Clin Infect Dis, 2005 Jan 15, 40(2), 280 - 5 Epub 2004 Dec 21. Inducible clindamycin resistance in Staphylococci: should clinicians and microbiologists be concerned? Lewis JS 2nd, Jorgensen JH. The increasing incidence of a variety of infections due to Staphylococcus aureus--and, especially, the expanding role of community-associated methicillin-resistant S . aureus (MRSA)--has led to emphasis on the need for safe and effective agents to treat both systemic and localized staphylococcal infections . Unlike most previously noted strains of health care-associated MRSA, community-acquired MRSA isolates are often susceptible to several non- beta -lactam drug classes, although they are usually not susceptible to macrolides . Several newer antimicrobial agents and a few older agents are available for treatment of systemic staphylococcal infections, but use may be limited by the relatively high cost of these agents or the need for parenteral administration . Inexpensive oral agents for treatment of localized, community-acquired MRSA infection include clindamycin, trimethoprim-sulfamethoxazole, and newer tetracyclines . Clindamycin has been used successfully to treat pneumonia and soft-tissue and musculoskeletal infections due to MRSA in adults and children . However, concern over the possibility of emergence of clindamycin resistance during therapy has discouraged some clinicians from prescribing that agent . Simple laboratory testing (e.g., the erythromycin-clindamycin "D-zone" test) can separate strains that have the genetic potential (i.e., the presence of erm genes) to become resistant during therapy from strains that are fully susceptible to clindamycin. Curr Opin Otolaryngol Head Neck Surg, 2005 Feb, 13(1), 39 - 44 Superantigen hypothesis for the early development of chronic hyperplastic sinusitis with massive nasal polyposis; Bernstein JM et al.; PURPOSE OF REVIEW: The pathogenesis, pathophysiology, and immunobiology of chronic hyperplastic sinusitis with massive nasal polyposis are starting to become unraveled . Allergy, viral infection, bacterial infection, fungal infection, and environmental pollution have all been suggested as possible initial triggers that may upregulate inflammation of the lateral wall of the nose to develop nasal polyposis . The purpose of this review is to present data from our laboratory that suggest that one of the possible early events in the development of inflammation of the lateral wall of the nose in chronic hyperplastic sinusitis with massive nasal polyposis is the production of exotoxins from Staphylococcus aureus . The exotoxins may act as superantigens and cause activation and clonal expansion of lymphocytes with specific Vbeta regions, resulting in massive cytokine production . RECENT FINDINGS: Recent published studies suggest that S . aureus is the most common organism isolated from the mucus adjacent to massive nasal polyposis . Staphylococci produce exotoxins . These exotoxins, sometimes known as enterotoxins, include SEA, SEB, and TSST-1 . These exotoxins are capable of acting as superantigens and therefore, reacting with T lymphocytes with specific Vbetas in the lateral wall of the nose . Thereafter, it is possible that these lymphocytes are stimulated to produce both TH1 and TH2 cytokines, which have also been demonstrated in the nasal polyp . The consequence of these findings may be the upregulation and increased survival of eosinophils in the nasal polyp . SUMMARY: Staphylococcus aureus is present in the mucin adjacent to nasal polyps in about 60 to 70% of cases of massive nasal polyposis . These organism, as studied up to the present, always produce exotoxins, which may act as superantigens, causing activation and clonal expansion of lymphocytes with specific Vbeta region in the lateral wall of the nose . The present review suggests that activation of these lymphocytes produce both TH1 and TH2 cytokines . The potential damage to the nasal mucosa from eosinophils is briefly discussed . Theoretically, topical antibiotics to suppress the colonization of S . aureus may be a logical approach to downregulate the production of superantigen in the lateral wall of the nose after appropriate endoscopic sinus surgery. Indian J Med Res, 2004 Dec, 120(6), 546 - 52 In vitro activity of linezolid & quinupristin/dalfopristin against Gram-positive cocci; Tunger A et al.; BACKGROUND & OBJECTIVES: Since the incidence of vancomycin- and methicillin-resistant Grampositive infections continue to increase, novel antimicrobials such as linezolid and streptogramin may provide new options to treat patients . The aim of this study was to investigate in vitro susceptibility of Enterococcus faecium resistant to glycopeptides, coagulase negative staphylococci and S . aureus resistant to methicillin isolated mainly from blood and also rectal swab cultures of patients against quinupristin/dalfopristin and linezolid . METHODS: The in vitro susceptibility to linezolid and quinupristin/dalfopristin for a total of 332 isolates of Gram-positive cocci {127 methicillin-resistant Staphylococcus aureus, 109 methicillinresistant coagulase negative staphylococci (71 S . epidermidis, 38 S . haemolyticus) and 96 vanA genotype vancomycin-resistant Enterococcus faecium} was investigated by E test . RESULTS: All MRSA and MRCoNS isolates were susceptible to linezolid (MICs < 4.0 mg/l) . Ninety per cent of VRE isolates were inhibited by linezolid at concentration of 2.0 mg/l and presented similar activities to quinupristin/dalfopristin . MICs for quinupristin/dalfopristin against staphylococci were also low (MIC(90) = 1.0 mg/l for both MRSA and MRCoNS isolates) . INTERPRETATION & CONCLUSION: The results of the present study demonstrated that quinupristin/ dalfopristin and linezolid, have good in vitro activity against MRSA, MRCoNS and vancomycin resistant E . faecium in Turkey . These drugs could be promising therapeutic options in an era of rapidly growing antibiotic resistance in all parts of world. J Dairy Sci, 2005 Feb, 88(2), 577 - 9 Short communication: association between milk yield at dry-off and probability of intramammary infections at calving; Rajala-Schultz PJ et al.; The association between milk yield at dry-off and intramammary infections at calving was evaluated from 116 lactations in one herd . Duplicate quarter samples were collected within 3 d after calving to estimate prevalence of intramammary infection at calving; information on cows' parity, daily milk yields, weekly somatic cell counts, and dry-off and calving dates were available for the data analyses . Generalized linear models with logit link were used to model the probability of a cow or a quarter being infected at calving, accounting for the clustered data . Increasing milk production at dry-off was a significant risk factor for both a cow and a quarter being infected with environmental pathogens at calving, but infections caused by coagulase-negative staphylococci at calving were not associated with milk yield at dry-off . For every 5-kg increase in milk yield at dry-off above 12.5 kg, the odds of a cow having an environmental intramammary infection at calving increased at least by 77%. J Antimicrob Chemother . 2005 Jan 13; {Epub ahead of print} Evaluation of cefoxitin 5 and 10 {micro}g discs for the detection of methicillin resistance in staphylococci; Skov R et al.; OBJECTIVE: To evaluate cefoxitin 5 and 10 microg discs for detection of methicillin resistance in staphylococci . METHODS: Six hundred and forty-one Staphylococcus aureus (261 mecA-negative and 380 mecA-positive) and 344 coagulase-negative staphylococci (CoNS) (132 mecA-negative and 212 mecA-positive) were investigated . The CoNS represented nine species, Staphylococcus epidermidis being the most frequent (n=231) . All isolates were tested using semi-confluent growth on Iso-Sensitest agar (ISA), and Mueller-Hinton agar (MH) using a 5 and a 10 microg cefoxitin disc and overnight incubation in ambient air at 35-37 degrees C . RESULTS: For S . aureus, both cefoxitin discs performed with high accuracy on both media . The sensitivity and specificity for the following proposed interpretive zone diameters were: ISA 5 microg, R < 14 mm (99.5% and 98.1%); ISA 10 microg, R < 22 mm (99.5% and 98.1%); MH 5 microg, R < 12 mm (99.7% and 98.1%); and MH 10 microg, R < 18 mm (99.5% and 98.9%), respectively . All four variants were superior to oxacillin using the former SRGA methodology . In CoNS, a substantial overlap was seen for all variants . However, by avoiding primary interpretation in the overlapping interval, highly accurate results could be obtained for 81%, 80%, 91% and 97% of the isolates, respectively . CONCLUSION: For S . aureus, cefoxitin 5 and 10 microg discs performed with high accuracy on both ISA and MH using semi-confluent growth and standard incubation conditions . With the introduction of a defined interval in which primary interpretation should be avoided, the method could also be used for CoNS. J Antimicrob Chemother . 2005 Jan 13; {Epub ahead of print} Enhanced resistance to erythromycin is conferred by the enterococcal msrC determinant in Staphylococcus aureus; Reynolds E et al.; The msrC gene, found on the chromosome of Enterococcus faecium, shares a high degree of similarity with the staphylococcal erythromycin resistance determinant msr(A) . The enterococcal determinant was cloned into Staphylococcus aureus to determine whether msrC could confer antibiotic resistance in staphylococci . A shuttle vector comprising pBluescript and pSK265 was used to introduce multiple copies of msrC into S . aureus RN4220 . The integration vector pCL84 was employed to insert a single copy of msrC into the S . aureus chromosome . MICs were determined by the broth microdilution method . Expression of msrC from both chromosomal and plasmid loci in erythromycin-susceptible S . aureus RN4220 (MIC 0.25 mg/L) gave rise to enhanced protection against erythromycin, with an MIC of 32-64 mg/L for S . aureus RN4220 containing msrC in multiple copies and an MIC of 16-64 mg/L with msrC inserted as a single copy in the S . aureus chromosome . MsrC mediates high-level resistance to erythromycin in S . aureus. J Microbiol Methods, 2005 Mar, 60(3), 413 - 6 Rapid detection of methicillin resistance in teicoplanin-resistant coagulase-negative staphylococci by a penicillin-binding protein 2' latex agglutination method; Knausz M et al.; Thirty-five clinical isolates of coagulase-negative staphylococci with decreased glycopeptide sensitivity were examined by a penicillin-binding protein (PBP2') latex agglutination (LA) test and were compared to the detection of the mecA gene by PCR, and oxacillin susceptibility determined minimum inhibitory concentrations . The latex test demonstrated high sensitivity and specificity for detecting methicillin resistance in coagulase-negative staphylococci after PBP2' induction with oxacillin. Heart Lung, 2005 Jan-Feb, 34(1), 69 - 71 Methicillin-resistant Staphylococcus aureus prosthetic aortic valve endocarditis with paravalvular abscess treated with daptomycin; Mohan SS et al.; Prosthetic valves have been used extensively for severe cardiac valvular dysfunction for the past 3 decades . Prosthetic cardiac valves may be infected with organisms causing bacteremia, particularly gram-positive cocci . Staphylococcus epidermidis (coagulase negative staphylococci) and Staphylococcus aureus , both methicillin-susceptible S . aureus and methicillin-resistant S . aureus (MRSA) strains, are the most frequent pathogens causing prosthetic valve endocarditis (PVE) . Vancomycin has been the cornerstone of therapy for serious MRSA infections including bacteremia and endocarditis . Clinicians have noted that MRSA bacteremias treated with vancomycin often fail to clear even with prolonged therapy . Persistent or prolonged MRSA bacteremia unresponsive to vancomycin therapy has led to the treatment of these infections by other agents, that is, quinupristin, dalfopristin, linezolid, or daptomycin . These antibiotics have been found particularly useful in treating MRSA bacteremias unresponsive to vancomycin therapy . We report a case of a patient who presented with MRSA PVE complicated by perivalvular aortic abscess with persistent MRSA bacteremia unresponsive to vancomycin therapy . The patient's MRSA bacteremia was cleared with daptomycin therapy (6 mg/kg/d) . Because the patient refused surgery, daptomycin therapy was continued in hopes of curing the endocarditis and sterilizing the perivalvular aortic abscess . Transesophageal echocardiogram revealed a decrease in abscess in the aortic perivalvular abscess after 1 week of daptomycin therapy . The patient made an uneventful recovery . The cure of PVE and perivalvular abscesses usually requires removal of the prosthetic device and abscess drainage . In this case, in which surgery was not an option, medical therapy of PVE and a decrease in size of the aortic perivalvular abscess were accomplished with daptomycin therapy . Daptomycin is an alternative to vancomycin therapy in patients with prolonged or persistent MRSA bacteremia secondary to endocarditis or abscess. Int J Hematol, 2004 Dec, 80(5), 467 - 9 Second transplantation from HLA 2-loci-mismatched mother for graft failure due to hemophagocytic syndrome after cord blood transplantation; Tanaka T et al.; A 7-year-old girl with acute myelogenous leukemia with multilineage dysplasia received unrelated cord blood transplantation but developed hemophagocytic syndrome (HPS) after sepsis with methicillin-resistant coagulase-negative staphylococci before engraftment . Bone marrow aspiration on day 20 revealed a markedly increased number of activated macrophages showing