Diagn Microbiol Infect Dis, 2003 Jan, 45(1), 81 - 4 Real-time PCR for the rapid detection of vanA and vanB genes; Palladino S et al.; A real-time PCR assay suitable for use on the Roche LightCycler platform was developed to replace an existing gel-based PCR assay for the simultaneous detection of the vanA & vanB genes in enterococcal isolates . Novel Fluorescence Resonance Energy Transfer (FRET) hybridization probes were designed . The multiplex real-time PCR assay and the existing gel-based assay were 100% concordant and both correctly detected the vanA or vanB genes in 4/4 VanA E . faecium and 25/25 VanB E . faecium . Additionally, 1/1 VanC1 E . gallinarum, 1/1 VanC2 E . casseliflavus and 47/47 vancomycin susceptible enterococci were negative for the vanA and vanB genes in both PCR assays . Results were available within 1.5 h for the real-time PCR assay compared to up to 5.5 h for the conventional PCR assay. Biometals, 2003 Mar, 16(1), 137 - 43 The Enterococcus hirae paradigm of copper homeostasis: copper chaperone turnover, interactions, and transactions; Lu ZH et al.; The cop operon is a key element of copper homeostasis in Enterococcus hirae . It encodes two copper ATPases, CopA and CopB, the CopY repressor, and the CopZ metallochaperone . The cop operon is induced by copper, which allows uncompromised growth in up to 5 mM ambient copper . Copper uptake appears to be accomplished by the CopA ATPase, a member of the heavy metal CPx-type ATPases and closely related to the human Menkes and Wilson ATPases . The related CopB ATPase extrudes copper when it reaches toxic levels . Intracellular copper routing is accomplished by the CopZ copper chaperone . Using surface plasmon resonance analysis, it was demonstrated that CopZ interacts with the CopA ATPase where it probably becomes copper loaded . CopZ in turn can donate copper to the copper responsive repressor CopY, thereby releasing it from DNA . In high copper, CopZ is proteolyzed . Cell extracts were found to contain a copper activated proteolytic activity that degrades CopZ in vitro . This post-translational control of CopZ expression presumably serves to avoid the accumulation of detrimental Cu-CopZ levels. Clin Infect Dis, 2003 Feb 15, 36(4), 440 - 6 Epub 2003 Jan 24. Changes in the prevalence of vancomycin-resistant enterococci in response to antimicrobial formulary interventions: impact of progressive restrictions on use of vancomycin and third-generation cephalosporins; Lautenbach E et al.; This study sought to assess the impact of restricting use of vancomycin and third-generation cephalosporins on vancomycin-resistant enterococci (VRE) prevalence . All clinical enterococcal isolates identified at a large academic medical center during a 10-year period were analyzed . Changes in VRE prevalence after sequential restrictions on use of vancomycin and third-generation cephalosporins were evaluated . The correlation between antibiotic use and VRE prevalence was also investigated . Vancomycin use initially decreased by 23.9% but returned to preintervention levels by the end of the study . Third-generation cephalosporin use decreased by 85.8% . However, VRE prevalence increased steadily from 17.4% to 29.6% during the 10-year period (P<.001) . Clindamycin use was significantly correlated with VRE prevalence . Restricting the use of vancomycin and third-generations cephalosporins had little impact on VRE prevalence . The association between clindamycin use and the prevalence of VRE suggests that restriction of this and perhaps other antianaerobic agents might be an important component of future antimicrobial interventions. J Infect Dis, 2003 Feb 1, 187(3), 508 - 12 Epub 2003 Jan 08. A potential virulence gene, hylEfm, predominates in Enterococcus faecium of clinical origin; Rice LB et al.; An open reading frame (hyl(Efm)) with homologies to previously described hyaluronidase genes has been identified in nonstool isolates of Enterococcus faecium . E . faecium isolates (n=577) from diverse sources were screened for the presence of hyl(Efm) and esp(Efm), a putative virulence gene associated with epidemic E . faecium strains . The presence of esp(Efm) was roughly twice that of hyl(Efm), but both were found primarily in vancomycin-resistant E . faecium isolates in nonstool cultures obtained from patients hospitalized in the United States . These data suggest that specific E . faecium strains may be enriched in determinants that make them more likely to cause clinical infections . Differences in the prevalence of these strains may help explain variations in the clinical importance of multiresistant E . faecium across different continents. Antimicrob Agents Chemother, 2003 Feb, 47(2), 786 - 9 Nonconjugative transposition of the vanB-containing Tn5382-like element in Enterococcus faecium; Dahl KH et al.; The vanB2 operon encoding glycopeptide resistance is an integral part of the putative conjugative transposon Tn5382 . Characterization of clinical glycopeptide resistant derivatives from an epidemic ampicillin-resistant Enterococcus faecium strain showed precise chromosomal or plasmid insertions of a vanB2-containing Tn5382-like element . Conjugative transposition of the Tn5382-like element was not demonstrated in retransfer studies. J Microbiol Methods, 2003 Mar, 52(3), 341 - 51 Evaluation of a novel method based on amplification of DNA fragments surrounding rare restriction sites (ADSRRS fingerprinting) for typing strains of vancomycin-resistant Enterococcus faecium; Krawczyk B et al.; In the search for an effective DNA-typing technique for use in hospital epidemiology, the performance and convenience of a novel assay based on the fingerprinting of bacterial genomes by amplification of DNA fragments surrounding rare restriction sites (ADSRRS fingerprinting) was tested . A large number of vancomycin-resistant Enterococcus faecium (VREM) isolates from haematological ward patients of the Clinical Hospital in Gdansk were examined . We found that ADSRRS fingerprinting analysis is a rapid method that offers good discriminatory power . The method demonstrated also excellent reproducibility . The usefulness of the ADSRRS fingerprinting method for molecular typing was compared with pulsed field gel electrophoresis (PFGE) method, which is currently considered the gold standard for molecular typing of isolates recovered from patients and the environment in the course of investigation and control of nosocomial outbreaks . Clustering of ADSRRS fingerprinting data matched pulsed field gel electrophoresis data.The features of ADSRRS fingerprinting technique is discussed in comparison with conventional methods . Data presented here demonstrate the complexity of the epidemiological situation concerning VREM that may occur in a single medical ward. Microb Drug Resist, 2002 Winter, 8(4), 369 - 74 Streptogramin resistance among Enterococcus faecium isolated from production animals in Denmark in 1997; Jensen LB et al.; The genetic background for streptogramin resistance was examined in Enterococcus faecium isolated from pigs (n = 55) and broilers (n = 207) in 1997 in Denmark . Fifty-one percent and 67%, respectively, of the isolates were resistant to streptogramins . Among streptogramin-resistant E . faecium (SREF), the genetic background for streptogramin A resistance could be determined in 96% of the isolates from broilers, compared with 14% among SREF from pigs . For broiler isolates 89% of SREF contained the vat(E) gene and 10% the vat(D) gene . Three of these isolates contained both resistance genes . Among SREF from pigs two isolates contained the vat(E) gene and two others the vat(D) gene . The genetic background for streptogramin B was most often identified as the erm(B) gene encoding macrolide, lincosamide, and streptogramin B (MLSB) resistance . Among SREF, 84% and 86% of isolates from broilers and pigs, respectively, contained the erm(B) . In SREF from broilers, the erm(B) gene was physically linked to the vat(E) gene in 62% of the vat(E)-positive isolates and 79% of the isolates containing vat(D) . erm(A) was detected in two SREF of broiler origin . Both isolates also contained the erm(B) gene . No SREF contained the vgb(A) gene encoding streptogramin B resistance . On the basis of genetic characterization, streptogramin-resistant isolates from broiler were divided into subgroups, according to the presence of the streptogramin A genes, to determine possible co-resistance to antimicrobials, especially glycopeptides . Twenty-five percent of the SREF from broilers were glycopeptide resistant (MIC > 16 microg/ml) . None of the isolates containing the streptogramin A gene vat(D) was resistant to glycopeptide, whereas isolates containing the vat(E) gene had a lower prevalence to glycopeptide resistance than the streptogramin-sensitive isolates. Microb Drug Resist, 2002 Winter, 8(4), 355 - 61 Persistence of vancomycin-resistant enterococci (VRE) in broiler houses after the avoparcin ban; Heuer OE et al.; The glycopeptide growth promoter avoparcin was banned from animal production in the EU in 1997 due to concern for the spread of vancomycin-resistant enterococci (VRE) from food animals to humans . In recent Norwegian and Danish studies, extensive occurrence of VRE on broiler farms and in broiler flocks after the avoparcin ban has been reported . The present study was undertaken to investigate the epidemiology of VRE on broiler farms in the absence of the selective pressure exerted by avoparcin . Environmental samples were obtained from five broiler houses after depopulation, cleaning, and disinfection of the houses between rotations, and two consecutive broiler flocks from each house were sampled by taking cloacal swabs from the broilers at the time of slaughter . A total of 69 vancomycin-resistant Enterococcus faecium isolates obtained from broiler flocks and broiler houses were subjected to molecular typing by pulsed-field gel electrophoresis (PFGE) . Forty-one PFGE-profiles were observed . VRE with indistinguishable or highly similar PFGE profiles were isolated from consecutive broiler flocks and from environmental samples from the houses in which the flocks were reared, whereas VRE-isolates from different broiler houses and from flocks reared in different houses appeared to be genetically unrelated . These findings indicated that VRE was transmitted between consecutive broiler flocks by clones of resistant bacteria surviving in the broiler houses despite cleaning and disinfection between rotations . Thus, the extensive occurrence of VRE in broiler flocks after the avoparcin ban may be explained by persistence of VRE in the broiler house environment. J Gastroenterol, 2002, 37(12), 1062 - 7 A pancreatic abscess 7 years after a pancreatojejunostomy for calcifying chronic pancreatitis; Suzuki A et al.; We present herein a case of a 75-year-old Japanese man who had developed a pancreatic abscess 7 years after a longitudinal pancreatojejunostomy for chronic pancreatitis . The patient, a heavy drinker of alcohol, underwent surgical decompression of a ductal obstruction to relieve persistent abdominal pain due to severely calcifying chronic pancreatitis . After the surgery, he stopped drinking alcohol and was treated with insulin to control secondary diabetes mellitus . Thereafter, his symptoms disappeared . Seven years after the surgery, however, he was hospitalized due to obstructive jaundice, high-grade fever, and right hypochondria pain . Ultrasound and computed tomographic scans of the abdomen both disclosed a cystic mass, approximately 6 cm in size, in the pancreatic head . Magnetic resonance imaging strongly suggested a pancreatic abscess with necrotic fluid and debris . First, percutaneous transhepatic cholangiodrainage (PTCD) was done to treat the progressively obstructive jaundice . Subsequently, fine-needle aspiration of the pancreatic abscess was performed under ultrasound guidance . Enterococcus avium and Klebsiella oxytoca were revealed by culture of abscess aspirates . He was successfully cured by treatment with both appropriate antibiotic and continuous PTCD for the obstructive jaundice. Appl Environ Microbiol, 2003 Jan, 69(1), 92 - 6 Enterococci as indicators of Lake Michigan recreational water quality: comparison of two methodologies and their impacts on public health regulatory events; Kinzelman J et al.; The frequency of poor-water-quality advisories issued in Milwaukee and Racine, Wisconsin, in the absence of identifiable sources of contamination brought into question the reliability of the present indicator organism, Escherichia coli . Enteroccoci have been suggested as an alternative to E . coli for freshwater monitoring due to their direct correlation to swimmer-associated gastroenteritis . The purpose of this research was threefold: (i) to explore enterococci as an alternative to E . coli for monitoring freshwater Lake Michigan beaches, (ii) to evaluate the impact of the two indicators on regulatory decisions, and (iii) to compare membrane filtration m-enterococcus agar with indoxyl-beta-D-glucoside to a chemical substrate technique (Enterolert) for the recovery of enterococci . Recreational water samples from Milwaukee (n = 305) and Racine (n = 153) were analyzed for the enumeration of E . coli and enterococci using IDEXX Colilert-18 and