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The influence of penicillin and quinaldofur on the growth of Staphylococcus epidermidis and Streptococcus faecalis in milk depends on aerobic conditions and the quality of milk.
We report the complete coding sequence and the partial amino acid sequence (determined by chemical sequencing) of Staphylococcus epidermidis extracellular cysteine (Ecp) and serine (Esp) proteases. The first enzyme shows an extended sequence similarity to Staphylococcus aureus cysteine protease (staphopain) and the second one resembles the serine protease produced by that species. The region directly upstream of the sequence coding for the mature protein in both enzymes displays significant homology to the profragments encoded by sspB and sspA, respectively, thus suggesting that the characterised enzymes may also be produced as proproteins. Furthermore, we report some biological properties of the cysteine protease, contributing to a better understanding of its role as a possible virulence factor. The proteolytic activity of this enzyme was rapidly and efficiently inhibited by human alpha-2-macroglobulin; however, human kininogen as well as cystatins (A, C and D) were not inhibitory. Moreover, the protease was capable of inactivating, by limited proteolysis, both alpha-1-antitrypsin and HMW-kininogen, but neither alpha-1-antichymotrypsin nor antithrombin III.
The role of renal failure in the pathogenesis of the Staphylococcus epidermidis (S. epidermidis) peritonitis presented by end-stage renal disease patients treated with continuous ambulatory peritoneal dialysis was investigated in a mouse model of surgically-induced renal failure. Six weeks after the surgery, an i.p. inoculum of 10(6) colony forming units S. epidermidis was administered to renal failure mice and their sham-operated and normal controls, and assessment of bacterial clearance and inflammatory response was conducted over the next 72 hours. Peritoneal clearance of S. epidermidis was complete in most animals; however, the process was significantly delayed in renal failure mice compared to sham-operated controls. Viable bacteria invariably remained associated with the peritoneum after peritoneal washings had become culture negative. Peritoneal inflammatory response was markedly diminished in renal failure mice, the early polymorphonuclear cell response being particularly affected. Peripheral response consisted of a prompt and short-lived polymorph increase which was similar in renal failure and sham-operated mice. The factors responsible for the observed impairment of local inflammatory response in association with delayed bacterial clearance in renal failure mice following i.p. challenge with S. epidermidis remain to be defined.
The importance of coagulase-negative staphylococci, especially Staphylococcus epidermidis in clinical and nosocomial infection are recognized increasingly in recent years. A rapid and accurate identification of S. epidermidis is therefore important and necessary. A new test, susceptibility to desferrioxamine, coupled with trehalose fermentation has been recommended for the identification of this organism. However, the medium and method used are different from what has been recommended by the NCCLS. To investigate the feasibility of using the desferrioxamine susceptibility test in conjunction with the routinely used disc agar diffusion test, we employed 111 staphylococcal strains (including 51 S. epidermidis isolates, 15 S. hominis and 45 other coagulase-negative staphylococci) as test organisms, and followed the procedures recommended by the NCCLS in which Mueller-Hinton agar and standard inoculum were used. Results indicated that all strains of S. epidermidis and S. hominis were susceptible to 1 mg desferrioxamine (the diameter of the inhibition zone were 28-37 mm). The minimum inhibitory concentrations of desferrioxamine to S. epidermidis and S. hominis isolates were determined to be 125 micrograms/ml. Further differentiation of S. hominis and S. epidermidis can be made by their ability to ferment trehalose, the former could while the latter could not. We conclude that the desferrioxamine susceptibility test of coagulase-negative staphylococci can be used in conjunction with the routine disc agar diffusion method. S. epidermidis can be identified rapidly and accurately by its susceptibility to 1 mg desferrioxamine and inability to ferment trehalose.
To determine whether a relationship exists between biofilm formation and hemagglutination in Staphylococcus epidermidis, 20 skin isolates and 19 prosthetic valve endocarditis isolates were characterized for biofilm formation, hemagglutination, and the presence of a 357-bp polymerase chain reaction product within icaA. A strong association existed between biofilm formation, which has been linked to strains that produce polysaccharide intercellular adhesin (PIA), and hemagglutination. Strains that produced biofilm were significantly (P<.001) more likely to mediate hemagglutination (16 biofilm-positive/hemagglutination-positive strains and 19 biofilm-negative/hemagglutination-negative strains) within the 39 clinical strains tested. In addition, Staphylococcus carnosus TM300, a biofilm-negative, hemagglutination-negative strain, carrying the ica operon-containing plasmid pCN27, produced significant biofilm on glass and mediated hemagglutination (>/=1/128). It was concluded that production of PIA and hemagglutination are strongly associated and that PIA, at least in part, mediates hemagglutination in S. epidermidis.
