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PubMed1975

[Addition of gentamicine to polymethyl methacrylate for therapy of infectious bone diseases. Expeimental in vivo tests].

In experimental in vivo tests the therapeutic effect of Gentamicin added to Palacos is detected in a standardized osteomyelitic infection caused by Staph. aureus haemolyticus. The number of germ populations is reduced significantly by the antibiotic which is released in a microbiologically active concentration. On the other hand the number of germ populations in control trials (without adding an antibiotic) is remaining on a high level. During the time of the tests the appearance of other bacterial is registrated.

Animals↗

PubMed1989

The acquisition of antibiotic resistant coagulase-negative staphylococci by aortic graft recipients.

The staphylococcal flora of the groin and perianal area was studied in 12 patients undergoing aortic grafts and 3 patients having repair of ventral abdominal hernias. A total of 892 isolates of coagulase-negative staphylococci (CNS) were identified and their antibiotic resistance determined. The species most frequently isolated were S. epidermidis and S. haemolyticus with the latter showing the highest incidence of antibiotic resistance. Five of 15 patients had CNS resistant to methicillin and gentamicin on admission. The effect of pre-operative bathing with chlorhexidine was studied in 6 patients and appeared to reduce the acquisition of multi-resistant CNS in the week after operation.

Anti-Bacterial Agents↗

PubMedDate not supplied

[Multicenter study of isolated micro-organisms resistant to antimicrobials in 10 Portuguese hospitals in 1994].

In 1994, Microbiology Laboratories of ten Portuguese hospitals analysed isolated microorganisms found in blood and urine samples and studied antimicrobial susceptibilities of the most frequent bacterial pathogens. From 63780 blood samples, the most frequent were Staphylococcus spp. and from 69189 urine samples significant numbers of Escherichia coli, Enterococcus spp., Pseudomonas aeruginosa and Candida spp. were isolated. Escherichia coli strains (c.7000) revealed a low percentage of resistance to antibiotics with the exceptions of ampicillin (48%) and co-trimoxazol (25%). Klebsiella pneumoniae isolates (c.2000) revealed important resistance to ampicillin (98%), cephalotin (31%), co-trimoxazol (38%) and gentamicin (28%), while values for 3rd generation cephalosporins varied among hospitals, with several strains showing phenotype of extended-spectrum beta-lactamase. A great variation in resistance values of P. aeruginosa (c.4000) was found in relation to the antibiotics as well as to the hospitals. Resistance to methicillin in S. aureus (c.6000) was high, reaching an average of 47%, and it was even higher with S. epidermidis (c.3000) and S. haemolyticus (c.650). Only vancomycin was always active against these strains. In E. faecalis (c.2500) resistance was of 2% to ampicillin, 35% to gentamicin, 45% to streptomycin and 1% to vancomycin. E. faecium isolates (c.300) showed the most worrying results with 70% resistance to ampicillin, 42% to gentamicin, 59% to streptomycin and 9% (30 strains isolated in 5 hospitals) to vancomycin. Vancomycin resistant strains were also resistant to all other antibiotics.

Anti-Bacterial Agents↗

PubMed1993

[Comparison of 2 methods of detecting slime production by coagulase-negative staphylococci].

In the series of 212 coagulase-negative staphylococci strains isolated from blood cultures, in which the slime production was tested by means of two methods--a method using agar with Congo red and a Christensen method--S. epidermidis, S. haemolyticus and S. hominis were identified most frequently (in 69.3%, 18.4% and 6.6%, respectively). The slime production was detected by both methods in 28 strains (13%) of four species, most often in S. epidermidis (in 17%). By comparing the Congo red agar method with the Christensen method the sensitivity and specificity of the former was determined (85% and 99%, respectively). Thus the Congo red agar method for the detection of slime production by staphylococci seems to be a reliable tool.

Bacteriological Techniques↗

PubMed1988

Studies on clinical isolates of coagulase-negative staphylococci resistant to methicillin. Evidence of cross-resistance between methicillin and cephalothin.