hemophagocytosis . The presence of donor-type chimera in the bone marrow was confirmed at that time . We therefore quickly started immunosuppressive and antibacterial treatment . Although her condition gradually improved, the patient suffered graft failure due to HPS . She received peripheral blood stem cell transplantation from her HLA 2-loci-mismatched mother on day 54 and continued in complete remission 12 months after the second transplantation . The results in this case suggested that because of fetomaternal microchimerism it may be useful to select an HLA-haploidentical mother as a backup donor for stem cell transplantation. Int J Clin Pract, 2004 Dec, 58(12), 1112 - 4 Slime factor positivity in coagulase negative staphylococci isolated from nasal samples of haemodialysis patients; Altoparlak U et al.; Polysaccharide slime seems to be the most important factor by which coagulase negative staphylococci (CNS) strains adheres and colonises catheters . The aim of this study was to determine the prevalence of slime-producing CNS strains isolated from nasal samples of patients on haemodialysis and of healthy persons as a control . Nasal cultures were performed for 131 haemodialysis patients and 56 healthy persons . CNS strains were isolated from 86 of 131 patients (72.9%), and 46 of 56 healthy persons (82.1%) . Twenty-four (27.9%) of the 86 CNS strains tested in the patient group and four (8.7%) of the 46 CNS strains in the control group were slime factor positive . There was a significant difference (p<0.01) in slime production between CNS strains isolated from haemodialysis patients and from control group . The detection of slime-producing coagulase-negative staphylococcus carriage in haemodialysis patients may prevent dialysis catheter-related coagulase-negative staphylococcal infections. Oral Health Prev Dent, 2003, 1(1), 29 - 35 Propolis extract as an adjuvant to periodontal treatment; Gebaraa EC et al.; PURPOSE: The aim of this study was to evaluate the effect of subgingival irrigation with propolis extract by clinical and microbiological parameters . MATERIALS AND METHODS: Twenty patients diagnosed with chronic periodontitis presenting three non-adjacent teeth with deep pockets were selected . After scaling and root planing, the selected periodontal sites were submitted to one of the following treatments: irrigation with a hydro alcoholic solution of propolis extract twice/week for two weeks (group A); irrigation with a placebo twice/week for two weeks (group B); or no additional treatment (C) . Subgingival plaque sampling and scaling and root planing were performed two weeks after clinical data recording . Two weeks later irrigation procedures were started (Baseline) . Microbiological and clinical data were collected at baseline, and after 4, 6 and 24 weeks . RESULTS: A decrease in total viable counts of anaerobic bacteria (p=0.007), an increase in the proportion of sites with low levels (< or = 10(3) cfu/mL) of Porphyromonas gingivalis (p=0.005), and a decrease in the number of sites with detectable presence of yeasts (p=0.000) were observed in group A sites when compared to group B and C sites . Propolis treatment did not lead to an increase in organisms such as coagulase positive Staphylococci and Pseudomonas spp . 24 weeks after treatment there was an increased proportion of sites showing probing depth (PD) < or = 3 mm in Group A sites . CONCLUSION: Subgingival irrigation with propolis extract as an adjuvant to periodontal treatment was more effective than conventional treatment both by clinical and microbiological parameters. APMIS, 2004 Dec, 112(11-12), 815 - 37 Genetic methods for detection of antimicrobial resistance; Sundsfjord A et al.; Sundsfjord A, Simonsen GS, Haldorsen BC, Haaheim H, Hjelmevoll SO, Littauer P, Dahl KH . Genetic methods for detection of antimicrobial resistance . APMIS 2004;112:815-37.Accurate and rapid diagnostic methods are needed to guide antimicrobial therapy and infection control interventions . Advances in real-time PCR have provided a user-friendly, rapid and reproducible testing platform catalysing an increased use of genetic assays as part of a wider strategy to minimize the development and spread of antimicrobial-resistant bacteria . In this review we outline the principal features of genetic assays in the detection of antimicrobial resistance, their advantages and limitations, and discuss specific applications in the detection of methicillin-resistant Staphylococcus aureus, glycopeptide-resistant enterococci, aminoglycoside resistance in staphylococci and enterococci, broad-spectrum resistance to beta-lactam antibiotics in gram-negative bacteria, as well as genetic elements involved in the assembly and spread of antimicrobial resistance. J Clin Microbiol, 2005 Jan, 43(1), 179 - 85 First report of vancomycin-resistant staphylococci isolated from healthy carriers in Brazil; Palazzo IC et al.; Reduced susceptibility or resistance to vancomycin has been reported among clinical isolates of staphylococci in previous studies . In the present study we report on the isolation of four vancomycin-resistant staphylococcal strains from healthy carriers inside and outside the hospital environment . These carriers did not receive treatment with any antibiotic . All coagulase-negative staphylococcal strains showed variable levels of resistance to several antimicrobial agents, including oxacillin, and unstable resistance to vancomycin, with decreased vancomycin MICs (<4 mg/liter) after 10 days of passage in a nonselective medium . However, exposure of these revertants to vancomycin selected staphylococcal strains resistant to vancomycin at very high frequencies (10(-2) and 10(-3)) . The vancomycin resistance in these staphylococcal strains was not mediated by the van gene . The cell wall of the staphylococcal strains studied became thickest after culture in medium containing vancomycin, and the differences in cell wall thickness were statistically significant (P < 0.001) . Thus, the thickening of the cell wall in these staphylococcal strains may be an important contributor to vancomycin resistance. Vnitr Lek, 2004 Oct, 50(10), 786 - 92 {Kikuchi-Fujimoto disease (histiocytic necrotizing lymphadenitis) . Case study and a literature review}; Olejarova M et al.; Kikuchi-Fujimoto disease is a rare feverish disease characterised by lymphadenopathy, the most frequently cervical, exanthem, arthralgias and arthritis . It affects especially young women . Patients have high erythrocyte sedimentation rate (ES) and leucopenia, antibodies are missing . Course of the disease is usually very benign and can subside spontaneously . However, clinical picture is usually very dramatic and can suggest infectious, autoimmune or malign systemic disease; also association with some of autoimmune diseases was described . Its occurrence is sporadic in all the world, the most of cases were seen in Asia, in the Czech Republic it has not been yet described . Therapy consists in antibiotics administration followed with corticoid therapy and usually can restore patients to perfect health . However, exacerbations have also been described . We describe a case of a 60 year old man, a past top sportsman, who has never been seriously ill except sport traumas and prosthesis implantation for coxarthritis reasons . The last two years he has suffered from exanthem and leucopenia of an unclear origin . In May 2002 he become feverish and arthritis, lymphadenopathy, splenomegalia and exanthem progression, high ES rate and high serum level of C-reactive protein (CRP) appeared in him . His condition was first evaluated as septic condition (founded staphylococci in two blood cultures), however, cause of potential sepsis has not been identified . The patient was treated with antibiotics with improvement of his total health condition after second treatment regiment . A neck node biopsy was done because of suspicion on lymphoprolipherative disease and histiocytic necrotizing lymphadenitis of Kikuchi type was found . Autoantibodies assessment was completely negative . After antibiotic and corticoid therapies his clinical condition quite quickly standardized and ES rate and serum CPR level decreased . 4 month after lowering the dose of prednisolon a temporary exacerbation of the disease appeared and again disappeared after increasing the dose of corticoid. J Infect Dis, 2005 Feb 1, 191(3), 435 - 43 Epub 2004 Dec 28. Intravital Fluorescence Microscopy: A Novel Tool for the Study of the Interaction of Staphylococcus aureus with the Microvascular Endothelium In Vivo; Laschke MW et al.; Background . The ability of Staphylococcus aureus to adhere to endothelial cells is a major prerequisite for the tissue-invasive stage of bacterial infection.Methods . To develop a model for the study of endothelial attachment and detachment kinetics of S . aureus within the host's microvasculature in vivo, we labeled inactivated staphylococci with fluorescein isothiocyanate and investigated their interaction with the vascular endothelium of arterioles, capillaries, and venules in the dorsal skin-fold chamber of untreated and tumor necrosis factor (TNF)- alpha -treated hamsters by use of intravital fluorescence microscopy.Results . During the first 20 min after injection, >99% of the bacteria were removed from the microvascular bloodstream . In parallel, single bacteria and bacterial clusters adhered to the endothelial lining of postcapillary venules and to nutritive capillaries . Bacterial adherence to the endothelium of arterioles was only rarely observed . TNF- alpha treatment significantly accelerated bacterial clearance and resulted in a significant increase of venular, but not arteriolar and capillary, bacterial adherence, indicating the venular endothelium to be the target structure for bacterial recruitment.Conclusion . The insights into host-pathogen interaction gained with this new in vivo model offer highly promising novel aspects of the understanding of infections caused by S . aureus. Zhonghua Yi Xue Za Zhi, 2004 Nov 17, 84(22), 1867 - 71 {Comparison of pharmacokinetics/pharmacodynamics of cefdinir, cefpodoxime proxetil and cefaclor against common bacteria of community acquired infections.}; Xiao YH et al.; OBJECTIVE: To compare the pharmacokinetics/pharmacodynamics property of cefdinir, cefpodoxime proxetil and cefaclor against common bacteria of community acquired infections and evaluate the recommended regimens . METHODS: The antibacterial activities of 3 agents against 238 clinical isolates were determined by standard agar dilution test and the pharmacokinetics of these antibiotics in male healthy volunteers were conducted in Latin-square manner . The time over MIC (T > MIC) of serum antibiotic concentrations were calculated with pharmacokinetic equation and MIC . RESULTS: The value of MIC(90)s cefdinir against these bacterial strains except penicillin non-sensitive pneumococci were 0.031 - 1 mg/L . Cefpodoxime held similar antibacterial activity with cefdinir, but was less potent against staphylococci . Cefaclor had much higher MIC values than other two drugs . After oral administration of 250 mg cefaclor, the drug concentration quickly reached peak concentration of 4.95 mg/L +/- 2.41 mg/L and the eliminative half time was 0.69 h +/- 0.6 h; the Tmax, Cmax and T(1/2)beta of cefdinir and cefpodoxime after oral administration of 100 mg were 2.5 h +/- 0.48 h, 0.81 mg/L +/- 0.19 mg/L, 1.73 h +/- 0.3 h and 2.38 h +/- 0.43 h, 1.12 mg/L +/- 0.28 mg/L, 1.92 h +/- 0.55 h, respectively . T > MIC of cefdinir in thrice daily administration were longer than 40% of medication interval against most of the tested isolates; no T > MIC period was found in cefpodoxime against staphylococci and the T > MICs of cefaclor after 250 mg oral administration were shorter than expected values against most bacteria . CONCLUSION: With powerful antibacterial activity, the T > MICs of cefdinir after 100 mg oral administration can meet with the clinical requirement in most infections; PK/PD value of cefpodoxime proxetil against staphylococci is lower than expectancy and 250 mg cefaclor 3 times daily is not enough to the treatment of common community acquired infections, the regimens of cefpodoxime proxetil and cefclor should be furtherly optimized. J Occup Environ Hyg, 2004 Oct, 1(10), 639 - 47 Evaluation and determinants of airborne bacterial concentrations in school classrooms; Bartlett KH et al.; A survey of 39 elementary schools was undertaken to determine indoor air concentrations of bioaerosols within a coastal, temperate climatic zone in British Columbia, Canada . This article reports the results for airborne bacteria . Determinants of exposure were grouped into environmental (outdoor temperature, relative humidity, season, weather), ventilation and comfort parameter (indoor relative humidity, temperature, indoor CO2 concentration, indoor fungal concentration), and occupancy (number of occupants, activity levels, occupancy patterns) variables . Regression modeling was used to evaluate the association between these factors and measured concentrations of indoor mesophilic bacteria . Naturally ventilated rooms (47% of the total rooms surveyed) had higher bacterial counts than mechanically ventilated rooms (geometric mean 325 vs . 