PURPOSE: To evaluate Staphylococcus epidermidis adherence to human amniotic membrane (HAM) and compare it with S epidermidis adherence to human, rabbit, and cat conjunctiva in vitro. SETTING: Research laboratory, Loyola University Medical Center, Maywood, Illinois, USA. METHODS: Commercially available HAM (N = 3) was used. Conjunctival specimens from humans, rabbits, and cats (n = 3 each) were processed similarly to HAM. The tissues were exposed to S epidermidis (3 x 10(8) colony-forming units per milliliter) for 0, 5, 30, and 90 minutes, rinsed in sterile saline, and processed for light, scanning (SEM), and transmission (TEM) electron microscopy. Scanning electron microscopy (x2000) was used to quantify adherent bacteria/mm(2) of tissue (SEM photographs = 144). RESULTS: The following mean levels (+/- SD) of adherent S epidermidis/mm(2) were found at 0, 5, 30, and 90 minutes: HAM, 3833 +/- 1570, 9060 +/- 2512, 15,431 +/- 10,752, and 30,315 +/- 14,803, respectively; human conjunctiva, 1493 +/- 672, 7218 +/- 3179, 17,273 +/- 7168, and 19,861 +/- 9624, respectively; rabbit conjunctiva, 3385 +/- 5074, 14,386 +/- 14,569, 15,283 +/- 13,679, and 20,113 +/- 24,016, respectively; and cat conjunctiva, 4032 +/- 2240, 12,345 +/- 3413, 8512 +/- 4032, and 19,214 +/- 5584, respectively. No statistically significant differences were found at any time point (P>.16). CONCLUSION: There was no statistically significant difference in the adherence of S epidermidis to HAM and to human, rabbit, and cat conjunctiva. Bacterial adherence to HAM may be clinically significant.
On the chromosome of Staphylococcus epidermidis RP62A the lipase gene (gehSE1) is immediately flanked by the icaAA'BC operon, which is involved in biofilm formation. Since lipase production might play a role in staphylococcal skin colonisation as well, we studied the biochemical properties of the staphylococcal lipases more closely. The DNA sequence and the deduced protein sequence revealed that gehSE1 is very similar to the lipase sequence of S. epidermidis strain 9. Like other staphylococcal lipases, gehSE1 is organised as a preproenzyme. The part of gehSE1 coding for the mature lipase was cloned and overexpressed as a fusion protein with an N-terminal histidine tag in Escherichia coli. The lipase was purified to homogeneity using a combination of precipitation techniques, metal-affinity chromatography and gel filtration. Biochemical characterisation showed that this lipase is closely related to the lipase from Staphylococcus aurelis NCTC8530. Both enzymes have a pH optimum around 6, are very stable at low pH, and need calcium as a cofactor for catalytic activity. The preferred substrates are small triacylglycerols, with a maximum activity toward tributyrylglycerol. Comparison of the substrate selectivity with those of other microbial lipases showed that phospholipids are generally poor substrates for lipases. An exception is the lipase from Staphylococcus hyicus, which prefers phospholipids as a substrate, distinguishing this staphylococcal lipase from other microbial lipases. These results are discussed in view of the structure/function relationships of staphylococcal lipases, and the possible involvement of these enzymes in biological processes such as skin colonisation and pathogenesis.
Because of their biofilm-forming capacity, invasive Staphylococcus epidermidis isolates, which cause the majority of nosocomial catheter-related bloodstream infections (BSIs), are thought to be selected at the time of catheter insertion from a population of less virulent commensal strains. This fact allows the prediction that invasive and contaminating strains can be differentiated via detection of virulence-associated genes. However, the hospital environment may pave the way for catheter-related infections by promoting a shift in the commensal bacterial population toward strains with enhanced virulence. The distribution of virulence-associated genes (icaADBC, aap, atlE, bhp, fbe, embp, mecA, IS256, and IS257), polysaccharide intercellular adhesin synthesis, and biofilm formation were investigated in S. epidermidis strains from independent episodes of catheter-related BSIs in individuals who have received bone marrow transplantation (BMT). The results were compared with those obtained for commensal S. epidermidis isolates from hospitalized patients after BMT and from healthy individuals, respectively. The clonal relationships of the strains were investigated by pulsed-field gel electrophoresis. icaADBC, mecA, and IS256 were significantly more prevalent in BSI isolates than in commensal isolates from healthy individuals. However, the prevalence of any of the genes in clonally independent, endogenous commensal strains from BMT patients did not differ from that in invasive BSI strains. icaADBC and methicillin resistance, factors important for the establishment of catheter-related infections, already ensure survival of the organisms in their physiological habitat in the hospital environment, resulting in a higher probability of contamination of indwelling medical devices with virulent S. epidermidis strains. The dynamics of S. epidermidis populations reveal that detection of icaADBC and mecA is not suitable for discriminating invasive from contaminating S. epidermidis strains.
