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PubMed1983

Characterization of coagulase-positive Staphylococcus intermedius and Staphylococcus aureus isolated from veterinary clinical specimens.

Staphylococci were the most frequent isolates from clinical specimens submitted from a large referral and teaching veterinary hospital. In this study a total of 160 isolates were examined by a wide range of biochemical tests and modifications of basic procedures. An attempt was made to test the validity of these procedures for use in characterization of clinical isolates of coagulase-positive staphylococci. Of the isolates examined, some 27 were Staphylococcus aureus, 115 were Staphylococcus intermedius, and the rest were coagulase-negative staphylococci and were not characterized further. The most useful discriminatory tests were acid production from maltose incubated overnight on maltose purple agar (W. E. Kloos and K. H. Schleifer, J. Clin. Microbiol., 1:82-88, 1975), acetoin production detected by the Barritt method, and detection of hyaluronidase activity. These gave accurate and fast results. Supplemented with the tellurite reduction test and the direct staphylocoagulase assay using Chromozym TH (Engels et al.; J. Clin. Microbiol. 14:496-500, 1981), these tests should eliminate the possibility of false identifications of these two species.

Animals↗

PubMed1986

Evaluation of laboratory tests for detection of methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis.

Few studies evaluating susceptibility testing of methicillin-resistant staphylococci have included isolates of Staphylococcus epidermidis, a known pathogen in many types of serious infections. We tested 175 S. epidermidis and 95 Staphylococcus aureus isolates to determine the most sensitive procedures for detecting methicillin-resistant staphylococci. Reference procedures included agar dilution with methicillin and 4% NaCl in the agar and broth microdilution with methicillin and 2% NaCl in cation-supplemented Mueller-Hinton broth. After 24 h of incubation, the results from both methods correlated well and were within 1 log2 dilution for all isolates tested. Only one-half of all resistant isolates (92 of 183) were detected at 18 h by using the standard disk diffusion technique with 5-micrograms methicillin disks, and even fewer were detected with 10-micrograms methicillin disks and newly recommended zone-size criteria. However, the standard disk diffusion method with 4% NaCl in the agar increased the sensitivity and specificity for identification of the proper phenotype to greater than 92%. The spread plate and new spot techniques, both using agar with 4% NaCl, were also sensitive methods. Of 47 S. epidermidis isolates tested against oxacillin, 6 (13%) were oxacillin susceptible but methicillin resistant. Two automated systems, the Automicrobic system (Vitek Systems) and MicroScan (American MicroScan), as well as two broth screening systems available from Remel and Austin Biological Laboratories, failed to detect several resistant isolates, depending on the species.

Culture Media↗

PubMed1990

Comparison of the staphylocoagulase activities of Staphylococcus aureus and Staphylococcus intermedius on Chromozym-TH.

The staphylocoagulases of Staphylococcus intermedius (39 strains from clinical samples from dogs and 1 strain from a pigeon) and Staphylococcus aureus (40 strains from nine different animal species) were compared by using the chromogenic methods of Engels et al. (W. Engels, M. Kemps, and C. P. A. van Boven, J. Clin. Microbiol. 14:496-500, 1981). S. intermedius produced staphylocoagulase which resembled that of S. aureus in its rate and method of action on prothrombin, but S. intermedius produced it in lesser amounts. Therefore, chromogenic substrate tests such as Chromozym-TH (Boehringer GmbH, Mannheim, Federal Republic of Germany) (as described by Engels et al.) can be used for the detection of staphylocoagulase in both of these species. However, to detect accurately the presence of S. intermedius staphylocoagulase by this method, preconcentration of the extracellular proteins or an extension of the reaction time of the test would be required. The method described by Engels et al. was designed specifically for clinical laboratories that handle human samples. Under those circumstances the method could be regarded as specific, enabling an identification of S. aureus only. However, as both S. aureus and S. intermedius give positive results in this test, in veterinary diagnostic laboratories, heavy reliance must not be placed on this test for determination of organisms to the species level.

Animals↗

PubMed1991

Comparison of the affinities to bovine and human prothrombin of the staphylocoagulases from Staphylococcus intermedius and Staphylococcus aureus of animal origin.

