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Bacterial growth and killing in chronic ambulatory peritoneal dialysis fluids.

We determined the ability of Staphylococcus epidermidis, Staphylococcus aureus, and Escherichia coli to survive and grow in peritoneal dialysis fluids from patients undergoing chronic ambulatory peritoneal dialysis. Staphylococci did not survive in commercially available dialysis solutions but grew readily in peritoneal effluents obtained from patients after the dialysis dwell time. The number of CFU doubled 6 and 13 times in 24 h for S. epidermidis and S. aureus, respectively. E. coli grew well in both the pre- and postdialysis peritoneal fluid. Peritoneal macrophages as well as peripheral blood leukocytes inhibited bacterial growth in peritoneal dialysis fluid. However, 10(6) phagocytes per ml were minimally required to obtain a bacteriostatic effect. The addition of serum to peritoneal dialysis fluid increased the antibacterial activity of macrophages and blood leukocytes. The capacity of the aminoglycoside antibiotic tobramycin to reduce bacterial CFU in peritoneal dialysis fluid was only 10% of its bactericidal capacity in standard Mueller-Hinton brush. Peritoneal dialysis fluid had no effect on the antibacterial activity of imipenem.

Blood Bactericidal Activity↗

Bacterial adhesion to intravenous cannulae: influence of implantation in the rabbit and of enzyme treatments.

Comparison was made of the adhesion of Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli, Klebsiella aerogenes and Pseudomonas aeruginosa to six types of intravascular cannula material. Adhesion to materials removed from rabbit tissues did not differ significantly between types of material or between bacterial species. In contrast, major differences were found when unimplanted materials were examined; the overall rank order of adhesiveness of bacteria to unimplanted materials (S. epidermidis greater than P. aeruginosa greater than S. aureus much greater than K. aerogenes greater than E. coli) was highly significant (F = 13.0, P less than 0.0005), and although no single material was consistently least attractive to all micro-organisms, FEP-Teflon and PTFE-Teflon showed significantly lower overall affinity for bacteria than other materials (P less than 0.001); all species showed a significant preference for a silicone polymer (P less than 0.0005). The nature of the bacterial surface structures responsible for adhesion were investigated by the actions of pronase and mixed glycosidase, which produced significant respective decreases and increases in adhesion of staphylococci to unimplanted materials; their effects on the Gram-negative bacilli were less consistent.

Animals↗

[Study of the virulence of coagulase-negative staphylococci in experimental infections].

The pathogenicity and virulence of 3 strains of Staphylococcus epidermidis, Staphylococcus haemolyticus and Staphylococcus saprophyticus, respectively were studied in BALB/c mice by intraperitoneal (ip) challenge using 4 bacterial suspensions of different colony forming units (CFU) of each strain. Strains were isolated from wound, blood, and urine of inpatients. On the base of the lethality rates, S. saprophyticus proved to be the most virulent (LD50 = 2.7-2.9 x 10(7) CFU/g body wt), while the S. epidermidis species was the least virulent (LD50 = 6-8 x 10(7) CFU/g body wt). The lethality rate of male mice was higher than that of the female ones at the same challenge bacterium concentration. Mice of higher body weight were generally more sensitive to a quality of bacteria calculated to 1 g of mice than the lighter mice. The 245 mice surviving the challenge were dissected at the 10th day of infection. Splenomegaly was found to be the most frequent macroscopic pathological alteration. There appeared kidney abscesses, liver abscesses, and rarely peritoneal abscess. The frequency of pathological findings were directly proportional to the amount of bacteria injected. The results indicate that clinical strains of coagulase-negative staphylococci (CNS) examined were pathogenic and virulent for BALB/c mice they are invasive after ip injection and can cause macroscopic pathological changes in parenchymal organs. Thus, ip CNS challenge in mice may be a model to imitate and study infections caused by CNS in human.

Animals↗

Peritonitis in an urban peritoneal dialysis program: an analysis of infecting pathogens.

