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PubMed1996

Different susceptibility of coagulase-positive and coagulase-negative staphylococci to ciprofloxacin.

Ciprofloxacin susceptibility was evaluated in 573 clinical staphylococcal isolates (Staphylococcus aureus 230, methicillin-resistant (MR) 36%; coagulase-negative strains 343, MR 66%) collected from 1989 to 1995. Resistance to ciprofloxacin for MR Staphylococcus aureus was 25% until 1991 when ciprofloxacin was introduced into the hospital formulary, and rose progressively to 90% in 1994-1995. MR Staphylococcus haemolyticus showed a ciprofloxacin resistance of 46% until 1991 and of 73% from 1992 to 1995. In contrast MR Staphylococcus epidermidis and other coagulase-negative strains showed a constant susceptibility to this agent (80%). Ciprofloxacin has limited usefulness against MR Staphylococcus aureus but can be still used to treat Staphylococcus epidermidis infections.

Ciprofloxacin↗

PubMed1987

Phenotypic expression and genetic heterogeneity of lincosamide inactivation in Staphylococcus spp.

We examined the resistance phenotype and the genetic basis of lincosamide modification in 25 clinical isolates of Staphylococcus spp. inactivating lincomycin and clindamycin. The strains were resistant to high levels of lincomycin but remained susceptible to clindamycin. However, MBCs and inoculum effects showed that the activity of clindamycin was impaired. The distribution in these strains of nucleotide sequences related to linA and linA', the genes encoding lincosamide nucleotidylation in Staphylococcus haemolyticus BM4610 and S. aureus BM4611, respectively, was studied by dot blot hybridization. The genes responsible for lincosamide inactivation in Staphylococcus spp. were found to constitute a family of related sequences which are not species specific.

Anti-Bacterial Agents↗

PubMed1998

European Glycopeptide Susceptibility Survey of gram-positive bacteria for 1995. European Glycopeptide Resistance Survey Study Group.

In the European Glycopeptide Susceptibility Survey 7078 Gram-positive isolates collected in 1995 from 70 centers in 9 countries of Western Europe were examined, using a standardized, quantitative susceptibility testing method. Of the 7078 isolates, 6824 (96.4%) were tested by the national coordinating centers. Teicoplanin (mode MIC 0.5 microgram/mL) was generally twice as active as vancomycin (mode MIC 1 microgram/mL) against Staphylococcus aureus (n = 2852). All isolates were susceptible to vancomycin (MIC < or = 4 micrograms/mL) and all but four to teicoplanin (MIC < or = 8 micrograms/mL); these four isolates were of intermediate susceptibility (MIC 16 micrograms/mL). With coagulase-negative staphylococci (n = 1444), the distribution of MIC of teicoplanin was wider than for vancomycin. Two and two-tenths percent of coagulase-negative staphylococci excluding Staphylococcus haemolyticus required 16 micrograms/mL teicoplanin for inhibition (intermediate) and 0.4% > or = 32 micrograms/mL (resistant). Among isolates of S. haemolyticus, 4.4% were of intermediate susceptibility (MIC 16 micrograms/mL) and 3.3% were resistant (MIC > or = 32 micrograms/mL) to teicoplanin. However, this species represented only 6.3% of the isolates of coagulase-negative Staphylococcus spp. Generally, teicoplanin (mode MIC < or = 0.12 microgram/mL) was four to eight times more active than vancomycin (mode MIC < or = 0.5 microgram/mL) against the 770 streptococcal isolates. Glycopeptide-susceptible Enterococcus spp. (n = 1695) were generally four times more susceptible to teicoplanin (mode MIC 0.25 microgram/mL) than to vancomycin (mode MIC 1 microgram/mL). Combined vancomycin and teicoplanin (VanA phenotype) resistance was observed more frequently (9.3%) in isolates of Enterococcus faecium than in Enterococcus faecalis (0.8%). Four isolates of unspeciated enterococci (1.4%) also expressed this resistance phenotype. Four isolates of E. faecium and four of E. faecalis expressed the VanB-type (low-level, vancomycin only) resistance. Spain was the only country not to submit resistant E. faecium strains while resistant E. faecalis isolates came only from Spain and Italy.