Staphylocoagulases of Staphylococcus aureus (40 strains originally isolated from horses, dogs, cats, cows, sheep, opossums, pigs, humans, and a goat) and Staphylococcus intermedius (19 isolates from dogs and 1 pigeon strain) were tested for their affinity to prothrombins of either bovine or human origin. The tests used were the coagulase tube test (using human, bovine, or equine fibrinogen with either bovine or human prothrombin as the source of coagulase-reacting factor) and a chromogenic assay which enabled quantification of the amidolytic activity of the staphylocoagulase-prothrombin complex. S. intermedius showed weak specificity for human prothrombin, with 15% of the coagulases clotting human fibrinogen, 25% clotting equine fibrinogen, and 40% clotting bovine fibrinogen. However, 65% of coagulases clotted equine fibrinogen, 75% of coagulases clotted human fibrinogen, and 100% of coagulases clotted bovine fibrinogen when bovine prothrombin was used. The animal isolates of S. aureus displayed more diverse specificity toward prothrombin than S. intermedius strains. While 85% of coagulase preparations clotted human fibrinogen when human prothrombin was used, only 45% of preparations clotted bovine fibrinogen when bovine prothrombin was used. However, 62.5% of coagulases clotted human fibrinogen when bovine prothrombin was used and 85% of coagulases clotted bovine fibrinogen when human prothrombin was used. This may be a reflection of the diversity of the animal origins of S. aureus isolates.

Animals↗

PubMed1997

Typing of Staphylococcus aureus and Staphylococcus epidermidis strains by PCR analysis of inter-IS256 spacer length polymorphisms.

IS256 elements are present in multiple copies in the staphylococcal genome, either flanking the transposon Tn4001 or independent of it. PCR-based analysis of inter-IS256 spacer polymorphisms was developed for typing of methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus epidermidis strains. Using SmaI macrorestriction analysis resolved by pulsed-field gel electrophoresis (PFGE) as the reference method for MRSA typing, excellent reproducibility (100%), discriminatory power (97%), and in vivo stability were observed. Good concordance of the results with those of other molecular typing methods was found for two MRSA collections. Inter-IS256 PCR analysis of a U.S. collection of MRSA strains (n = 36), previously characterized by 15 typing methods, showed more limited discrimination. Agreement was 78% with PFGE analysis and 83% with ribotyping (HindIII). Analysis of a second set of Belgian MRSA strains (n = 17), categorized into two widespread epidemic clones by PFGE analysis, showed 65% agreement. For typing of S. epidermidis strains (n = 26), inter-IS256 PCR showed complete typeability (100%) and good discriminatory power (85%). Inter-IS256 PCR analysis is proposed as an efficient molecular typing assay for epidemiological studies of MRSA or S. epidermidis isolates.

Bacterial Typing Techniques↗

PubMed1999

Detection of Staphylococcus aureus and Staphylococcus epidermidis in clinical samples by 16S rRNA-directed in situ hybridization.

Staphylococcus epidermidis and Staphylococcus aureus are the most common causes of medical device-associated infections, including septicemic loosenings of orthopedic implants. Frequently, the microbiological diagnosis of these infections remains ambiguous, since at least some staphylococci have the capacity to reduce their growth rate considerably. These strains exhibit a small-colony phenotype, and often they are not detectable by conventional microbiological techniques. Moreover, clinical isolates of S. aureus and S. epidermidis adhere to polymer and metal surfaces by the generation of thick, multilayered biofilms consisting of bacteria and extracellular polysaccharides. This study reports improved detection and identification of S. aureus and S. epidermidis by an in situ hybridization method with fluorescence-labeled oligonucleotide probes specific for staphylococcal 16S rRNA. The technique has proven to be suitable for the in situ detection of staphylococci, which is illustrated by the identification of S. epidermidis in a connective tissue sample obtained from a patient with septicemic loosening of a hip arthroplasty. We also show that this technique allows the detection of intracellularly persisting bacteria, including small-colony variants of S. aureus, and the differentiation of S. epidermidis from other clinically relevant staphylococci even when they are embedded in biofilms. These results suggest that the 16S rRNA in situ hybridization technique could represent a powerful diagnostic tool for the detection and differentiation of many other fastidious microorganisms.

Genes, rRNA↗

PubMed1999

Comparison of susceptibility testing methods with mecA gene analysis for determining oxacillin (methicillin) resistance in clinical isolates of Staphylococcus aureus and coagulase-negative Staphylococcus spp.

