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In vitro postantibiotic effect of daptomycin (LY146032) against Enterococcus faecalis and methicillin-susceptible and methicillin-resistant Staphylococcus aureus strains.

The suppression of bacterial growth that persists after brief exposure to antimicrobial agents has been termed the postantibiotic effect (PAE). This pharmacodynamic interaction varies for each microorganism-antimicrobial agent combination. Daptomycin (LY146032) is a new lipopeptide antibiotic with activity against gram-positive organisms. We studied the in vitro bactericidal activities and PAEs of the following drugs: daptomycin compared with penicillin G and vancomycin, without and with gentamicin against Enterococcus faecalis strains; daptomycin compared with nafcillin and vancomycin against methicillin-susceptible Staphylococcus aureus strains; and daptomycin compared with vancomycin against methicillin-resistant S. aureus strains. Daptomycin, alone and when used in combination with gentamicin, exhibited greater bactericidal activity and in general produced a longer PAE than standard effective regimens against the organism strains studied.

Anti-Bacterial Agents↗

Restriction maps of the regions coding for methicillin and tobramycin resistances on chromosomal DNA in methicillin-resistant staphylococci.

Chromosomal BamHI DNA fragments containing both the mecA gene encoding the penicillin-binding protein responsible for methicillin resistance and the aadD gene encoding 4',4"-adenylyltransferase responsible for tobramycin resistance were cloned from three methicillin- and tobramycin-resistant strains of Staphylococcus aureus and one strain of Staphylococcus epidermidis. Physical maps of the fragments were similar, suggesting their unique origin.

Blotting, Southern↗

Cloning and characterization of the fmt gene which affects the methicillin resistance level and autolysis in the presence of triton X-100 in methicillin-resistant Staphylococcus aureus.

In methicillin-resistant Staphylococcus aureus (MRSA) strains, Triton X-100 reduced the oxacillin resistance level, although the degree of reduction varied from strain to strain. To study the responses of MRSA strains to Triton X-100, we isolated a Tn551 insertion mutant of the COL strain that became more susceptible to oxacillin in the presence of 0.02% Triton X-100. The Tn551 insertion of the mutant was transduced back to the parent strain, other MRSA strains (strains KSA8 and NCTC 10443), and methicillin-susceptible strain RN450. All transductants of MRSA strains had reduced levels of resistance to oxacillin in the presence of 0.02% Triton X-100, while those of RN450 did not. Tn551 mutants of KSA8 and NCTC 10443 also had reduced levels of resistance in the absence of 0.02% Triton X-100. The autolysis rates of the transductants in the presence of 0.02% Triton X-100 were significantly increased. Amino acid analysis of peptidoglycan and testing of heat-inactivated cells for their susceptibilities to several bacteriolytic enzymes showed that there were no significant differences between the parents and the respective Tn551 mutants. The Tn551 insertion site mapped at a location different from the previously identified fem and llm sites. Cloning and sequencing showed that Tn551 had inserted at the C-terminal region of a novel gene designated fmt. The putative Fmt protein showed a hydropathy pattern similar to that of S. aureus penicillin-binding proteins and contained two of the three conserved motifs shared by penicillin-binding proteins and beta-lactamases, suggesting that fmt may be involved in cell wall synthesis.

Bacteriolysis↗

Mupirocin prophylaxis against methicillin-susceptible, methicillin-resistant, or vancomycin-intermediate Staphylococcus epidermidis vascular-graft infection.

A rat model was used to investigate the efficacy of mupirocin in the prevention of vascular prosthetic graft infection due to Staphylococcus epidermidis strains with different susceptibility patterns (methicillin susceptible, methicillin resistant, and with intermediate resistance to vancomycin). The effect of mupirocin-soaked Dacron was compared to that of perioperative intraperitoneal prophylaxis with vancomycin. Graft infections were established in the back subcutaneous tissue of adult male Wistar rats by implantation of Dacron prostheses (1 cm(2)) followed by topical inoculation with 5 x 10(7) CFU of one staphylococcal strain. The study included a control group (no graft contamination), three contaminated groups that did not receive any antibiotic prophylaxis, three contaminated groups that received mupirocin-soaked grafts, three contaminated groups in which perioperative intraperitoneal vancomycin prophylaxis (10 mg/kg of body weight) was administered, and three contaminated groups that received mupirocin-soaked grafts and perioperative intraperitoneal vancomycin prophylaxis (10 mg/kg). The grafts were sterilely removed 7 days after implantation, and the infection was evaluated by using sonication and quantitative agar culture. Data analysis showed the efficacy of mupirocin against all three strains, with growth of the strains in treated rats significantly different than that in the untreated control. In addition, mupirocin was more effective than vancomycin against the strain with intermediate susceptibility to the glycopeptide. Finally, the combination of mupirocin and vancomycin produced complete suppression of the growth of all of the strains.