Anti-Bacterial Agents↗

PubMedDate not supplied

Mechanisms of action of quinolones against staphylococci and relationship with their in vitro bactericidal activity.

Ciprofloxacin and norfloxacin exhibited mechanism A (requires cell division as well as bacterial protein and RNA synthesis to kill bacteria) and C (active against nondividing bacteria but requires protein and RNA synthesis) against the reference strain Staphylococcus aureus ATCC 25923, yet only mechanism A was exhibited by these fluoroquinolones when tested against three clinical isolates: S. aureus Sa-215, Staphylococcus epidermidis Se-81 and Staphylococcus haemolyticus Sx-1. On the contrary, fleroxacin exerted mechanism A and C against the three clinical isolates but only mechanism A against the reference strain. Ofloxacin displayed mechanism A against S. epidermidis Se-81, mechanism A and C against S. haemolyticus and mechanism A and B (active against nondividing bacteria and does not require protein and RNA synthesis) against the two S. aureus tested. Sparfloxacin showed mechanism A and C against the four Staphylococcus species studied, and temafloxacin was the only fluoroquinolone tested that exhibited mechanism A and B against the four bacterial strains assayed. No correlation was found between the in vitro bactericidal activity (expressed as minimum inhibitory concentration and optimal bactericidal concentration) and the mechanisms of action exhibited by these fluoroquinolones.

Anti-Bacterial Agents↗

PubMed1993

[Study on mec gene in methicillin-resistant staphylococci].

Methicillin-resistant Staphylococcus aureus (MRSA), methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus epidermidis and methicillin-resistant Staphylococcus haemolyticus strains which were isolated in 1992 were surveyed for distribution of mecA, mecR1 (putative mecA gene inducer gene) and mecI gene (putative mecA gene repressor gene) by the method of polymerase chain reaction (PCR). All the 30 strains (100%) of MRSA carried mecA gene and mecR1 gene-1 (32-357 bp, closer to mecA gene). In contrast, mecI gene (putative mecA gene repressor gene) and mecR1 gene-2 (987-1221 bp, closer to mecI gene) were detected in only 23 (73%) of the MRSA strains. By RT-PCR, the mecI gene existing in these highly resistant MRSA strains was demonstrated to remain inactivated. In 28 strains of MSSA and two strains of methicillin with MIC of 6.25 micrograms/ml, all of the mec genes (mecA, mecR1 and mecI) were not detected. In contrast, in the other three strains, which were sensitive to methicillin in spite of the presence of mecA gene and defined as MSSA, the mec genes other than mecA were detected except for one strain. Methicillin-resistant (with MIC of 12.5 micrograms/ml or more) coagulase-negative staphylococci (15 S. epidermidis and 9 S. haemolyticus) strains were surveyed as well. As results, all the mec genes mentioned above were detected in all the strains except for one of S. epidermidis, but only mecA gene in the strains of S. haemolyticus. Thus, it was found that the composition of the mec genes of methicillin-resistant S. haemolyticus were more variant from that of methicillin-resistant strains of S. aureus and S. epidermidis.

Genes, Bacterial↗

PubMed1995

In-vitro activity of three fluoroquinolones, cefotaxime and clindamycin against staphylococci.

The in-vitro activity of three fluoroquinolones (ciprofloxacin, ofloxacin, temafloxacin) against 200 well-defined clinical isolates of staphylococci was investigated. The in-vitro activity of the fluoroquinolones tested was equal, with a strong indication of cross resistance. A clear distribution over two populations of different quinolone susceptibility--"naturally susceptible" and "naturally resistant" strains--could be found in penicillin-resistant, methicillin-susceptible Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus haemolyticus.

