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Correlation of agar dilution and VITEK2 system for detection of resistance to macrolides, lincosamides and pristinamycin among Staphylococcus aureus and Staphylococcus epidermidis: association with genotypes.

The performance of the VITEK2 system was evaluated against the agar dilution reference procedure for testing susceptibility of Staphylococcus aureus and Staphylococcus epidermidis to macrolides, lincosamides and streptogramins (MLS). Eighty clinical isolates were selected according to their resistance phenotype and genotype. Results for erythromycin and clindamycin showed 100% agreement; results for lincomycin showed agreement of 78%, with one very major error and 17 minor errors; and results for pristinamycin showed agreement of 46%, with one major error and 43 minor errors. Most isolates resistant to lincomycin and streptogramin A (L SgAr phenotype) were falsely susceptible to lincomycin, and intermediately-resistant or resistant to pristinamycin, with the VITEK2 system. No resistance gene was detected. Most (80%) isolates resistant constitutively to MLS (MLS(r)BC phenotype) were falsely intermediately-resistant to pristinamycin with the VITEK2 system. The erm(A) gene was more common than erm(C) in MLS(r)BC strains. Resistance to pristinamycin alone (SgA SgB PTr phenotype), or associated with either lincomycin resistance (L SgA SgB PTr phenotype) or constitutive MLS(B) resistance (MLS(BC) SgA PTr phenotype), was well-characterised without discordant results. Resistance to pristinamycin was always associated with resistance to streptogramin A, encoded by the vga(A), vga(B), vgb(A) and vat(A) genes in association with the erm(A) or erm(C) genes.

Agar↗

Differentiation of strains of Staphylococcus epidermidis biotypes 1 and 4 by aminopeptidase profiles.

Seven different strains of Staphylococcus epidermidis and one strain of S. aureus were catagorized by their aminopeptidase activity. Eight different profiles were obtained: five profiles for S. epidermidis biotype 4 (each representing a different known serotype), two profiles for S. epidermidis biotype 1 (one for each strain used) and one for the strain of S. aureus used. Despite some similarities, distinct patterns of the hydrolysis of 22 different amino acid-beta-naphthylamides were evident for each individual strain. The evaluation of aminopeptidase profiles provides another means for distinguishing between strains of the same species and can, therefore, be used advantageously and in combination with serotyping as an epidemiological tool.

2-Naphthylamine↗

Comparative in vitro and in vivo efficacy of roxithromycin and erythromycin against a strain of methicillin-susceptible Staphylococcus epidermidis.

The in vitro and in vivo efficacy of roxithromycin was compared with that of erythromycin, against a methicillin-susceptible strain of Staphylococcus epidermidis. We performed standard in vitro testing (MIC, MBC, and time-kill kinetics) for roxithromycin, erythromycin, and rifampin. Both macrolides were bacteriostatic in vitro. There was no significant difference in microbial survival between erythromycin and roxithromycin groups in the time-kill kinetics (p = 0.3). For the in vivo experiments, using the rabbit experimental endocarditis model, roxithromycin was found to be inferior to erythromycin in decreasing the microbial burden of the endocardial vegetations (p < 0.05). Rifampin was highly effective, both in vitro and in vivo. In conclusion, the efficacy of roxithromycin was poor and inferior to erythromycin against a strain of methicillin-susceptible S. epidermidis.

Analysis of Variance↗

Typing of Staphylococcus epidermidis strains by random amplification of polymorphic DNA.

The polymerase chain reaction was used to obtain randomly amplified polymorphic DNA profiles for typing of Staphylococcus epidermidis strains. Epidemiologically unrelated S. epidermidis isolates were screened with randomly amplified polymorphic DNA analysis. The discriminating ability of 45 randomly designed 10-mer primers was assessed. The highest discriminatory power was obtained with the 10-mer oligonucleotide OPAM-12. In typing a total of 13 unrelated S. epidermidis strains with OPAM-12, 11 different banding profiles were obtained reproducibly by agarose gel electrophoresis. The discriminatory power of the method with OPAM-12 was estimated using the D value of Hunter and Gaston (1988) to be 0.961. A reproducibility index of 1 was obtained after typing a total of 40 cultures including 12 triplicates and one quadruplicate of the 13 unrelated strains. Following the described procedure, the randomly amplified polymorphic DNA method provided a rapid, simple and reproducible alternative to other S. epidermidis typing systems.

Base Sequence↗

Induction of Staphylococcus epidermidis biofilm formation via proteolytic processing of the accumulation-associated protein by staphylococcal and host proteases.