Ninety-nine clinical staphylococcal isolates (58 coagulase-negative Staphylococcus spp. [CoNS] and 41 Staphylococcus aureus isolates) were evaluated for susceptibility to oxacillin. The following susceptibility testing methods, media, and incubation conditions were studied: agar dilution by using Mueller-Hinton (MH) medium (Difco) supplemented with either 0, 2, or 4% NaCl and incubation at 30 or 35 degrees C in ambient air for 24 or 48 h; disk diffusion by using commercially prepared MH medium (Difco) and MH II agar (BBL) and incubation at 35 degrees C in ambient air for 24 or 48 h; and agar screen (spot or swab inoculation) by using commercially prepared agar (Remel) or MH agar (Difco) prepared in-house, each containing 4% NaCl and 6 microg of oxacillin/ml (0.6-microg/ml oxacillin was also studied with MH agar prepared in-house for the agar swab method and CoNS isolates) and incubation at 35 degrees C in ambient air for 24 or 48 h for swab inoculation and at 30 or 35 degrees C in ambient air for 24 or 48 h for spot inoculation. The results for these methods were compared to the results for mecA gene detection by a PCR method. Given the ability to support growth and the results for susceptibility testing (the breakpoint for susceptible isolates was </=2 microg/ml), the best methods for CoNS isolates were (i) agar dilution by using MH medium supplemented with 4% NaCl and incubation at 35 degrees C for 48 h (no growth failures were noted, and sensitivity was 97.6%) and (ii) agar screen (swab inoculation) by using MH medium prepared in-house supplemented with 4% NaCl and containing 0.6 microg oxacillin/ml and incubation at 35 degrees C for 48 h (one isolate that did not carry the mecA gene did not grow, and the sensitivity was 100%). All but one (agar dilution without added NaCl and incubation at 30 degrees C for 48 h) of the methods tested revealed all oxacillin-resistant S. aureus isolates, and no growth failures occurred with any method. If the breakpoint for susceptibility was lowered to </=1 microg/ml for agar dilution methods, more CoNS isolates with oxacillin resistance related to the mecA gene were detected when 0 or 2% NaCl agar supplementation was used. Only one CoNS isolate with mecA gene-associated resistance was not detected by using agar dilution and MH medium supplemented with 4% NaCl with incubation for 48 h. When the breakpoint for susceptibility was decreased 10-fold (from 6.0 to 0.6 microg of oxacillin per ml) for the agar swab screen method, fully 100% of the CoNS isolates that carried the mecA gene were identified.

Bacterial Proteins↗

PubMed2004

Clinical isolates of Staphylococcus intermedius masquerading as methicillin-resistant Staphylococcus aureus.

Staphylococcus intermedius is a zoonotic organism that can be associated with human disease. We report two separate cases of S. intermedius infection in which a false-positive rapid penicillin binding protein 2a latex test in conjunction with the phenotypic properties of beta-hemolysis and coagulase positivity allowed the clinical isolates to masquerade as methicillin-resistant Staphylococcus aureus. 16S rRNA gene sequencing and the absence of mecA revealed the strains to be methicillin-susceptible S. intermedius.

Adult↗

PubMed2005

The pls gene found in methicillin-resistant Staphylococcus aureus strains is common in clinical isolates of Staphylococcus sciuri.

pls, a gene found in type I staphylococcal cassette chromosome mec (SCCmec) regions of methicillin-resistant Staphylococcus aureus strains, was present in 12 of the 15 human clinical Staphylococcus sciuri isolates studied. Pls was expressed in the S. sciuri isolates, although at a lower level than in S. aureus. Other parts of SCCmec could also be found in the S. sciuri genome.

Bacterial Proteins↗

PubMed2005

Comparison of conventional susceptibility testing, penicillin-binding protein 2a latex agglutination testing, and mecA real-time PCR for detection of oxacillin resistance in Staphylococcus aureus and coagulase-negative Staphylococcus.

Penicillin-binding protein (PBP) 2a latex agglutination was compared with conventional susceptibility testing and mecA real-time PCR for the detection of oxacillin resistance in Staphylococcus aureus. Inoculum volume and induction with oxacillin were PBP 2a testing variables. For coagulase-negative Staphylococcus, an increased inoculum volume of 10 microl greatly reduced the number of isolates requiring induction.

Bacterial Proteins↗

PubMedDate not supplied

In vitro antibacterial activity of TOC-50, a new parenteral cephalosporin against methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis.