The in vitro susceptibility to cephalothin and cefuroxime of 195 isolates of methicillin-resistant coagulase-negative staphylococci was determined by the agar-diffusion test, using 7.5% NaCl-supplemented agar. The distribution of the inhibition zone diameters for isolates of S. epidermidis (S. biotype 1) as well as for S. haemolyticus (S. biotype 4) was trimodal. While 4% of the isolates were found susceptible to cefuroxime, 39% of the S. epidermidis/S. hominis (S. biotype 1) isolates and 34% of the S. haemolyticus (S. biotype 4) isolates were found susceptible to cephalothin by this method. Eight of these isolates (six S. epidermidis, two S. haemolyticus) were selected for susceptibility testing by the tube-dilution method, together with four isolates (three S. haemolyticus, one S. epidermidis) found resistant to cephalothin by the agar-diffusion test. The first-mentioned isolates were all found susceptible to cephalothin with MICs less than or equal to 2 micrograms/l, while the last-named all were resistant with MICs greater than or equal to 16 micrograms/ml. Population analyses revealed sub-populations of highly resistant bacteria in all methicillin-resistant isolates of S. epidermidis (S. biotype 1), as well as in all isolates of S. haemolyticus (S. biotype 4). We thus concluded that methicillin-resistance in isolates of coagulase-negative staphylococci implies resistance to cephalosporins and that the difference between S. epidermidis and S. haemolyticus as regards cephalosporin-susceptibility is quantitative and not qualitative. Eighty-nine per cent of the 195 methicillin-resistant isolates in this study were resistant to penicillin and at least one more antibiotic. We therefore think that resistance to penicillin and one or more non-beta-lactam antibiotics strongly suggests methicillin-resistance and that such isolates should be further tested on hypertonic media.

Anti-Bacterial Agents↗

PubMed2001

Isolation and characterisation of a 17-kDa staphylococcal heparin-binding protein with broad specificity.

A previous study reported the ability of staphylococci to bind heparin and heparin-dependent host growth factors. The present study isolated and identified heparin- and basic fibroblast growth factor (bFGF)-binding surface components of S. epidermidis strain RP12 and S. haemolyticus strain SM 131. The staphylococcal heparin-binding component(s) were purified by affinity chromatography on heparin-Sepharose and a major heparin-binding protein, here designated HBP, was identified by immunoblot in these two coagulase-negative staphylococcal (CNS) species. The HBP was shown to be acidic with an approximate pI of 4.6 and a molecular mass around 17 kDa. The binding of heparin to HBP was inhibited by heparin, fucoidan, pentosan polysulphate and various other sulphated polysaccharides, but not by non-sulphated compounds. However, the purified HBP from both S. epidermidis and S. haemolyticus revealed broad specificity, and also bound bFGF, thrombospondin, von Willebrand factor and, weakly, fibrinogen. The N-terminal sequences of the 17-kDa HBP from S. epidermidis and S. haemolyticus showed only limited identity. Comparison of the first 15 amino acid residues derived from either strain with known sequences in the protein databases revealed no close similarities. Taken together, these results suggest that the adhesion of at least some CNS to host sulphated glycosaminoglycans may be mediated by a previously uncharacterised group of surface proteins.

Amino Acid Sequence↗

PubMed1992

In vitro activity of teicoplanin and vancomycin against gram-positive bacteria from human clinical and veterinary sources.