Because of its biofilm forming potential Staphylococcus epidermidis has evolved as a leading cause of device-related infections. The polysaccharide intercellular adhesin (PIA) is significantly involved in biofilm accumulation. However, infections because of PIA-negative strains are not uncommon, suggesting the existence of PIA-independent biofilm accumulation mechanisms. Here we found that biofilm formation in the clinically significant S. epidermidis 5179 depended on the expression of a truncated 140 kDa isoform of the 220 kDa accumulation-associated protein Aap. As expression of the truncated Aap isoform leads to biofilm formation in aap-negative S. epidermidis 1585, this domain mediates intercellular adhesion in a polysaccharide-independent manner. In contrast, expression of full-length Aap did not lead to a biofilm-positive phenotype. Obviously, to gain adhesive function, full-length Aap has to be proteolytically processed through staphylococcal proteases as demonstrated by inhibition of biofilm formation by alpha(2)-macroglobulin. Importantly, also exogenously added granulocyte proteases activated Aap, thereby inducing biofilm formation in S. epidermidis 5179 and four additional, independent clinical S. epidermidis strains. It is therefore reasonable to assume that in vivo effector mechanisms of the innate immunity can directly induce protein-dependent S. epidermidis cell aggregation and biofilm formation, thereby enabling the pathogen to evade clearance by phagocytes.

Adhesins, Bacterial↗

[A patient with acute leukemia and meningitis caused by Staphylococcus epidermidis treated with fosfomycin].

In a 17-year-old male patient with acute lymphoblastic leukaemia, who was being treated with chemotherapy, a Staphylococcus epidermidis infection with several septicaemias developed during a period of protracted neutropenia. The patient was treated with vancomycin and fusidic acid, but blood cultures remained positive. The patient also developed staphylococcal meningitis. After the antibiotic regimen was supplemented by fosfomycin, the blood cultures became sterile. Combination treatment with vancomycin and fosfomycin was continued for two months without apparent toxicity. In individual cases of infection with multiresistant S. epidermidis fosfomycin may be included in the antibiotic regimen. This is the first report of parenteral use of fosfomycin in the Netherlands.

Adolescent↗

[Relationship between an offensive smell given off from human foot and Staphylococcus epidermidis].

The bacteria isolated from foot skins of 17 volunteers by the swab sampling method were mostly gram-positive cocci, which were identified as Staphylococcus epidermidis by the ID-kit SP-18 (Nissui Co., Ltd). After incubation of S. epidermidis on agar plates containing oleic acid and Tween 80 for 24 h at 35 C, the smell noticed was similar to an offensive smell of human pes. However, under the same conditions, the smell of another staphylococcal species was different from that of S. epidermidis. Except for the staphylococcal species, the colonies isolated from the skins were mostly those of yeast (unidentified) and gave off no offensive smell. From these results, it was considered that the smell of human pes might be given off by S. epidermidis, and if this species is inhibited, the smell would also be inhibited. A selective bactericide for gram-positive bacteria, which is a lotion containing deoxycholic acid, was applied to the feet of the 17 volunteers. The experiments showed that the application obviously decreased the counts of colonies of S. epidermidis and inhibited the smell as compared with controls.

Adult↗

The inhibitory effect of Staphylococcus epidermidis slime on the phagocytosis of murine peritoneal macrophages is interferon-independent.

The extracellular slime produced by Staphylococcus epidermidis has been shown to interfere with several human neutrophil functions in vitro, such as chemotaxis, degranulation and phagocytosis. Slime production has been suggested as a useful marker for clinically significant infections with coagulase-negative Staphylococcus. Since the main role of macrophages in defense mechanisms is phagocytosis, the effect of slime on the phagocytic activity of macrophages was investigated. The phagocytic activity of murine peritoneal macrophages treated with slime in vitro decreased in a dose-dependent fashion. A similar decrease was also observed in macrophages isolated from mice that had previously received intraperitoneal injection of slime. To investigate whether interferon also plays a role in this process, mice were treated with interferon or an interferon inducer, polyinosinic-polycytidylic acid (poly I:C), together with slime before macrophage isolation. The slime-suppressed phagocytic activity of macrophages was partially relieved by both agents, and the recovery effect of poly I:C in slime-suppressed phagocytosis of macrophages in vivo might be attributed to the increased interferon level in peritoneal fluid and sera. However, when slime was given to poly I:C-pretreated mice, the phagocytic activity remained suppressed. Thus, it appears that slime is able to suppress the phagocytic activity of macrophages regardless of the state of macrophage activation by poly I:C. The results suggest that the inhibition of phagocytosis by S. epidermidis slime may be independent from the activation of interferon.

Animals↗

[Genetic nature of the bacteriocinogenicity factor of Staphylococcus epidermidis strain No. 259].