Aminoglycosides↗

Insights on evolution of virulence and resistance from the complete genome analysis of an early methicillin-resistant Staphylococcus aureus strain and a biofilm-producing methicillin-resistant Staphylococcus epidermidis strain.

Staphylococcus aureus is an opportunistic pathogen and the major causative agent of numerous hospital- and community-acquired infections. Staphylococcus epidermidis has emerged as a causative agent of infections often associated with implanted medical devices. We have sequenced the approximately 2.8-Mb genome of S. aureus COL, an early methicillin-resistant isolate, and the approximately 2.6-Mb genome of S. epidermidis RP62a, a methicillin-resistant biofilm isolate. Comparative analysis of these and other staphylococcal genomes was used to explore the evolution of virulence and resistance between these two species. The S. aureus and S. epidermidis genomes are syntenic throughout their lengths and share a core set of 1,681 open reading frames. Genome islands in nonsyntenic regions are the primary source of variations in pathogenicity and resistance. Gene transfer between staphylococci and low-GC-content gram-positive bacteria appears to have shaped their virulence and resistance profiles. Integrated plasmids in S. epidermidis carry genes encoding resistance to cadmium and species-specific LPXTG surface proteins. A novel genome island encodes multiple phenol-soluble modulins, a potential S. epidermidis virulence factor. S. epidermidis contains the cap operon, encoding the polyglutamate capsule, a major virulence factor in Bacillus anthracis. Additional phenotypic differences are likely the result of single nucleotide polymorphisms, which are most numerous in cell envelope proteins. Overall differences in pathogenicity can be attributed to genome islands in S. aureus which encode enterotoxins, exotoxins, leukocidins, and leukotoxins not found in S. epidermidis.

Biofilms↗

Temperature-related variations in the lipid composition of methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains.

Growth of methicillin-susceptible and -resistant strains of Staphylococcus aureus at 28 and 37 degrees C led to changes in the relative amounts of phospholipids recovered. The major phospholipids present in these strains were phosphatidylglycerol, lysophosphatidylglycerol, and diphosphatidylglycerol. Phosphatidylglycerol tended to occur in lower amounts in the resistant strains, but it was the major phospholipid in both methicillin-susceptible and -resistant S. aureus strains. Growth patterns and total lipid concentrations were similar in all strains.

Methicillin↗

Correlation between esterase electrophoretic types and capsular polysaccharide types 5 and 8 among methicillin-susceptible and methicillin-resistant strains of Staphylococcus aureus.

The relationship between capsular polysaccharide types 5 and 8 and esterase electrophoretic types (zymotypes) in 160 French clinical isolates of Staphylococcus aureus was studied. Methicillin-susceptible strains of capsular types 5 and 8 were represented by 11 zymotypes, indicating a high polymorphism. Methicillin-resistant strains were mainly distributed in only two distinct populations. The predominant population was represented by strains of zymotype 6 and capsular type 5, and the second population was represented by strains of zymotype 14 and capsular type 8.

Bacterial Capsules↗

Partial excision of the chromosomal cassette containing the methicillin resistance determinant results in methicillin-susceptible Staphylococcus aureus.

We report a detailed characterization of methicillin-susceptible Staphylococcus aureus isolates from five French hospitals negative for both the mecA and the ccrAB loci but positive for the IS431::pUB110::IS431::dcs structure, present in some Staphylococcus cassette chromosome mec (SCCmec) types. The presence of SCCmec-associated elements suggests that this unusual resistant phenotype is due to a partial excision of SCCmec from epidemic methicillin-resistant S. aureus. The hypothesis of a genetic relatedness is strengthened by common sequence and spa types and similar susceptibility patterns.

Chromosomes, Bacterial↗

A comparison of clinical features and mortality among methicillin-resistant and methicillin-sensitive strains of Staphylococcus aureus endocarditis.