Enterolert . Correlation between the indicators was low (R(2) = 0.60 and 0.69) . Based on U.S . Environmental Protection Agency bacterial indicator threshold levels of risk for full body immersion, using enterococci would have resulted in 56 additional unsafe-recreational-water-quality advisories compared to the total from using E . coli and the substrate-based methods . A comparison of the two enterococcal methods (n = 124) yielded similar results (R(2) = 0.62) . This was further confounded by the frequent inability to verify enterococci from those wells producing fluorescence by the defined substrate test using conventional microbiological methods . These results suggest that further research is necessary regarding the use of defined substrate technology interchangeably with the U.S . Environmental Protection Agency-approved membrane filtration test for the detection of enterococci from fresh surface water. Folia Microbiol (Praha), 2002, 47(5), 573 - 8 Treatment of experimental adjuvant arthritis with the combination of methotrexate and lyophilized Enterococcus faecium enriched with organic selenium; Rovensky J et al.; The efficacy of combination therapy with methotrexate (MTX) and probiotic bacteria Enterococcus faecium enriched with organic selenium (EFSe) in rats with adjuvant arthritis was determined . Rats with adjuvant arthritis were given MTX (0.3 mg/kg 2-times weekly, orally); lyophilized E . faecium enriched with Se (15 mg/kg, 5 d per week, orally); and a combination of MTX plus EFSe for a period of 50 d from the immunization . Levels of serum albumin, serum nitrite/nitrate concentrations, changes in hind paw swelling, arthrogram score, bone erosions, whole body bone mineral density (BMD) and bone mineral content (BMC) were assayed in the rats as variables of inflammation and destructive arthritis-associated changes . Treatment with MTX and with the combination MTX + EFSe significantly inhibited markers of both inflammation and arthritis . Significant differences in favor of combination therapy with MTX + EFSe as compared to MTX alone were seen in serum albumin concentration, hind paw swelling and arthrogram score . Reductions in radiographic scores were also more pronounced in the combination therapy group . Combination therapy, but not MTX alone, inhibited the reduction of BMD and BMC; treatment with lyophilized EFSe alone had no significant effect on adjuvant arthritis in rats . The potent therapeutic effect of low dosage MTX therapy in combination with lyophilized EFSe on adjuvant arthritis in rats was shown. Microbiol Res, 2002, 157(4), 293 - 303 Lectin-like binding and antibiotic sensitivity of enterococci from wild herbivores; Styriak I et al.; Fifty eight enterococcal isolates from wild herbivores were tested for their antibiotic sensitivity pattern and lectin-like binding of extracellular matrix (ECM) and serum proteins . Kanamycin resistance was very frequent; many multiresistant strains were also isolated . All isolates were sensitive to rifampicin . Resistance to gentamicin, novobiocin, and tetracycline was widely distributed in the microflora of wild herbivores breeded in zoological garden in Kosice . No autoaggregating strains were detected among these 58 enterococcal isolates . Various degrees of binding of mucins, fetuin, heparin, fibrinogen, and fibronectin were observed in individual strains . However, bovine lactoferrin binding by enterococci from deers and chamoises was either negative (0) or strongly positive (3) . With regard to influence of growth media, TH agar was found to be better for the expression of lectin-like binding than blood agar, TH broth and Nutrient broth . A significant effect (P < 0.001 or P < 0.05) of proteolytic treatment was observed in six selected strains . However, there is a difference between the effect of trypsin and pronase P . Pronase treatment more effectively decreased binding of some strains (1H, 6A, EF 1111, EC 1292), while trypsin treatment decreased more binding of other enterococcal strains (EF 953 and 1E) . Significant (P < 0.001) influence of metaperiodate, which cleaves the C-C bond between vicinal groups of sugars, on collagen I binding by three selected strains (1E, 1H, 6A) and bovine lactoferrin binding (by EF 1111, EC 1292, EF 953) was also observed . However, its influence was very different . In two strains (1H and EC 1292), ECM binding was decreased, while in four other strains (1E, 6A, EF 1111, EF 953) it was increased. Antimicrob Agents Chemother, 2003 Jan, 47(1), 7 - 18 VanD-type vancomycin-resistant Enterococcus faecium 10/96A; Depardieu F et al.; VanD type Enterococcus faecium 10/96A is constitutively resistant to vancomycin and to low levels of teicoplanin by nearly exclusive synthesis of peptidoglycan precursors terminating in D-alanyl-D-lactate (L . M . Dalla Costa, P . E . Reynolds, H . A . Souza, D . C . Souza, M . F . Palepou, and N . Woodford, Antimicrob . Agents Chemother . 44:3444-3446, 2000) . A G(184)S mutation adjacent to the serine involved in the binding of D-Ala1 in the D-alanine:D-alanine ligase (Ddl) led to production of an impaired Ddl and accounts for the lack of D-alanyl-D-alanine-containing peptidoglycan precursors . The sequence of the vanD gene cluster revealed eight open reading frames . The organization of this operon, assigned to a chromosomal location, was similar to those in other VanD type strains . The distal part encoded the VanH(D) dehydrogenase, the VanD ligase, and the VanX(D) dipeptidase, which were homologous to the corresponding proteins in VanD-type strains . Upstream from the structural genes for these proteins was the vanY(D) gene; a frameshift mutation in this gene resulted in premature termination of the encoded protein and accounted for the lack of penicillin-susceptible D,D-carboxypeptidase activity . Analysis of the translated sequence downstream from the stop codon, but in a different reading frame because of the frameshift mutation, indicated homology with penicillin binding proteins (PBPs) with a high degree of identity with VanY(D) from VanD-type strains . The 5' end of the gene cluster contained the vanR(D)-vanS(D) genes for a putative two-component regulatory system . Insertion of ISEfa4 in the vanS(D) gene led to constitutive expression of vancomycin resistance . This new insertion belonged to the IS605 family and was composed of two open reading frames encoding putative transposases of two unrelated insertion sequence elements, IS200 and IS1341. J Am Coll Surg, 2002 Dec, 195(6), 804 - 13 Portal versus systemic drainage of small bowel allografts: comparative assessment of survival, function, rejection, and bacterial translocation; Berney T et al.; BACKGROUND: Portal venous drainage of small bowel grafts is theoretically more physiologic than systemic drainage, but is technically more demanding . Comparisons in animal models have not demonstrated a clear advantage of one technique over the other, but clinical data are lacking . STUDY DESIGN: Clinical records of 36 patients who underwent 37 small bowel transplantation procedures from January 1995 to August 2001 were reviewed . Portal drainage was performed in 19 patients (PD group) . Systemic drainage was performed in 18 patients (SD group) . Median followup was 531 days . RESULTS: PD and SD patients had similar ICU stays (median 7 versus 9 days) and endotracheal intubation durations (median 3 versus 5 days) . All current survivors, with the exception of one patient in each group, are independent from parenteral nutrition . Liver function tests were similar in both groups . There was a twofold increase in tacrolimus dosage in the PD group to achieve similar trough levels indicating a "first-pass" hepatic clearance effect . Cumulative incidence of acute rejection episodes and OKT3-requiring rejection episodes were similar in both groups . To the contrary, a lower incidence of gram-negative rods of Enterococcus sp . in blood or bronchoalveolar lavage suggested that the clearance of translocated intestinal bacteria was more efficient in the PD group . Graft and patient survival rates were similar in both groups . CONCLUSIONS: Systemic venous drainage of small bowel transplants is a dependable technique, associated with similar results as portal venous drainage, in terms of overall mortality, morbidity, rejection, function, and patient and graft survival . But attention should be paid to an impaired clearance of intestinal bacterial translocation after systemic drainage. Chest, 2002 Dec, 122(6), 2259 - 62 Bacterial endocarditis and functional mitral stenosis: a report of two cases and brief literature review; Tiong IY et al.; Mitral valve endocarditis typically results in mitral regurgitation . However, endocarditis leading to functional mitral stenosis is uncommon and, when present, fungal organisms are typically implicated . Thus, obstructive-type bacterial endocarditis due to large vegetations blocking the mitral valve orifice is a rare occurrence, with approximately 20 reported cases in the literature . We report on two patients with bacterial endocarditis and severe functional mitral stenosis requiring emergent surgery . Additionally, this is the first report of vancomycin-resistant enterococcus causing endocarditis and functional mitral stenosis . The discussion emphasizes the hemodynamic instability of these patients and need for early surgical intervention. J Hosp Infect, 2002 Dec, 52(4), 292 - 6 Emergence of vancomycin-resistant Enterococcus faecium at a tertiary care hospital in Karachi, Pakistan; Khan E et al.; Vancomycin-resistant enterococcus (VRE) has not been reported previously in Pakistan . This is the first report where in vancomycin-resistant Enterococcus faecium was isolated from the clinical specimens of six patients admitted to the intensive care unit (ICU) and neonatal intensive care unit (NICU) of the Aga Khan University Hospital, Karachi . To identify the extent of the outbreak, rectal swabs were obtained from all the patients admitted to the ICU and NICU at that time . A total of 10 strains of vancomycin-resistant Enterococcus faecium were isolated . All the strains showed high-level resistance to both glycopeptides (vancomycin and teicoplanin) with a vancomycin minimum inhibitory concentration greater than 256 mg/L . All isolates had the vanA gene detected by polymerase chain reaction . The contour-clamped homogeneous electric field (CHEF) pattern demonstrated that all but one of the isolates were of a single clone, suggesting that they were derived from common source . Use of vancomycin and prolonged hospitalization were common features in all cases investigated . Biochem J, 2003 Apr 1, 371(Pt 1), 191 - 7 Biochemical and genetic characterization of a novel enzyme of pentitol metabolism: D-arabitol-phosphate dehydrogenase; Povelainen M et al.; An enzyme with a specificity that has not been described previously, D-arabitol-phosphate dehydrogenase (APDH), has been purified from cell lysate of Enterococcus avium . SDS/PAGE indicated that the enzyme had a molecular mass of 41+/-2 kDa, whereas a molecular mass of 160+/-5 kDa was observed under non-denaturing conditions, implying that the APDH may exist as a tetramer with identical subunits . Purified APDH was found to have a narrow substrate specificity, converting only D-arabitol 1-phosphate and D-arabitol 5-phosphate into xylulose 5-phosphate and ribulose 5-phosphate, respectively, in the oxidative reaction . Both NAD(+) and NADP(+) were accepted as cofactors . Based on the partial protein sequences, the APDH gene was cloned . Homology comparisons place APDH within the medium-range dehydrogenase family . Unlike most members of this family, APDH requires Mn(2+) but no Zn(2+) for enzymic activity . The DNA sequence surrounding the gene suggests that it belongs to an operon that also contains several components of phosphotransferase system . Both biochemical evidence and protein sequence homology comparisons indicate that similar enzymes are widespread among the Gram-positive bacteria . Their apparent biological role is to participate in arabitol catabolism via the 'arabitol phosphate route', similar to the ribitol and xylitol catabolic routes described previously. Fish Shellfish Immunol, 2002 Nov, 13(5), 351 - 65 Effect of hypoxia on the immune response of giant freshwater prawn Macrobrachium rosenbergii and its susceptibility to pathogen Enterococcus; Cheng W et al.; Giant freshwater prawns Macrobrachium rosenbergii (14-19 g) were challenged with Enterococcus (3 x 10(5) cfu prawn(-1)) previously incubated in TSB medium for 24 h, then placed in water having concentrations of dissolved oxygen (DO) at 7.75, 4.75, 2.75 and 1.75 mg l(-1) . Onset of mortality occurred after 6 h exposure to 1.75 mg l(-1) DO, and after 12 h exposure to 2.75 mg 1(-1) DO . Cumulative mortality of prawns at 1.75 mg l(-1) DO was significantly higher than that at 4.75 and 2.75 mg l(-1) DO, and cumulative mortality of prawns at 4.75 and 2.75 mg l(-1) DO was significantly higher than that at 7.75 mg l(-1) DO after 96 h . The prawns (20-30 g) which had been placed in water for 0 to 120 h at 7.75, 4.75, 2.75 and 1.75 mg l(-1) DO were examined for the THC (total haemocyte counts), DHC (differential haemocyte counts), phenoloxidase activity, respiratory burst, percentage