166 CFU/m3, respectively, p < 0.001) . In univariate analyses, bacterial counts were negatively correlated with supply and exhaust flow rates, air exchange rates, and the percentage of the day that occupants spent quietly sitting at their desks . Analysis of bacterial groups indicated various sources of the bacterial aerosol, with micrococci and staphylococci closely associated with occupancy variables, Bacillus with site and occupancy, and coryneform bacteria with site variables only . Approximately 60% of the variance in the outcome measurement of total bacteria was accountedfor by indoor CO2, lower air exchange rate, the age of the building, signs of current or old moisture stains, room volume, indoor relative humidity, and occupant activity level in a multiple linear regression model, with ventilation factors accounting for 40% of the variance. Diagn Microbiol Infect Dis, 2005 Jan, 51(1), 57 - 62 Evaluation of alternative disk diffusion methods for detecting mecA-mediated oxacillin resistance in an international collection of staphylococci: Validation report from the SENTRY antimicrobial surveillance program; Pottumarthy S et al.; To validate the current National Committee for Clinical Laboratory Standards recommendations of the cefoxitin disk as a preferred surrogate marker to detect oxacillin resistance in staphylococcal isolates, 304 staphylococcal isolates originating from 49 sites in 16 countries in the SENTRY Antimicrobial Surveillance Program (2003) were tested . Two hundred three Staphylococcus aureus and 101 coagulase-negative staphylococci (CoNS), of which >95% were bloodstream isolates, were evaluated by comparing the results of the National Committee for Clinical Laboratory Standards broth microdilution method for oxacillin with those of the disk diffusion test using oxacillin, cefoxitin and ceftizoxime disks . Discrepancies were resolved using the PBP2a latex agglutination test . For S . aureus, the cefoxitin disk performed without interpretive error followed by the ceftizoxime disk (1% major and 0.5% minor errors; >/=20 mm = susceptible); use of the oxacillin disk test had the highest error rates with 4.4% major and 1.5% minor errors, whereas the oxacillin minimal inhibitory concentration (MIC) test was 99.0% accurate . For CoNS, the oxacillin disk test had the highest error rate with 4.0% major errors, followed by the cefoxitin (3.0% major error rate) and the ceftizoxime (1% very major and 1% minor error: >/=20 mm = susceptible) disk tests . The oxacillin MIC test was also 99.0% accurate for CoNS testing . Modification of the ceftizoxime disk diffusion breakpoints for CoNS resulted in complete intermethod categorical agreement . The overall accuracy of the four tests was as follows: modified ceftizoxime disk (99.3%) > oxacillin MIC = cefoxitin disk (99.0%) > current ceftizoxime disk (98.4%) > oxacillin disk (94.7%) . In conclusion, these results confirm the superior performance characteristics of cefoxitin and ceftizoxime disk tests as surrogate markers to detect oxacillin resistance; by using an international collection of clinically significant staphylococcal isolates, we also demonstrate its wide global application. Diagn Microbiol Infect Dis, 2005 Jan, 51(1), 13 - 7 Rapid detection of mecA and nuc genes in staphylococci by real-time multiplex polymerase chain reaction; Costa AM et al.; A multiplex real-time polymerase chain reaction (RT-PCR) targeting the mecA and nuc genes was developed for the detection of methicillin resistance and identification of Staphylococcus aureus . Novel mecA and nuc primers and fluorescence resonance energy transfer hybridization probes specific for the mecA and nuc genes were evaluated . The assay was performed using the LightCycler system (Roche Molecular Biochemicals, Mannheim, Germany) and evaluated against the traditional gel-based multiplex PCR (PCR-gel) method currently used at Royal Perth Hospital . Clinical isolates (n = 222) and isolates from a culture collection library (n = 206) were tested by both assays in parallel . The RT-PCR assay was 100% sensitive and specific for the detection of methicillin resistance and for the identification of S . aureus when compared with the PCR-gel assay . Results from the RT-PCR assay showed 5 isolates with lower efficiency fluorescence curves for the nuc gene PCR fragment . DNA sequencing showed mutations within the region of the probe-binding sites compared with the reference strain . The results of the RT-PCR assay were available within 2 h . This rapid mecA/nuc RT-PCR assay is a suitable and practical tool for the routine detection of methicillin resistance and identification of S . aureus, which can be easily incorporated into the diagnostic molecular microbiology laboratory work flow. Rhinology, 2004 Dec, 42(4), 213 - 8 Microbiology of chronic hyperplastic sinusitis; Kostamo K et al.; BACKGROUND: Patients with chronic hyperplastic sinusitis (CHS) form a heterogeneous group with similar symptoms and similar treatment despite of possible different mechanisms behind the disease . In the present study we focused on the microbiological findings in CHS and compared these results to the patient history in order to find out a possible explanation for the aetiology and chronicity of CHS . METHODS: In 30 patients the sinus mucus was collected under endoscopic sinus surgery . Samples from 20 healthy volunteers were collected by nasal lavage . Eosinophil staining, bacterial culturing and fungal staining and culturing were done . Histological samples were obtained from all patients . RESULTS: Bacterial cultures were positive in 93% of the patients compared to 70% in controls . Staphylococcus aureus and coagulase-negative Staphylococci were the two most common findings in both groups . A total of seven patients had positive fungal finding . The only fungal genus found was Aspergillus . In the control group no samples were positive for fungi . CONCLUSIONS: Microbiological findings do not seem to explain the chronic course of CHS, but fungi may play some part in the pathophysiology of the disease . These results may be more a reflection of a change in the environment in the paranasal sinuses and a change in normal flora than the actual cause of CHS. Ann Pharmacother . 2004 Dec 28; {Epub ahead of print} Antibiotic Lock Technique: A Review of the Literature (February); Segarra-Newnham M et al.; OBJECTIVE: To review the literature on the use of the antibiotic lock technique (ALT) as a treatment option for patients with highly needed catheters . DATA SOURCES: MEDLINE and International Pharmaceutical Abstracts were searched (1980-August 2004) . Search terms included antibiotic lock, catheter infection, and topical treatment . STUDY SELECTION AND DATA EXTRACTION: Articles describing use of ALT in the treatment of catheter infections in humans and studies evaluating in vitro stability of antibiotics were included . DATA SYNTHESIS: ALT has been used in patients with highly needed catheters, usually for parenteral nutrition, cancer chemotherapy, or dialysis . Catheters are considered highly needed when removal is not feasible or desirable due to lack of alternative injection sites for required therapy . Success rates in saving the infected catheter have been variable and may depend on the infecting organism . In addition, there are conflicting data in terms of compatibility of antibiotics with heparin solutions . CONCLUSIONS: Consensus appears to be that the ALT can be tried for patients with highly needed catheters when infection with coagulase-negative staphylococci is documented and no systemic signs of sepsis, such as hypotension, are evident . Most of these patients are likely to need systemic therapy as well . Infection of the catheter associated with systemic gram-negative bacteremia or fungemia will most likely require removal of the catheter to prevent systemic complications . Additional research with the ALT is warranted given unanswered questions. Int J Food Microbiol, 2005 Jan 15, 98(1), 73 - 9 Coagulase-positive Staphylococci and Staphylococcus aureus in food products marketed in Italy; Normanno G et al.; Staphylococcus aureus is a very common organism capable of producing several enterotoxins (SEs) that cause intoxication symptoms of varying intensity in humans when ingested through contaminated food . This paper reports the results of an investigation on the presence of Coagulase-Positive Staphylococci (CPS) and S . aureus in several food products marketed in Italy and on food contact surface swabs sampled from the food industry . A total of 11,384 samples were examined and 1971 of them (17.3%) were found to contain CPS . The assays performed on 541 CPS strains led to the identification of 537 S . aureus strains on which characterization of type A, B, C and D staphylococcal enterotoxins (SEA, SEB, SEC and SED) was performed . A total of 298 S . aureus strains (55.5%) produced one or more SEs: 33.9% of the strains produced SEC, 26.5% SEA, 20.5% SEA+SED, 13.4% SED, 2.7% SEB, 1.7% SEA+SEB, 0.7% SEC+SED and 0.3% produced SEA+SEC and SEB+SEC . The investigation highlighted that these organisms are very common and constitute a potential risk for consumers' health. Lett Appl Microbiol, 2005, 40(1), 24 - 9 The killing effect of cryptolepine on Staphylococcus aureus; Sawer IK et al.; Abstract i.k . sawer, m.i . berry and j.l . ford . 2004.Aim: This study was undertaken to further examine the antimicrobial actions of the alkaloid cryptolepine . Methods and Results: The minimum inhibitory concentration (MIC) of cryptolepine against Staphylococcus aureus was determined using the broth dilution method . Time-kill kinetics and scanning electron microscopy (SEM) techniques were employed to monitor the survival characteristics and the changes in morphologies respectively of staphylococci in the presence of cryptolepine . A notable antistaphylococcal activity was recorded for cryptolepine (MIC against S . aureus NCTC 10788 = 5 mug ml(-1)) . Cryptolepine appears to have a lytic effect on S . aureus as seen in SEM photomicrographs following 3, 6 or 24 h treatment with 4X MIC, i.e . 20 mug ml(-1) of cryptolepine . The surface morphological appearance of the staphylococcal cells was also altered . The lytic effect appeared to coincide with low viable counts recorded in survival curves following treatment with cryptolepine . Significance and Impact of the Study: These findings demonstrate that lysis occurs when susceptible organisms are exposed to cryptolepine. J Vet Med B Infect Dis Vet Public Health, 2004 Dec, 51(10), 449 - 54 Microbiological Features of Staphylococcus schleiferi subsp . coagulans, Isolated from Dogs and Possible Misidentification with Other Canine Coagulase-positive Staphylococci; Zdovc I et al.; Summary Staphylococcus schleiferi subsp . coagulans has only rarely been isolated and identified from the external auditory meatus of dogs suffering from external otitis . Its morphological and basic biochemical characteristics are of relatively little value for identification, as it phenotypically resembles another coagulase-positive staphylococci (CPS) and, consequently, may be easily misidentified as S . intermedius or even as S . aureus . In the present work, differentiation of S . schleiferi ssp . coagulans was therefore based on specific biochemical and genetic methods . All the strains were evaluated with the following commercial methods: Api Staph System (bioMerieux, Marcy l'Etoil, France), BBL Crystal Identification Systems (Gram-Positive ID Kit and Rapid Gram-Positive ID Kit; Becton Dickinson), and GEN-PROBE((R)) AccuProbe, Staphylococcus aureus identification test (bioMerieux) . Gram-Positive ID System/GP database includes the broadest range of staphylococcal species and correctly identifies the majority of strains important in veterinary medicine . Therefore, it is an acceptable alternative to conventional methods for identification of canine staphylococcal isolates . Reliable differentiation of S . aureus from S . schleiferi ssp . coagulans and S . intermedius was feasible with AccuProbe for S . aureus, which gave positive results only for S . aureus; all other CPS tested were negative. J Small Anim Pract, 2004 Dec, 45(12), 591 - 7 Methicillin-resistant Staphylococcus aureus in dogs and cats: an emerging problem? Duquette RA, Nuttall TJ. There is concern over transmission of methicillin-resistant Staphylococcus aureus (MRSA) between animals and humans . The spread of hospital-acquired and community-acquired MRSA is a major challenge in human medicine . MRSA is rarely isolated from animals but methicillin resistance occurs in staphylococci that are more prevalent in animals . MRSA infections in animals are uncommon and most are associated with exposure to medical hospitals, extensive wounds, prolonged hospitalisation and immunosuppression . The risk to human health appears to be small but a survey of methicillin-resistant staphylococci in animals is required . Thorough investigation of possible zoonotic infections to establish linkage is encouraged . Medical and veterinary staff should appreciate that animals can carry MRSA, cooperate in eliminating infections and monitor animals in medical environments . Veterinary clinics should implement guidelines for dealing with MRSA . Responsible antibiotic use should minimise the spread of antibiotic resistance but a UK monitoring scheme is desirable. Pathol Biol (Paris), 2004 Dec, 52(10), 575 - 578 Laurain C, Durand PY, Albert M, Weber M, Kessler M, Chanliau J, Dailloux M. The purpose of this study was to analyse the microbiological characteristics of infectious peritonitis in patients undergoing continuous ambulatory peritoneal dialysis . This study was conducted at the CHU Nancy from 1999 to 2002 . The diagnosis of peritonitis was based on cloudy peritoneal effluent (>100 cells per mm(3)) with an elevated leukocyte count (>50%), on isolation of bacteria or fungi and on symptoms such as abdominal discomfort or pain . The majority of infections associated with continuous ambulatory peritoneal dialysis were caused by Gram-positive bacteria (68%), Gram-negative