Our objective was to assess the clinical factors that would reliably distinguish methicillin-resistant S. aureus (MRSA) from methicillin-susceptible S. aureus (MSSA) endocarditis. A retrospective cohort study of clinical features and mortality in patients with MRSA and MSSA endocarditis between March 1986 and March 2004 was performed in a 750-bed, tertiary care teaching hospital. A total of 32 patients (10 MRSA [31.3%] vs 22 MSSA [68.7%]) were evaluated. Their mean age and sex ratio (male/female) were as follows: 30.8 +/- 16.0 vs 24.4 +/- 19.6 years old and 6/4 vs 13/9, for MRSA and MSSA infective endocarditis (IE), respectively. Univariate and multivariate analyses revealed that persistent bacteremia was significantly more prevalent in MRSA IE (OR, 10.0 [1.480- 67.552]; p, 0.018). There was a higher mortality trend for MRSA IE (50.0%) than for MSSA IE (9.1%) (p=0.019). However, persistent bacteremia was not associated with higher mortality (p > 0.05). These results indicate that if persistent bacteremia is documented, the likelihood of MRSA endocarditis should be viewed as high, and the patient's anti- staphylococcal therapy should be prolonged and/or changed to a more "potent" regimen.

Adult↗

Prevalence of methicillin-resistant and methicillin-sensitive Staphylococcus aureus nasal colonization among patients at the time of admission to the hospital.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is an important agent of hospital-acquired infection. The mode of entry of MRSA in the hospital might be on admission of patients with MRSA infection or nasal colonization. The present study was undertaken to determine the prevalence of MRSA nasal colonization among patients on admission to hospital. METHODS: Six hundred patients were screened for nasal colonization of MRSA on admission to hospital. Nasal swabs were cultured on salt mannitol agar and blood agar. Age, sex, previous admission to hospital and antibiotic therapy were recorded. RESULTS: S. aureus was isolated from the nasal swabs of 122 patients (20.2%) on admission to hospital. MRSA was isolated from 7 patients (1.1%) and methicillin-sensitive S. aureus (MSSA) from 115 patients (19.1%). Nasal colonization of S. aureus was higher in younger and elderly patients and significantly higher colonization was observed among females. The MRSA strains isolated from nasal swabs had a different antibiotic susceptibility pattern than those isolated from patients having hospital-acquired MRSA infection. Previous admissions to hospital, underlying disease antibiotic therapy were not risk factors for MRSA nasal colonization. CONCLUSION: MRSA nasal colonization of patients on admission to hospital is low in this region. The screening of every new admission would not be cost effective, but patients transferred form other institutions should be screened for MRSA. Standard infection control precautions should be strictly implemented to prevent the spread and control of MRSA infections.

Adolescent↗

[Antimicrobial activity of fosfomycin against beta-lactamase-producing methicillin-sensitive Staphylococcus aureus and methicillin-sensitive coagulase-negative staphylococci].

Antimicrobial activity of fosfomycin (FOM), cefazolin (CEZ), cefmetazole (CMZ), cefotiam (CTM) and piperacillin (PIPC) against clinical isolates of methicillin-sensitive Staphylococcus aureus (MSSA) (beta-lactamase-producing or non-producing) and methicillin-sensitive coagulase-negative Staphylococci (MSCNS) (beta-lactamase-producing or non-producing) were determined to make clear the differences in antimicrobial activity of FOM and beta-lactam antibiotics. The antimicrobial activity of PIPC against beta-lactamase-producing strains of MSSA was lower than that against non-producing ones, judging from the distribution patterns of susceptibility of the strains to PIPC. There were no differences in the antimicrobial activity of FOM, CEZ and CMZ for the producing and non-producing strains. The activity of FOM against MSCNS was comparable to that against MSSA, although those of CEZ, CMZ, CTM and PIPC were decreased. FOM, CEZ, CMZ and CTM showed bactericidal activity against TH4278 (MIC [microgram/ml]: FOM, 1; CEZ, 0.5; CMZ, 1; CTM, 0.5; PIPC, 1) of beta-lactamase-producing MSSA at 1 microgram/ml for 6 h, but PIPC did not at the same condition. FOM and CMZ at MIC suppressed regrowth of the strain, but CEZ, CTM and PIPC did not. In conclusion, FOM, which is not affected by beta-lactamase, demonstrated strong bactericidal activity at low concentration against the beta-lactam-resistant strains due to beta-lactamase production.

Anti-Bacterial Agents↗

Latex agglutination and hemagglutination tests for the rapid identification of methicillin sensitive and methicillin resistant Staphylococcus aureus.