phagocytosis and clearance efficiency . No significant difference in semi-granular cells and granular cells of prawns was observed among four treatments . The prawns following 120 h exposure to 2.75 mg l(-1) DO decreased significantly the hyaline cells and THC by 39% and 36%, respectively . Phenoloxidase activity and respiratory burst decreased significantly by 33% and 11% when the prawns were exposed to 2.75 mg l(-1) DO after 24 h, respectively . Percentage phagocytosis and clearance efficiency to Enterococcus decreased significantly by 44% and 54% for the prawns following 12 h exposure to 2.75 mg l(-1) DO, respectively . It is concluded that DO as low as 2.75 mg l(-1) and 4.75 mg l(-1) causes depression in immune system of M . rosenbergii, and increases its susceptibility to Enterococcus infection. Int J Antimicrob Agents, 2002 Dec, 20(6), 432 - 7 Epidemiology of multiresistant Enterococcus avium isolates in a Greek tertiary care hospital; Papaparaskevas J et al.; A retrospective survey of the isolation rate of Enterococcus avium during the period March 1994-February 2000 conducted in Laikon General Hospital using the WHONET software, revealed a peak in the isolation rates of this species during March 1995-February 1996 . The ten strains isolated during this time were studied further . No glycopeptide resistance was detected but resistance to ampicillin, ciprofloxacin, erythromycin, gentamicin (high-level) and streptomycin (high-level) was present in nine, ten, nine, three and seven of the isolates, respectively . The genes aac(6')-Ie+aph(2")-Ia and ant(6)-I, encoding for high-level gentamicin and streptomycin resistance, respectively, were detected only in the isolates with the corresponding phenotypes . Beta-lactamase production and haemolysis were not detected . There was evidence of ward-, floor- and building-specific distribution among the different aminoglycoside resistance phenotypes . DNA fingerprinting by PFGE grouped six of the ten isolates in a single cluster with 83% similarity, even though they expressed various resistance phenotypes . These results suggest dissemination of resistance genes among both genetically related and unrelated strains . Appl Environ Microbiol, 2002 Dec, 68(12), 6457 - 61 Vancomycin-resistant enterococci in humans and imported chickens in Japan; Ozawa Y et al.; The phenotypes and genotypes of 22 VanA-type vancomycin-resistant enterococci that had been isolated in Japan were examined . The VanA resistance determinant was plasmid mediated in each of the 22 strains . Of the 22 strains, 8 were isolated from different patients and 11 and 3 were obtained from different samples of chickens imported from Thailand and France, respectively . Three of the strains that were isolated from patients and the 11 strains isolated from the Thai chickens showed high-level vancomycin resistance (MICs, 512 to 1,024 micro g/ml) and low-level teicoplanin resistance (MICs, 0.5 to 4 micro g/ml) . Each of these strains had three amino acid substitutions in the N-terminal region of the deduced VanS sequence . L50 was converted to V, E54 was converted to Q, and Q69 was converted to H compared to the vanS gene sequence of Tn1546. J Int Med Res, 2002 Sep-Oct, 30(5), 529 - 34 A comparison of high-level aminoglycoside resistance in vancomycin-sensitive and vancomycin-resistant Enterococcus species; Yazgi H et al.; The aim of this study was to investigate whether there was a significant difference in high-level aminoglycoside resistance (HLAR) between vancomycin-sensitive enterococci (VSE) and vancomycin-resistant enterococci (VRE) . Vancomycin resistance was determined in 116 Enterococcus isolates using brain-heart infusion agar containing 6 micrograms/ml vancomycin . HLAR was determined by both standard agar screening and disk diffusion methods . Streptomycin and gentamicin were used as predictors of HLAR . Vancomycin resistance and HLAR were found in 17 (14.7%) and 41 (35.3%) of the Enterococcus strains, respectively . HLAR was found in 11 of 17 VRE and 30 of 98 VSE strains . HLAR in VRE strains was significantly higher than in VSE . More enterococcal strains were found to be resistant to both gentamicin and streptomycin (29) than to gentamicin (one) or streptomycin (11) alone . The HLAR rate in VRE was two-fold higher than in VSE . The synergistic bactericidal effect of aminoglycosides and beta-lactam or glycopeptide antibiotics is lost if there is high-level resistance to aminoglycosides. Water Res, 2002 Dec, 36(20), 4963 - 74 Occurrence and levels of indicator bacteriophages in bathing waters throughout Europe; Contreras-Coll N et al.; Somatic coliphages, F-specific RNA bacteriophages, bacteriophages infecting Bacteroides fragilis, Escherichia coli and enterococci were counted in bathing waters in the late spring and summer . We tested fresh and marine bathing waters from North, South, East and West Europe expected to contain between 100 and 500 E . coli per 100 ml, although wider ranges were sometimes found . Bacteriophages were counted after concentration, since a preliminary study proved that this step was necessary to obtain positive counts . During monitoring, a first-line quality control with reference materials for bacteria and bacteriophages was performed by all the laboratories participating in the study . The same microbes were also counted in raw sewage samples from various areas in Europe, where the bacterial indicators and the three groups of bacteriophages were detected in roughly the same numbers . All groups of bacteriophages were detected in both fresh and marine bathing waters throughout Europe . Reliable and complete results from 147 samples showed that for log-transformed values, E . coli and bacteriophages were slightly correlated . However, the slope of the regression line changed according to E . coli concentration and the correlation diminished when this concentration was close to zero per 100 ml . The ratios between E . coli and phages in bathing waters differed significantly from those in sewage . The relative amounts of bacteriophages, mainly somatic coliphages and phages infecting Bact . fragilis RYC2056, increased in bathing waters with low E . coli concentration, especially in seawater samples containing < 100 E . coli per 100 ml . The relationship of bacteriophages with respect to enterococci paralleled that of bacteriophages with respect to E . coli . Somatic coliphages and bacteriophages infecting Bact . fragilis are useful to predict the presence of some pathogens with the same origin as present bacterial indicators but with higher survival rates. Nippon Yakurigaku Zasshi, 2002 Oct, 120(4), 245 - 52 {Antimicrobial and clinical effect of linezolid (ZYVOX), new class of synthetic antibacterial drug}; Irinoda K et al.; Linezolid (ZYVOX), a novel synthesized antibacterial drug, was first approved in April 2001, as an antibacterial against vancomycin (VCM)--resistant enterococci in Japan . LZD has a wide spectrum of antibacterial activity against gram-positive bacteria with MIC90 of 0.5-4 mcg/mL . These antibacterial activities of LZD are similar to those of vancomycin (VCM) . LZD also has similar antibacterial activities against drug-resistant bacteria including VRE and MRSA . Protein-synthesis inhibitors, e.g., macrolides, tetracycline, aminoglycosides, and chloramphenicol, are known to bind the 30S and 50S subunits of ribosomes and inhibit the elongation cycle of protein synthesis . In contrast, LZD was found to inhibit the process of formation of the 50S, 30S-mRNA, and fMet-tRNA complex in the ribosome cycle, but not the elongation cycle . Due to this novel mechanism of action, LZD does not have a cross-resistance to drug-resistant bacteria and development of its resistance is quite slow . The antibacterial activity of LZD against VRE is bacteriostatic . In vivo antibacterial activity of orally administered LZD was demonstrated in a mouse model of systemic infection by VRE . When administered orally prior to the abscess formation in a mouse model of soft tissue infection by VRE, LZD showed similar antibacterial activity against VRE infection to that against VCM-susceptible enterococci . LZD is rapidly absorbed following oral administration and bioavailability when compared with intravenous administration is almost 100% . LZD administered orally twice-daily showed excellent efficacy in clinical trials with VRE-infected patients. Int J Food Microbiol, 2003 Feb 25, 81(1), 1 - 10 Identification and antibiotic susceptibility of bacterial isolates from probiotic products; Temmerman R et al.; In the present study, a total of 55 European probiotic products were evaluated with regard to the identity and the antibiotic resistance of the bacterial isolates recovered from these products . Bacterial isolation from 30 dried food supplements and 25 dairy products, yielded a total of 268 bacterial isolates selected from several selective media . Counts of food supplements showed bacterial recovery in 19 (63%) of the dried food supplements ranging from 10(3) to 10(6) CFU/g, whereas all dairy products yielded growth in the range of 10(5)-10(9) CFU/ml . After identification of the isolates using whole-cell protein profiling, mislabeling was noted in 47% of the food supplements and 40% of the dairy products . In six food supplements, Enterococcus faecium was isolated whereas only two of those products claim this species on their label . Using the disc diffusion method, antibiotic resistance among 187 isolates was detected against kanamycin (79% of the isolates), vancomycin (65%), tetracycline (26%), penicillinG (23%), erythromycin (16%) and chloramphenicol (11%) . Overall, 68.4% of the isolates showed resistance against multiple antibiotics including intrinsic resistances . Initially, 38% of the isolated enterococci was classified as vancomycin resistant using the disc diffusion method, whereas additional broth dilution and PCR assays clearly showed that all E . faecium isolates were in fact vancomycin susceptible. J Hosp Infect, 2002 Nov, 52(3), 206 - 11 Cleaning efficacy of nine different cleaners in a washer-disinfector designed for flexible endoscopes; Zuhlsdorf B et al.; Studies on processing endoscopes usually involve the combined cleaning and disinfecting activity . We compared nine cleaning agents designed for automatic processing for cleaning efficacy alone using soft and hard water as controls in 12 different processes in a washer-disinfector . Experiments were performed according to the German Endoscopy Working Group recommendations using transparent Teflon tubes (internal diameter 2mm, length 2m) as test pieces . For each test three pieces contaminated with a blood/test soil containing Enterococcus faecium were used; two for the test and one as a control; each test was repeated three times . Tests were run according to the manufacturer's instructions . Test pieces were assessed visually and microbiologically {log(10) reduction factors (RF) vs . untreated controls} . Soft water alone gave poor visible cleanliness and an RF of 0.3 (SD 0.2), while hard water produced adequate visible cleanliness and an RF of 1.2 (SD 1.0) . Five processes gave better visible cleanliness than soft water, but only three were better than hard water . Six processes were worse than soft water and five worse than hard water . Nine processes gave a better microbiological reduction factor than soft water, but the difference was only statistically significant in three . Only one process yielded a significantly higher RF than hard water; three were significantly worse . None of the cleaning processes reached the RF of 4 specified in the US regulations . This study confirms the variability of cleaning processes to dissolve blood residues and reduce the bioburden . We do not recommend abandoning cleaning agents, but suggest that further research is needed to clarify the relationships between washer-disinfectors, cleaning agents, and cleaning performance . J Biochem (Tokyo), 2002 Nov, 132(5), 789 - 94 Nucleotide-binding sites in V-type Na+-ATPase from Enterococcus hirae; Murata T et al.; Enterococcus hirae V-ATPase, in contrast to most V-type ATPases, is resistant to N-ethylmaleimide (NEM) . Alignment of the amino acid sequences of NtpA suggests that the NEM-sensitive Cys of V-type ATPases is replaced by Ala in E . hirae V-ATPase . Consistent with this prediction, the V-ATPase became sensitive upon substitution of the Ala with Cys . The three-dimensional structure of the NtpB subunit of V-ATPase was modeled based on the structure of the corresponding subunit (alpha subunit) of bovine F(1)-ATPase by homology modeling . Overall, the 3D structure of the subunit resembled that of alpha subunit of bovine F(1)-ATPase . The NtpB subunit, which lacks the P-loop consensus sequence for nucleotide binding, was predicted to bind a nucleotide at the modeled nucleotide-binding site . Experimental data supported the prediction that the E . hirae V-ATPase had about six nucleotide-binding sites. Rev Port Cardiol, 2002 Sep, 21(9), 989 - 99 Perioperative mortality and long-term outcome of infective endocarditis; Moura L et al.; INTRODUCTION AND OBJECTIVES: Infective endocarditis (IE) is a disease with a high morbidity and mortality during the active phase and a considerable risk of complications during follow-up . The aim of this study is to describe the clinical features of IE associated with perioperative mortality in patients undergoing surgery and the short- and long-term prognosis of this disease after surgical treatment . PATIENTS AND METHODS: The clinical files of 150 patients (pts) admitted to our institution between 1989 and 2001 and whose final diagnosis was IE (Duke criteria) were retrospectively reviewed . Thus all patients included underwent at least 1 transthoracic examination, and a transesophageal examination if indicated . The study population was 69 patients with IE, 62 of whom (90%) underwent prosthetic valve replacement and 7 (10%) underwent explanation of pacemaker electrocatheter . In the statistical analysis, quantitative variables are expressed as means +/- SD and qualitative variables as proportions (percentages) . Differences in survival were determined using the Kaplan-Meier log rank method . Associations were considered statistically significant when the p value was < 0.05 . RESULTS: The mean age was 56 +/- 15 years . Thirty-five cases (51%) had prosthetic valve endocarditis, 30 (39%) had native valve IE and 7 (10%) pacemaker electrocatheter IE . Fifty-two patients (75%) had positive blood cultures . The most frequent agents were S . epidermidis (14.5%), S . aureus (14.5%), Enterococci (13%) and S . viridans (10%) . Total perioperative mortality was 17.4% (n = 12), and surgical mortality was 19.4% . Our study shows a statistical tendency for higher mortality in diabetic patients (50% vs . 