bacteria (31%), and Candida (1%) . The coagulase-negative staphylococci were the most common cause of peritonitis . The antibiotic sensitivity of species corresponded to community-acquired isolation. J Infect Dis, 2005 Jan 1, 191(1), 138 - 44 Epub 2004 Nov 30. Corticoids normalize leukocyte production of macrophage migration inhibitory factor in septic shock; Maxime V et al.; BACKGROUND: A regulatory loop between macrophage migration inhibitory factor (MIF) and glucocorticoids has been characterized in animal models . Renewed interest in glucocorticoid treatment for septic shock offers an opportunity to analyze this regulatory loop in humans . METHODS: We investigated the ex vivo release of MIF by peripheral blood mononuclear cells (PBMCs) sampled from glucocorticoid-treated and -untreated patients with septic shock . Blood was obtained, before glucocorticoid treatment, and within the first day of treatment, from patients with septic shock who required treatment with moderate doses of hydrocortisone and fludrocortisone . RESULTS: PBMCs from patients contained significantly higher amounts of MIF than cells from healthy control subjects . In culture, spontaneous release of MIF and release induced by lipopolysaccharide (LPS), heat-killed staphylococci, and red blood cell lysates were significantly higher in patients than in control subjects . PBMCs from patients treated with glucocorticoids showed a lower release of MIF in response to LPS, heat-killed Escherichia coli, and peptidoglycan than did PBMCs from untreated patients and showed levels similar to PBMCs from healthy control subjects . CONCLUSION: To our knowledge, MIF is the first proinflammatory cytokine in which ex vivo release by circulating cells is enhanced during sepsis . Glucocorticoid treatment normalized the release of MIF by circulating PBMCs from patients with septic shock. J Med Microbiol, 2004 Dec, 53(Pt 12), 1195 - 9 Evaluation of phenotypic methods for methicillin resistance characterization in coagulase-negative staphylococci (CNS); Caierao J et al.; Coagulase-negative staphylococci (CNS) are the major cause of nosocomial infections . Methicillin-resistant strains are particularly important because they narrow therapeutic options . Detecting methicillin resistance among CNS has been a challenge for years . The objective of this study was to determine the accuracy of an agar screening test (0.6 and 4 microg oxacillin ml(-1)), disc diffusion and the automated MicroScan system to characterize methicillin resistance among CNS . One hundred and seventy five strains were analysed: 41.1 % Staphylococcus epidermidis and 59.9 % other species; 69.1 % were mecA-positive . The results showed that the methods have optimal correlation with the detection of mecA gene for S . epidermidis, Staphylococcus hominis and Staphylococcus haemolyticus . However, accuracy of the tests is impaired when less common species are analysed . The only 100 % accurate test was agar screening with 4 microg oxacillin ml(-1). Pharmacotherapy, 2004 Dec, 24(12 Pt 2), 203S - 15S Resistance to antimicrobial agents: an update; Rybak MJ; Decades of overuse and misuse of antibiotics have turned the resistance problem into a crisis . Preventive and corrective measures are urgently needed . This article presents a broad overview of the status of antimicrobial resistance . Resistance in key bacterial pathogens such as enterococci, staphylococci, pneumococci, and gram-negative bacilli; resistance to fluoroquinolones, third-generation cephalosporins, and the carbapenems; and mechanisms of resistance are discussed . The article also reviews the adverse impact of resistance on outcomes for patients with nosocomial infections . Finally, a brief overview of how the medical community can face its moment of truth is presented. Biomaterials, 2005 May, 26(15), 2247 - 54 Characterisation of antibacterial copper releasing degradable phosphate glass fibres; Abou Neel EA et al.; Phosphate-based glass fibres (PGF) of the general formula Na(2)O-CaO-P(2)O(5) are degradable in an aqueous environment, and therefore can function as antibacterial delivery systems through the inclusion of ions such as copper . In this study, PGF with varying amounts of copper oxide (CuO) were developed for potential uses in wound healing applications . PGF with 0, 1, 5 and 10mol% CuO were produced with different diameters and characterised in terms of structural and antibacterial properties . The effect of CuO and fibre pulling speed on the glass properties were investigated using rapid differential scanning calorimetry, differential thermal analysis and X-ray diffraction . The effect of two fibre diameters on short-term (3h) attachment and killing against Staphylococcus epidermidis were investigated and were related to their rate of degradation in deionised water, as well as copper ion release measured using ion chromatography . Thermal analysis showed that there was a significant increase in the PGF glass transition temperature as the CuO content increased . There was a significant decrease in the rate of degradation with increasing CuO content and an increase in fibre diameter . Over 6h, both the amount and rate of copper ions released increased with CuO content, as well as a reduction in fibre diameter thus increasing the surface area to volume ratio . There was a decrease in the number of viable staphylococci both attached to the CuO-containing fibres and in the surrounding environment. J Clin Microbiol, 2004 Dec, 42(12), 5739 - 44 Simultaneous detection of Staphylococcus aureus and coagulase-negative staphylococci in positive blood cultures by real-time PCR with two fluorescence resonance energy transfer probe sets; Sakai H et al.; A real-time PCR assay that uses two fluorescence resonance energy transfer probe sets and targets the tuf gene of staphylococci is described here . One probe set detects the Staphylococcus genus, whereas the other probe set is specific for Staphylococcus aureus . One hundred thirty-eight cultured isolates, which contained 41 isolates of staphylococci representing at least nine species, and 100 positive blood cultures that contained gram-positive cocci in clusters were tested . This assay was 100% sensitive and 100% specific for the detection of the Staphylococcus genus and of S . aureus. Acta Microbiol Immunol Hung, 2004, 51(3), 371 - 84 Different staphylococcal strains elicit different levels of production of T-helper 1-inducing cytokines; Buzas K et al.; Cytokine production has been implicated in the pathogenic mechanisms of infections caused by the staphylococci, since these bacteria may act as strong cytokine inducers . To gain deeper insight into the Th1 immune response activated by these bacteria, we have analyzed the interferon (IFN), interleukin-12 (IL-12) and IL-18-inducing activities of different Staphylococcus aureus (S . aureus), S . epidermidis and S . saprophyticus strains in human monocytes and murine bone marrow macrophages . A large majority of the S . aureus strains elicited the simultaneous production of IL-12 p70 and IFN-alpha in the human monocytes, while the S . epidermidis and S . saprophyticus strains induced only a low level of production, if any, of these cytokines . Furthermore, a majority of the S . aureus strains induced significantly higher IL-12 p70 and IL-18 titers in the murine bone marrow macrophages than did the S . epidermidis and S . saprophyticus strains . As IL-12, IL-18 and IFN-alpha stimulate Th1 differentiation synergistically, we suggest that S . aureus strains bias the immune response toward a Th1 phenotype, whereas S . epidermidis and S . saprophyticus strains provide a weaker stimulus for the production of Th1-inducing cytokines, and accordingly possibly elicit a less extensive Th1-associated adaptive immunity. East Mediterr Health J, 2003 Jan-Mar, 9(1-2), 185 - 90 Use of thermonuclease testing to identify Staphylococcus aureus by direct examination of blood cultures; Kaplan NM; Blood cultures submitted to the Clinical Microbiology Laboratory, Queen Alia Military Hospital, Amman during 1999-2001 were examined to evaluate thermonuclease testing for identifying Staphylococcus aureus in blood culture broths growing gram-positive cocci . Of 170 cultures studied, 129 yielded gram-positive staphylococci and 41 yielded other gram-positive cocci . Toluidine blue-deoxynucleic acid agar plates were used to test for thermonuclease activity . Standard tube coagulase tests were performed on the isolates . Direct detection of thermonuclease activity in 76 blood culture broths containing gram-positive staphylococci showed 100% correlation with subsequent tube coagulase tests . The thermonuclease test provides a fast, specific and reliable confirmation of S . aureus bacteraemia by direct examination of blood culture broths that contain gram-positive cocci . This allows for timely, optimal antibiotic therapy. Antimicrob Agents Chemother, 2004 Dec, 48(12), 4754 - 61 Antistaphylococcal activity of WCK 771, a tricyclic fluoroquinolone, in animal infection models; Patel MV et al.; WCK 771, the arginine salt of S-(-)-nadifloxacin, was evaluated in animal models of staphylococcal infection and in vitro . For 302 methicillin-susceptible strains the MIC at which 50% of isolates are inhibited (MIC50) and the MIC90 of WCK 771 were 0.03 and 0.03 microg/ml, respectively, and for 198 methicillin-resistant strains the MIC50 and the MIC90 were 0.5 and 1.0 microg/ml, respectively . All methicillin-susceptible staphylococci were quinolone susceptible, and almost all methicillin-resistant staphylococci were quinolone resistant . WCK 771 was more potent than moxifloxacin, trovafloxacin, levofloxacin, and ciprofloxacin and had potency comparable to that of clinafloxacin . Only WCK 771 and clinafloxacin demonstrated strong potencies against vancomycin-intermediate Staphylococcus aureus strains (MICs = 1 microg/ml) . WCK 771 is not a substrate of the NorA pump, as evident from the lack of an effect of reserpine on the MICs and similar protective doses against infections caused by efflux-positive and -negative staphylococci . WCK 771 was effective by both the oral and the subcutaneous routes in mice infected intraperitoneally with quinolone-susceptible methicillin-susceptible S . aureus (MSSA) strains . For infections caused by quinolone-resistant methicillin-resistant S . aureus (MRSA) strains, the activity of WCK 771 administered subcutaneously was superior to those of trovafloxacin and sparfloxacin, with a 50% effective dose range of 27.8 to 46.8 mg/kg of body weight . The activity of WCK 771 was superior to those of moxifloxacin, vancomycin, and linezolid in a mouse cellulitis model of infection caused by one MSSA and two MRSA strains, with effective doses of 2.5 and 5 mg/kg for the MSSA strain and 10-fold higher effective doses for MRSA strains . WCK 771, like vancomycin and linezolid, eradicated MRSA from mouse liver, spleen, kidney, and lung when it was administered subcutaneously at a dose of 50 mg/kg for four doses . These studies have demonstrated the effectiveness of WCK 771, administered orally and parenterally, for the treatment of diverse staphylococcal infections in mice, including those caused by quinolone-resistant strains. Clin Orthop, 2004 Oct, (427), 28 - 36 Eradication by surfactant irrigation of Staphylococcus aureus from infected complex wounds; Huyette DR et al.; Treatment of infected orthopaedic hardware usually requires the removal of the appliance . When the device is removed and immediately replaced, persistent infection frequently complicates this exchange procedure . We modeled the exchange procedure in rats by passing a wire suture through a posterior spinous process and then contaminating the wound with Staphylococcus aureus . We then investigated whether a sequence of surfactant enriched irrigation solutions (Castile soap followed by benzalkonium chloride, sequential surfactant irrigation) had a greater capacity to eradicate Staphylococcus aureus from the experimental wound than did the standard wound irrigant, normal saline . When we left the wire in place through the 2-week course of the study, sequential surfactant irrigation showed only a modest advantage over normal saline (staphylococci recovered from 39% versus 58% of wound cultures respectively) . Simple removal of the wire 24 hours after implantation and bacterial contamination prevented wound infection in most animals (with the wire removed, 38% of the animals remained infected versus 85% with the wire left in place), without regard to the irrigation solution . Alternatively, when we removed the wire after 24 hours, irrigated the wound, and then placed a fresh wire back into the wound, sequential surfactant irrigation showed a significant advantage over NS (54% of the animals irrigated with sequential surfactants remained infected versus 100% of the animals irrigated with normal saline) . Our findings confirm the importance of a contaminated medical device for promoting foreign body infection; our findings also show that sequential surfactant irrigation has therapeutic value in a rat model of orthopaedic device infection; this irrigation protocol should be studied further as a potential agent for the treatment of infected orthopaedic wounds. Clin Orthop, 2004 Oct, 1(427), 28 - 36 Eradication by Surfactant Irrigation of Staphylococcus aureus from Infected Complex Wounds; Huyette DR et al.; Treatment of infected orthopaedic hardware usually requires the removal of the appliance . When the device is removed and immediately replaced, persistent infection frequently complicates this exchange procedure . We modeled the exchange procedure in rats by passing a wire suture through a