We have previously found that race, level of education, and peritoneal dialysis system are factors that significantly and independently influence peritonitis rates in our patient population. We now extend these observations by assessing the pathogens responsible for peritonitis in these subgroups. Between January 1, 1981, and May 15, 1993, 248 peritoneal dialysis patients underwent dialysis at our facility. The rate of peritonitis by pathogen was determined in these patients using the fixed effects Poisson model. Total peritonitis rates in black patients (1.89 episodes/patient-year) were significantly greater compared with white patients (1.11 episodes/patient-year; P < 0.0001). Increased infection rates in black patients were significant for Staphylococcus epidermidis, Staphylococcus aureus, and gram-negative pathogens. The level of education had a negative correlation with peritonitis rates (< or = 8 years, 2.00 episodes/patient-year; 9 to 12 years, 1.64 episodes/patient-year; and > or = 13 years, 1.24 episodes/patient-year) with patients having > or = 13 years of education at the start of dialysis demonstrating a significantly lower total peritonitis rate compared with patients with 9 to 12 years (P = 0.001) or < or = 8 years (P < 0.001) of education. This was accounted for by a significant decrease in infection rates for S epidermidis, polymicrobial, and gram-negative organisms. Finally, patients on automated peritoneal dialysis had significantly lower total peritonitis rates (0.59 episodes/patient-year) compared with patients on either a connect (2.11 episodes/patient-year) or disconnect (1.46 episodes/patient-year) system.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Pathogenicity and virulence of coagulase negative staphylococci in relation to adherence, hydrophobicity, and toxin production in vitro.

AIMS: To study the pathogenicity and virulence characteristics of Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus sapro-phyticus. METHODS: BALB/c mice were challenged intraperitoneally with graded doses of three strains belonging to each species. LD50s were measured for each strain. Haemolysin (alpha- and delta-) and enzyme (DNAase, lipase, and esterase) production in vitro were measured qualitatively and quantitatively. Adhesion to plastic was measured and related to cell surface hydrophobicity among the strains. RESULTS: S saprophyticus proved the most virulent (LD50 = 2.7-2.9 x 10(7) cfu/g body weight) while S epidermidis was the least virulent (LD50 = 6-8 x 10(7) cfu/g body weight). An enlarged spleen was the most common macroscopic pathological feature. Kidney, liver, and more rarely peritoneal abscesses were also seen in the infected animals. No direct correlation was found between adherence in vitro, cell surface hydrophobicity, or toxin/enzyme biosynthesis and virulence in mice. CONCLUSION: The results show that coagulase negative staphylococci are pathogenic in BALB/c mice. It is clear that these bacteria can cause invasive disease. However, the in vitro characteristics of coagulase negative staphylococci are not related to the pathogenicity of the organisms in mice.

Animals↗

Regulation of agr-dependent virulence genes in Staphylococcus aureus by RNAIII from coagulase-negative staphylococci.

Many of the genes coding for extracellular toxins, enzymes, and cell surface proteins in Staphylococcus aureus are regulated by a 510-nucleotide (nt) RNA molecule, RNAIII. Transcription of genes encoding secreted toxins and enzymes, including hla (alpha-toxin), saeB (enterotoxin B), tst (toxic shock syndrome toxin 1), and ssp (serine protease), is stimulated, while transcription of genes encoding cell surface proteins, like spa (protein A) and fnb (fibronectin binding proteins), is repressed. Besides being a regulator, RNAIII is also an mRNA coding for staphylococcal delta-lysin. We have identified RNAIII homologs in three different coagulase-negative staphylococci (CoNS), i.e., Staphylococcus epidermidis, Staphylococcus simulans, and Staphylococcus warneri. RNAIII from these CoNS turned out to be very similar to that of S. aureus and contained open reading frames encoding delta-lysin homologs. Though a number of big insertions and/or deletions have occurred, mainly in the 5' half of the molecules, the sequences show a high degree of identity, especially in the first 50 and last 150 nt. The CoNS RNAIII had the ability to completely repress transcription of protein A in an RNAIII-deficient S. aureus mutant and the ability to stimulate transcription of the alpha-toxin and serine protease genes. However, the stimulatory effect was impaired compared to that of S. aureus RNAIII, suggesting that these regulatory functions are independent. By creating S. epidermidis-S. aureus RNAIII hybrids, we could also show that both the 5' and 3' halves of the RNAIII molecule are involved in the transcriptional regulation of alpha-toxin and serine protease mRNAs in S. aureus.

Amino Acid Sequence↗

In vitro and ex vivo activities of minocycline and EDTA against microorganisms embedded in biofilm on catheter surfaces.