The minimum inhibitory concentration (MIC) of teicoplanin and vancomycin was determined by the agar dilution method for 186 Gram-positive bacteria from human clinical and veterinary sources. Teicoplanin MIC values were less than or equal to 4 micrograms/ml for 94% of staphylococci (group A, n = 52) and less than or equal to 2 micrograms/ml for all streptococci, enterococci, aerococci and pediococci (group B, n = 75). Seventy-eight percent of Gram-positive rods, Rhodococcus and Leuconostoc spp. (group C, n = 59) were inhibited by 4 micrograms/ml. Teicoplanin resistance (MIC greater than or equal to 16 micrograms/ml) was demonstrated for all Nocardia strains and for some strains of Lactobacillus, E. rhusiopathiae, Leuconostoc, and S. haemolyticus. Cross-resistance between teicoplanin and vancomycin was observed for all Nocardia strains and for some strains of Lactobacillus, E. rhusiopathiae, and Leuconostoc. Three methicillin-resistant S. haemolyticus strains were either resistant or intermediately susceptible to teicoplanin and susceptible to vancomycin. Eight strains (motile enterococci four, E. rhusiopathiae three and Leuconostoc sp. one) were susceptible to teicoplanin and resistant to vancomycin. Teicoplanin disc diffusion on Danish Blood Agar with NeoSensitabs (Rosco), PDM AB Biodisc and locally prepared discs revealed a wide range of zone diameters in groups B and C. The relation between MIC values and zone diameters for teicoplanin was analysed by the error-rate bounded method. Zone size interpretive criteria as suggested by the manufacturers (greater than or equal to 15 mm) produced 2.7% (95% confidence limits 0.9-6.2%) and 1.6% (95% confidence limits 0.3-4.6%) very major errors for NeoSensitabs and PDM-disc, respectively. Using a zone size breakpoint for susceptibility of greater than or equal to 25 mm for NeoSensitabs and greater than or equal to 20 mm for PDM-disc, the proportions of very major errors were 0.5% (95% confidence limits 0.0-3.0%) at the expense of 5.9% (95% confidence limits 3.0-10.3%) indeterminate strains that belonged to E. rhusiopathiae, Leuconostoc, Lactobacillus and S. haemolyticus. However, using these zone size breakpoints five major errors (beta-haemolytic streptococci, group B three, S. aureus one, Leuconostoc sp. one) were observed for NeoSensitabs and two major errors (beta-haemolytic streptococcus, group B one, Leuconostoc sp. one) were observed for PDM-disc. Susceptibility testing against teicoplanin among these taxa should therefore include a determination of MIC.

Animals↗

PubMed1990

Gel electrophoretic analysis of penicillin-binding proteins of coagulase negative staphylococci.

We analyzed gel electrophoretic banding patterns of penicillin-binding proteins (PBPs) of 16 type strains of coagulase-negative staphylococci (CNS). S. epidermidis, S. haemolyticus, S. saprophyticus, S. hominis, S. xylosus, S. simulans, S. warneri, S. capitis, S. saccharolyticus, S. auricularis, S. caseolyticus, S. gallinarum, S. hycus subsp. hycus, S. cohnii, S. caprae, and S. sciuri subsp. sciuri. The PBP profile of each CNS species was found to be unique and was clearly distinguishable from those of the rest of the species. Together with the previous work of other researchers, this study substantiates the applicability of the PBP profile analysis to the identification of clinical CNS strains.

Bacterial Proteins↗

PubMed1982

Clinical significance of coagulase-negative staphylococci.

Although coagulase-negative staphylococci (C-NS) have been implicated in certain human infections, they are generally regarded as contaminants, and their clinical significance is questioned. To assess their role as pathogens, we studied 205 isolates of C-NS from wounds and body fluids (blood, urine, pleural and peritoneal fluids, etc.). Patient's charts were reviewed, and, by using strict criteria, a determination was made regarding the clinical significance of these isolates. The organisms were then identified to determine whether certain species of C-NS were associated with specific infections. S epidermidis sensu stricto accounted for 81% of the C-NS isolated. The frequencies of other species were: S. haemolyticus (6%), S. hominis (5%), S. capitis (4%), S. warneri (3%), and others (1%). Only two isolates were novobiocin resistant; neither was identified as S. saprophyticus. By using our criteria, 22% of the C-NS were considered to be clinically significant, and the majority of these (93%) was S. epidermidis. The most common source of the clinically relevant C-NS isolates was wounds. These data suggest that identification of C-NS species other than S. epidermidis may be of limited value in predicting clinical significance.

Cerebrospinal Fluid↗

PubMed1994

Comparison of genotypic and phenotypic methods for species identification of coagulase-negative staphylococcal isolates from blood cultures.