posterior spinous process and then contaminating the wound with Staphylococcus aureus . We then investigated whether a sequence of surfactant enriched irrigation solutions (Castile soap followed by benzalkonium chloride, sequential surfactant irrigation) had a greater capacity to eradicate Staphylococcus aureus from the experimental wound than did the standard wound irrigant, normal saline . When we left the wire in place through the 2-week course of the study, sequential surfactant irrigation showed only a modest advantage over normal saline (staphylococci recovered from 39% versus 58% of wound cultures respectively) . Simple removal of the wire 24 hours after implantation and bacterial contamination prevented wound infection in most animals (with the wire removed, 38% of the animals remained infected versus 85% with the wire left in place), without regard to the irrigation solution . Alternatively, when we removed the wire after 24 hours, irrigated the wound, and then placed a fresh wire back into the wound, sequential surfactant irrigation showed a significant advantage over NS (54% of the animals irrigated with sequential surfactants remained infected versus 100% of the animals irrigated with normal saline) . Our findings confirm the importance of a contaminated medical device for promoting foreign body infection; our findings also show that sequential surfactant irrigation has therapeutic value in a rat model of orthopaedic device infection; this irrigation protocol should be studied further as a potential agent for the treatment of infected orthopaedic wounds. Med Dosw Mikrobiol, 2004, 56(2), 127 - 31 {Determination of methicillin resistance in clinical strains of Staphylococcus aureus in routine microbiological diagnostics}; Slomka M et al.; The aim of this study was to compare the results of two methods: disc-diffusion test and ATB STAPH 5 (version 2000) test applied for determination of methicillin-resistant staphylococci . One hundred and twenty S . aureus strains isolated from clinical specimens in microbiological laboratory of Medical University in Warsaw were investigated . Identification and drug resistance were determined using automatic ATB Expression System . We compared the results of methicillin-resistant S . aureus detection by disc-diffusion method (oxacillin 1 microg) and ATB STAPH 5; version 2000 (bioMerieux sa) . In the case of 116 strains S . aureus identical results were obtained in both methods, 4 strains gave inconsistent results in these two methods . Results that were obtained show that the ATB STAPH 5 (version 2000) test is as useful for routine determination of MRSA and MSSA strains as the disc-diffusion method, and may be used alternatively. Diagn Microbiol Infect Dis, 2004 Nov, 50(3), 223 - 5 Improvement of a latex agglutination test for the evaluation of oxacillin resistance in coagulase-negative staphylococci; Corso A et al.; The "Slidex MRSA Detection" test (Denka Seiken, Japan) is a latex agglutination assay able to detect PBP2a . We evaluated its ability to differentiate mecA-positive from mecA-negative coagulase-negative staphylococci . We included 100 coagulase-negative staphylococci clinical isolates belonging to 9 species, 54 mecA positive and 46 mecA negative, as characterized by PCR . The specificity achieved using the manufacturer's instructions was 100%, but the sensitivity was only 57% . To increase sensitivity, we introduced modifications into the standard protocol . Using either large inocula or oxacillin induction before test performance, we achieved 100% sensitivity. Acta Vet Scand, 2004, 45(1-2), 37 - 45 Antimicrobial resistance in Staphylococcus spp., Escherichia coli and Enterococcus spp . in dogs given antibiotics for chronic dermatological disorders, compared with non-treated control dogs; Rantala M et al.; The aim of this study was to evaluate antimicrobial resistance in canine staphylococci, Escherichia coli and enterococci, which were isolated from 22 dogs with pyoderma and a history of previous antibiotic treatment, compared to bacterial isolates from 56 non-treated control dogs . Two isolates of each bacterial species per dog were investigated, if detected . Staphylococcal isolates from dogs with pyoderma (35 isolates) were more resistant to sulphatrimethoprim than the isolates from controls (56 isolates) (57% vs . 25%, p < 0.004) . Multiresistance in staphylococci was also more common in dogs with pyoderma (29% vs . 9%, p = 0.02) . A similar trend among isolates of E . coli was detected (24 and 74 isolates from treated and control dogs, respectively), but the differences were not significant . Resistance for macrolide-lincosamides was approximately 20% among staphylococci in both groups . Resistance to ampicillin among enterococci was 4%-7% . The age of the dogs might have an impact on resistance: multiresistance among staphylococcal isolates from younger dogs (< or = 5 years) was more common than in older dogs (26 years) (24%, vs . 0%, 63 and 27 isolates, respectively, p = 0.02) . Staphylococci in younger dogs were more resistant to tetracycline (48% vs . 11%, p < 0.001) and sulphatrimethoprim (48% vs . 15%, p < 0.01) than those in older dogs . In contrast, the isolates of E . coli from older dogs tended to be more resistant, although a significant difference was detected only in resistance to tetracycline (13% vs . 2% of 40 and 50 isolates respecthely, p = 0.04)) . The results of this small study indicate that resistance in canine staphylococci in the capital area of Finland is comparable with many other countries in Europe . Resistance in indicator bacteria, E . coli and enterococci, was low. J Antimicrob Chemother, 2004 Dec, 54(6), 1127 - 9 Epub 2004 Nov 05. Comparison of assays for detection of agents causing membrane damage in Staphylococcus aureus; O'neill AJ et al.; OBJECTIVES: To develop a novel beta-galactosidase leakage assay for Staphylococcus aureus and to evaluate this alongside other simple methods for detection of agents that cause membrane damage in staphylococci . METHODS: Using a PCR-based approach, a derivative of S . aureus RN4220 was constructed carrying the Escherichia coli lacZ gene under the control of the strong staphylococcal promoter, cap1A . Leakage of beta-galactosidase (BG) from this strain was examined after exposure for 10 min to various membrane-damaging agents at 4xMIC, using a fluorescence assay and the substrate 4-methylumbelliferyl-beta-d-galactoside . Other assays for membrane damage involving protoplast lysis (PL), leakage of material absorbing at 260 nm (OD) and ATP release as well as the BacLight (BL) assay were carried out using established methods . RESULTS: All the assays, with the exception of the PL assay, detected membrane damage induced by cetyltrimethylammonium bromide, nisin, clofazimine and protegrin IB-367 . However, the ability to detect membrane damage induced by these agents differed between the assay systems . The assays also varied considerably in their signal-to-noise ratio, with the ATP assay providing values for nisin approaching 100-fold that of the control . CONCLUSIONS: The PL assay is unsuitable for detection of membrane-damaging agents in S . aureus . The other assays, including the BG assay, detect membrane damage . The OD assay should be sufficient for most purposes since it is effective, rapid and cheap to perform . Studies requiring maximum sensitivity and discrimination should employ the ATP assay. Folia Microbiol (Praha), 2004, 49(4), 353 - 86 Molecular diagnostics of clinically important staphylococci; Stepan J et al.; Bacterial species of the genus Staphylococcus known as important human and animal pathogens are the cause of a number of severe infectious diseases . Apart from the major pathogen Staphylococcus aureus, other species until recently considered to be nonpathogenic may also be involved in serious infections . Rapid and accurate identification of the disease-causing agent is therefore prerequisite for disease control and epidemiological surveillance . Modern methods for identification and typing of bacterial species are based on genome analysis and have many advantages compared to phenotypic methods . The genotypic methods currently used in molecular diagnostics of staphylococcal species, particularly of S . aureus, are reviewed . Attention is also paid to new molecular methods with the highest discriminatory power . Efforts made to achieve interlaboratory reproducibility of diagnostic methods are presented. J Clin Microbiol, 2004 Nov, 42(11), 5245 - 8 Usefulness of routine epicardial pacing wire culture for early prediction of poststernotomy mediastinitis; Mekontso-Dessap A et al.; Poststernotomy mediastinitis (PSM) is one of the most serious complications of cardiac surgery, and its associated morbidity and mortality demand early recognition for emergency therapy . In this study, we investigated the usefulness of epicardial pacing wire (EPW) cultures for the prediction of PSM . Among 2,200 patients who underwent a cardiac surgical procedure at our hospital between 1 January 1999 and 31 December 2001, 82 (3.7%) had PSM; Staphylococcus aureus was the organism (45.1%) most frequently isolated at the time of surgical debridement . EPWs from 1,607 (73.0%) patients, 73 (4.5%) of whom developed PSM, were cultured . EPW cultures from 466 (29.0%) were positive, most often (74.9%) for coagulase-negative Staphylococci . EPW cultures were truly positive in 26 cases, truly negative in 1,106 cases, falsely positive in 428 cases, and falsely negative in 47 cases (with sterile cultures in 35 cases and a culture positive for an organism different from that isolated at the time of debridement in 12 cases) . EPW culture had a positive predictive value of only 5.7% and a high negative predictive value (95.9%) for the diagnosis of PSM, with an accuracy of 70.4% . However, the likelihood ratio of positive (1.27) and negative (0.89) tests indicated only small changes in pretest-to-posttest probability . Therefore, a strategy of routine culture of EPWs to predict PSM seems questionable. J Clin Microbiol, 2004 Nov, 42(11), 5041 - 6 Evaluation of the BD PHOENIX automated microbiology system for detection of methicillin resistance in coagulase-negative staphylococci; Horstkotte MA et al.; The new BD PHOENIX automated microbiology system (Becton Dickinson Diagnostic Systems, Sparks, Md.) is designed for automated rapid antimicrobial susceptibility testing and identification of clinically relevant bacteria . In our study, the accuracy and speed of the BD PHOENIX oxacillin MIC determination for detecting methicillin resistance was evaluated for 200 clinical isolates of coagulase-negative staphylococci (CoNS) . Compared to mecA PCR, the BD PHOENIX system detected methicillin resistance with a sensitivity of 99.2% . According to the actual NCCLS oxacillin MIC breakpoint of > or =0.5 microg/ml, the specificity was only 64.9%, attributable to false-positive results in 26 mecA-negative strains, including 16 non-Staphylococcus epidermidis strains . Alternative oxacillin breakpoints of > or =1, > or =2, and > or =4 microg/ml resulted in increased specificities of 83.8, 94.6, and 100% and high sensitivities of 99.2, 99.2, and 96.7%, respectively . Similarly, NCCLS broth microdilution oxacillin MICs exhibited a sensitivity of 100% but a low degree of specificity . However, the previous oxacillin MIC breakpoint of > or =4 microg/ml performed with a sensitivity of 98.4% and a specificity of 98.7% . BD PHOENIX oxacillin MIC results were available after 9 h for 40.5% of the examined CoNS strains and were completed after 17 h . Our results revealed the high reliability of the BD PHOENIX system as a phenotypic method for detection of resistance to oxacillin in mecA-positive CoNS . However, for the improvement of specificity, reevaluation of the optimal oxacillin MIC breakpoint for CoNS appears to be necessary. J Clin Microbiol, 2004 Nov, 42(11), 4988 - 95 Development and evaluation of a quality-controlled ribosomal sequence database for 16S ribosomal DNA-based identification of Staphylococcus species; Becker K et al.; To establish an improved ribosomal gene sequence database as part of the Ribosomal Differentiation of Microorganisms (RIDOM) project and to overcome the drawbacks of phenotypic identification systems and publicly accessible sequence databases, both strands of the 5' end of the 16S ribosomal DNA (rDNA) of 81 type and reference strains comprising all validly described staphylococcal (sub)species were sequenced . Assuming a normal distribution for pairwise distances of all unique staphylococcal sequences and choosing a reporting criterion of > or =98.7% similarity for a "distinct species," a statistical error probability of 1.0% was calculated . To evaluate this database, a 16S rDNA fragment (corresponding to Escherichia coli positions 54 to 510) of 55 clinical Staphylococcus isolates (including those of the small-colony variant phenotype) were sequenced and analyzed by the RIDOM approach . Of these isolates, 54 (98.2%) had a similarity score above the proposed threshold using RIDOM; 48 (87.3%) of the sequences gave a perfect match, whereas 83.6% were found by searching National Center for Biotechnology Information (NCBI) database entries . In contrast to RIDOM, which showed four ambiguities at the species level (mainly concerning Staphylococcus intermedius versus Staphylococcus delphini), the NCBI database search yielded 18 taxon-related ambiguities and showed numerous matches exhibiting redundant or unspecified entries . Comparing molecular results with those of biochemical procedures, ID 32 Staph (bioMerieux, Marcy I'Etoile, France) and VITEK 2 (bioMerieux) failed to identify 