The opsonic requirements of 65 strains of Staphylococcus epidermidis were compared in fresh and in heated normal human serum. The strains were isolated from patients with CAPD peritonitis (n = 26), neonatal septicaemia (n = 24) and nasal cultures (n = 15). A wide variation was observed in opsonic requirements between the different strains, both with fresh and with heated serum. Opsonization in heated serum proceeded less efficiently and higher concentrations (mean three-fold compared to fresh serum) were needed for adequate phagocytosis. However, a highly significant correlation was found between the minimal opsonic concentrations of fresh and of heated serum (r = 0.84, P less than 0.0005). In addition, S. epidermidis can become opsonized in agammaglobulinaemic serum. Thus, opsonization of S. epidermidis can be mediated by antibodies alone and by complement alone. Slime-producing strains and encapsulated strains did not require higher concentrations of serum to become opsonized. Opsonic requirements were highly significantly correlated with surface hydrophobicity. Enzymatic treatment rendered the strains more hydrophilic and decreased their opsonic requirements. Isolates from nasal cultures required significantly higher concentrations of both fresh and heated serum to become adequately phagocytozed, whereas isolates from CAPD peritonitis required higher concentrations of heated serum only compared to blood isolates. The uptake of S. epidermidis preopsonized in heated serum as determined in our direct phagocytosis assay did not result in a comparable chemiluminescence response.
Methicillin resistance (Mecr) was transduced into a methicillin-susceptible variant of the Mecr donor Staphylococcus epidermidis BS10. UV irradiation of phage stimulated Mecr transduction frequency. If loss of Cd and Hg ion resistance occurred in this recipient, or if the three markers Mecr, Cdr, and Hgr were co-eliminated from the donor, neither strain acted as a recipient for Mecr.
Three hundred sixty-five isolates of Staphylococcus epidermidis (coagulase-negative) obtained over a 12-month period from 10 selected wards at New York University Medical Center-Goldwater Memorial Hospital, Roosevelt Island, N.Y., were typed serologically by slide agglutination. The isolates were first biotyped by the Baird-Parker scheme and then subtyped as proteinase-positive or proteinase-negative so that the selection of strains for immunologic study was based on biochemical and enzymatic factors rather than on random choice. The biotyping scheme of Baird-Parker and the differentiation between proteolytic and nonproteolytic strains were of limited in initial categorization of isolates of S. epidermidis. Antisera to proteinase-positive and proteinase-negative S. epidermidis biotypes 1 and 4 were absorbed with different permutations of homologous and heterologous thermostable (TS) and thermolabile (TL) antigens, and four distinct TL and two distinct TS antigenic groups, representing five different serotype patterns (i.e., A1-2, B1-2, C-D1, C-D1-2, and E1), were detected. The TL antigens also appeared to be more widespread and were shared more frequently by strains of S. epidermidis than were TS antigens, but the use of both TS and TL antigens provided a better serologic system than use of either TS or TL antigens alone.
Staphylococcus epidermidis, a major component of the skin flora, is usually considered a contaminant when recovered from diagnostic cultures. Since 1974 infections caused by gram-negative bacilli and S. aureus occurring among patients with granulocytopenic cancer have remained constant; infections due to S. epidermidis occurring before 1977 at a rate of 2.0 per 1000 days of hospitalization of patients with acute leukemia, increased to 14.6 per 1000 days in 1979. All S. epidermidis infections before 1977 originated from skin sites; since 1977 the respiratory tract and alimentary canal have become the predominant sites of origin. Predisposing factors for infection were profound granulocytopenia (less than 100/microL), a diagnosis of acute leukemia, and an oral nonabsorbable antibiotic regimen lacking vancomycin. The increased incidence of infection was not related to a nosocomial source or the more frequent use of long-term indwelling catheters. Methicillin resistance among isolates was common (40%). Intravenous vancomycin therapy provided the best therapeutic results. Diagnostic cultures positive for S. epidermidis in the setting of infection and profound granulocytopenia warrant appropriate antibiotic therapy.