Coagulase-negative staphylococci are often isolated from blood cultures. Simple methods are needed for correct identification of those species most frequently found. In this study, PCR methods were developed for the identification of S. epidermidis and S. haemolyticus, based on DNA sequences in their 16S rDNA. The results obtained by these methods were compared with those obtained using a number of phenotypic methods, including two commercial kits API Staph and Staphzyme. Fifteen type collection strains and 133 blood culture isolates were tested. The sensitivity of the PCR for identification of S. epidermidis was 99% compared with API Staph, but the specificity was lower, 94%, because of positive results also for S. capitis. The results by the PCR for identification of S. haemolyticus correlated closely with the Staphzyme results, 13 isolates being identified by Staphzyme and 16 by the PCR. API Staph, however, identified only four clinical isolates as S. haemolyticus, probably too few. Among the individual phenotypic tests performed, a trehalose-mannitol agar method and a desferrioxamine disc diffusion test for the identification of S. epidermidis were found to be very accurate. Anaerobic growth after overnight incubation could be used to distinguish S. epidermidis from S. hominis. The conclusion is that a majority of all Gram-positive, catalase-positive and coagulase-negative blood culture isolates can be typed as regards species level using only a few genotypic and/or phenotypic tests.

Bacterial Typing Techniques↗

PubMed2002

Oxacillin susceptibility testing of coagulase-negative staphylococci using the disk diffusion method and the Vitek GPS-105 card.

One hundred and ninety-three isolates of coagulase-negative staphylococci (CoNS) were tested against oxacillin by agar dilution, disk diffusion, and Vitek (GPS-105 card), and the presence of the mecA gene determined by multiplex PCR. The results obtained by all testing methods were in agreement for 190 isolates. Two mecA-negative isolates (S. lugdunensis and S. haemolyticus) had MICs of < or = 0.25 microg/ml by agar dilution and Vitek but were resistant by disk diffusion. One mecA-positive isolate was resistant by Vitek and disk diffusion but had an agar dilution MIC of < or = 0.25 microg/ml. For the species of CoNS tested, oxacillin susceptibility results obtained with the Vitek GPS-105 card and disk diffusion correlated well with results obtained by National Committee for Clinical Laboratory Standards agar dilution and with the presence of the mecA gene.

Bacterial Proteins↗

PubMed1977

Classification of coagulase-negative staphylococci in the diagnostic laboratory.

One hundred and ninety-eight coagulase-negative staphylococci isolated from urines, blood cultures, and pus samples were classified by means of two identification schemes, and their wall teichoic acids were determined serologically. S. epidermidis, S. saprophyticus, and S. cohnii were identified reliably by the use of five criteria: acid aerobically from sucrose, trehalose, and mannitol, phosphatase production, and sensitivity to novobiocin. Further species, notably S. haemolyticus and S. hominis, could be identified when haemolysis on blood agar plates was included in the criteria group. The investigation shows, that a considerable number of coagulase-negative staphylococci isolated from human specimens belong to species other than S. epidermidis and S. saprophyticus. These staphylococci can cause human infections and should be identified in the diagnostic laboratory. S. epidermidis and S. saprophyticus were found to contain the teichoic acids previously identified in these species. S. cohnii contained the same teichoic acids as S. saprophyticus. No characteristic teichoic acid was demonstrated in the other species, but several strains contained poly C (beta-N-acetylglucosaminylglycerol teichoic acid).

Bacteriological Techniques↗

PubMed2006

Simple and economical method for speciation and resistotyping of clinically significant coagulase negative staphylococci.

An attempt was made to speciate 102 clinically significant isolates of coagulase negative staphylococci (CoNS) by a practical scheme adapted from various references. This scheme utilizes slide and tube coagulase test, urease test ornithine decarboxylase, novobiocin susceptibility and aerobic acid from mannose for assigning species group. Inclusion of one or two additional tests in a species group could identify the isolates to species level. Ninety eight (97%) isolates were conveniently identified as S. epidermidis (41%), S. saprophyticus (16.6%), S. haemolyticus (14.7%), S. hominis (14.7%), S. lugdunensis (4.9%), S. schleiferi (1.9%) and S. capitis (1.9%). Only four isolates were not identified to the species level, two of which were probably S. capitis subspecies ureolyticus / S. warneri / S. simulans . Antibiotic susceptibility testing showed maximum resistance to ampicillin (89%) followed by cefotaxime (59%) with no resistance to vancomycin. The increasing recognition of pathogenic potential of CoNS and emergence of drug resistance amongst them denotes the need to adopt simple laboratory procedures to identify and understand the diversity of staphylococci isolated from clinical material.