Genetic nature of the bacteriogenicity factor of the Staphylococcus epidermidis No. 259 strain was studied by acting upon the strain with ultraviolet irradiation, acridine orange at a temperature of 37 and 44 degrees C, and cultivation of the strain at a temperature of 44 degrees C for 24 and 96 hours. The most effective elimination of the bacteriocinogenicity factor was reached with the action of acridine orange at 44 degrees C and the strain passage at 44 degrees C for 96 hours. A study of 212 subcultures which lost the bacteriocinogenicity factor under the effect of various factors demonstrated that they failed to differ by properties from the initial strain and retained the bacteriocin resistance of the initial strain. A conclusion was drawn that the bacteriocinogenicity factor was an extrachomosomic agent, and was not associated with any other strain properties.

Acridines↗

Transduction and plasmid deoxyribonucleic acid analysis in a multiply antibiotic-resistant strain of Staphylococcus epidermidis.

A genetic analysis of a multiply antibiotic-resistant strain of Staphylococcus epidermidis was performed. Experiments designed to show reversion of organisms to antibiotic susceptibility, as well as studies of the influence of ultraviolet irradiation of phage on the transduction frequencies of the resistance markers, indicated that determinants of chloramphenicol (cml), tetracycline (tet), and neomycin (neo) resistance are present on separate plasmids, but the streptomycin marker is chromosomal. In 2 to 6% of tetracycline-resistant transductants, co-transduction of cml was also observed. By using CsCl-dye density gradients followed by neutral sucrose gradients, the plasmids carrying cml, tet, and neo could be isolated and their molecular weights could be determined. The tetracycline plasmid is shown to be incompatible with one of the cryptic plasmids of a recipient strain.

Anti-Bacterial Agents↗

Epidemiologic features and phage types associated with nosocomial infections caused by Staphylococcus epidermidis.

An apparent problem with increased numbers of nosocomial infections caused by Staphylococcus epidermidis at a large hospital was studied in a clinical-epidemiological investigation. Thirty-six cases of S. epidermidis infection were confirmed on the thoracic surgery, general surgery, nursery and pediatric services during a 3-year period. Nine cases were fatal and six of these occurred in patients following cardiovascular surgery with implanted prothesis. The majority of cases (25/36) occurred following cardiovascular surgery. The median onset of infection was six days from the date of surgery, suggesting infection during the intraoperative period. Although the number of cases studied remains small, two phage types, 71/108/275a/459 and 407-2, did predominate among these clinical infections. Interestingly, during this same time interval these same types predominated among surgical staff members closely associated with these patients. Resistance to antimicrobials was high among isolates tested, with more than 50% of the strains resistant to six or more antimicrobial agents.

Adolescent↗

Transduction of penicillinase production and other antibiotic-resistance markers in Staphylococcus epidermidis.

Transduction of resistance from a multiply antibiotic-resistant strain of Staphylococcus epidermidis sub-group II was studied using the typing phage 108. The effect of increasing doses of ultraviolet radiation on the transducing phage was used to indicate the chromosomal or plasmid nature of the genes. Tetracycline and chloramphenicol resistance behaved as plasmid genes and streptomycin resistance as a chromosomal marker. It was also possible to transduce penicillin resistance (Pc) due to penicillinase production (bla+) using a low level of benzylpenicillin (0.03 microgram ml-1) for recovery. Approximately 10(-5) transductant colonies per phage input were obtained and ultraviolet kinetics indicated that Pc was plasmid carried. Pc transductants fell into two categories. In one group PC was stable as in the donor strain and transductants had the same phage sensitivity as the recipient. In the other, Pc was unstable at 37 degrees C and the instability was enhanced by growth at approximately 43.5 degrees C; these transductants also gained genes for restriction and modification of certain phages. Transductants that subsequently lost bla+ also lost the restriction and modification characters.

Anti-Bacterial Agents↗

A comparative study of Staphylococcus epidermidis isolates from clinical material and healthy carriers.

A total of 190 Staphylococcus epidermidis isolates from healthy carriers and clinical material were biotyped according to the Baird-Parker scheme, phage typed by a provisional set of 18 typing phages and tested for antimicrobial susceptibility. The majority of the strains from both groups were found to belong to biotype 1. A considerable number of strains were lysed at 100 x RTD by at least one phage. The strains from healthy carriers were distributed in 24 phage types and about half of them were clustered among three major phage types. Nine of the 30 clinical isolates were of the phage type 130/456 and another nine strains were of eight different phage types. The strain 130/456 represented only three per cent of the phage types from healthy carriers. The clinical strains exhibitbd a multiple resistance pattern to 13 antibiotics. Statistically significant was the difference in the resistance to Penicillin, Streptomycin, Carbenicillin, Erythromycin, Lincomycin, Ampicillin and Kanamycin between the groups of strains from clinical material and healthy carriers. No correlation was observed between biotypes, phage types and antimicrobial susceptibility in both groups of the isolates.