13 (23.6%) and 19 (34.5%) isolates, respectively, due to incorrect identification and/or categorization below acceptable values . In contrast to phenotypic methods and the NCBI database, the novel high-quality RIDOM sequence database provides excellent identification of staphylococci, including rarely isolated species and phenotypic variants. J Clin Microbiol, 2004 Nov, 42(11), 4947 - 55 New quadriplex PCR assay for detection of methicillin and mupirocin resistance and simultaneous discrimination of Staphylococcus aureus from coagulase-negative staphylococci; Zhang K et al.; Major challenges in diagnostic molecular microbiology are to develop a simple assay to distinguish Staphylococcus aureus from the less virulent but clinically important coagulase-negative staphylococci (CoNS) and to simultaneously determine their antibiotic resistance profiles . Multiplex PCR assays have been developed for the detection of methicillin- and mupirocin-resistant S . aureus and CoNS but not for the simultaneous discrimination of S . aureus from CoNS . We designed a new set of Staphylococcus genus-specific primers and developed a novel quadriplex PCR assay targeting the 16S rRNA (Staphylococcus genus specific), nuc (S . aureus species specific), mecA (a determinant of methicillin resistance), and mupA (a determinant of mupirocin resistance) genes to identify most staphylococci, to discriminate S . aureus from CoNS and other bacteria, and to simultaneously detect methicillin and mupirocin resistance . Validation of the assay with 96 ATCC control strains and 323 previously characterized clinical isolates, including methicillin- and mupirocin-sensitive and -resistant S . aureus and CoNS isolates and other bacteria, demonstrated 100% sensitivity, specificity, and accuracy . This assay represents a simple, rapid, accurate, and reliable approach for the detection of methicillin- and mupirocin-resistant staphylococci and offers the hope of preventing their widespread dissemination through early and reliable detection. Med Dosw Mikrobiol, 2004, 56(1), 19 - 27 {Coagulase-negative staphylococci in municipal waste}; Sobis-Glinkowska M et al.; The total of 259 coagulase-negative staphylococci isolates gained from aerosols created during dumping and utilisation of municipal waste were investigated . Species and subspecies identification allowed to determine predominance of novobiocin-resistant species which primarily colonise rodents or other animals . However, about 30% of isolates were species originating from humans . Metabolic properties of the studied group of isolates were examined, as well as their sensitivity to a set of 8 commonly used antibiotics: amoxicillin, erythromycin, tetracycline, clindamycin, chloramphenicol, gentamicin, trimethoprim/sulfamethoxazole and ciprofloxacin . Many isolates resistant to erythromycin (19%), tetracycline (13%) and a considerably smaller group resistant to clindamycin (6.2%), chloramphenicol (2%) and trimethoprim/sulfamethoxazole (1%) were found . It appeared that processes of occurring at the time of dumping and utilisation of waste affect the prevalence of only some features . The examined metabolic features were found to be relatively stable and no changes were observed, which would indicate the tendency of adaptation to existence in the inanimate matter environment . Only isolates with active nitrate reductase were more frequently detected . The increase in frequency of occurrence of isolates resistant to tetracycline and chloramphenicol and simultaneous elimination of isolates resistant to other antibiotics were observed. Med Dosw Mikrobiol, 2004, 56(1), 11 - 7 {Drug resistant coagulase-positive staphylococci strains in food}; Majczyna D et al.; From the 1572 food samples, examined in Microbiology Department of Frozen Food Industry Research Laboratory in Lodz, 79 (5.0%) coagulase-positive staphylococci strains were isolated . All the strains were sensitive to vancomycin, gentamycin and chloramphenicol . Only individual staphylococci strains were resistant to erythromycin (1.3%), lincomycin (2.5%) and ciprofloxacin (2.5%) . 20.3% strains, isolated mainly from raw meat, were resistant to doxycycline and 6.3% to oxacillin . 38.0% of coagulase-positive staphylococci strains had positive results of cefinase test . One strain isolated from minced meat was resistant to methicillin and at the same time it was producing beta-lactamases. Med Dosw Mikrobiol, 2004, 56(1), 1 - 9 {Molecular analysis of methicillin-resistant staphylococci isolated from hospital patients and in the out-patient clinic}; Malecka-Mikosz O et al.; In this study, a molecular analysis of the methicillin-resistant coagulase-negative staphylococci strains was performed . The obtained results of the biochemical and drug resistant pattern investigations were insufficient to assess the relationship between the strains . Therefore genotyping by the restriction fragment length polymorphism analysis (PCR-RFLP) method was performed . Analyzed strains characterized presence of the mecA gene-PCR products . The PCR products were digested with DraI and TasI, and the fragments separated by agarose gel electrophoresis . Typing of the methicillin-resistant gene using PCR-RFLP showed that all MRCNS strains possess an identical restriction pattern of the mecA gene . This identical restriction pattern of the mecA gene in investigated strains may suggest an easy transfer of this gene between different staphylococci species and lead to the spreading of methicillin-resistant among hospital strains . Furthermore performing the comparison of different phenotype and genotype methods has shown that the PCR-RFLP method is quick and reliable, enabling the detection and estimation of the relationship between MRCNS strains. FEMS Microbiol Lett, 2004 Nov 15, 240(2), 225 - 8 Rapid detection of Panton-Valentine leukocidin from clinical isolates of Staphylococcus aureus strains by real-time PCR; Deurenberg RH et al.; To allow rapid identification of Panton-Valentine leukocidin (PVL)-producing Staphylococcus aureus strains, a real-time PCR assay for detection of PVL was developed . This assay is convenient, since it can be applied directly on bacterial suspensions and does not require previous DNA purification . Furthermore, the assay was found to be highly reproducible, robust and specific, since positive results were generated exclusively with PVL-positive S . aureus strains, and neither with PVL-negative strains nor staphylococci other than S . aureus. Rhinology, 2004 Sep, 42(3), 131 - 6 The pathogenicity and antibiotic resistance of coagulase-negative Staphylococci isolated from the maxillary and ethmoid sinuses; Aral M et al.; AIM OF THE STUDY: To investigate the pathogenicity and antibiotic resistance of coagulase-negative staphylococci (CNS) isolated from the maxillary and ethmoid sinuses of patients undergoing endoscopic sinus surgery for chronic sinusitis . PATIENTS AND METHODS: Ninety-three patients (63 males, 30 females) aged between 19 - 68 years, who had undergone functional endoscopic sinus surgery (FESS) for chronic sinusitis, were included in the study . Nasal mucosa, skin and adjacent structures were cleansed with povidone-iodine solution before surgery to prevent a probable contamination . In all patients, nasal swabs were taken before and after the application of povidone-iodine solution . Colonies isolated and identified as Staphylococci in cultures were further investigated for pathogenicity and antibiotic susceptibility . Slime test was used to determine the pathogenicity of CNS . The relationship between antibiotic resistance of pathogenic and non-pathogenic CNS was compared by chi2 analysis . RESULTS: While bacterial growth rate was 62.3% in nasal swab cultures taken before the application of povidone-iodine solution, it decreased to 12.9% after the application of solution . Microorganisms were isolated in 95.6% of cultures taken from maxillary sinuses and in 91.3% of cultures obtained from ethmoid sinuses during the FESS . The most frequently isolated microorganism in each of the sinuses was CNS . Slime test was carried out in 30 CNS isolated . Twelve of these were slime positive and 18 were slime negative . While 83.3% of CNS isolated was resistant to penicilin, all of CNS were sensitive to vancomycin and teikoplanine . The difference between slime positive and slime negative CNS for gentamicin and ciprofloxacin resistance was statistically significant (p<0.05) . CONCLUSION: We consider that the pathogenicity tests like slime production and antimicrobial susceptibilities of CNS frequently isolated from the patients with chronic sinusitis should be investigated and also these microorganisms should be kept in mind in the selection of empiric treatment. Arch Pediatr, 2004 Nov, 11(11), 1314 - 8 {A routine prospective survey process to detect nosocomial bacterial colonization in a neonatal unit: risk factors for acquisition.}; Campeotto F et al.; A systematic analysis of weekly nasal and rectal swabs was carried out in a neonatal unit in order to detect colonization with multiresistant bacteria (MRB) . PATIENTS AND METHODS: During a 6-month period, rectal and nasal samples were taken in 187 consecutively hospitalized newborns, the day of the admission (day 0) and every week until discharge, in order to detect MRB, mainly methicillin-resistant coagulase negative staphylococci (MRCoNS), Staphylococcus aureus and multi-resistant Gram-negative bacilli . RESULTS: Among 187 infants, 50 were already colonized at entrance and excluded from the study . In others, 49 (35%) were colonized by at least one MRB, with a total of 71 strains isolated . The most frequent was MRCoNS, especially Staphylococcus epidermidis (66.1%) . Gram-negative bacilli accounted for 9.8% . Colonization began earlier with MRCoNS than with Gram-negative bacilli, 7.8 +/- 6 vs . 15.5 +/- 16 days, P=0.004, and finished earlier 22.7 +/- 15 vs . 38.5 +/- 16 days, P=0.03 . Colonized children exhibited by univariate analysis a lower birth weight, more frequent parenteral nutrition or previous hospitalization in a neonatal unit and a younger age at admission . Odds ratio for colonization were 4.06 for prematurity and 43.83 for a previous hospitalization . MRCoNS at days 15 (P <0.05) and 22 (P <0.05) were correlated with the empiric use of antibiotics . No nosocomial infection occurred during the study . CONCLUSION: A high rate of newborns were colonized with MRB in our unit, especially MRCoNS, acquired earlier than Gram-negative bacilli, with a favoring action of empiric antibiotherapy. Eur J Cancer, 2004 Nov, 40(16), 2452 - 8 Port-A-Cath infections in children with cancer; Hengartner H et al.; Implanted subcutaneous (s.c.) central venous port accesses including Port-A-Cath (PAC) facilitate the administration of chemotherapy or blood products and are frequently used in children with cancer . The incidence of PAC-related infections was determined in 155 consecutive paediatric cancer patients with PAC followed for a total of 134,773 days (median, 738; range, 25-2080) . Overall, 48 bloodstream infections occurred in 26 patients . 12 (25%) of these infections and 3 local infections at the insertion site were treatment-resistant and demanded removal of the PAC . Coagulase-negative staphylococci were involved in 12 of these 15 episodes . The rate of clearly PAC-related infections in this so far largest reported series was 0.11 episodes per 1000 PAC days, one of the lowest in the literature . Although catheter-related infections demanded PAC removal in 8% of our patients, the long periods PAC were in use and their benefits argue for continued PAC use in the paediatric cancer population. Int J Antimicrob Agents, 2004 Nov, 24(5), 444 - 9 Bactericidal activity of quinupristin-dalfopristin against strains of Staphylococcus aureus with the MLS(B) phenotype of resistance according to the erm gene type; Clarebout G et al.; The bactericidal activity of quinupristin-dalfopristin was assessed by time-kill experiments against Staphylococcus aureus strains with characterized phenotypes and genotypes of MLS(B) resistance . A set of laboratory strains composed of isogenic pairs of S . aureus RN4220 derivatives containing or not the erm(A), erm(B) or erm(C) genes constitutively expressed and of 13 clinical isolates containing these genes inducibly or constitutively expressed were studied . Three of the clinical isolates with erm(B) or erm(A) genes had an unusual inducible MLS(B) cross resistance . The early bactericidal activity of quinupristin-dalfopristin was altered against strains expressing constitutive quinupristin resistance regardless of the erm(A), erm(B) or erm(C) type of gene . We conclude that the bactericidal activity of quinupristin-dalfopristin against staphylococci was dependent on the activity of quinupristin rather than on the erm genotype of the strain. Antimicrob Agents Chemother, 2004 Nov, 48(11), 4240 - 5 Evolution of the antimicrobial resistance of Staphylococcus spp . in Spain: five nationwide prevalence studies, 1986 to 2002; Cuevas O et al.; Data regarding the evolution of Staphylococcus resistance in a whole country have a definite influence on the design of empirical treatment regimens . Nevertheless, incidence studies over long periods of time are expensive and very difficult to carry out . In order to ascertain the present situation of the antimicrobial resistance of Staphylococcus in Spain and the change of this resistance over time, we performed five point prevalence studies (1986 to 2002) in a large group of Spanish hospitals (from 68 institutions in 1986 to 143 in 2002) collecting all Staphylococcus strains isolated on a single selected day . All