Aminoglycosides↗

PubMedDate not supplied

Comparative in vitro activity of teicoplanin and vancomycin against United States teicoplanin clinical trial isolates of gram-positive cocci.

In this study, the in vitro activity of teicoplanin and vancomycin was directly compared against 503 Gram-positive cocci isolated during the U.S. teicoplanin clinical trials. Both antibiotics were equally active against oxacillin-sensitive Staphylococcus aureus, oxacillin-sensitive and oxacillin-resistant Staphylococcus epidermidis, and other coagulase-negative staphylococci, except Staphylococcus haemolyticus. Teicoplanin was fourfold more active than vancomycin against oxacillin-resistant S. aureus (MIC90, 0.5 vs. 2.0 micrograms/ml), whereas vancomycin was more active than teicoplanin (MIC90, 2.0 vs. 8.0 micrograms/ml) against oxacillin-resistant S. haemolyticus. Teicoplanin was two- to eightfold more active than vancomycin against the streptococci and enterococci tested.

Anti-Bacterial Agents↗

PubMed1999

The distribution of mecA, mecR1 and mecI and sequence analysis of mecI and the mec promoter region in staphylococci expressing resistance to methicillin.

The presence and sequences of genes that regulate the expression of methicillin resistance was investigated in 42 isolates of Staphylococcus aureus and 102 isolates of coagulase-negative staphylococci (CNS). PCR was used to detect mecA and the regulatory genes mecR1 and mecI. In a selected group of isolates, the sequences of mecI and the mec promoter region were also determined and compared with the sequences obtained from pre-MRSA strain N315. The genetic diversity of the collection was assessed by pulsed-field gel electrophoresis (PFGE). mecA was present in 21 S. aureus and 44 CNS. mecR1 was associated with mecA in all S. aureus and in all CNS, except two isolates of Staphylococcus haemolyticus. mecI was present in 48% of mecA-positive S. aureus and 50% of mecA-positive CNS. In six S. aureus isolates, mecI contained a termination codon at nucleotide 202 which would truncate the MecI protein. No mutation was found in the mecI gene of the four other S. aureus and 15 CNS sequenced. Seven isolates of Staphylococcus simulans had a single nucleotide substitution in the mec promoter region. Expression of methicillin resistance could be explained for all mecA-positive staphylococci with mutations within mecI or in the mec promoter region or in which mecI was deleted. However, the 'wild type' sequences observed in four S. aureus and eight CNS suggest that there is another mechanism for overcoming the repression of resistance caused by mecI.

Bacterial Proteins↗

PubMed1997

Comparative in-vitro activities of quinupristin-dalfopristin against Gram-positive bloodstream isolates.

The in-vitro activity of quinupristin-dalfopristin was compared with those of vancomycin, teicoplanin, erythromycin, clarithromycin, rifampicin, imipenem, meropenem, ciprofloxacin and sparfloxacin against 414 bloodstream isolates of Gram-positive cocci. Quinupristin-dalfopristin inhibited strains of Streptococcus pyogenes and Streptococcus agalactiae at 0.12 mg/L, methicillin- and/or erythromycin-resistant Staphylococcus aureus and Staphylococcus epidermidis at 0.5 mg/L, Staphylococcus haemolyticus, Enterococcus faecium, Streptococcus pneumoniae, Streptococcus mitis, Streptococcus bovis, Streptococcus sanguis and Streptococcus anginosus at 1 mg/L and Enterococcus faecalis at 8 mg/L.

Anti-Bacterial Agents↗

PubMed2006

Automated systems in the identification and determination of methicillin resistance among coagulase negative staphylococci.

