399 patients (0.18%) admitted during the 8-year study. There was no correlation between the annual rates of MRSA and methicillin-sensitive Staphylococcus aureus (MSSA) infections (P = .66). The frequency of both nosocomial and non-nosocomial cases increased significantly over the last 4 years of the study (P < .001 for trend). The ratio of patients who had acquired MRSA nosocomially to those admitted who already were infected or colonized decreased significantly during the study period (P = .002 for trend). There was a significant increase in the frequency of patients with MRSA being transferred from nursing homes and other chronic care facilities (P = .011). A cost-benefit analysis suggested that surveillance cultures of patients transferred from other healthcare facilities would save between $20,062 and $462,067 and prevent from 8 to 41 nosocomial infections. CONCLUSIONS: An increase in the incidence of nosocomial MRSA infection was associated with an increased frequency of transfer of colonized patients from nursing homes and other hospitals. The lack of correlation between rates of MRSA and MSSA infections suggested that MRSA infections significantly increased the overall rate of staphylococcal infection. Screening cultures of transfer patients from facilities with a high prevalence of MRSA may offer significant benefit by preventing nosocomial infections and reducing patient days spent in isolation.

Cost-Benefit Analysis↗

Methicillin-resistant Staphylococcus aureus control in hospitals: the French experience. Association des Pays de la Loire pour l'Eviction des Infections Nosocomiales.

The first cases of isolation of Staphylococcus aureus resistant to methicillin in France were published in 1962. However, until recently, very few epidemiological studies or attempts to control the epidemic have been done in France. In this article, we present the results of a prospective study performed during a 3-month period in 27 hospitals of the Région des Pays de la Loire. Among the 94,605 hospitalized patients included in the study, 0.45% (427) developed methicillin-resistant Staphylococcus aureus (MRSA) infections, the incidence rate ranging from 0% to 1.2%. Thirty-four percent of MRSA-infected patients were 80 years old or older, 30% had been transferred from another service and 19% from another hospital, 56% were hospitalized at least once during the previous year, MRSA had been isolated at least once previously in 18% of MRSA-infected patients, 19% died, 16% were transferred to another service and 11% to another hospital, and only 32% were discharged to their homes. A poor compliance to contact isolation precautions was observed in all hospitals: 46% of MRSA-infected patients were hospitalized in a private room; gloves, masks, and gowns were worn for the care of 63.4%, 14%, and 42.5% of MRSA-infected patients, respectively; and handwashing was feasible in the rooms of 52% of the patients.

Cross Infection↗

Genetic linkage between resistance to quaternary ammonium compounds and beta-lactam antibiotics in food-related Staphylococcus spp.

Little is known about the occurrence of antimicrobial resistance determinants in staphylococci isolated from food and food processing industries. Quaternary ammonium compound (QAC)-resistant coagulase-negative staphylococci (CNS) isolated from food and food-processing industries were investigated for the presence of genetic determinants (qacA/B and qacC/smr) encoding resistance to the QAC benzalkonium chloride (BC), several antibiotic resistance genes, and staphylococcal insertion sequences IS257 and IS256. Six qacA/B-harboring strains were resistant to penicillin and hybridized to a blaZ probe. The qacA/B and blaZ probes hybridized to plasmids of similar size in three isolates. Molecular and genetic characterization of the 23-kb plasmid (pST6) of Staphylococcus epidermidis St.6 revealed the presence of qacB adjacent to an incomplete beta-lactamase transposon Tn552 encoding the gene cluster blaZ, blaR, and blaI. Sequence analysis of flanking regions and the intergenic region between blaZ and qacB revealed the presence of IS257 downstream of blaZ as well as sin and binR between blaZ and qacB. In the three other BC and penicillin-resistant strains, the qacA/B and blaZ genes were located on separate plasmids. A qacC harboring S. epidermidis strain (St.17) also hybridized to tetK (tetracycline resistance) and ermB (erythromycin resistance) genes. The individual genes were located on separate plasmids, suggesting no linkage between QAC and antibiotic resistance determinants. Plasmid-free Staphylococcus aureus RN4220 allowed uptake of the pST6 plasmid DNA, indicating that the resistance genes could potentially be transferred to pathogens under selective stress. In conclusion, presence of both resistance determinants could lead to co-selection during antimicrobial therapy or disinfection in hospitals or in food industries.

Anti-Bacterial Agents↗

The epidemiology of methicillin-resistant Staphylococcus aureus at a Saudi university hospital.

To determine changes in the epidemiology of methicillin-resistant Staphylococcus aureus (MRSA), a prospective study was conducted for a 30-month period in a university hospital in the eastern province of Saudi Arabia. Of 1,096 Staphylococcus aureus isolates identified, 92 (8.4%) were MRSA. Seventy-two (78%) of the 92 isolates were from nosocomial cases and 20 (22%) isolates were from community cases. Thirteen (14%) of 20 patients with community-acquired MRSA had no discernible risk factors. The community-acquired MRSA from patients without identified risk were susceptible to a greater number of antimicrobials compared with isolates obtained from patients with community-acquired MRSA with identified risk or with nosocomially acquired MRSA. No strain resistant to vancomycin was recorded in this study. The impact of these organisms could be substantial if they become more frequent or widespread.

Community-Acquired Infections↗

Restriction fragment fingerprint and genome sizes of Staphylococcus species using pulsed-field gel electrophoresis and infrequent cleaving enzymes.

Large restriction fragments of genomic DNA from Staphylococcus species were separated by pulsed-field gel electrophoresis (PFGE). Five different strains of S. aureus (ISP8, SAU3A, PS96, ATCC 6538, ATCC 15564) and three representative strains of S. haemolyticus SM102, S. warneri MCS4, S. cohnii LK478 from human hosts, and one strain of S. aureus (ATCC 8432) from an avian host were used in this study. Since Staphylococcus is A + T rich (approximately 67%), restriction fragments were obtained by digesting chromosomal DNA with endonucleases that recognize GC-rich sequences. Five enzymes Csp I, Sma I, Ecl XI, Ksp I, or Sac II were used for generation of few (7 to 16) distinctly separated fragments, with average sizes in the range of 200-300 kb. The size distribution of restriction fragments for each enzyme for each strain produced a strain-identifying fingerprint, and the genome size of each strain was determined from such restriction fragments separated by PFGE.

DNA Fingerprinting↗