Minocycline-EDTA (M-EDTA) flush solution has been shown to prevent catheter-related infection and colonization in a rabbit model and in hemodialysis patients. We undertook this study in order to determine the activities of M-EDTA against organisms embedded in fresh biofilm (in vitro) and mature biofilm (ex vivo). For the experiment with the in vitro model, a modified Robbin's device (MRD) was used whereby 25 catheter segments were flushed for 18 h with 10(6) CFU of biofilm-producing Staphylococcus epidermidis, Staphylococcus aureus, and Candida albicans per ml. Subsequently, each of the catheter segments was incubated in one of the following solutions: (i) streptokinase, (ii) heparin, (iii) broth alone, (iv) vancomycin, (v) vancomycin-heparin, (vi) EDTA, (vii) minocycline (high-dose alternating with low-dose), or (viii) M-EDTA (low-dose minocycline alternating with high-dose minocycline were used to study the additive and synergistic activities of M-EDTA). All segments were cultured quantitatively by scrape sonication. For the experiment with the ex vivo model, 54 catheter tip segments removed from patients and colonized with bacterial organisms by roll plate were longitudinally cut into two equal segments and exposed to either saline, heparin, EDTA, or M-EDTA (with high-dose minocycline). Subsequently, all segments were examined by confocal laser electron microscopy. In the in vitro MRD model, M-EDTA (with a low concentration of minocycline) was significantly more effective than any other agent in reducing colonization of S. epidermidis, S. aureus, and C. albicans (P < 0.01). M-EDTA (with a high concentration of minocycline) eradicated all staphylococcal and C. albicans organisms embedded in the biofilm. In the ex vivo model, M-EDTA (with a high concentration of minocycline) reduced bacterial colonization more frequently than EDTA or heparin (P < 0.01). We concluded that M-EDTA is highly active in eradicating microorganisms embedded in fresh and mature biofilm adhering to catheter surfaces.

Anti-Bacterial Agents↗

Persistence of bacteria on antibiotic loaded acrylic depots. A reason for caution.

Bacterial growth on the surface of antibiotic loaded acrylic cement was examined in an in vitro model. Tobramycin or vancomycin impregnated discs were incubated in broth containing either Staphylococcus epidermidis or Staphylococcus aureus organisms. At 24, 48, and 96 hours, the broth and the surface of the acrylic discs were examined for viable organisms. In the broth, only 6% (8 of 136) of samples contained viable organisms at 24 hours, and all samples were sterile by 96 hours. This is in contrast to the surface of the discs, which revealed the presence of viable organisms at all study time periods. Growth was noted in 99% and 30% of the Staphylococcus epidermidis and Staphylococcus aureus discs, respectively, at 24 hours. Viable organisms were found on both types of discs through 96 hours (20% of Staphylococcus epidermidis and 15% of Staphylococcus aureus). The surface of bone cement is therefore a suitable substrate for bacterial growth, even in the presence of antibiotics. In clinical practice, antibiotic loaded acrylic cement should therefore be used with caution and clear indications.

Anti-Bacterial Agents↗

In vitro susceptibilities of four species of coagulase-negative staphylococci.

The in vitro susceptibilities of 260 strains of coagulase-negative staphylococci to penicillin G, oxacillin, nafcillin, methicillin, cephalothin, and seven non-beta-lactam antimicrobial agents were determined and compared with the susceptibilities of 54 strains of Staphylococcus aureus with known patterns of susceptibility. Penicillin G susceptibility for S. aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis was readily determined by using beta-lactamase tests with induced cells and with a standardized microdilution test. MIC criteria for susceptibility used for S. aureus were applicable to the coagulase-negative species. Percentages of organisms susceptible were as follows: S. epidermidis, 7%; S. haemolyticus, 5%; and S. hominis, 47%. Oxacillin susceptibility for these four species was readily determined by using a modification of the microdilution test. MIC criteria for susceptibility used for S. aureus were applicable to S. haemolyticus and S. hominis, but alternate criteria were necessary for S. epidermidis. Percentages of organisms susceptible were as follows: S. epidermidis, 29%; S. haemolyticus, 36%; and S. hominis, 97%. Staphylococcus saprophyticus differed from the other staphylococcal species; all strains were beta-lactamase negative and were penicillin susceptible but had higher penicillin G MICs than did susceptible strains of the other species. There was total cross resistance among the penicillinase-resistant penicillins and cephalothin for the coagulase-negative staphylococci as well as for S. aureus; oxacillin MICs were more reliable than MICs of the other drugs or a standardized disk diffusion test for distinguishing resistant from susceptible strains. Vancomycin, rifampin, and ciprofloxacin were consistently active against all staphylococci. Erythromycin, clindamycin, gentamicin, and trimethoprim-sulfamethoxazole were more active against oxacillin-susceptible staphylococci than against oxacillin-resistant staphylococci.