The in vitro activity of TOC-50, a new parenteral cephalosporin, was assessed against methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus epidermidis (MRSE). TOC-50 showed excellent activity, which was stronger than that of methicillin, cloxacillin, the cephalosporins tested, imipenem, gentamycin, minocycline, ofloxacin and ciprofloxacin against MRSA and had a minimum inhibitory concentration (MIC) comparable to that of vancomycin (the MICs of TOC-50 and vancomycin for growth inhibition of 90% of the strains tested were 3.13 and 1.56 micrograms/ml, respectively). Against MRSE, TOC-50 exhibited excellent activity, which was stronger than that of methicillin, ampicillin, the cephalosporins tested and imipenem, and was twice as active as vancomycin. In terms of the bactericidal effect against MRSA, TOC-50 was superior to vancomycin.

Anti-Bacterial Agents↗

PubMed1990

Topical vancomycin for the treatment of Staphylococcus epidermidis and methicillin-resistant Staphylococcus aureus conjunctivitis.

Staphylococcus aureus and Staphylococcus epidermidis are organisms that frequently cause conjunctivitis or blepharoconjunctivitis. We describe a patient with methicillin-resistant S. aureus and S. epidermidis conjunctivitis who was treated successfully using an extemporaneously prepared topical ophthalmic solution of vancomycin hydrochloride 31 mg/mL. Studies describing the preparation, stability, and comfort of this solution, as well as reports pertaining to efficacy, are reviewed. Controlled clinical trials evaluating the safety and efficacy of vancomycin ophthalmic solution have not yet been performed.

Administration, Topical↗

PubMed2003

Comparative in vitro activity of linezolid and five other antimicrobials against nosocomial isolates of methicillin-resistant Staphylococcus aureus, methicillin-resistant Staphylococcus epidermidis and vancomycin-resistant Enterococcus faecium.

The activity of linezolid in comparison to vancomycin, teicoplanin, oxacillin, clindamycin and gentamicin was tested against 60 strains of methicillin-resistant Staphylococcus aureus, 60 strains of methicillin-resistant Staphylococcus epidermidis isolated from patients with nosocomial infections and 24 strains of vancomycin-resistant Enterococcus faecium isolated from feces of hospitalized patients. Minimum Inhibitory Concentrations (MICs) were determined by the Epsilometer test method. All tested strains were sensitive to linezolid and specifically all methicillin-resistant S. aureus had MIC range 0.25-3.00, MIC50 = 0.75, MIC90 = 1.5, all methicillin-resistant S. epidermidis had MIC range 0.125-1.5, MIC50 = 0.5, MIC90 = 1 and all vancomycin-resistant E. faecium had MIC range 0.5-1.5, MIC50 = 1, MIC90 = 1. Linezolid is the first of a novel antimicrobial class, the oxazolidinones, which is a promising treatment for serious Gram-positive infections, including multiresistant strains.

Acetamides↗

PubMed1984

Multiple antibiotic resistance in Staphylococcus aureus and Staphylococcus epidermidis: plasmids in strains associated with nosocomial infection.

The plasmid DNA profiles were compared to phenotypically-similar, antibiotic-resistant strains of Staphylococcus aureus and Staphylococcus epidermidis associated with nosocomial infections in a Melbourne hospital. Whereas resistance to gentamicin, tobramycin and kanamycin was encoded by one of 3 plasmids [pSK1, 18 megadalton (Md); pSK4, 22 Md; pSK9, 17 Md] in S. aureus, no similar plasmids were detected in S. epidermidis. Mediated exclusively by the chromosome in S. aureus, tetracycline resistance was encoded either by the chromosome or by a 2.8 Md plasmid in strains of S. epidermidis. The inability to detect common resistance plasmids in strains of S. aureus and S. epidermidis recovered from this outbreak is in contrast to recent observations with staphylococci from other geographic areas; nevertheless, on the basis of restriction endonuclease analyses of 3 Md chloramphenicol resistance plasmids, it is suggested that a common gene pool does exist within isolates of S. aureus and S. epidermidis from Melbourne hospitals.

Aminoglycosides↗

PubMed2005

Subclinical mastitis and antimicrobial susceptibility of Staphylococcus caprae and Staphylococcus epidermidis isolated from two Italian goat herds.