Coagulase-negative staphylococci (CoNS) are an important cause of nosocomial bacteremia, specially in patients with indwelling devices or those submitted to invasive medical procedures. The identification of species and the accurate and rapid detection of methicillin resistance are directly dependent on the quality of the identification and susceptibility tests used, either manual or automated. The objective of this study was to evaluate the accuracy of two automated systems--MicroScan and Vitek--in the identification of CoNS species and determination of susceptibility to methicillin, considering as gold standard the biochemical tests and the characterization of the mecA gene by polymerase chain reaction, respectively. MicroScan presented better results in the identification of CoNS species (accuracy of 96.8 vs 78.8%, respectively); isolates from the following species had no precise identification: Staphylococcus haemolyticus, S. simulans, and S. capitis. Both systems were similar in the characterization of methicillin resistance. The higher discrepancies for gene mec detection were observed among species other than S. epidermidis (S. hominis, S. saprophyticus, S. sciuri, S. haemolyticus, S. warneri, S. cohnii), and those with borderline MICs.

Bacterial Proteins↗

PubMed1989

Comparative virulence of human isolates of coagulase-negative staphylococci tested in an infant mouse weight retardation model.

Human infections caused by coagulase-negative staphylococci have steadily increased in numbers and severity. Causes may be the use of artificial prostheses, immunocompromising chemotherapy and radiation therapy, and sophisticated surgical techniques, to name a few. Although the infectivity of coagulase-negative staphylococci as a group has been well documented for humans, attempts to study the pathogenesis of infections caused by individual species of coagulase-negative staphylococci have been hampered by the lack of an animal model that is not refractory to infection by these organisms. In the study reported here, a 2-day-old-mouse weight retardation test was used to assay the virulence of 60 clinical and reference strains of coagulase-negative staphylococci. These strains represented eight species of coagulase-negative staphylococci. The most virulent strains were demonstrated to be of the species Staphylococcus haemolyticus, S. saprophyticus, and S. epidermidis. The data further suggest that production of slime is a marker of virulence in S. epidermidis and that intraspecies differences in virulence occur.

Animals↗

PubMed2001

Genomic rearrangement of the mec regulator region mediated by insertion of IS431 in methicillin-resistant staphylococci.

Genomic diversification of the mec regulator region mediated by IS431 was investigated for clinical isolates of methicillin-resistant staphylococci. A single rearranged form of the mecR1 gene due to IS431 insertion was detected in the three staphylococcal species, while another type of mecR1 truncation with IS431 and an IS431 located downstream of mecI were found only in Staphylococcus haemolyticus. Genetic differentiation of IS431 and staphylococcal isolates suggested transmission of mecDNA with IS431-mediated rearrangement among different staphylococcal species.

Bacterial Proteins↗

PubMed1990

In vitro activity of LY264826, a new glycopeptide antibiotic, against gram-positive bacteria isolated from patients with cancer.

The in vitro activity of LY264826, a novel glycopeptide antibiotic produced by Amycolatopsis orientalis, was compared with those of vancomycin, teicoplanin, and oxacillin against 311 gram-positive clinical isolates from patients with cancer, LY264826 had lower MICs for 90% of isolates (MIC90) than vancomycin for all species tested. It was active against oxacillin-resistant isolates including Staphylococcus aureus (MIC90, 0.5 micrograms/ml), Staphylococcus haemolyticus (MIC90, 2.0 micrograms/ml), Enterococcus spp. (MIC90, 0.5 micrograms/ml), Bacillus cereus (MIC90, 0.25 micrograms/ml), and Corynebacterium jeikeium (MIC90, 0.12 micrograms/ml). For S. aureus, including oxacillin-resistant isolates, the MICs of LY264826 were similar to those of teicoplanin. For coagulase-negative staphylococci, however, LY264826 had MICs that were 4- to 32-fold lower than those of teicoplanin. Against most streptococcal species the activities of LY264826 and teicoplanin were similar. Bactericidal activity against Staphylococcus spp. and most Streptococcus pyogenes isolates was less than or equal to 1 dilution of the MIC. One isolate of S. pyogenes and all Enterococcus faecalis strains tested were tolerant of LY264826, with MBCs greater than or equal to 32-fold greater than the MICs. The addition of 50% human serum resulted in a significant increase in activity only against Staphylococcus epidermidis. Variations in pH from 6.4 to 8.4 and in inoculum from 10(3) to 10(7) CFU/ml did not significantly affect the activity of LY264826.