Anti-Bacterial Agents↗

Inhibition of neutrophil elastase prevents cathelicidin activation and impairs clearance of bacteria from wounds.

The host defense roles of neutrophil elastase in a porcine skin wound chamber model were explored. Analysis of wound fluid by acid-urea polyacrylamide gel electrophoresis, Western blot, and bacterial overlay confirmed that the neutrophil-derived protegrins constituted the major stable antimicrobial polypeptide in the wound fluid. The application to the wound of 0.10 and 0.25 mM N-methoxysuccinyl-alanine-alanine-proline-valine (AAPV) chloromethyl ketone, a specific neutrophil elastase inhibitor (NEI), blocked the proteolytic activation of protegrins and diminished the associated antimicrobial activity as detected by radial diffusion assay against Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans or by bacterial gel overlay against S epidermidis and E coli. The application of the related cathepsin G inhibitor (CGI), benzyloxycarbonyl-glycine-leucine-phenylalanine (ZGLF) chloromethyl ketone, had no effect. In wound chambers that received 10(6) colony-forming unit (CFU)/mL of S epidermidis, the presence of NEI significantly decreased the 24-hour clearance of bacteria from the wound compared to wounds treated with CGI or solvent only. Neither inhibitor, at 0.10 or 0.25 mM concentration, affected leukocyte accumulation or degranulation in the wound chambers. The in vitro microbicidal decrement due to NEI was restored by an amount of the specific protegrin (PG-1), which was equivalent to the measured difference of protegrin between control and inhibited chambers. Administration of 1 microg/mL exogenous PG-1 4 hours after chamber preparation was sufficient to normalize in vivo antimicrobial activity. Although pharmacologic NEIs are promising candidates as anti-inflammatory drugs, they may impair host defense in part by inhibiting the activation of cathelicidins by neutrophil elastase.

Animals↗

Influence of the incubation atmosphere on the production of biofilm by staphylococci.

Biofilm formation by Staphylococcus epidermidis, Staphylococcus hemolyticus, Staphylococcus sciuri and Staphylococcus aureus in aerobic, anaerobic and CO2 incubation atmospheres was quantified by the modified microtiter plate test. The S. epidermidis and S. aureus strains showed significantly lower biofilm production when grown in a CO2-rich environment compared to that exhibited in aerobic incubation. The amount of biofilm produced by these strains under anaerobic conditions did not differ significantly from the biofilm formation detected in the aerobic incubation. The incubation atmosphere did not affect S. sciuri biofilm formation. Biofilm production by S. hemolyticus isolates was very low regardless of the experimental conditions used.

Aerobiosis↗

In vitro evaluation of antibiotic release from and bacteria growth inhibition by antibiotic-loaded acrylic bone cement spacers.

The antibiotic release from and the bacteria growth inhibition by antibiotic-loaded acrylic bone cement hip spacers were studied. The cement used was Palacos R, and it was loaded with either one antibiotic powder (gentamicin, vancomycin, teicoplanin, or synercid) [monoantibiotic case] or two antibiotic powders (gentamicin + vancomycin or gentamicin + teicoplanin) [biantibiotic case] and then tested against Staphylococcus epidermidis, Staphylococcus aureus, Enterococcus faecalis, and methicillin-resistant Staphylococcus aureus (MRSA). Antibiotic elution and bacteria growth were measured every 24 h simultaneously by fluorescence polarization immunoassay and photometrically, respectively. The gentamicin + vancomycin combination achieved the longest growth inhibition on S. epidermidis and MRSA (mean of 20 and 14 days, respectively). Gentamicin + teicoplanin-loaded spacers were capable of inhibiting growth on E. faecalis and S. aureus for the longest period (11 and 16 days, respectively). The highest concentrations of gentamicin and vancomycin could be assayed during the first 4 days. Teicoplanin concentrations could be detected only during the first 72 h, synercid was not detected at all, possibly because of the limitation of the detection technique used. A greater percentage of the gentamicin was released than of the vancomycin. The aminoglycosid-glycopeptid combination showed a synergistic effect on the release of gentamicin, but not on vancomycin or teicoplanin. Biantibiotic-impregnated hip spacers proved to be superior to monoantibiotic ones. Because of important differences between the conditions used for the present tests and the in vivo environment, any recommendation with regard to the use of monoantibiotic- and biantibiotic-loaded acrylic bone cement spacers must await the results of further investigations.