A total of 156 goats from 2 commercial dairy goat farms were monitored for intramammary infections during an entire lactation. Most of the infections (80.7%) observed were due to coagulase-negative staphylococci (CNS) species. In herd 1, nearly all of the infections (96%) were due to CNS species, with Staphylococcus caprae (SCAP) being the most common specific pathogen observed, accounting for about 43% of the infections. In herd 2, the proportion of the infections due to CNS was 67% and Staphylococcus epidermidis (SEPI) was the most common pathogen (48% of infections) and SCAP was not present. Linear somatic cell scores (SCS) were greater in milk from infected udder halves, with an average difference of 0.78 SCS. The SCS for infected udder halves was greater than noninfected for all CNS species, although differences among species were observed. The ranking across CNS species was SCAP > other CNS > SEPI > no infection. However, infections by SEPI tended to be more persistent. Increased SCS was associated with a statistically significant decrease in milk yield, but no effect was observed for intramammary infections (IMI). Ninety-seven isolates of CNS (53 SCAP and 44 SEPI) were investigated for in vitro susceptibility to several antimicrobial agents. Benzylpenicillin was the most effective antimicrobial agent against SCAP and SEPI. A concentration of 0.05 microg/mL was sufficient to inhibit growth of 90% of SCAP colonies, and 0.10 microg/mL yielded a similar effectiveness for SEPI. Amoxicillin and the combination of amoxicillin and clavulanic acid were only slightly less effective. Tetracycline (62.5 microg/mL) and tilmicosin (500 microg/mL) were the least effective treatments for SEPI and SCAP, respectively.

Amoxicillin↗

PubMed1992

Plasmids encoding for erythromycin ribosomal methylase of Staphylococcus epidermidis and Staphylococcus simulans.

Two 1.7 Md plasmids of Staphylococcus epidermidis and three ones of Staphylococcus simulans determining inducible macrolide-lincosamide resistance are identical as judged by restriction endonuclease fingerprinting. These plasmids designated pEI2101, pEI9105, pE1107, pEI1108 and pEI6104, respectively, belong to the incompatibility group 12. Dot-blot hybridization by photobiotin-labelled gene probe developed from S. aureus erythromycin ribosomal methylase gene showed cross hybridization between methylase-coding reference plasmids and the tested ones. The examined plasmids proved to be no transmissible in mating experiments into S. aureus recipients.

Drug Resistance, Microbial↗

PubMed1990

Neutrophil chemotactic activity of peptidoglycan. A comparison between Staphylococcus aureus and Staphylococcus epidermidis.

The ability of peptidoglycan from Staphylococcus epidermidis and Staphylococcus aureus to generate in human serum chemoattractant for peripheral blood neutrophils was studied. It was shown that PG from the two bacteria was able to induce chemotactic activity in normal human serum. Sonication of PG was required to generate this activity. Very little or no activity was generated in heat-treated or C5-deficient human serum by PG, indicating that PG treatment of serum resulted in generation of chemoattractants by activation of complement. Kinetics studies employing C2-deficient or MgEGTA-chelated serum revealed that S. epidermidis induced chemotactic activity by activating the alternative complement pathway. The alternative complement activation induced by S. epidermidis occurred rapidly and was completed after 15 min, whereas S. aureus activated the alternative pathway much more slowly, with activation reaching a maximum at 60 min. The rapid activation of the alternative complement pathway by S. epidermidis PG may partly explain why this bacterium does not normally cause infections in healthy individuals.

Antigens, Bacterial↗

PubMed1985

Colonization of hospitalized patients by Staphylococcus aureus, Staphylococcus epidermidis and enterococci.

Patients admitted to a medical intensive care unit (29), a surgical intensive care unit (29) and a general medical ward (34) were studied to determine the frequency of colonization by Staphylococcus aureus, Staph. epidermidis and enterococci. Cultures of anterior nares, pharynx, axilla, periurethral tissues, and rectum were done within 24 h of admission and every 48-72 h thereafter for the duration of hospitalization. We found Staph. aureus colonization of pharynx, axilla, periurethral tissues and rectum to be equally frequent in patients with and without nasal colonization. Staphylococcus epidermidis was commonly recovered from all sites except the pharynx. The 49 strains of this organism acquired in the hospital were significantly more often resistant to nafcillin, cephalothin, chloramphenicol, trimethoprim-sulfamethoxazole and gentamicin than were 199 isolates recovered from patients at the time of admission. Enterococci were commonly isolated from rectal swabs and, less often, from other sites. Acquisition of enterococcal colonization during hospitalization was correlated significantly with antibiotic administration.

Anti-Bacterial Agents↗