14%, p = 0.052) and in women (29% vs . 11%, p = 0.065) . In multivariate analysis, the presence of heart failure was an independent predictor of perioperative mortality (OR = 11.4; 95% CI: 2.0-215.2; p = 0.024) . Accumulated mortality in the first year was 28% and 5-year mortality was 48% . CONCLUSIONS: Prosthetic valve replacement in patients with IE is associated with high perioperative mortality (17.4%) . Despite good early postoperative results, the mortality rate increases during the first year as well as the need for reoperation. Clin Infect Dis, 2002 Nov 15, 35(10), 1269 - 72 Epub 2002 Oct 28. Risk factors associated with the development of infection with linezolid- and vancomycin-resistant Enterococcus faecium; Pai MP et al.; This retrospective cohort study revealed that linezolid resistance in vancomycin-resistant Enterococcus faecium was dependent on prior linezolid exposure and duration of linezolid therapy . These strains of E . faecium were resistant to the entire class of oxazolidinones. Clin Infect Dis, 2002 Nov 15, 35(10), 1139 - 46 Epub 2002 Oct 17. Risk factors for development of vancomycin-resistant enterococcal bloodstream infection in patients with cancer who are colonized with vancomycin-resistant enterococci; Zaas AK et al.; Vancomycin-resistant Enterococcus faecium (VRE) is a common nosocomial isolate, especially among patients with cancer . VRE infections have substantial attributable mortality among patients with cancer . The purpose of this study was to identify risk factors for developing bloodstream infection with VRE in patients with cancer who are colonized with VRE . VRE colonization was prospectively identified in 197 patients with cancer during 4-year period, of whom 179 (91%) had complete records for evaluation . Of these 179 patients, 24 (13.4%) developed hospital-acquired VRE bloodstream infections . Risk factors for VRE bloodstream infection included vancomycin use (relative risk {RR}, 1.98; 95% confidence interval {CI}, 1.25-3.14), diabetes mellitus (RR, 3.91; 95% CI, 1.20-12.77), gastrointestinal procedures (RR, 4.56; 95% CI, 1.05-19.7), and acute renal failure (RR, 3.10; 95% CI, 1.07-8.93) . Strategies for preventing VRE bloodstream infection in VRE-colonized patients with cancer should include limiting vancomycin use and, perhaps, gastrointestinal procedures. Epidemiol Infect, 2002 Oct, 129(2), 421 - 4 Nucleotide substitutions in vanC-2 gene of Enterococcus casseliflavus isolates obtained from chickens; Murase T et al.; DNA sequencing of the vanC-2 gene was partially carried out on 10 isolates of Enterococcus casseliflavus obtained from 8 samples of imported chickens in Japan between July 1999 and June 2001 to evaluate the variation in the gene . Forty nucleotide substitutions in 36 codons were identified within 345 base pairs when compared with the vanC-2 sequence of the reference strain E . casseliflavus ATCC25788 . Identical nucleotide substitutions were commonly found in the isolates recovered from chickens imported from both Brazil and China . Pulsed-field gel electrophoresis (PFGE) patterns of NotI-digested chromosomal DNA of these strains were distinguished by two, or more than six, band differences . These observations suggest that sequencing of the vanC-2 gene may be helpful for epidemiological investigation in combination with the PFGE analyses of the isolates, although particular genotypes are unlikely to be restricted to each of the countries that exported chickens. J Microbiol Methods, 2003 Jan, 52(1), 123 - 31 Application of the fluorogenic probe technique (TaqMan PCR) to the detection of Enterococcus spp . and Escherichia coli in water samples; Frahm E et al.; A recent PCR detection technique (TaqMan) based on the 5'-3'-exonuclease activity of the Taq DNA polymerase was applied to the detection of indicator organisms in water samples . In this technique, an increasing fluorescence signal is measured online which enables direct assessment of results after PCR without additional detection steps . The test is completed within about 5 h . Two sets of primers and probes were designed and tested: a genus-specific assay for the detection of Enterococcus spp . based on 23S rRNA sequence and an Escherichia coli-specific assay based on the uidA gene sequence . Specificity of the assays was confirmed by testing strains of target bacteria and potential interfering microorganisms . Application of the tests to 55 natural water samples showed the need of an overnight enrichment step to achieve compliance with detection limits of existing regulations . Compared with a parallel microbiological examination of the samples, agreement was 96% with the Enterococcus assay and 98% with the E . coli assay . The rapidity and feasibility of the method point to benefits in drinking water analysis, particularly in emergency situations and, thus, to improved public health management. Fitoterapia, 2002 Oct, 73(6), 532 - 5 Antibacterial and antioxidant activities of Artemisia annua essential oil; Juteau F et al.; The essential oil of Artemisia annua aerial parts, consisting of camphor (44%), germacrene D (16%), trans-pinocarveol (11%), beta-selinene (9%), beta-caryophyllene (9%) and artemisia ketone (3%), was screened for its antimicrobial activity . The essential oil remarkably inhibited the growth of tested Gram-positive bacteria Enterococcus hirae and both tested fungi . This oil has shown an antioxidant activity equivalent to 18% of the reference compound (alpha-tocopherol) . Antimicrob Agents Chemother, 2002 Nov, 46(11), 3676 - 8 Sequential emergence of antibiotic resistance in enterococcal bloodstream isolates over 25 years; Murdoch DR et al.; We determined the antibiotic susceptibilities of 1,785 enterococcal bloodstream isolates collected over 25 years . Antibiotic resistance emerged at a greater rate in Enterococcus faecium than in other enterococcal species, and E . faecium isolates became proportionally more common over time . Our findings confirm the pattern of emerging antibiotic resistance among enterococci and highlight the increasing importance of E . faecium as a cause of bloodstream infection. Antimicrob Agents Chemother, 2002 Nov, 46(11), 3657 - 9 Mutations in ribosomal protein L16 and in 23S rRNA in Enterococcus strains for which evernimicin MICs differ; Zarazaga M et al.; Mutations in ribosomal protein L16 and in 23S rRNA were investigated in 22 Enterococcus strains of different species and for which the MICs of evernimicin differ (MICs, 0.023 to 16 micro g/ml) . Amino acid changes (Arg56His, Ile52Thr, or Arg51His) in protein L16 were found in seven strains, and a nucleotide G2535A mutation in 23S rRNA was found in 1 strain among 13 for which the MICs are > or =1 micro g/ml. Infection, 2002 Oct, 30(5), 262 - 6 Glycopeptide use at four university hospitals in southern Germany; Kern WV et al.; BACKGROUND: Excessive use of glycopeptide antibiotics may enhance the risk of glycopeptide resistance among enterococci and staphylococi, but there is little data on the use of these antibiotics in German hospitals . METHODS: Hospital pharmacy records for the years 1992 to 1994 were evaluated . The number of defined daily doses (DDD) per 100 patient days was calculated to compare glycopeptide use between four state university hospitals . At one of the hospitals with comparatively low glycopeptide usage but an active antimicrobial management program, data were prospectively evaluated for 1995 to 2000 to assess the variation of glycopeptide use over time . RESULTS: The 3-year averages in glycopeptide use for the four hospitals ranged between 1.03 and 3.14 DDD/100 patient days . In all four hospitals, glycopeptide use was higher in the medical service (range, 1.59-7.26) than in the surgical service (range, 0.66-4.39) . Active antimicrobial management in one of the hospitals was associated with containment of glycopeptide consumption in the medical and surgical service at < 1.5 DDD/100 patient days in the last 3 years . CONCLUSION: Glycopeptide use differs considerably at tertiary care hospitals in southern Germany, but use of < 1.5 DDD/100 patient days in both surgical as well as medical tertiary care hospital departments appears achievable. Int J Food Microbiol, 2003 Jan 25, 80(2), 145 - 52 Effect of enterococcin A 2000 on biological and synthetic phospholipid membranes; Pantev A et al.; Lactic acid bacterium isolated from Bulgarian cheese and identified as Enterococcus faecium produces a small hydrophobic peptide substance (enterococcin A 2000) with broad spectrum of antimicrobial activity . The wide range of enterococcin antibacterial activity of this compound against Gram-positive, as well as against some Gram-negative bacteria, suggests a single mechanism of action . The mode of action of enterococcin A 2000 was studied in intact liver mitochondria and synthetic phospholipid liposomes used as model systems . Enterococcin A 2000 stimulated the ATPase activity in intact mitochondria . The kinetic curve of ATP hydrolysis differed from that obtained in presence of dinitrophenol (DNP) and showed a character similar to the ATP hydrolysis in the presence of classic ionophores . Enterococcin A 2000, when bound to synthetic phospholipid liposomes, permeabilized liposomes liberating the marker carboxyfluorescein (CF). Int J Food Microbiol, 2002 Dec 15, 79(3), 143 - 51 Vancomycin-resistant enterococci in shellfish, unchlorinated waters, and chicken; Wilson IG et al.; Vancomycin-resistant enterococci (VRE) have been a cause of increasing concern chiefly regarding the infection of hospital patients . There is suspicion, but limited evidence, that food and environmental spread may be important . Biomonitoring by examination of bivalve shellfish was used to assess the occurrence of VRE entering the environment . Using pre-enrichment and Lewisham and Slanetz and Bartley agars, 2/125 (1.6%) of shellfish were found to contain enterococci resistant to high levels of vancomycin . Lewisham agar allows relatively rapid identification of VRE . In a second phase of the work using pre-enrichment and Slanetz and Bartley agar, 4/151 (2.7%) shellfish and 5/27 (18.5%) raw chickens contained VRE . Using filtration and pre-enrichment, no VRE were found in 54 unchlorinated water samples . The study shows that environmental prevalence of VRE is low, and that raw chickens are frequently contaminated. Acta Anaesthesiol Scand, 2002 Oct, 46(9), 1075 - 81 Antibiotic prescription practices, consumption and bacterial resistance in a cross section of Swedish intensive care units; Walther SM et al.; BACKGROUND: The purpose of this work was to study usage of antibiotics, its possible determinants, and patterns of bacterial resistance in Swedish intensive care units (ICUs) . METHODS: Prospectively collected data on species and antibiotic resistance of clinical isolates and antibiotic consumption specific to each ICU in 1999 were analyzed together with answers to a questionnaire . Antibiotic usage was measured as defined daily doses per 1000 occupied bed days (DDD1000) . RESULTS: Data were obtained for 38 ICUs providing services to a population of approximately 6 million . The median antibiotic consumption was 1257 DDD1000 (range 584-2415) and correlated with the length of stay but not with the illness severity score or the ICU category . Antibiotic consumption was higher in the ICUs lacking bedside devices for hand disinfection (2193 vs . 1214 DDD1000, p=0.05) . In the ICUs with a specialist in infectious diseases responsible for antibiotic treatment the consumption pattern was different only for use of glycopeptides (58% lower usage than in other ICUs: 26 vs . 