Bacteriological Techniques↗

PubMed1994

[Determination of the susceptibility of staphylococci and enterococci to plycopeptides by the E-test method].

E-test was evaluated by comparison with the reference agar dilution method on 15 enterococci and 15 staphylococci with variable resistance to glycopeptides and isolated from clinical samples: 15 enterococci with 3 glycopeptide-resistant (2 E. facalis and 1 E. faecium), 1 glycopeptide intermediate E. gallinarum and 11 glycopeptide-sensible E. faecalis; 15 staphylococci with 4 teicoplanine-resistant (1 S. aureus, 1 S. epidermidis and 2 S. hominis), 3 teicoplanin-intermediate (1 S. epidermidis, 2 S. haemolyticus) and 8 teicoplanin sensible strains (6 S. aureus, 1 S. epidermidis and 1 S. hominis). Repeatability (10 determinations by strain) and exactitude were tested for all strains. Our data shows very good results for repeatability and exactitude with the reference method except in the case of coagulase negative staphylococci and teicoplanin. Some technical problems appear in our experience: for staphylococci, a 48 h incubation is necessary to detect teicoplanin-intermediate strains but 24 h is sufficient for enterococci. E-test is expensive, require a strict methodology, but is a good method to detect glycopeptide resistance for staphylococci and enterococci except for coagulase negative staphylococci and teicoplanine.

Bacteriological Techniques↗

PubMed2001

[Coagulase negative staphylococci isolated on blood cultures].

224 coagulase-negative strains of staphylococci (CNS) isolated from haemocultures of hospitalized patients were classified into 11 types. The most frequent one was S. epidermis (73.2%), S. haemolyticus (7.6%), S. lugdunensis (6.7%) and S. hominis (3.6%). In different strains virulence factors were assessed: production of mucus and delta toxin. Based on these properties the strains were classified into biotypes. Of 164 strains of S. epidermis mucus was produced by 64.6% and delta toxin by 75.0%. Of 17 strains of S. haemolyticus mucus was produced by 8 and delta toxin by 10 strains. Of 15 strains of S. lugdunensis 9 strains produced both virulence factors. Of 8 strains of S. hominis 5 strains produced mucus and 3 delta toxin. In strains S. capitis, S. sciuri, S. auricularis, S. caprae these factors were not detected. The most frequent biotypes were: subtype 3 (43.3%), 1a (26.8%) and 1b (10.3%). The sensitivity of CNS to 11 antibacterial substances was assessed quantitatively by estimating the MIC (mg/l). The most effective antibiotic was vancomycin (100%) and tetracycline (76.8%). Only 26.3% strains were sensitive to oxacillin and cefalotine. Of 224 CNS (70%) were resistant to more than 8 antibacterial substances.

Coagulase↗

PubMed1995

Susceptibility of staphylococci and enterococci to glycopeptides comparison of 3 test methods.

The significance of grampositive bacteria, especially staphylococci and enterococci, as nosocomial pathogens has increased in the last decade. Furthermore, resistance to commonly used antibiotics like beta-lactams has also become more common and even resistance to glycopeptides has been observed. We evaluated the susceptibility of 150 staphylococcal clinical isolates (52 S. epidermidis, 52 S. haemolyticus, 10 S. saprophyticus, 10 S. hominis, 4 S. warneri, 4 S. simulans, 4 S. capitis and 14 S. aureus) and of 50 enterococci (49 E. faecalis, 1 E. faecium) to the glycopeptides, vancomycin and teicoplanin. The data from the agar dilution test used as reference method were compared with the results from the E test and the agar disk diffusion test. Concerning vancomycin, no resistance among all the staphylococcal isolates was observed whereas one single enterococcal strain (E. faecium) proved to be resistant. The overall resistance of staphylococcal isolates against teicoplanin was about 10.7% (15 S. haemolyticus, 1 S. epidermidis) being mainly due to the high proportion of S. haemolyticus strains (52 out of 150) among the staphylococcal isolates. Teicoplanin resistance among the enterococci was not detected. For vancomycin, a very close correlation between the MICs from the agar dilution test and the E test was noticed. As concerns teicoplanin, the MICs from the E-test were usually somewhat lower than those obtained by the agar dilution test. No correlation was found between the MICs of resistant and intermediate staphylococcal strains and the results from the teicoplanin agar disk diffusion test. For routine teicoplanin susceptibility testing of staphylococci, the determination of the MIC (e.g., by the E test) is much more reliable for detecting resistant strains than the agar disk diffusion test.