Anti-Bacterial Agents↗

PubMed1994

Dissemination among staphylococci of DNA sequences associated with methicillin resistance.

DNA probes consisting of pUC19 containing cloned Staphylococcus aureus chromosomal fragments were constructed from two methicillin-resistant S. aureus strains with different DNA sequences 5' to mecA, the gene that mediates methicillin resistance. The probe from one strain, BMS1, contained a portion of the regulatory sequences (the terminal 641 bp of mecR1 and all of mecI) associated with the induction and repression of mecA transcription (pGO195). The second probe, from strain COL (pGO198), contained DNA not found in strain BMS1. This DNA was within the sequences added at the site of a mecR1 deletion. Genomic digests of 14 S. aureus isolates recovered between 1961 and 1969 all hybridized with pGO198. In contrast, 78% (36 of 46) of the S. aureus organisms isolated since 1988 hybridized with pGO195 but not with pGO198; the remainder hybridized with pGO198. No S. aureus isolates hybridized with both probes. Staphylococcus epidermidis digests hybridized with pGO198 (46%), pGO195 (14%), or both probes (35%); all 20 Staphylococcus haemolyticus isolates hybridized with pGO198. The restriction fragment length polymorphism patterns of all pGO198-hybridizing regions in S. aureus were identical to those in strain COL. In addition, the mecR1 deletion junction nucleotide sequences of eight S. aureus and six S. epidermidis isolates were identical. However, 21 of 23 S. epidermidis and all 20 S. haemolyticus isolates had from 5 to more than 20 additional chromosomal bands that hybridized with pGO198; none of 21 S. aureus isolates had additional hybridizing bands. These data suggest that the additional DNA responsible for the mecR1 deletion was part of a repetitive, and possibly mobile, element resident in coagulase-negative staphylococci but not in S. aureus. These data also support a hypothesis that the deletion event occurred in a coagulase-negative staphylococcus with subsequent acquisition of the interrupted sequences by S. aureus.

Base Sequence↗

PubMed1998

Clinical importance of identifying coagulase-negative staphylococci isolated from blood cultures: evaluation of MicroScan Rapid and Dried Overnight Gram-Positive panels versus a conventional reference method.

We evaluated the clinical usefulness of species identification of blood isolates of coagulase-negative staphylococci as a predictor of the clinical significance of the isolates. In addition, we compared results of species identification obtained with MicroScan Rapid Gram-Positive Identification panels and Dried Overnight (Conventional) Gram-Positive Identification panels with those obtained by a tube reference method. Two hundred eighty-five blood isolates were tested, including 92 judged to represent true bacteremia and 193 judged to represent contamination. The most common species detected were Staphylococcus epidermidis, Staphylococcus hominis, and Staphylococcus haemolyticus. These three species accounted for nearly 98% of the clinically significant isolates and 89% of the contaminants. The isolation of other species almost always represented contamination. However, identification of the three most common species did not help distinguish pathogens from contaminants. Both the Rapid and the Dried Overnight Gram-Positive panels identified S. epidermidis strains accurately, but the panels performed less well for the other species. Analysis revealed that S. hominis was frequently misidentified due to the presence of a previously unknown subspecies. Based on the initial results, revised investigational Dried Overnight Gram-Positive Identification panels (CPID-2) were prepared and tested. The CPID-2 panels identified 85 to 95% of S. epidermidis strains, 76 to 86% of S. hominis strains, and 88 to 92% of S. haemolyticus strains with high probability (>85%) and, overall, represented a significant improvement over the other panels for identification of these staphylococcal species.

Bacteremia↗

PubMed1988

Characteristics of coagulase-negative staphylococci that help differentiate these species and other members of the family Micrococcaceae.