microorganisms were identified in the five studies at the same laboratory, and antimicrobial susceptibility testing was performed against 17 antimicrobial agents by the agar dilution method and a microdilution method . During this period, there was an overall increase in resistance to most antimicrobials among Staphylococcus aureus/coagulase-negative staphylococci, mainly to oxacillin (1.5%/32.5% in 1986 versus 31.2%/61.3% in 2002) (P < 0.001), erythromycin (7%/41.1% in 1986 versus 31.7%/63% in 2002) (P < 0.001), gentamicin (5.2%/25.4% in 1986 versus 16.9%/27.8% in 2002) (P < 0.001; P = 0.5), and ciprofloxacin (0.6%/1.1% in 1986 versus 33.9%/44.9% in 2002) (P < 0.001) . All of the isolates were uniformly susceptible to glycopeptides, linezolid, and quinupristin/dalfopristin . Resistance of S . aureus to trimethoprim/sulfamethoxazole was very low (from 0.5% to 2.1%) (P = 0.152) . Periodic performance of prevalence studies is a useful, inexpensive, and easy tool to know the nationwide situation of a microorganism and its resistance to antimicrobials; it also helps us assess the emergence and spread of antimicrobial resistance. Turk J Pediatr, 2004 Jul-Sep, 46(3), 251 - 5 Pediatric blood culture: time to positivity; Kara A et al.; The aim of this study was to determine how long it takes blood culture to become positive using a blood culture system that can be monitored continuously in pediatric patients . Data were collected prospectively on 1,000 positive blood culture results from a tertiary pediatric university hospital from April 2000 to May 2002 . The laboratory used the BACTEC 9120 fluorescent blood culture system . Patient's age ranged from less than a day to 20 years of age (mean 3 years) . Five hundred and four cultures (50.4%) out of 1,000 yielded coagulase negative staphylococcus (CNS), 81 (8.1%) S . aureus, 53 (5.3%) . Pseudomonas and 50 (5.0%) Klebsiella species . Of the 504 coagulase negative staphylococcal blood culture isolates, 314 (62.3% of CNS) were regarded as skin contaminants . Of the 1,000 cultures, 9.6% were reported as positive in the first day, 27.8% in the second day, 54.7% in the third day, 77.0% in the fourth and 89.4% in the fifth day . There was no association between previous antibiotic usage and the period required for isolate recovery . The clinician can expect to get results of positive blood cultures with susceptibility data, at a rate of 77.1% by day four and almost 90% by day five of sampling in the bacteriemic patient . Blood cultures yielding coagulase negative staphylococci in the first three days almost always show bacteremia with those microorganisms. Res Microbiol, 2004 Nov, 155(9), 755 - 60 Comparative evaluation of coagulase-negative staphylococci (CoNS) adherence to acrylic by a static method and a parallel-plate flow dynamic method; Cerca N et al.; The main goal of this work was to evaluate the adhesion to acrylic of several clinical strains of Staphylococcus epidermidis and Staphylococcus haemolyticus using both static and dynamic adhesion methods, and to compare the results obtained with these two methods . Adhesion was evaluated using the static slide method with different washing procedures, and the parallel plate flow chamber method . The extent of S . epidermidis adhesion, assessed by both methods, was greater than that of S . haemolyticus . The number of bacteria which adhered using the static method was lower than that using the dynamic method . It was found that the simple static method, when performed with an accurate washing procedure, can be as effective as the dynamic flow method for assessing differences in the adherence capacity of strains . Although the dynamic flow method yielded more overall information, its greater complexity and cost may not always justify its use for certain experimental comparisons . This investigation has shown that simple static adhesion methods, when performed accurately, can be used to evaluate differences in adhesion capacity. Clin Infect Dis, 2004 Nov 1, 39(9), 1285 - 92 Epub 2004 Oct 11. A randomized clinical trial to compare fleroxacin-rifampicin with flucloxacillin or vancomycin for the treatment of staphylococcal infection; Schrenzel J et al.; BACKGROUND: Oral combination therapy with fluoroquinolones plus rifampicin is a promising alternative to standard parenteral therapy for staphylococcal infections . METHODS: In a multicenter, randomized trial, we compared the efficacy, safety, and length of hospital stay for patients with staphylococcal infections treated either with an oral combination of a fluoroquinolone (fleroxacin) plus rifampicin or with standard parenteral treatment (flucloxacillin or vancomycin) . Patients were included if cultures showed the presence of bacteremia or deep-seated infections with Staphylococcus aureus (104 patients) or catheter-related bacteremia due to drug-susceptible, coagulase-negative staphylococci (23 patients) . RESULTS: The cure rate in the intention-to-treat analysis was 78% for the fleroxacin-rifampicin group (68 patients) and 75% for the standard therapy group (59 patients; 47 received flucloxacillin, and 12 received vancomycin); in the population of clinically evaluable patients (n=119), the cure rate was 82% and 80%, respectively; and in the population of microbiologically evaluable patients (n=103), the cure rate was 86% and 84%, respectively . Clinical and bacteriological failures after S . aureus infections were documented in similar proportions of patients . The median length of hospital stay after study entry was 12 days in the fleroxacin-rifampicin group, compared with 23 days in the standard treatment group (P=.006) . More adverse events probably related to the study drug were reported in the fleroxacin-rifampicin group than in the standard therapy group (15 of 68 vs . 5 of 59 patients; P=.05) . CONCLUSIONS: This study suggests that an oral regimen containing a fluoroquinolone plus rifampicin may be effective for treating staphylococcal infections, allowing earlier discharge from the hospital. Ann Hematol, 2005 Jan, 84(1), 40 - 6 Epub 2005 Jan. Bacterial and fungal bloodstream isolates from 796 hematopoietic stem cell transplant recipients between 1991 and 2000; Ortega M et al.; To examine shifts in the etiology, incidence, evolution, susceptibility, and patient mortality of bacterial and fungal bloodstream isolates (BSIs) from hematopoietic stem cell transplantation (HSCT) recipients, we reviewed the BSIs of 796 patients who underwent an HSCT in our institution during a 10-year period . Four hundred eighty-nine episodes of bacterial and fungal BSI were detected in 330 patients (41%) . Three hundred ten isolates (63%) were gram-positive bacteria, 142 (29%) were gram-negative, and 18 and 19 isolates were different species of anaerobic organism and Candida spp . (both 4%) . Coagulase-negative staphylococci (CoNS), with 210 isolates, were the organism most frequently isolated in each year of study and during the three phases of immune recovery after HSCT . The ratio of gram-positive to gram-negative has declined from 3.3 (1991-1992) to 1.8 (1999-2000) . Crude mortality occurred in 47 cases of 489 BSI episodes (10%) . Mortality according to groups was gram-negative, 7%; gram-positive, 9%; and anaerobic bacteria, 11% . Candida spp . was the group that accounted for the highest crude mortality, with 42% . Gram-positive microorganisms were isolated more often than gram-negative organisms, but the trend is reversing . CoNS were the leading pathogen during the 10 years of study and during the three phases of immune recovery after HSCT . Crude mortality of HSCT patients with BSI was low except for infections caused by Candida spp. Ann Rheum Dis, 2004 Nov, 63(11), 1450 - 2 The pustular skin lesions in Behcet's syndrome are not sterile; Hatemi G et al.; BACKGROUND: The pustular skin lesions of Behcet's syndrome (BS) are clinically and histopathologically similar to ordinary acne, but BS patients get lesions at sites not commonly involved in acne, such as the legs and arms . The microbiology of these lesions has not been studied adequately . OBJECTIVE: To make a detailed study of the microbiology of BS lesions . METHODS: Subjects were patients with BS and acne vulgaris . Material was extracted from pustular lesions and directly plated to aerobic and anaerobic media by sterile swab . Anaerobic bacteria were identified using a commercial kit (API 20A) . Aerobic bacteria were defined by standard procedures . RESULTS: 58 BS patients and 37 acne patients were studied . Pustules were cultured from the following sites: BS patients (70 pustules): face (17), back (30), chest (2), arm (4), leg (17); acne patients (37 pustules): face (27), back (6), chest (1), arm (2), leg (1) . At least one type of microorganism was grown from each pustule . Staphylococcus aureus (41/70, 58.6%, p = 0.008) and Prevotella spp (17/70, 24.3%, p = 0.002) were significantly more common in pustules from BS patients, and coagulase negative staphylococci (17/37, 45.9%, p = 0.007) in pustules from acne patients . CONCLUSIONS: The pustular lesions of BS are not usually sterile . The microbiology of these lesions is different from ordinary acne . It remains to be determined whether the infection is secondary or has any pathogenic implications. Am J Vet Res, 2004 Sep, 65(9), 1265 - 8 Methicillin resistance of staphylococci isolated from the skin of dogs with pyoderma; Kania SA et al.; OBJECTIVE: To determine the methicillin-resistant profile of staphylococcal isolates from the skin of dogs with pyoderma . ANIMALS: 90 dogs with pyoderma . PROCEDURE: Staphylococci isolated from dogs with pyoderma were tested for susceptibility to methicillin by use of a standard disk diffusion test with oxacillin disks . The DNA extracted from the isolates was tested for the mecA gene that encodes the penicillin-binding protein 2a (PBP2a) by use of a polymerase chain reaction (PCR) assay . The expression of PBP2a was determined with a commercial latex agglutination assay . Species of staphylococcal isolates were identified by use of morphologic, biochemical, and enzymatic tests . RESULTS: Most of the isolated staphylococci were methicillin-susceptible, coagulase-positive Staphylococcus intermedius isolates . Whereas only 2 of 57 S . intermedius isolates were resistant to methicillin, approximately half of the isolates had the mecA gene and produced PBP2a . Staphylococcus schleiferi was the second most common isolate . Widespread resistance to methicillin was found among S . schleiferi isolates . More coagulase-negative S . schleiferi isolates were identified with mecA gene-mediated resistance to methicillin, compared with coagulase-positive S . schleiferi isolates . CONCLUSIONS AND CLINICAL RELEVANCE: The latex agglutination assay for the detection of PBP2a expression coupled with the PCR assay for the mecA gene may provide new information about emerging antimicrobial resistance among staphylococcal isolates. Haematologica, 2004 Oct, 89(10), 1232 - 7 Quality control of bacterial contamination in autologous peripheral blood stem cells for transplantation; Larrea L et al.; BACKGROUND AND OBJECTIVES: Microbiological follow-up is part of quality control of peripheral blood stem cell (PBSC) manipulation . DESIGN AND METHODS: We prospectively studied microbiological cultures performed in 865 consecutive untreated autologous PBSC harvests from 348 patients . Our aim was to know the rate of microbiological contamination, the optimum moment to evaluate the sample and the clinical significance of the positive findings . RESULTS: Fifty-nine of the 852 samples (6.9%) yielded a positive culture after PBSC collection (sample 1) and 62 samples also yielded positive results before cryopreservation (7.2%) (sample 2) . At the time of the analysis, a total of 520 aphereses had been infused and the number of positive cultures after thawing (sample 3) and after washing (sample 4; 82 aphereses) was 5.4% and 2.3%, respectively . Most of the positive cultures were due to coagulase-negative staphylococci (48 isolates) . After thawing 15 coagulase-negative staphylococci and 2 enterococci isolates were recovered . Comparison between samples using a marginal homogeneity test showed no differences in the rate of contamination observed at the different sampling points . INTERPRETATION AND CONCLUSIONS: Positive microbiological findings in collected PBSC are not due to contamination within the laboratory . Cryopreservation using DMSO does not eradicate bacteria and manipulation does not seem to affect results . To simplify the procedure it would be possible to eliminate the microbiological controls performed immediately before cryopreservation. Phytother Res, 2004 Aug, 18(8), 674 - 6 Antimicrobial activity of Mahonia aquifolium crude extract and its major isolated alkaloids; Slobodnikova L et al.; The crude extract of Mahonia aquifolium (Pursh) Nutt . stem bark and its two main protoberberine alkaloids, berberine and jatrorrhizine, were tested for their in vitro antimicrobial activity . Twenty strains of coagulase-negative staphylococci and 20 strains of Propionibacterium acnes isolated from skin lesions of patients with a severe form of acne, and 20 strains of Candida sp . isolated from chronic vulvovaginal candidoses were tested for their susceptibility to crude extract and two isolated alkaloids . The minimum inhibitory concentrations obtained in this study illustrate the varying degrees of antibacterial and antifungal activity of the tested agents . The results indicate a rational basis for the traditional use of Mahonia aquifolium for localized skin and mucosal infection therapy, as well as for the possible development of a preparation for supportive