Anti-Bacterial Agents↗

Bacterial peritonitis in continuous ambulatory peritoneal dialysis: effect on dialysis on host defense mechanisms.

We have used an experimental model of continuous ambulatory peritoneal dialysis (CAPD) to investigate the effect of dialysis on host defense mechanisms in the peritoneal cavity. Laboratory rats undergoing regular dialysis were challenged with a single bolus of Staphylococcus epidermidis, Staphylococcus aureus, Candida albicans, and Pseudomonas aeruginosa. Intraperitoneal challenges with all four microbial strains underwent a rapid clearance during the first 4 hours, irrespective of whether the host was undergoing dialysis. Studies using S epidermidis showed that the rapid clearance was not mediated by the diaphragmatic stomata as might have been expected. Dialysis did, however, cause delays over the following 48 hours with all of the pathogens except C albicans. Bacterial clearance over a 10-day period was strain dependant, and microbial persistence was related to adhesion formation around the cannula. Again C albicans was an exception. Our conclusions are that dialysis does not affect the innate defense mechanisms involved in the early clearance of a bolus challenge to the peritoneal cavity but can cause delays in eradicating microorganisms that survive this phase. Because these experiments were carried out using a bolus inoculum, it may be necessary to reevaluate the effect of dialysis on host defenses in the peritoneal cavity under conditions in which the challenge more closely approximates a touch contamination event.

Animals↗

Study of the efficacy of Coated VICRYL Plus Antibacterial suture (coated Polyglactin 910 suture with Triclosan) in two animal models of general surgery.

OBJECTIVES: To evaluate the efficacy in vitro and in vivo of a new antibacterial suture (PGAB) compared with a traditional braided suture (PG). Our primary goals were to study microbiological effectiveness and impact on wound healing of PGAB vs PG. Secondary goal was to analyze influence on inflammatory response. METHODS: In vitro study: clinical samples of Staphylococcus epidermidis, Staphylococcus aureus, S. hominis, Staphylococcus haemolyticus, Staphylococcus auricularis, Enterococcus faecalis, Corynebacterium spp. and Escherichia coli were studied. We also implanted a flat mesh in 10 minipigs, four incisions each (two PG and two PGAB) two contaminated with S. epidermidis and two not contaminated. Finally, we performed four colic anastomosis in each of 10 minipigs, two contaminated with E. coli and two not contaminated (two PG and two PGAB). We studied the inflammatory and wound healing processes in both models. RESULTS: We observed a bactericidal efficacy of PGAB against grampositive, and bacteriostatic effect against E. coli. Mesh study: recovered CFU were lower in the group PGAB vs PG. In the group PGAB, inflammatory mediators' concentrations were lower. In the group PGAB, concentrations of wound healing mediators were normal. Colic anastomosis: recovered CFU were lower in the group PGAB vs the group PG. In the group PGAB we observed a reduction of inflammatory mediators. In the group PGAB we observed normalized concentrations of wound healing mediators. CONCLUSIONS: This study demonstrates microbiological efficacy of PGAB, that normalizes wound healing process, and an anti-inflammatory effect.

Anastomosis, Surgical↗

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

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

Bacteremia↗

Inhibition of adhesion of yeasts and bacteria by poly(ethylene oxide)-brushes on glass in a parallel plate flow chamber.