Anti-Bacterial Agents↗

Evaluation of trehalose-mannitol broth for differentiation of Staphylococcus epidermidis from other coagulase-negative staphylococcal species.

Trehalose-mannitol agar and various broth formulations thereof were evaluated for the differentiation of Staphylococcus epidermidis from other common coagulase-negative staphylococcal species isolated from human blood cultures. While this differentiation was accurately made on trehalose-mannitol agar in 18 h, modification of the medium to a broth formulation containing 2% each trehalose and mannitol in 0.2 ml of purple broth base provided greater than 95% accuracy when a heavy inoculum was incubated on a rotary shaker for at least 4 h.

Culture Media↗

Utilizing whole genome sequencing to characterize central line-associated bloodstream infections due to Staphylococcus epidermidis.

Whole genome sequencing (WGS) and clinical review were used to characterize 14 cases of central line-associated bloodstream infection (CLABSI) due to Staphylococcus epidermidis. WGS, which demonstrated disparate strains, suggested that 42.9% of S. epidermidis CLABSI cases were due to contamination, while clinical review suggested that 57.1% were contamination events.

Humans↗

Resistance plasmids for inducible macrolide-lincosamide resistance in Staphylococcus simulans and Staphylococcus epidermidis.

Localization and genetic nature of inducible macrolide resistance determinants in some coagulase negative staphylococci (CNS) were studied and compared with those in Staphylococcus aureus. An 1.7-megadalton R-plasmid, pEI 1107, mediating inducible resistance to macrolides and lincosamides was present in part of Staphylococcus simulans and Staphylococcus epidermidis strains isolated in the Central Hospital for Infectious Diseases. Two other plasmids were harboured by S. simulans LK 6108 strain. One of these plasmids, pEI 6108 (molecular mass 5.2 Md) was encoded for inducible macrolide-lincosamide resistance, and the second one, pE 6108 (molecular mass 4.9 Md) for constitutive marcolide resistance. The similarity in size and phenotype of small plasmids in CNS and S. aureus suggest a common origin of these resistance determinants.

DNA, Bacterial↗

The lipid composition of a halotolerant species of Staphylococcus epidermidis.

Studies were carried out on the lipid composition of a halotolerant Staphylococcus epidermidis isolated in pure culture from a growth medium for extreme halophiles containing 25% NaCl. The four major polar lipid components in this bacterium were found to be: (a) glycerophosphoryl diglucosyl diglyceride (10% by weight) with structure 3(1)-O-(-sn-glycerol-1-phosphoryl-6'-O=(beta-D glucopyranosyl-(1 leads to 6)- O-beta-D-glucopyranosyl)-1(3),2-diacyl-sn-glycerol; (b) diglucosyl diglyceride (15% by weight) with structure 3(1)-O-(beta-D-glucopyranosyl (1 leads to 6)-O-beta-D-glucopyranosyl)-1(3),2-diacyl-sn-glycerol; (c) monoglucosyl diglyceride (3% by weight) with structure 3(1)-O-(beta-D-glucopyranosyl)-1(3),2-diacyl-sn-glycerol, and (d) phosphatidylglycerol (60% by weight) with structure 1,2 diacyl-sn-glycero-3-phosphoryl-1'-sn-glycerol. Phosphatidic acid, cardiolipin, lysophosphatidylglycerol and three unidentified phospholipids were also detected in small amounts. Each lipid component had essentially the same fatty acid composition namely, anteiso-15:0 (60-75%), anteiso-17:0 (18-24%), iso-17:0 (8--10%), and small amounts of palmitic and stearic acids (2-5%). The fatty acids were non-randomly distributed in phosphatidylglycerol, the shorter chain anteiso 15:0 fatty acid being exclusively esterified to the 2-position and the longer chain anteiso- and iso-17:0 fatty acids at the 1-position. The fatty acid composition was not affected by increaseing NaCl content in the medium in the rande 0--15% but the proportion of anteiso-15:0 increased greatly when the salt concentration was increased to 25%. The proportions of ionic polar lipids were modified to give an increased net negative charge per mol ionic lipids when NaCl in the medium was increased from 15 to 25%, but the proportions of neutral glycolipids remained fairly constant.

Cardiolipins↗

Typing of Staphylococcus epidermidis colonizing in human nares by pulsed-field gel electrophoresis.

From the nares of 11 healthy adults, 253 strains of coagulase negative staphylococcus were isolated and 88% of them were identified as Staphylococcus epidermidis using the API STAPH system. Chromosomal DNA fingerprinting of the isolated strains revealed that each person carried multiple types of S. epidermidis in his or her nares. The colonization of the strains was not stable; the types of the isolates changed in the first and the second examinations 5 months apart. The results contrasted with previous findings in which only one strain of S. aureus colonized persistently in the nares of healthy adults.

Adult↗