therapy of the diseases mentioned above. Diagn Microbiol Infect Dis, 2004 Oct, 50(2), 125 - 30 Clinical glycopeptide-intermediate staphylococci tested against arbekacin, daptomycin, and tigecycline; LaPlante KL et al.; We examined the activity of arbekacin, daptomycin, tigecycline, and vancomycin against various Staphylococci isolates with glycopeptide-intermediate (n = 25) and heterogeneous susceptibilities (n = 22) (GISS and hGISS) . The minimum inhibitory concentrations (MIC) of each antimicrobial was evaluated in time-kill experiments by using 4 randomly selected GISS isolates tested at 2 and 4 times their respective MIC . The MIC(90) microg/mL ranges for arbekacin, daptomycin, tigecycline, and vancomycin were 2 (0.25-4), 1 (0.0625-2), 0.5 (0.0625-2), and 8 (4-8), respectively . Time kill at 2 times the MIC demonstrated a mean log(10) colony forming units (CFU)/mL change of -2.98 +/- 0.708, -3.6 +/- 0.509, -2.48 +/- 0.647, and +1.14 +/- 0.1 arbekacin, daptomycin, tigecycline, and vancomycin, respectively . At 4 times the MIC, significant activity for all compounds was noted with a log(10) CFU/mL decrease range from 3.68 to 2.74 +/- 0.66 . Overall, all the antimicrobials tested (with the exception of vancomycin) exhibited significant in vitro activity against GISS . These compounds may offer therapeutic options for the treatment of GISS. Epidemiol Infect, 2004 Oct, 132(5), 921 - 5 Molecular epidemiology of catheter-related bloodstream infections caused by coagulase-negative staphylococci in haematological patients with neutropenia; Muller-Premru M et al.; Catheter-related bloodstream infection (CRBSI) caused by coagulase-negative staphylococci (CNS) is common in haematological patients with febrile neutropenia . As the clinical signs of CRBSI are usually scarce and it is difficult to differentiate from blood culture contamination, we tried to confirm CRBSI by molecular typing of CNS isolated from paired blood cultures (one from a peripheral vein and another from the central venous catheter hub) . Blood cultures were positive in 59 (36%) out of 163 patients . CNS were isolated in 24 (40%) patients; in 14 from paired blood cultures (28 isolates) and in 10 from a single blood culture . CNS from paired blood cultures were identified as Staphylococcus epidermidis . Antimicrobial susceptibility was determined and bacteria were typed by pulsed-field gel electrophoresis (PFGE) of bacterial genomic DNA . In 13 patients, the antibiotic susceptibility of isolates was identical . The PFGE patterns from paired blood cultures were identical or closely related in 10 patients, thus confirming the presence of CRBSI . In the remaining four patients they were unrelated, and suggested a mixed infection or contamination . Since CNS isolates from three patients had identical PFGE patterns, they were probably nosocomially spread amongst them. Clin Infect Dis, 2004 Oct 1, 39(7), 1010 - 5 Epub 2004 Sep 10. Antimicrobial resistance to linezolid; Meka VG et al.; Acquired resistance to linezolid, the first approved oxazolidinone, has been selected in laboratory experiments and has been observed in clinical isolates of gram-positive cocci . This resistance has typically been associated with single-nucleotide changes in varying numbers of copies of the genes encoding 23S ribosomal RNA . In the current environment of increasingly prevalent resistance to standard antibiotics, linezolid is an important drug because of its activity against a number of clinically significant gram-positive cocci, including multidrug-resistant staphylococci and enterococci . Although resistance to linezolid remains uncommon, the development of resistance by clinical isolates should prompt increased attention to susceptibility testing for this agent and should be taken into account in consideration of the therapeutic use of this drug. J Clin Microbiol, 2004 Oct, 42(10), 4846 - 9 icaA is not a useful diagnostic marker for prosthetic joint infection; Frank KL et al.; A collection of 99 staphylococcal isolates associated with prosthetic joint infection and 23 coagulase-negative staphylococci isolated from noninfected arthroplasty-associated specimens were screened in order to determine whether the presence of icaA could be used to distinguish between pathogens and nonpathogens . All Staphylococcus aureus prosthetic joint infection isolates (n = 55) were icaA positive . A total of 46% (20 out of 44) of coagulase-negative staphylococcal prosthetic joint infection isolates were icaA positive, and 30% (7 out of 23) of arthroplasty-associated non-prosthetic joint infection-associated coagulase-negative staphylococcal isolates were icaA positive (P = 0.23) . Certain coagulase-negative Staphylococcus species appeared more likely to be isolated as either arthroplasty-associated non-prosthetic joint infection-associated isolates (e.g., Staphylococcus warneri and Staphylococcus hominis) or pathogens (e.g., Staphylococcus lugdunensis) . The presence of icaA in a coagulase-negative staphylococcal isolate associated with an arthroplasty is not a useful diagnostic indicator of pathogenicity. J Antimicrob Chemother, 2004 Nov, 54(5), 889 - 896 Epub 2004 Oct 7. Phenotypic and genotypic aminoglycoside resistance in blood culture isolates of coagulase-negative staphylococci from a single neonatal intensive care unit, 1989-2000; Klingenberg C et al.; OBJECTIVES: To investigate the prevalence of aminoglycoside resistance and genes encoding aminoglycoside-modifying enzymes (AME) in blood culture isolates of coagulase-negative staphylococci (CoNS) from neonates . Materials and methods: A total of 180 isolates from 148 patients collected in a single neonatal unit over a 12 year period were examined for susceptibility to gentamicin, tobramycin, netilmicin, amikacin and arbekacin by Etest and/or disc diffusion . AME genes were detected by PCR . RESULTS: The overall non-susceptibility rates to gentamicin, tobramycin, netilmicin, amikacin and arbekacin were 66%, 68%, 52%, 38% and 1%, respectively . Gentamicin non-susceptibility rates were 4% and 91% in methicillin-susceptible and -resistant isolates, respectively . aac(6')-Ie-aph(2'')-Ia, aph(3')-IIIa and/or ant(4')-Ia were encountered in 125 (69%), 1 (0.5%) and 30 (16.6%) isolates, respectively . Forty-six (26%) isolates negative for AME genes were susceptible to all aminoglycosides . In contrast, 115 (92%), 91 (73%) and 66 (53%) of aac(6')-Ie-aph(2'')-Ia positive isolates were non-susceptible to gentamicin, netilmicin and amikacin, respectively . Only one isolate showed arbekacin resistance . However, aac(6')-Ie-aph(2'')-Ia positive isolates and isolates with gentamicin MIC >/=128 mg/L displayed a significant reduction in arbekacin inhibition zones . CONCLUSIONS: A high prevalence of aminoglycoside resistance was detected and associated with methicillin resistance . Discrepancies between phenotypic and genetic detection of aminoglycoside resistance were discerned . Gentamicin was the preferred substrate for phenotypic detection of aac(6')-Ie-aph(2'')-Ia . Arbekacin showed favourable antibacterial activity even in aac(6')-Ie-aph(2'')-Ia-positive isolates . We suggest including arbekacin in future clinical trials of empirical treatment of late onset neonatal sepsis. Antibiot Khimioter, 2004, 49(4), 16 - 20 {Experimental basis for the use of new dosage forms of doxorubicin for correction of its hepatotoxic, prooxidant and immunosuppressory effects} {Investigation of cell wall components in actinomycin D resistant Staphylococcus aureus} {No authors listed} Cell walls in 2 strains of Staphylococcus aureus 209P, i.e . actinomycin D susceptible and resistant ones were comparatively investigated . The resistant cells contained much more wall material per a unit of the biomass weight vs the susceptible strain cells, that conformed to thickening of the resistant cell walls detected by electron microscopy and a sharp increase of their electron density . Investigation of peptidoglycans and teichoic acids did not reveal any significant alterations in the structure of the wall components in the actinomycin D resistant cells . Only some increase of glucosamine in the peptidoglycan fraction of the resistant cells vs the susceptible ones was observed . It was shown that preparations of the resistant cell walls and peptidoglycan isolated from the resistant cells were able to bind somewhat lower quantities of actinomycin D vs the analogous preparations of the susceptible cells . The significant decrease of the antibiotic binding by live cells of the resistant strain probably slightly depended on the structure characteristics of the main wall components . The barrier properties of the walls in resistant staphylococci are most likely defined by the wall thickening and consolidation while adapting to actinomycin D. J Dairy Sci, 2004 Jun, 87(6), 1719 - 26 Changes in milk composition as affected by subclinical mastitis in goats; Leitner G et al.; The mechanism of the effects of subclinical mastitis (SM) at the glandular level in dairy goats on milk yield and its composition as expressed in curd yield (Yc) was studied . Twenty-five Israeli goats of various crossbreeds were chosen; one udder half was naturally infected with identified coagulase-negative staphylococci, and the contralateral gland was free of bacteria . The milk yield of the infected halves was significantly lower than that of the uninfected ones . Somatic cell count and N-acetyl-beta-D-glucosaminidase activity were significantly higher in the infected halves . The lactose concentration in the infected glands was significantly lower than that in the uninfected ones, casein concentrations did not differ, and the whey protein and albumin concentrations were significantly higher in the infected glands . Plasmin activity was significantly higher in the infected glands, whereas plasminogen activity was undetectable . Concentrations of Ca2+ did not differ, whereas Ca2+ activity was significantly lower in the infected glands . The proteose peptone concentration was 1.5 times as great in the infected glands as in the uninfected ones . The Yc was significantly lower in the infected halves, and clotting time was significantly longer . The mechanisms of the effects of SM on milk yield and Yc in goats and sheep are discussed and compared. Curr Opin Microbiol, 2004 Oct, 7(5), 477 - 87 Staphylococci in colonization and disease: prospective targets for drugs and vaccines; Gotz F; Pathogenic staphylococci are now regarded in the scientific community as antibiotic resistant 'superbugs' because they have an amazing capacity to acquire resistance traits . Surprisingly, antibiotic development has decelerated . Promising targets for drug development are enzymes involved in the biosynthesis of cell envelope structures such as peptidoglycan, teichoic acids, membrane lipids, or cell wall associated adhesins . Compounds that inactivate or neutralize the most aggressive toxins such as the superantigens and the pore forming toxins have also been considered . In the past decade, global regulatory systems have been studied that contribute to virulence and might be candidates for target development . Targets that are particularly promising include all enzymatic reactions that are unique to bacteria and that are involved in central metabolism, such as methionine-tRNA(fMet) formyltransferase or the peptide deformylase, which have been successfully used for designing new inhibitors . There are also several known antibiotics that have roused new interest especially if they are active against multi-resistant staphylococci . Various cell wall components are promising candidates for active and passive immunization strategies such as capsule, slime, teichoic acids or cell wall bound adhesins . Several new targets for drugs or vaccines will arise from the functional analysis of the staphylococcal genomes that contain many hitherto unknown targets. J Pak Med Assoc, 2004 Jul, 54(7), 357 - 60 Emerging bacterial resistance patterns in febrile neutropenic patients: experience at a tertiary care hospital in Pakistan; Khan MA et al.; OBJECTIVE: To look at the clinical presentations, spectrum and site of isolation of the organisms, sensitivity patterns of the organisms and the antibiotic prescribing practices for the treatment of febrile neutropenic patients at our hospital . METHODS: The data were collected retrospectively from the records of all neutropenic patients with an absolute neutrophil count (ANC) of less than 500/ml admitted during the period of 3 years from August 1999 to July 2002 at AKUH . RESULTS: Out of the total of 404 patients, 65% had hematological malignancies and around half of them had leukaemia, 86% of the patients presented with fever . A total of 124 bacterial organisms were isolated from 96 patients among which 47% were gram positive and 53% were gram negative organisms; 16.1% of the patients had septicaemia . Coagulase Negative Staphylococci (CoNS) were the most common gram positive and E . coli was the most commonly isolated gram negative organism . Most of the gram positive organisms were isolated from blood (67%) . There was emerging resistance to all commonly used antibiotics including imipenem, cloxacillin, vancomycin and amikacin . The average duration of neutropenia was 6.4 days . The mortality rate was 6% . CONCLUSION: There is increasing trend of gram negative organisms developing resistance to commonly used antibiotics . Gram positive bacteria including Enterococcus spp . and CoNS are also showing emerging resistance to vancomycin. Antimicrob Agents Chemother, 2004 Oct, 48(10), 4037 - 9 Antistaphylococcal activity of CB-181963 (CAB-175), an experimental parenteral cephalosporin; Hoellman