11 DDD1000,P=0.02) . Only 21% of the ICUs had a written guideline on the use of antibiotics, 57% received information on antibiotic usage at least every 3 months and 22% received aggregated resistance data annually . Clinically significant antimicrobial resistance was found among Enterbacter spp . to cephalosporins and among Enterococcus spp . to ampicillin . CONCLUSIONS: Availability of hand disinfection equipment at each bed and a specialist in infectious diseases responsible for antibiotic treatment were factors that correlated with lower antibiotic consumption in Swedish ICUs, whereas patient-related factors were not associated with antibiotic usage. Microb Drug Resist, 2002 Fall, 8(3), 161 - 70 Stability, persistence, and evolution of plasmid-encoded VanA glycopeptide resistance in enterococci in the absence of antibiotic selection in vitro and in gnotobiotic mice; Johnsen PJ et al.; Long-term persistence of VanA glycopeptide-resistant enterococci (GRE) has been observed in the absence of antibiotic selection . In the present study, we examined fitness parameters of a glycopeptide-susceptible Enterococcus faecium parent strain and its plasmid-mediated, VanA-resistant derivative before and after 1,000 generations in serial transfer broth cultures with or without antibiotic selection . With the exception of the vanA-containing plasmid, the strains were otherwise isogenic . The stability of the plasmid-encoded vanA resistance determinant was also investigated in vitro and in gnotobiotic mice . Competition experiments revealed that GRE with newly acquired VanA resistance had a 4% reduction in fitness relative to their susceptible parental counterpart . The relative difference in competitive fitness between resistant and susceptible strains was not significantly changed after 1,000 generations . Environmental adaptation was observed in all strains and exceeded the biological cost of resistance . Thus, the evolved VanA-resistant E . faecium populations out-numbered their unevolved ancestral susceptible E . faecium strain in mixed cultures, but remained less competitive than the evolved parent . The glycopeptide resistance determinant was similarly stably maintained during long-term colonization in gnotobiotic mice without antibiotic selection . In vivo vanA plasmid transfer was observed . The results suggest that environmental adaptation, in vivo gene transfer, and plasmid maintenance system(s) favor long-term VanA GRE persistence without antibiotic selection and compensate for the biological costs of possessing the resistance genes. Clin Orthop, 2002 Oct, (403), 38 - 44 Osteomyelitis attributable to vancomycin-resistant enterococci; Holtom PD et al.; Vancomycin-resistant enterococcus first was described in 1988, and has become a major problem in nosocomial infections . This is a retrospective review of 10 patients, seen at the authors' hospital during a 2-year period, with confirmed vancomycin-resistant enterococcal osteomyelitis: four patients had total joint arthroplasty infections, one patient had an infected tibial nail, three patients had infections associated with external fixators, and two patients had osteomyelitis of the femur . Four of the 10 patients had underlying medical illnesses (diabetes mellitus, systemic lupus erythematosus, human immunodeficiency virus infection); four of the 10 patients were intravenous drug users . Two patients had vancomycin-resistant enterococci on admission, and the other eight patients were admitted to the hospital for a mean of 21.3 days (range, 3-73 days) before vancomycin-resistant enterococci were identified in the bone . Eight of the 10 patients had monomicrobial infections with vancomycin-resistant enterococci . Patients were treated by surgical debridement, removal of hardware, and antibiotics (chloramphenicol in eight patients, quinupristin and dalfopristin (Synercid) in two patients) . All patients initially improved with therapy, but one patient had a recurrence of vancomycin-resistant enterococcal osteomyelitis and died of bacteremia . Bone infections with vancomycin-resistant enterococcus still may be uncommon, but with time and selective antibiotic pressures, vancomycin-resistant enterococci may become a more prominent entity in orthopaedic infections. J Antimicrob Chemother, 2002 Oct, 50(4), 569 - 76 Evaluation of antimicrobial therapy management of 120 consecutive patients with secondary peritonitis; Sotto A et al.; OBJECTIVES: To evaluate antimicrobial therapy management of secondary peritonitis in a University Hospital . PATIENTS AND METHODS: All patients admitted to the intensive care unit of the University Hospital of Nimes from 1 January 1997 to 31 July 1999 with a diagnosis of secondary peritonitis were retrospectively included . Patients' medical records were collected from the data recordings of the Department of Critical Care and Emergency and the Departments of Surgery . Acute Physiology and Chronic Health Evaluation II (APACHE II) was calculated for each patient at the time of admission . Antimicrobial treatment management before and after the diagnosis of peritonitis was studied . RESULTS: One hundred and twenty patients were included . Results concerning mortality, aetiology of peritonitis and microbiological data were in accordance with previous studies . APACHE II score (P = 0.005), age (P = 0.002), presence of Enterococcus in the peri-operative samples (P = 0.02) and period between diagnosis and surgery (P = 0.04) were predictive of death within 30 days after diagnosis of peritonitis . No significant difference was shown in the mortality rate in patients whose post-operative antibiotic treatment was changed following results of intra-operative peritoneal cultures versus patients having inappropriate treatment (P = 0.96) . The same observations were noted for anti-enterococcal treatment . CONCLUSION: This study emphasizes the importance of prompt surgical treatment and shows the modest impact of adapting antibiotic treatment . The morbidity and mortality associated with the presence of Enterococcus, which was not influenced by antibiotic treatment, would seem to suggest the pro-inflammatory role of Enterococcus . However, prospective randomized studies are needed to evaluate the real contribution of enterococcal antibiotic coverage in this context. Clin Infect Dis, 2002 Oct 15, 35(8), 935 - 42 Epub 2002 Sep 19. Effect of gastrointestinal bleeding and oral medications on acquisition of vancomycin-resistant Enterococcus faecium in hospitalized patients; Cetinkaya Y et al.; There has been minimal investigation of medications that affect gastrointestinal function as potential risk factors for the acquisition of vancomycin-resistant enterococci (VRE) . We performed a retrospective case-control study, with control subjects matched to case patients by time and location of hospitalization . Strict exclusion criteria were applied to ensure that only case patients with a known time of acquisition of VRE were included . Control patients were patients with > or =1 culture negative for VRE . The risk factors identified were use of vancomycin (odds ratio {OR}, 3.2; 95% confidence interval {CI}, 1.7-6.0; P=.0003), presence of central venous lines (OR, 2.2; 95% CI, 1.04-4.6; P=.04), and use of antacids (OR, 2.9; 95% CI, 1.5-5.6; P=.002) . Two protective factors included gastrointestinal bleeding (OR, 0.26; 95% CI, 0.08-0.79; P=.02) and use of Vicodin (Knoll Labs; hydrocodone and acetaminophen; OR, 0.93; 95% CI, 0.90-0.97; P=.0003) . Changes in gastrointestinal function, whether due to bleeding or to the effects of oral medications, may affect whether patients become colonized with VRE. FEMS Microbiol Lett, 2002 Sep 10, 214(2), 159 - 63 Influence of growth media on vancomycin resistance of Enterococcus isolates and correlation with resistance gene determinants; Nayak R et al.; The effect of Mueller-Hinton (MH), MH+blood or brain heart infusion medium (agar or broth) on 13 Enterococcus isolates was determined, when testing their antibiotic susceptibility . Disk diffusion and Vitek methods were used to determine vancomycin resistance, while broth dilution and E-test methods were used to measure the minimum inhibitory concentration . The data were correlated with the presence of vancomycin resistance genes . A definite correlation pattern could not be established between the presence of van genes and vancomycin resistance in any plating medium, when tested by the disk diffusion assay . The broth dilution, irrespective of the plating medium, and Vitek methods were more reliable than the E-test method in testing isolates with vanA or vanB genes . However, for vanC2/C3 genotypes, the E-test method, irrespective of the plating medium, tested better than the broth dilution assay. J Biol Chem, 2002 Nov 29, 277(48), 46779 - 84 Epub 2002 Sep 18. RNase 7, a novel innate immune defense antimicrobial protein of healthy human skin; Harder J et al.; We analyzed healthy human skin for the presence of endogenous antimicrobial proteins that might explain the unusually high resistance of human skin against infections . A novel 14.5-kDa antimicrobial ribonuclease, termed RNase 7, was isolated from skin-derived stratum corneum . RNase 7 exhibited potent ribonuclease activity and thus may contribute to the well known ribonuclease activity of human skin . RNase 7 revealed broad spectrum antimicrobial activity against many pathogenic microorganisms and remarkably potent activity (lethal dose of 90% < 30 nm) against a vancomycin-resistant Enterococcus faecium . Molecular cloning from skin-derived primary keratinocytes and purification of RNase 7 from supernatants of cultured primary keratinocytes indicate that keratinocytes represent the major cellular source in skin and that RNase 7 is secreted . RNase 7 mRNA expression was detected in various epithelial tissues including skin, respiratory tract, genitourinary tract, and at a low level, in the gut . In addition to a constitutive expression, RNase 7 mRNA was induced in cultured primary keratinocytes by interleukin-1beta, interferon-gamma, and bacterial challenge . This is the first report demonstrating RNases as a novel class of epithelial inducible antimicrobial proteins, which may play an important role in the innate immune defense system of human epithelia. Antimicrob Agents Chemother, 2002 Oct, 46(10), 3273 - 5 In vitro activity of the new oxazolidinone AZD2563 against Enterococci; Eliopoulos GM et al.; The activity of a new oxazolidinone antimicrobial, AZD2563, was assessed against >500 clinical isolates of enterococci representing six species . All isolates, including those resistant to other antibiotic classes, were inhibited by AZD2563 at concentrations </=2 micro g/ml, except for four strains resistant to linezolid . In most cases, AZD2563 was twofold more active than linezolid against enterococci. Antimicrob Agents Chemother, 2002 Oct, 46(10), 3253 - 6 Mutations in the aph(2")-Ic gene are responsible for increased levels of aminoglycoside resistance; Lee HK et al.; Random PCR mutagenesis of the enterococcal aph(2")-Ic gene followed by selection for mutant enzymes that confer enhanced levels of aminoglycoside resistance resulted in mutants of APH(2")-Ic with His-258-Leu and Phe-108-Leu substitutions, all of which conferred rises in the MICs of several aminoglycosides . The mutated residues are located outside conserved regions of aminoglycoside phosphotransferases. Antimicrob Agents Chemother, 2002 Oct, 46(10), 3125 - 32 Biochemical and genetic characterization of the vanC-2 vancomycin resistance gene cluster of Enterococcus casseliflavus ATCC 25788; Dutta I et al.; The vanC-2 cluster of Enterococcus casseliflavus ATCC 25788 consisted of five genes (vanC-2, vanXY(C-2), vanT(C-2), vanR(C-2), and vanS(C-2)) and shared the same organization as the vanC cluster of E . gallinarum BM4174 . The proteins encoded by these genes displayed a high degree of amino acid identity to the proteins encoded within the vanC gene cluster . The putative D,D-dipeptidase-D,D-carboxypeptidase, VanXY(C-2), exhibited 81% amino acid identity to VanXY(C), and VanT(C-2) displayed 65% amino acid identity to the serine racemase, VanT . VanR(C-2) and VanS(C-2) displayed high degrees of identity to VanR(C) and VanS(C), respectively, and contained the conserved residues identified as important to their function as a response regulator and histidine kinase, respectively . Resistance to vancomycin was expressed inducibly in E . casseliflavus ATCC 25788 and required an extended period of induction . Analysis of peptidoglycan precursors revealed that UDP-N-acetylmuramyl-L-Ala-delta-D-Glu-L-Lys-D-Ala-D-Ser could not be detected until several hours after the addition of vancomycin, and its appearance coincided with the resumption of growth . The introduction of additional copies of the vanT(C-2) gene, encoding a putative serine racemase, and the presence of supplementary D-serine in the growth medium both significantly reduced the period before growth resumed after addition of vancomycin . This suggested that the availability of D-serine plays an important role in the induction process. Pediatr Infect Dis J, 2002 Aug, 21(8), 798 - 800 Linezolid treatment of vancomycin-resistant Enterococcus faecium ventriculitis; Graham PL et al.; The successful treatment of a 7-month-old infant with shunt-associated ventriculitis caused by vancomycin-resistant Enterococcus faecium is presented . Linezolid was administered intravenously every 8 h; children have a greater volume of distribution and total body clearance than adults and therefore require more frequent dosing . The patient tolerated the therapy without adverse effects. Perit Dial Int, 2002 May-Jun, 22(3), 339 - 44 Comparison of vancomycin versus cefazolin as initial therapy for peritonitis in peritoneal dialysis patients; Khairullah Q et al.; The incidence of peritonitis ranges from 1 episode every 24 patient treatment months to 1 episode every 60 patient treatment months {Keane WF, et al . ISPD Guidelines/Recommendations . Adult peritoneal dialysis-related peritonitis treatment recommendations: 2000 update . Perit Dial Int 2000; 20:396-411.