Ten latex agglutination (LA) and hemagglutination (HA) kits for the identification of Staphylococcus aureus were compared with reference methods for their reliability and performance. The ten commercial kits consisted of Accu-Staph, Bacto-Staph, Hemastaph, Staphaurex, Staph-Latex, Staphylochrome, Staphyloslide, Staph-Rapid, Sero-Stat and Veri-Staph. The conventional methods included slide coagulase test, tube coagulase test (4 hr, 24 hr), thermonuclease and growth on mannitol salt agar (MSA). A total of 583 clinical isolates of staphylococci were used and all the kits correlated well with the conventional methods (93.1-99.4% sensitivity) in their ability to identify both methicillin sensitive (MSSA) and methicillin resistant S. aureus (MRSA). Although all were rapid, easy to perform and simple to interpret, Staphaurex and Staphyloslide gave the best sensitivities and specificities.

Agglutination Tests↗

In vitro activity of sulbactam/ampicillin and ampicillin against methicillin-sensitive and methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis.

Minimum inhibitory concentrations (MICs) of ampicillin and ampicillin + sulbactam (1:1) against 165 strains of Staphylococcus aureus and 72 strains of Staphylococcus epidermidis have been evaluated. The activity of the combination was very good. A concentration of 16 micrograms/ml + 16 micrograms/ml inhibited 96.9% of S. aureus and the 100% of S. epidermidis strains (at the same concentration ampicillin alone inhibited only 55.15% and 56.9% of S. aureus and S. epidermidis strains respectively). Activity against methicillin-resistant S. aureus (14.5%) was poor, whereas against methicillin-resistant S. epidermidis (67.2%) the combination maintained high efficacy.

Ampicillin↗

Synergistic effects of anacardic acids and methicillin against methicillin resistant Staphylococcus aureus.

The synergistic effects of 6-alk(en)ylsalcylic acids, also known as anacardic acids, in combination with methicillin against Staphylococcus aureus ATCC 33591 (MRSA) was investigated. The double bond in C15-anacardic acids is not essential in eliciting the antibacterial activity but is associated with increasing the activity. The synergistic effects decreased with increasing the number of double bonds in the alkyl chain. On the other hand, the antibacterial activity of anacardic acids possessing different alkyl chain lengths against the same MRSA strain was found to be a parabolic function of their lipophilicity and maximized with the alkyl chain length of C10 and C12. Notably, the synergistic effects were noted to increase with increasing the alkyl chain length.

Anacardic Acids↗

Decreased affinity of PBP3 to methicillin in a clinical isolate of Staphylococcus epidermidis with borderline resistance to methicillin and free of the mecA gene.

Analysis of the antibiotic binding capacity of penicillin-binding proteins (PBPs) in a group of clinical isolates of Staphylococcus epidermidis suggests that the increased level of resistance to methicillin (MIC 4.0 microg/ml) in an isolate free of the mecA gene is due to the decreased binding capacity of PBP3.

Bacterial Proteins↗

Examination of methicillin-resistant and methicillin-susceptible Staphylococcus aureus mutants with low-level fluoroquinolone resistance.

For Staphylococcus aureus, stepwise mutations result in high-level quinolone resistance. Methicillin-resistant and -susceptible quinolone-resistant, first-step mutants generated in vitro were obtained and found to be no different than those recovered from murine abscesses. Approximately 10% of all first-step mutants were resistant to ethidium bromide, and selected strains had mutations that mapped to flqB. NorA-mediated resistance among first-step mutants may be more prevalent than previously reported.

Animals↗

A novel methicillin-resistance cassette in community-acquired methicillin-resistant Staphylococcus aureus isolates of diverse genetic backgrounds.

Until recently, it has been unclear whether community-acquired (CA) methicillin-resistant Staphylococcus aureus (MRSA) isolates represent the spread of hospital MRSA isolates into the community. In 2 CA-MRSA isolates, a novel genetic element, designated staphylococcal cassette chromosome mec (SCCmec) type IV, was found; it differs from SCCmec types I-III in its small size and absence of non-beta-lactam genetic-resistance determinants. To study the prevalence of type IV SCCmec, polymerase chain reaction characterization of SCCmec was performed on DNA from 12 CA-MRSA isolates. The 12 CA-MRSA isolates were from diverse genetic backgrounds, as evidenced by their stratification into 5 pulsed-field gel electrophoresis types, 4 coagulase types, and 2 ribotypes. Eleven of the 12 isolates contained the novel SCCmec type IV element. Ten were resistant only to beta-lactam antibiotics. SCCmec type IV is present on the genome of CA-MRSA isolates. Its relatively small size and presence in isolates of diverse genetic backgrounds suggest that it may spread among S. aureus isolates.

Community-Acquired Infections↗