One hundred reference strains and 1,240 clinical isolates representing 26 species of the family Micrococcaceae were used to evaluate the potential of tests for synergistic hemolysis, adherence to glass, pyroglutamyl-beta-naphthylamide hydrolysis, and susceptibility to a set of five antimicrobial agents for differentiating these species and strains within the species. Sixty-eight percent of the clinical isolates exhibited synergistic hemolysis; 69% of the clinical staphylococci but none of the micrococci or stomatococci were adherence positive, and 92% of the strong positive adherence reactions were produced by strains of Staphylococcus epidermidis. Strains from 15 of the species were pyroglutamyl-beta-naphthylamide positive, but this test separated Staphylococcus xylosus from other novobiocin-resistant staphylococci and Staphylococcus intermedius from other coagulase-positive species. A polymyxin B disk helped differentiate S. epidermidis from most other coagulase-negative staphylococci, and a bacitracin disk (10 U) helped differentiate Staphylococcus haemolyticus from most other novobiocin-susceptible staphylococci. All strains that were susceptible to furazolidone and resistant to Taxo A disks (bacitracin, 0.04 U; BBL Microbiology Systems, Cockeysville, Md.) were staphylococci. We observed a 91% correlation between species identification obtained with the Staph-Ident system (Analytab Products, Plainview, N.Y.) and conventional methods; but the micrococci and stomatococci were incorrectly identified as staphylococci with Staph-Ident, and several isolates of S. epidermidis were misidentified as Staphylococcus hominis because they were alkaline phosphatase negative. Both these problems can be prevented by adding the simple tests we describe to those already recommended when the Staph-Ident system is used to identify isolates of gram-positive, catalase-positive cocci.

Bacitracin↗

PubMed1996

Plasmid analysis of 26 staphylococcal species by a rapid microscale technique.

OBJECTIVE: Evaluate a plasmid typing technique for a diverse group of staphylococci. DESIGN: In vitro testing on known isolates. SETTING: University hospital. INTERVENTION: The plasmid content of 195 isolates representing 26 staphylococcal species was analyzed by agarose gel electrophoresis following lysostaphin-alkaline-SDS lysis, with and without acetone treatment. RESULTS: Isolates yielded plasmid profiles with 0 to 7 extra-chromosomal bands (median, 1; 1.5); 171 (88%) had a profile with at least 1 band. Species with more than one isolate available for testing showed considerable diversity of plasmid profiles, except for Staphylococcus haemolyticus. CONCLUSIONS: The ease of the procedure and the diversity of plasmid profiles within each species examined suggests that plasmid profiling is an accessible and useful epidemiologic tool applicable to most staphylococcal species by epidemiologic units or clinical laboratories.

Bacterial Typing Techniques↗

PubMed1991

Nucleotide sequence and phylogeny of a chloramphenicol acetyltransferase encoded by the plasmid pSCS7 from Staphylococcus aureus.

The nucleotide sequence of the chloramphenicol acetyltransferase gene (cat) and its regulatory region, encoded by the plasmid pSCS7 from Staphylococcus aureus, was determined. The structural cat gene encoded a protein of 209 amino acids, which represented one monomer of the enzyme chloramphenicol acetyltransferase (CAT). Comparisons between the amino acid sequences of the pSCS7-encoded CAT from S. aureus and the previously sequenced CAT variants from S. aureus, Staphylococcus intermedius, Staphylococcus haemolyticus, Bacillus pumilis, Clostridium difficile, Clostridium perfringens, Escherichia coli, Shigella flexneri, and Proteus mirabilis were performed. An alignment of CAT amino acid sequences demonstrated the presence of 34 conserved amino acids among all CAT variants. These conserved residues were considered for their possible roles in the structure and function of CAT. On the basis of the alignment, a phylogenetic tree was constructed. It demonstrated relatively large evolutionary distances between the CAT variants of enteric bacteria, Clostridium, Bacillus, and Staphylococcus species.

Amino Acid Sequence↗