} . Gram-positive organisms account for over 80% of continuous ambulatory peritoneal dialysis (PD)-associated peritonitis . Recent fear of vancomycin-resistant enterococci (VRE) has prompted suggestions of limiting vancomycin use . Fifty-one episodes of peritonitis in 30 patients studied over 2 years were evaluated . Cloudiness of the PD fluid and/or abdominal pain were considered suggestive of peritonitis and were confirmed by cell count and culture . Baseline cell count, Gram stain, and cultures were obtained, with periodic follow-up . Patients were randomized to receive either vancomycin 1 g/L intraperitoneally (IP) as loading dose, repeated on day 5 or day 8, depending on residual renal function, for 2 weeks, or cefazolin 1 g in the first PD bag and continued with 125 mg/L every exchange for 2 or 3 weeks, depending on culture results . All patients also received gentamicin 40 mg IP every day until the culture results were available . A similar randomized trial comparing vancomycin and cefazolin in the past used a lower concentration of cefazolin 50 mg/L {Flanigan MJ, Lim VS . Initial treatment of dialysis associated peritonitis: a controlled trial of vancomycin versus cefazolin . Perit Dial /nt 1991; 11:31-7.} . Peritoneal dialysate fluid cultures revealed 31(60.7%) gram-positive organisms, 7(13.7%) gram-negative organisms, and 2 (3.9%) cultured yeast; 11 (21.5%) cultures yielded no growth . The incidence of peritonitis at our center was 1 episode every 42 patient treatment months . No case of VRE was noted . There was no statistical difference in clinical response or relapse rate for the two protocols . It was the authors' and nurses' observation that patient compliance and satisfaction was better with vancomycin, and the cost per treatment was 23% less than cefazolin . Based on these data we believe vancomycin should still be considered for first-line treatment of PD-associated peritonitis. Biosci Biotechnol Biochem, 2002 Jul, 66(7), 1597 - 600 Isolation of Enterococcus hirae mutant deficient in low-affinity potassium uptake at alkaline pH; Kawano M et al.; We here isolated an Enterococcus hirae mutant unable to grow well at pH 10 . The influx rate calculated from steady-state 42K+/K+ exchange and the intracellular K+ concentration of the mutant were reduced to 53 and 55% of those of the wild-type, respectively . The activities of two high-affinity K+ uptake systems, KtrI and KtrII, were normal in the mutant, but the kinetics of net K+ uptake at pH 10 indicated that a low-affinity K+ uptake with a Km of about 20 mM (Kawano, M, Abuki, R, Igarashi, K, Kakinuma, Y . (2001) Arch . Microbiol . 175: 41-45), which were seen in the wild-type, was deficient in this mutant. Epidemiol Infect, 2002 Aug, 129(1), 57 - 64 The epidemiology of glycopeptide-resistant enterococci on a haematology unit--analysis by pulsed-field gel electrophoresis; Bradley SJ et al.; As part of an interventional study to determine glycopeptide-resistant enterococci (GRE) acquisition on a three-ward haematology unit, rectal swabs were taken weekly from 293 patients recruited to the study between June 1995 and December 1996 . The GRE isolates obtained from the first positive rectal swab from 120 colonized patients, the isolates from 7 patients with clinical infection and 43 isolates obtained from the ward environment were compared by pulsed-field gel electrophoresis (PFGE) . Sixty-three of 120 patients were colonized by one of strains A-H, while 49 were colonized by unique strains . The first 18 weeks were associated with the highest prevalence of GRE by rectal swab, with a single strain A responsible for 52% of acquisitions on ward 2, 22% on ward 3 and 36% on ward 4 . Other smaller ward associated clusters were evident . Environmental sampling of ward 2 during this time showed that all but 2 of 30 isolates were indistinguishable from strain A . As the GRE prevalence fell, rectal swab and environmental isolates became more heterogeneous, and strain A disappeared after week 55 . GRE prevalence rose again in the final 15 weeks of the study, and a new predominant strain B emerged on ward 2 responsible for 50% of new acquisitions . In the seven patients with clinical infection with GRE, the clinical isolates were compared with the contemporaneous rectal swab isolate, and were found to be the same in only two cases . An analysis of five long-term carriers colonized for a median of 19 weeks (range 11-34) showed colonization with at least two and in one case six distinct strains, raising the question of how many strains may be colonizing a patient at any one time, and suggesting that multiple colonies should be analysed . These data suggest that cross-infection was an important factor in the spread of GRE when the colonization rate was high. J Antimicrob Chemother, 2002 Sep, 50(3), 397 - 401 First outbreak of vancomycin-resistant enterococci in a tertiary hospital in Turkey; Colak D et al.; Twenty multidrug-resistant vancomycin-resistant Enterococcus faecium strains of the VanA phenotype were isolated over a 1 year period from five patients in the intensive care unit at the University Hospital of Antalya, Turkey . Molecular investigation showed that these strains belonged to five different pulsotypes and that the vanA gene was carried by a Tn1546-like transposon inserted onto a self-transferable plasmid of approximately 200 kb . One patient was infected by two different strains, suggesting horizontal gene transfer within that patient . This is the first documented outbreak of VRE in Turkey with concomitant spread of plasmid and strains. Biochem Soc Trans, 2002 Aug, 30(4), 688 - 91 Role of proteolysis in copper homoeostasis; Solioz M; The cop operon of Enterococcus hirae controls cytoplasmic copper levels . It encodes two copper ATPases, a repressor, and the CopZ metallochaperone . Transcription of these genes is induced by copper . However, at higher copper concentrations, CopZ is degraded by a copper-activated proteolytic activity . This specific proteolysis of CopZ can also be demonstrated in vitro with E . hirae extracts . Growth of the cells in copper increases the copper-inducible proteolytic activity in extracts . Zymography reveals the presence of a copper-dependent protease in crude cell lysates . Copper-stimulated proteolysis of CopZ appears to play an important role in copper homoeostasis by E . hirae. Emerg Infect Dis, 2002 Sep, 8(9), 961 - 5 First characterization of a cluster of VanA-type glycopeptide-resistant Enterococcus faecium, Colombia; Panesso D et al.; From August 1998 to October 1999, glycopeptide-resistant enterococci (GRE) were isolated from 23 infected patients at a teaching hospital in Medellin, Colombia . Identification at the species level and by multiplex polymerase chain reaction assay indicated that all isolates were Enterococcus faecium . The isolates were highly resistant to ampicillin, ciprofloxacin, gentamicin, penicillin, streptomycin, teicoplanin, and vancomycin; they were susceptible only to chloramphenicol, linezolid, and nitrofurantoin . Determination of glycopeptide genotype indicated the presence of the vanA gene in all isolates . Molecular typing by pulsed field gel electrophoresis showed that all isolates were closely related . This study is the first molecular characterization of GRE in Colombia. Arch Microbiol, 2002 Sep, 178(3), 172 - 9 Epub 2002 Jun 20. Promoter analysis of the sodium-responsive V-ATPase (ntp) operon in Enterococcus hirae; Yasumura K et al.; The Enterococcus hirae ntp operon encodes all subunits of the vacuolar-type ATPase (V-ATPase), which transports Na(+) or Li(+) . This operon is expressed preferentially in response to Na(+), but not to Li(+) . Deletion analysis of the ntp promoter region in plasmids indicated that the AT-tract between -198 and -132 is required for Na(+)-specific transcriptional regulation . In addition, lithium-tolerant (LTR) mutants were isolated in which functional V-ATPase levels were high even in Na(+)-depleted medium . Western blot and Northern blot experiments revealed an increase in basal Na(+)-independent transcription in one of the mutants (LTR1) . The nucleotide sequences of the ntp promoter region of the LTR mutants showed mutational conversion of single base-pairs between positions -23 and +1 . Na(+)-independent expression of a reporter gene linked to the ntp promoter in plasmids was elevated by base substitutions at -23 to +1, and promoter activity induced by these base substitutions was lost by deletion of the region between -198 and -132 . These results suggest that the AT-tract between -198 and -132 is indispensable for transcription of the ntp operon. Infect Control Hosp Epidemiol, 2002 Aug, 23(8), 471 - 4 Impact of infection by vancomycin-resistant Enterococcus on survival and resource utilization for patients with leukemia; Bach PB et al.; We estimated the impact of vancomycin-resistant Enterococcus (VRE) infection on the outcomes of patients with leukemia in a case-control study . Compared with their matched controls (n = 45), cases (n = 23) had 22% greater total charges and shorter survival (P = .04) . These findings substantiate the need for aggressive interventions to prevent VRE transmission. Infect Control Hosp Epidemiol, 2002 Aug, 23(8), 468 - 70 An outbreak of vancomycin-resistant enterococci in a hematology-oncology unit: control by patient cohorting and terminal cleaning of the environment; Sample ML et al.; We describe the impact of enhanced infection control interventions on controlling the spread of vancomycin-resistant enterococci (VRE) in our hematology-oncology unit . Between April and September 1998, 13 patients on this unit were identified as having VRE . In addition to contact precautions, other measures that were needed to control the outbreak included closure of the unit to new admissions, creation of a cohort of VRE-positive patients and staff, and thorough cleaning of patients' rooms with 0.5% sodium hypochlorite. Infect Control Hosp Epidemiol, 2002 Aug, 23(8), 436 - 40 Recurrence of vancomycin-resistant Enterococcus stool colonization during antibiotic therapy; Donskey CJ et al.; OBJECTIVE: To test the hypothesis that antibiotic therapy may promote recurrence of vancomycin-resistant Enterococcus (VRE) stool colonization in patients who have previously had three consecutive negative stool cultures obtained at least 1 week apart . DESIGN: One-year prospective cohort study examining the effect of antibiotic therapy on recurrence and density of VRE stool colonization in patients who have cleared colonization . Pulsed-field gel electrophoresis (PFGE) was performed to determine whether recurrent VRE strains were the same clone as the previous colonizing strain . SETTING: A Department of Veterans Affairs medical center including an acute care hospital and nursing home . PATIENTS: All patients with at least one stool culture positive for VRE who subsequently had three consecutive negative stool cultures obtained at least 1 week apart . RESULTS: Of the 16 patients who cleared VRE colonization, 13 received antibiotic therapy during the study period . Eight (62%) of the 13 patients who received antibiotics developed recurrent high-density VRE stool colonization (range, 4.9 to 9.1 log10 colony-forming units per gram) during a course of therapy . Five patients had VRE strains available for PFGE analysis; recurrent strains were unrelated to the prior strain in 3 patients, closely related in 1 patient, and indistinguishable in 1 patient . CONCLUSIONS: Antibiotic therapy may be associated with recurrent high-density VRE stool colonization in many patients who have previously had three consecutive negative stool cultures . These patients should be screened for recurrent stool colonization when antibiotic therapy is administered. Infect Control Hosp Epidemiol, 2002 Aug, 23(8), 429 - 35 Cost-effectiveness of perirectal surveillance cultures for controlling vancomycin-resistant Enterococcus; Muto CA et al.; BACKGROUND: Several hospitals opting not to use active surveillance cultures to identify carriers of vancomycin-resistant Enterococcus (VRE) have reported that adoption of other parts of the Centers for Disease Control and Prevention guideline for controlling VRE has had little to no impact . Because use of surveillance cultures and contact isolation controlled a large outbreak at this hospital, their costs were estimated for comparison with the excess costs of VRE bacteremias occurring at a higher rate at a hospital not employing these measures . SETTING: Two university hospitals . METHODS: Inpatients