The low-level resistance of three clinical isolates of Staphylococcus epidermidis to glycopeptide antibiotics was found to be constitutive, not inducible, and was not increased by passage in the presence of either vancomycin or teicoplanin. There was no loss of resistance on repeated passage in antibiotic-free broth. In contrast, the susceptibility to these antibiotics declined for S. epidermidis NCTC 6513 that been sequentially passaged in either vancomycin or teicoplanin whereas the variants reverted to being susceptible on further passage in antibiotic-free broth. Antibiotic activity was almost completely abolished when cultures of the resistant S. epidermidis strains were exposed overnight to sub-MIC concentrations. No evidence of drug-modifying activity was obtained. Experiments of antibiotic-binding activity indicated that the resistant strains exhibited an increased ability to sequester antibiotics which was particularly rapid in stationary phase cultures when most of the antibiotic activity disappeared from the growth medium within 30 min of exposure to the drugs. Teicoplanin was sequestered more efficiently than vancomycin and some loss of activity was also observed when stationary phase cultures of S. epidermidis NCTC 6513 were exposed to glycopeptides. These results suggest that glycopeptide-resistant isolates of S. epidermidis are able to bind large amounts of these antibiotics, possibly at sites unassociated with the D-alanyl-D-alanine target, and that teicoplanin is bound more avidly than vancomycin.
Medical device-associated infections, most frequently caused by coagulase-negative staphylococci, especially Staphylococcus epidermidis, are of increasing importance in modern medicine. The formation of adherent, multilayered bacterial biofilms is the most important factor in the pathogenesis of these infections, which regularly fail to respond to appropriate antimicrobial therapy. Progress in elucidating the factors functional in elaboration of S. epidermidis biofilms and the regulation of their expression with a special emphasis on the role of quorum sensing are reviewed. Significant progress has been made in recent years, which provides the rationale for developing better preventive, therapeutic and diagnostic measures.
To determine whether a slime-producing strain of Staphylococcus epidermidis was capable of producing acute infection of a prosthetic vascular graft, 5 cm segments of knitted Dacron were implanted in the infrarenal aortic position of dogs in three groups of animals. These included a control group (no graft contamination), a contaminated group that received a graft soaked in an S. epidermidis solution (untreated group), and a contaminated group in which perioperative antibiotics (three doses of cefamandole, 100 mg/kg) were administered (prophylaxis group). In all the animals reexploration and graft removal were performed at 10 days, with replacement of the defect being achieved with a new uncontaminated graft. These animals underwent exploration a third time after an additional 10-day period. S. epidermidis was not grown from the control animals (n = 7) but was cultured in 44% of the prophylaxis group (n = 9) and 88% of the untreated group (n = 16) during at least one of the operative procedures (chi 2 = 15.859; p less than 0.001). The pathologic features of acute S. epidermidis infection were best seen in the untreated animals and included anastomotic disruption (56%), periaortic hematoma, and lymphadenopathy (94%). Microscopic examination of the aortic tissues revealed extensive infiltrates of leukocytes, macrophages, and foreign body giant cells with aortic necrosis. These features were less prominent in the prophylaxis animals. We conclude that S. epidermidis is capable of producing acute graft infection with perigraft inflammation and anastomotic disruption. The administration of perioperative antibiotics reduced but did not abolish these effects of bacterial contamination of prosthetic vascular grafts.
This report concerns 60 children with documented Staphylococcus epidermidis sepsis. There were 34 boys and 26 girls, ages 2 weeks to 15 years. The primary diagnosis included malignancy (13), congenital (13) or acquired (11) gastrointestinal disorders, prematurity (7), cardiac defect (5), hydrocephalus (2) and miscellaneous (9). Clinical presentation included fever (54), tachycardia (15), lethargy (20), hypotension (8), irritability (6), increased gastric residuals (6) and apnea/bradycardia (3). A documented source of sepsis was noted in 56 patients, including percutaneous central venous catheters (23), Broviac catheters (17), umbilical arterial catheters (6), wound (3), V-P shunt (2), cardiac defect (2), cholangitis (1), chest tube (1) and peripheral arterial line (1). There were six sepsis-related deaths, four in premature infants. Two of six infected subclavian catheters were treated successfully with vancomycin. Infection was successfully cleared in 20 of 23 infected Broviac catheters with vancomycin through the line. However, six were eventually removed for tract infection (1), persistent fever (2), and Candida sp. infection (3). Although once considered a non-pathogenic skin contaminant, S. epidermidis has emerged as a serious pathogen in hospitalized, immunosuppressed, premature and malnourished pediatric patients. Indwelling catheters enhance the likelihood of infection in these patients. Aggressive antimicrobial therapy is vital in this potentially lethal infection. Vancomycin proved efficacious in this series.