Poly(ethylene oxide) (PEO)-brushes are generally recognized as protein-repellent surfaces, and although a role in discouraging microbial adhesion has been established for some strains and species, no study exists on the effects of PEO-brushes on a large variety of bacterial and yeast strains. In this paper, a PEO-brush has been covalently attached to glass and silica by reaction in a polymer melt. Subsequently, the presence of a PEO-brush was demonstrated using contact angle measurements, X-ray photoelectron spectroscopy and ellipsometry. For five bacterial (Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus salivarius, Escherichia coli and Pseudomonas aeruginosa) and two yeast strains (Candida albicans and Candida tropicalis), adhesion to PEO-brushes was compared with adhesion to bare glass in a parallel plate flow chamber. The initial deposition rates of Sta. epidermidis, Sta. aureus and Str. salivarius to glass were relatively high, between 2400 and 2600 cm(-2) s(-1), while E. coli and P. aeruginosa deposited much more slowly. The initial deposition rates of the yeasts to glass were 144 and 444 cm(-2) s(-1) for C. albicans GB 1/2 and C. tropicalis GB 9/9, respectively. Coating of the glass surface with a PEO-brush yielded more than 98 % reduction in bacterial adhesion, although for the more hydrophobic P. aeruginosa a smaller reduction was observed. For both yeast species adhesion suppression was less effective than for the bacteria and here too the more hydrophobic C. tropicalis showed less reduction than the more hydrophilic C. albicans. The PEO-brush had a thickness of 22 nm in water, as inferred from ellipsometry. Assuming that on bare glass the adhered micro-organisms are positioned only a few nanometers away from the surface and that the brush keeps them at a distance of 22 nm, it is calculated that the brush yields a sevenfold attenuation of the Lifshitz-Van der Waals attraction to the surface between the micro-organisms and the surface. Decreased Lifshitz-van der Waals attraction may be responsible for the suppression of the microbial adhesion observed.

Bacterial Adhesion↗

In vitro antistaphylococcal activity of pefloxacin alone and in combination with other antistaphylococcal drugs.

MICs of pefloxacin and nine antistaphylococcal drugs were determined for 200 isolates of Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis, and Staphylococcus saprophyticus. All the strains were susceptible to pefloxacin, vancomycin, and rifampin. Oxacillin-resistant strains were uniformly resistant to cephalothin and were more likely to be resistant to gentamicin, erythromycin, clindamycin, doxycycline, and trimethoprim-sulfamethoxazole than were oxacillin-susceptible strains. Time-kill studies with 23 strains of S. aureus, S. epidermidis, and S. haemolyticus indicated that the relative order of bactericidal activities was gentamicin greater than or equal to pefloxacin greater than oxacillin greater than vancomycin greater than rifampin. Pefloxacin combined with oxacillin or vancomycin killed staphylococci more rapidly than oxacillin or vancomycin alone but less rapidly than pefloxacin alone. Gentamicin combined with oxacillin, vancomycin, or pefloxacin resulted in the most rapid killing of gentamicin-susceptible strains. Rifampin combined with oxacillin, vancomycin, or pefloxacin reduced the bactericidal activities of those drugs, but rifampin resistance was not observed as it was with rifampin alone. Pefloxacin is a potentially useful antistaphylococcal agent.

Anti-Bacterial Agents↗

Bap-dependent biofilm formation by pathogenic species of Staphylococcus: evidence of horizontal gene transfer?

The biofilm-associated protein (Bap) is a surface protein implicated in biofilm formation by Staphylococcus aureus isolated from chronic mastitis infections. The bap gene is carried in a putative composite transposon inserted in SaPIbov2, a mobile staphylococcal pathogenicity island. In this study, bap orthologue genes from several staphylococcal species, including Staphylococcus epidermidis, Staphylococcus chromogenes, Staphylococcus xylosus, Staphylococcus simulans and Staphylococcus hyicus, were identified, cloned and sequenced. Sequence analysis comparison of the bap gene from these species revealed a very high sequence similarity, suggesting the horizontal gene transfer of SaPIbov2 amongst them. However, sequence analyses of the flanking region revealed that the bap gene of these species was not contained in the SaPIbov2 pathogenicity island. Although they did not contain the icaADBC operon, all the coagulase-negative staphylococcal isolates harbouring bap were strong biofilm producers. Disruption of the bap gene in S. epidermidis abolished its capacity to form a biofilm, whereas heterologous complementation of a biofilm-negative strain of S. aureus with the Bap protein from S. epidermidis bestowed the capacity to form a biofilm on a polystyrene surface. Altogether, these results demonstrate that Bap orthologues from coagulase-negative staphylococci induce an alternative mechanism of biofilm formation that is independent of the PIA/PNAG exopolysaccharide.

Adhesins, Bacterial↗