deemed high risk for VRE acquisition at this hospital underwent weekly perirectal surveillance cultures . Estimated costs of cultures and resulting isolation during a 2-year period were compared with the estimated excess costs of more frequent VRE bacteremias at another hospital of similar size and complexity not using surveillance cultures to control spread throughout the hospital . RESULTS: Of 54,052 patients admitted, 10,400 had perirectal swabs taken . Cultures and isolation cost an estimated $253,099 . VRE culture positivity was limited to 193 (0.38%) and VRE bacteremia to 1 (0.002%) as compared with 29 bacteremias at the comparison hospital . The estimated attributable cost of VRE bacteremia at the comparison hospital of $761,320 exceeded the cost of the control program at this hospital by threefold . CONCLUSIONS: The excess costs of VRE bacteremia may justify the costs of preventive measures . The costs of VRE infections at other body sites, of deaths from untreatable infections, and of dissemination of genes for vancomycin resistance also help to justify the costs of implementing an effective control program. Infect Control Hosp Epidemiol, 2002 Aug, 23(8), 424 - 8 A prospective study to determine whether cover gowns in addition to gloves decrease nosocomial transmission of vancomycin-resistant enterococci in an intensive care unit; Srinivasan A et al.; BACKGROUND: Vancomycin-resistant enterococci (VRE) remain a significant nosocomial pathogen . Current guidelines of the Hospital Infection Control Practices Advisory Committee (HICPAC) of the Centers for Disease Control and Prevention (CDC) recommend the use of gowns and gloves for some interactions with VRE-infected or -colonized patients to prevent nosocomial transmission of VRE . OBJECTIVE: To assess the effect of disposable cover gowns on preventing nosocomial transmission of VRE . DESIGN AND SETTING: Prospective study in a 16-bed medical intensive care unit of a university teaching hospital . PATIENTS: All patients who were at risk to acquire VRE, were admitted to the intensive care unit from August 1998 to January 1999, and had at least two perirectal cultures were included in the analysis of VRE acquisition . INTERVENTION: VRE isolation precautions were changed from gowns and gloves to gloves alone . MAIN OUTCOME risk factors for VRE acquisition . RESULTS: The VRE acquisition rate was 1.80 cases per 100 days at risk in the gown and gloves period compared with 3.78 in the gloves only period (P = .04) . In a proportional hazards model adjusted for length of stay, gloves only precautions with a hazard ratio of 2.5 (P = .02; 95% confidence interval, 1.2 to 5.3) were the only independent risk factor for VRE acquisition . CONCLUSION: Our data lend support to current HICPAC recommendations for the use of cover gowns to decrease nosocomial transmission of VRE. Antimicrob Agents Chemother, 2002 Sep, 46(9), 3088 - 90 Incidence of high-level evernimicin resistance in Enterococcus faecium among food animals and humans; Aarestrup FM et al.; Six high-level evernimicin-resistant Enterococcus faecium isolates were identified among 304 avilamycin-resistant E . faecium isolates from animals and 404 stool samples from humans with diarrhea . All four animal isolates, and one of the human isolates, were able to transfer resistance to a susceptible E . faecium strain . The resulting transconjugants all tested positive for the presence of emtA, a gene encoding a methyltransferase previously linked with high-level evernimicin resistance . The four transconjugants derived from animal isolates all carried the same plasmid, while a differently sized plasmid was found in the isolate from humans . This study demonstrated a low incidence of high-level evernimicin resistance mediated by the emtA gene in different E . faecium isolates of animal and human origin. J Antimicrob Chemother, 2002 Aug, 50(2), 253 - 6 Association between the vanB2 glycopeptide resistance operon and Tn1549 in enterococci from France; Umeda A et al.; Linkage between the vanB2 gene cluster and transposon Tn1549 was found in clonally unrelated VanB-type vancomycin-resistant Enterococcus spp . strains isolated in France . The transposon was chromosomally located or plasmid borne. Scand J Infect Dis, 2002, 34(6), 471 - 2 Enterococcus casseliflavus septicaemia in a preterm neonate; Ruess M et al.; Enterococcus casseliflavus is a yellow-pigmented, motile, Gram-positive coccus that is only rarely isolated from clinical specimens . We report the first case of neonatal sepsis due to E . casseliflavus in a premature neonate . Precise identification of Enterococcus species is difficult but is of significant clinical interest because of the intrinsic low-level resistance to vancomycin of species such as E . casseliflavus and E . gallinarum. Presse Med, 2002 Jun 8, 31(20), 933 - 4 {Enterococcal meningitis revealing endocarditis}; Tonnellier M et al.; INTRODUCTION: Enterococci are frequently responsible for endocarditis, but a rare cause of meningitis . OBSERVATION: A 55 years-old man presented with Enterococcus faecium meningitis . Systematic transoesophageal echocardiography (TOE), despite the absence of organic murmur and the negativity of the hemocultures, revealed a concomitant aortic endocarditis . CONCLUSION: Review of the literature suggests that the association of endocarditis with enterococcal meningitis is far from accidental . We suggest that a TOS be conducted systematically when faced with this disease . The therapeutic implications are important, notably regarding the duration of antibiotherapy. Appl Environ Microbiol, 2002 Aug, 68(8), 4127 - 9 Antimicrobial resistance among enterococci from pigs in three European countries; Aarestrup FM et al.; Enterococci from pigs in Denmark, Spain, and Sweden were examined for susceptibility to antimicrobial agents and copper and the presence of selected resistance genes . The greatest levels of resistance were found among isolates from Spain and Denmark compared to those from Sweden, which corresponds to the amounts of antimicrobial agents used in food animal production in those countries . Similar genes were found to encode resistance in the different countries, but the tet(L) and tet(S) genes were more frequently found among isolates from Spain . A recently identified transferable copper resistance gene was found in all copper-resistant isolates from the different countries. Zh Mikrobiol Epidemiol Immunobiol, 2002 May-Jun, (3), 100 - 6 {Mechanisms of enterococcal survival in the host}; Bukharin OV et al.; Information on virulence and persistence factors of enterococci as important nosocomial pathogens with ever increasing multiple drug resistance is updated . The importance of the virulent and persistent properties of enterococci in the processes of adhesion to host tissues, invasion, modulation of inflammatory responses, as well as their toxic action on the host body is considered . The pathogenicity factors of enterococci are relevant to the mechanisms of the bacterial survival in the host. J Mol Microbiol Biotechnol, 2002 Jul, 4(4), 357 - 65 Bacterial bioinformatics: pathogenesis and the genome; Paine K et al.; As the number of completed microbial genome sequences continues to grow, there is a pressing need for the exploitation of this wealth of data through a synergistic interaction between the well-established science of bacteriology and the emergent discipline of bioinformatics . Antibiotic resistance and pathogenicity in virulent bacteria has become an increasing problem, with even the strongest drugs useless against some species, such as multi-drug resistant Enterococcus faecium and Mycobacterium tuberculosis . The global spread of Human Immunodeficiency Virus (HIV) and Acquired Immune Deficiency Syndrome (AIDS) has contributed to the re-emergence of tuberculosis and the threat from new and emergent diseases . To address these problems, bacterial pathogenicity requires redefinition as Koch's postulates become obsolete . This review discusses how the use of bacterial genomic information, and the in silico tools available at present, may aid in determining the definition of a current pathogen . The combination of both fields should provide a rapid and efficient way of assisting in the future development of antimicrobial therapies. Microb Drug Resist, 2002 Summer, 8(2), 133 - 8 Vancomycin-resistant enterococci (VRE) in broiler flocks 5 years after the avoparcin ban; Heuer OE et al.; The glycopeptide growth promoter avoparcin was banned from animal production in Denmark in 1995 . In this study, we investigated the occurrence of vancomycin-resistant enterococci (VRE) in broiler flocks in the absence of the selective pressure exerted by the use of avoparcin . One hundred sixty-two broiler flocks from rearing systems with different histories of avoparcin exposure were investigated for the presence of VRE . Using a direct selective plating procedure, VRE were isolated from 104 of 140 (74.3%) broiler flocks reared in broiler houses previously exposed to avoparcin on conventional and extensive indoor broiler farms . In contrast, only 2 of 22 (9.1%) organic broiler flocks reared on free-range farms with no history of previous exposure to avoparcin were VRE-positive . Furthermore, the occurrence of VRE over time in flocks reared in broiler houses previously exposed to avoparcin was investigated . Results obtained by direct selective plating showed no significant decrease in the proportion of VRE-positive flocks during the study period (1998-2001) . This study demonstrated the extensive occurrence of VRE in broiler flocks more than 5 years after the avoparcin ban in Denmark, and indicates that VRE may persist in the absence of the selective pressure exerted by avoparcin . The results differ markedly from previously published Danish surveillance data on VRE in broilers . This may reflect differences in isolation procedures. Intensive Care Med, 2002 Jun, 28(6), 692 - 7 Epub 2002 Apr 12. Vancomycin-sensitive and vancomycin-resistant enterococcal infections in the ICU: attributable costs and outcomes; Pelz RK et al.; OBJECTIVES: To determine the economic and clinical outcomes associated with infection with vancomycin-resistant Enterococcus (VRE) and to compare these outcomes to those associated with infection with vancomycin-sensitive Enterococcus (VSE) . METHODS: During a 3-month, prospective, cohort study of 117 high-risk, critically ill patients we collected complete clinical and demographic and ICU cost data from all patients during their ICU stays . RESULTS: After adjusting for variables in a stepwise multiple regression model VRE infections were associated with a median attributable increased ICU cost per patient of $33,251 (38,088 euros) and an increased length of hospital stay (LOS) of 22 days, while VSE infections were associated with an increased cost of $21,914 (25,102 euros) and an increased LOS of 27 days . The effect of VRE and VSE infections were not significantly different . Over the entire cohort the attributable cost per ICU patient day associated with VRE infection was $304 (348 euros) . CONCLUSIONS: The attributable cost of ICU care associated with VRE infection is $33,251 (38,088 euros) and per ICU patient day was $304 (348 euros) . VRE and VSE infections do not differ in associated cost of ICU care, LOS, or mortality . Any VRE control strategy is be cost-effective if the overall cost per ICU patient-day is less than $304 (348 euros). Diagn Microbiol Infect Dis, 2002 Jul, 43(3), 183 - 8 Risk factors for acquisition of vancomycin-resistant enterococci among hematology-oncology patients; Suntharam N et al.; The incidence of VRE has increased dramatically and hematology-oncology patients are at high risk for acquisition of colonization and development of infection . Therefore, we performed a prospective cohort study to determine risk factors for VRE acquisition among hematology-oncology patients . Patients admitted to a single unit at Northwestern Memorial Hospital, which was predominantly comprised of patients with hematologic malignancies and recipients of hematopoietic stem cell transplants, were enrolled . Rectal or perianal swabs were obtained on hospital day 1, 4, 7 and then weekly thereafter . Data were collected by medical record review . We evaluated 155 study patients; 12 patients (7.7%) converted from VRE negative to positive . Among these 12 patients, 3 were positive on prior admissions, and 9 acquired VRE during the study . The median time to acquisition was 9 days . The median length of stay was significantly longer for patients with VRE compared to those who were VRE negative (31 vs . 