OBJECTIVE: To investigate the basic pharmacodynamic properties of teicoplanin in vitro for Staphylococcus aureus, Staphylococcus epidermidis and Enterococcus faecium. METHODS: The following experiments were performed: (1) bacterial killing by teicoplanin at different concentrations; (2) bacterial killing by teicoplanin at 8 x MIC against the same strains with inocula of 5 x 10(3), 5 x 10(5) and 5 x 10(7) CFU/mL; (3) studies of the postantibiotic effect (PAE) and the postantibiotic sub-MIC effect (PASME) of teicoplanin; (4) studies of the killing by teicoplanin in an in vitro kinetic model following exposure to simulated human serum pharmacokinetic concentrations (6 mg/kg OD at steady state). RESULTS: Concentration-dependent killing was noted against S. epidermidis, with a > 4 log10 difference in CFUs between 2 x MIC and 64 x MIC at 24 h. Also, against S. aureus there was slight concentration-dependent killing, which, however, did not reach 2 log10 CFU/mL. Teicoplanin exerted a similar killing rate at all inocula for S. epidermidis, except for slower initial killing up to 6 h at the highest inoculum. In contrast, overall slower killing at all inocula was seen for S. aureus, where an inoculum effect was noted at the highest inoculum. For E. faecium, only a bacteriostatic effect was noted at all concentrations and inocula. No or very short PAEs were noted for the investigated strains. However, when the strains in the postantibiotic phase were exposed to 0.1, 0.2 and 0.3 x MIC of teicoplanin (PASME), substantial prolongation of the PAEs was seen. Although no significant killing was achieved in our kinetic model for any of the strains, regrowth of S. epidermidis was noted first after 8 h, despite a T > MIC24 of only 5% (1.2 h), illustrating the long post-MIC effect for this strain. For S. aureus, T > MIC was 38%, and regrowth occurred later than for S. epidermidis. Neither killing nor regrowth was seen for E. faecium with a T > MIC24 of 27%. CONCLUSION: Teicoplanin exerted a concentration-dependent bactericidal effect against S. epidermidis, a less notable one against S. aureus, and a bacteriostatic effect against E. faecium. A reduced killing rate with increasing inocula was seen for S. aureus and also for S. epidermidis at the highest inoculum. No or very short PAEs were noted for the investigated strains, but were substantially prolonged with the addition of subinhibitory concentrations. When human pharmacokinetics was simulated (6 mg/kg OD at steady state) in the kinetic model, no net bactericidal effect was noted for any of the strains at 24 h.
BACKGROUND: The aim of this study was to constitute a valid graft infection model with Staphylococcus epidermidis in rats. METHODS: Rats were divided into seven groups. In groups 1 and 2, 2 cm x 2 cm polypropylene grafts were incubated with 10(8) c.f.u./mL slime-positive S. epidermidis at 37 degrees C for 2 and 24 h and were then placed subfascially to the groins of rats. In the third group, naive grafts were placed and 0.5 mL of 3 x 10(7) c.f.u. slime-positive S. epidermidis were injected on the inside of the wounds. Rifampicin (30 mg/kg) in group 4 and teicoplanin (20 mg/kg) in group 5 were applied i.p. to rats with 2-h incubated grafts for prophylaxis. The same prophylactic regimens were given to groups 6 and 7 in which rats were incubated for 24 h. At eighth day, rats were killed and wounds were assessed with macroscopic evaluation and cultures. RESULTS: No death occurred in any of the groups. In groups 1 and 2, 100% infection rates were achieved. However, graft infection was detected in only two (20%) of the rats in group 3 (P = 0.001). Prophylactic application of teicoplanin or rifampicin decreased the infection rates significantly in the short-incubation groups. CONCLUSION: Incubation of polypropylene grafts with slime-producing S. epidermidis for 2 and 24 h in the pre-application period achieved the occurrence of a standardized graft infection. Prophylactic use of teicoplanin and rifampicin decreased the infection rates. We propose to use this reproducible and reliable animal model of graft infection in future studies.