Anti-Bacterial Agents↗

PubMed2000

Binding of von Willebrand factor by coagulase-negative staphylococci.

Coagulase-negative staphylococci (CNS) are the most common infectious micro-organisms isolated from prosthetic devices. To determine whether von Willebrand factor (vWF) acts as an adhesin in bacterial recognition, bacterial binding of recombinant vWF (rvWF) was studied. Eleven CNS strains, belonging to S. epidermidis, S. haemolyticus and S. hominis species, bound soluble rvWF, but to a lesser extent than S. aureus. S. epidermidis strain H2-W bound 125I-labelled rvWF in a dose-dependent manner. The binding could be inhibited by unlabelled rvWF and thrombospondin, but not by fibrinogen, vitronectin or the carbohydrates N-acetylgalactoseamine, D-galactose, D-glucose, and D-fucose. Pre-incubation of rvWF with type I collagen and Arg-Gly-Asp-Ser (RGDS) peptides did not inhibit binding, whereas pre-incubation of rvWF with heparin decreased binding significantly. The interaction between CNS and rvWF was sensitive to proteinase treatment of bacterial cells. CNS strains bound to immobilised rvWF an extent greater or equal to the positive control strain S. aureus Cowan I. rvWF binding structures from bacterial cell wall were detected by immunoblot. Cowan I strain had 140-, 90- and 38-kDa binding molecules. S. haemolyticus strain SM131 and S. epidermidis strain H2-W had two (120 and 60 kDa) and five (120, 90, 60, 52 and 38 kDa) binding molecules, respectively. Similar binding structures were formed when cell wall extracts from these strains were incubated with thrombospondin. These results indicate that specific ligand-receptor interaction between CNS and rvWF may contribute to bacterial adhesion and colonisation on biomaterial surfaces. Heparin-binding domains of rvWF might be the crucial regions for bacterial attachment. rvWF and thrombospondin may recognise similar molecules in staphylococcal cell wall extracts.

Bacterial Adhesion↗

PubMed1989

[The occurrence and significance of plasma coagulase negative staphylococci from the genital tract of horses].

Classification based on biochemical characteristics of 389 strains of plasma-coagulase-negative (plc-) staphylococci isolated from the genital tract of mares and stallions resulted in the following distribution of species: St. sciuri 130 (33.4%), St. equorum 42 (10.8%), St. xylosus 16 (4.1%), St. epidermidis 35 (9.0%), St. simulans 24 (6.2%), St. haemolyticus 33 (8.5%), St. warneri 18 (4.6%), St. lentus 12 (3.1%), St. hyicus 11 (2.8%). Strains of St. cohnii, St. capitis, St. gallinarum, St. saprophyticus and St. hominis have only been found sporadically (a. 1%). 48 (12.3%) strains could not be classified. With regard to species distribution of isolates from stallions and mares. 63.7% of the isolates from stallions belonged to St. sciuri and 9.3% to St. lentus, whereas in isolates of mares these species numbered only 24.9% and 0.4%, respectively. On the other hand the species St. equorum (14.9% vs. 6.8%), St. epidermidis (14.5% vs. 1.7%), St. haemolyticus (14.0% vs. 1.7%), St. warneri (7.7% vs. 0.8%) and St. xylosus (5.9% vs. 2.5%) predominated in mares. St. simulans was found occurring equally in mares and stallions (7.7% vs. 5.9%). Comparing the staphylococcal species of healthy mares and of mares which have not become pregnant after copulation no indication was found for a significant role of certain plc- staphylococci in infertility. All of the 389 isolates were tested for production of protein A, i.e. Fc-fragment binding receptors, using a microenzyme-assay with peroxidase-labelled rabbit immunoglobulin G. With this method cell-bound or extracellular Fc-receptors could not be detected in anyone of the plc- staphylococcal strains.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