6 days, P < 0.01) . Patients with VRE were significantly more likely than those without VRE to have had an ICU admission within 3 months (P = 0.003), been admitted from an acute care facility (P = 0.001), or to have received amikacin (P = 0.02) . Antimicrobials were commonly prescribed to all of the patients as 87% received an antimicrobial prior to their first swab . The crude mortality rate for patients with VRE was 67% . Prolonged length of stay, prior hospitalization, previous ICU admission and receipt of amikacin were risk factors associated with VRE acquisition among hematology-oncology patients . Mortality among these patients was high, due to serious underlying disease. Semin Dial, 2002 May-Jun, 15(3), 162 - 71 National surveillance of dialysis-associated diseases in the United States, 2000; Tokars JI et al.; In December 2000, all U.S . dialysis centers were surveyed regarding selected patient care practices and dialysis-associated diseases . The results were compared with similar surveys conducted in previous years . During 1997-2000, the percentage of patients vaccinated against hepatitis B virus infection increased from 47% to 58% and the percentage of staff vaccinated increased from 87% to 88% . In 2000, an estimated 64% of patients were vaccinated for influenza and 27% for pneumococcal pneumonia . In 2000, routine testing for antibody to hepatitis C virus (anti-HCV) was performed on staff at 40% of centers and on patients at 58% of centers; anti-HCV was found in 1.7% of staff and 8.4% of patients . During 1995-2000, the percentage of patients who received dialysis through central catheters increased from 13% to 24%; this trend is worrisome because infections and antimicrobial use are higher in patients receiving dialysis through catheters . However, during the same period the percentage of patients receiving dialysis through fistulas increased from 22% to 28% . In 2000, 25% of catheters were used for new patients awaiting an implanted access, 28% for established patients with a failed access awaiting a new implanted access, 41% as an access of last resort, and 6% for other reasons, including patient preference . The percentage of centers reporting one or more patients infected or colonized with vancomycin-resistant enterococcus (VRE) increased from 11.5% in 1995 to 32.7% in 2000. Arch Phys Med Rehabil, 2002 Jul, 83(7), 899 - 902 Patient colonization and environmental contamination by vancomycin-resistant enterococci in a rehabilitation facility; Trick WE et al.; OBJECTIVES: To determine the frequency of environmental contamination in patient and common-use rooms and patient colonization by vancomycin-resistant enterococci (VRE) . DESIGN: Cross-sectional study . SETTING: A 146-bed rehabilitation facility . PARTICIPANTS: Rectal cultures were collected from 74 (80%) of 93 patients . Environmental cultures were obtained from surfaces in 15 patient rooms (5 floors) and common-use areas on 8 floors . INTERVENTIONS: Not applicable . MAIN OUTCOME MEASURES: Gastrointestinal colonization of patients and environmental contamination of surfaces by VRE . RESULTS: VRE was detected from 13 (18%) of 74 patients and 32 (10%) of 319 surfaces . The frequency of positive environmental cultures varied by location; cultures were more likely to be positive in patient rooms (15%), followed by common areas on patient floors (9%) and common areas separate from patient floors (1.3%) . Surfaces were more likely to be positive in rooms with a VRE-colonized patient (24%), compared with rooms in which patient colonization status was unknown (13%, P=.13) or the patient was not colonized (0%, P=.002) . Surfaces were more likely to be contaminated in a room that housed an incontinent compared with continent patients (22% vs 7%, P=.01) . CONCLUSIONS: Although environmental contamination by VRE was common in patient rooms, contamination of common-use areas separate from patient floors was infrequent . Despite use of common-use areas by colonized patients, isolation practices at this facility appear to have minimized environmental surface contamination in these areas . J Vasc Surg, 2002 Jul, 36(1), 180 - 3 Stent graft infection after abdominal aortic aneurysm repair: a case report; Baker M et al.; A 77-year-old man had clinical and radiologic signs of graft infection develop 1 year after stent grafting for abdominal aortic aneurysm . Blood cultures grew Bacteroides fragilis, and cultures of the aneurysm sac grew Enterococcus . The patient's condition was successfully managed with staged extraanatomic revascularization followed by graft excision . Although stent graft infection to date is extremely rare, some aspects peculiar to the placement of these devices potentially could increase their susceptibility to infection . Recognition and standard techniques in management can lead to successful outcome. Planta Med, 2002 Jun, 68(6), 541 - 3 Antibacterial activity of calozeyloxanthone isolated from Calophyllum species against vancomycin-resistant Enterococci (VRE) and synergism with antibiotics; Sakagami Y et al.; Calozeyloxanthone ( 1) was re-isolated from the root bark of Calophyllum moonii, an endemic species of Sri Lanka, and found to be active against vancomycin-resistant Enterococci (VRE) and vancomycin-sensitive Enterococci (VSE) with MIC values of 6.25 microg/ml and 12.5 microg/ml, respectively . Further, a marked synergism between 1 and vancomycin hydrochloride (VCM) against VRE was also observed . These findings suggest that 1 in combination with VCM against VRE may be useful in controlling VRE infections. J Am Soc Echocardiogr, 2002 Jul, 15(7), 702 - 7 Better prognosis of elderly patients with infectious endocarditis in the era of routine echocardiography and nonrestrictive indications for valve surgery; Zamorano J et al.; OBJECTIVE: It has been reported that endocarditis in the elderly may have a poor outcome . Our aim was to assess the different features and prognosis, if any, in the present time . METHODS: Of 103 patients with proven endocarditis, 31 were 65 years or older and 72 were younger than 65 years . Degenerative heart disease was seen more frequently in the elderly (22.5% vs 2.7%, P =.003) . Drug abuse and immunodeficiency virus infection were more common in the younger group, as was tricuspid endocarditis (26.3% vs 0%, P <.001) . At clinical presentation cardiac failure (41.9 vs 19.4%, P =.02) and leukocytosis (61.2% vs 40.2%, P =.049) were seen more frequently in the elderly . RESULTS: Despite other similar clinical features, it took longer to diagnose older patients (7.2 +/- 6.2 vs 3.2 +/- 3.5 days, P <.001) . Enterococcus infected the aged more often (32.2% vs 13.1%, P =.001) . During hospitalization, heart failure and embolization tended to be more common in the elderly and the younger group, respectively . There were no significant differences in the incidence of anatomic complications, the need for operation, and overall mortality . CONCLUSION: Although a worse prognosis has been reported in elderly patients with infective endocarditis, the early use of transesophageal echocardiographic examinations and equal therapeutic options provides a similar outcome when compared with younger subjects. J Clin Microbiol, 2002 Jul, 40(7), 2689 - 90 Native valve endocarditis due to Enterococcus hirae; Poyart C et al.; Enterococcus hirae is a rare isolate in clinical specimens . We describe a case of native aortic-valve endocarditis that was caused by Enterococcus hirae in a 72-year-old man . This is the first reported case of endocarditis due to this organism. Contemp Top Lab Anim Sci, 1999 Nov, 38(6), 39 - 41 Pyometra in a Siberian Polecat (Mustela eversmanni); Johnson JD et al.; A 2-year-old Siberian polecat (Mustela eversmanni) from a breeding colony presented for ultrasound evaluation for pregnancy . It was paired with a male for 2.75 months and had remained absent of pregnancy signs when it was anesthetized and clinically evaluated . Until this time, the animal had eaten well and shown no outward signs of debility . On palpation, the animal had a fluid-filled tubular structure in the caudal abdomen, consistent in location and size with the uterus . No sign of vaginal discharge was present . Ultrasonography revealed 10 fluid-filled evaginations (approximately 12 mm in diameter) of the uterine horns . A presumptive diagnosis of a fluid-filled reproductive tract and likely reproductive failure was made in light of the animal's history, its clinical signs, and the ultrasound findings . Euthanasia was performed because the animal was nonreproductive and might yield information relevant to the breeding colony as a whole . Necropsy of the polecat revealed a distended fluctuant uterus containing mildly odiferous, thick, yellow-green, purulent material . Histopathology confirmed the diagnosis of pyometra . A pure and heavy growth of Enterococcus fecalis was cultured from the uterine contents . In light of results from routine minimal inhibitory concentration antibiotic sensitivity screening, this isolate was resistant to all antibiotics tested in the standard teaching hospital screen. Clin Nephrol, 2002 Jun, 57(6), 468 - 73 Cerebral, myocardial and cutaneous ischemic necrosis associated with calcific emboli from aortic and mitral valve calcification in a patient with end-stage renal disease; Li Y et al.; We report the case of a 57-year-old diabetic male with chronic renal failure who developed secondary hyperparathyroidism and calcification of mitral and aortic valves and interatrial septum . Multiple ischemic lesions developed in the skin of hands, feet and penis, and in the brain, and these were presumed to be due to septic emboli from cardiac valvular infective endocarditis . Multiple blood cultures were negative, however, and despite antibiotic therapy the patient expired . Autopsy (limited to trunk) demonstrated multiple calcific emboli in the heart and spleen, apparently derived from the prominent calcific deformities in the aortic and mitral valves . These were associated with acute and organizing myocardial infarcts and acute splenic infarcts, suggesting that the multiple ischemic lesions in the brain were also due to calcific emboli . A possible contributory component of infective endocarditis, however, was indicated by postmortem cultures of aortic and mitral valves positive for Enterococcus faecium . Calcific embolism is a rarely recognized but potentially lethal complication of end-stage renal disease, and the clinical diagnosis and the preventive therapeutic options for the control of the product of calcium and phosphate and/or parathyroidectomy should be considered. J Biol Chem, 2002 Sep 27, 277(39), 35801 - 7 Epub 2002 Jun 19. Balance between two transpeptidation mechanisms determines the expression of beta-lactam resistance in Enterococcus faecium; Mainardi JL et al.; The d,d-transpeptidase activity of high molecular weight penicillin-binding proteins (PBPs) is essential to maintain cell wall integrity as it catalyzes the final cross-linking step of bacterial peptidoglycan synthesis . We investigated a novel beta-lactam resistance mechanism involving by-pass of the essential PBPs by l,d-transpeptidation in Enterococcus faecium . Determination of the peptidoglycan structure by reverse phase high performance liquid chromatography coupled to mass spectrometry revealed that stepwise selection for ampicillin resistance led to the gradual replacement of the usual cross-links generated by the PBPs (d-Ala(4) --> d-Asx-Lys(3)) by cross-links resulting from l,d-transpeptidation (l-Lys(3) --> d-Asx-Lys(3)) . This was associated with no modification of the level of production of the PBPs or of their affinity for beta-lactams, indicating that altered PBP activity was not required for ampicillin resistance . A beta-lactam-insensitive l,d-transpeptidase was detected in membrane preparations of the parental susceptible strain . Acquisition of resistance was not because of variation of this activity . Instead, selection led to production of a beta-lactam-insensitive d,d-carboxypeptidase that cleaved the C-terminal d-Ala residue of pentapeptide stems in vitro and caused massive accumulation of cytoplasmic precursors containing a tetrapeptide stem in vivo . The parallel dramatic increase in the proportion of l-Lys(3) --> d-Asx-Lys(3) cross-links showed that the enzyme was activating the resistance pathway by generating the substrate for the l,d-transpeptidase. Chem Biol Interact, 2002 May 20, 140(2), 185 - 98 The antimicrobial properties of milkfat after partial hydrolysis by calf pregastric lipase; Sun CQ et al.; Studies on the kinetic characteristics of calf pregastric lipase (EC 3.1.1.3) have shown that it preferentially releases short chain fatty acids (SCFAs) from bovine milkfat . The released fatty acids form mixed micelle structures . The aim of this investigation has been to test whether hydrolysed milkfat is antimicrobial, and how the state of the emulsion alters the bactericidal or bacteriostatic effects . Partial hydrolysis of milkfat by pregastric lipase was carried out in two types of emulsion systems, containing either Triton X-100 or casein/lecithin, plus milkfat in citrate/phosphate buffer (pH 5.0-6.0) . The concentrations and compositions of fatty acids were determined by gas chromatography . The minimum percentages of hydrolysed milkfat which affected growth and survival of selected Gram-positive and Gram-negative bacteria were measured . The bacterial experiments were repeated using pure fatty acids at similar concentrations . Lauric acid (C12:0) was found to be the most potent bactericidal fatty acid against Enterococcae (Gram-positive), and caprylic acid (C8:0) was the most potent against coliforms (Gram-negative) . Use of Triton X-100 for milkfat emulsification provided a more compatible medium for studying bacterial growth in the hydrolysed milkfat than did use of casein/lecithin . The results also show that the antimicrobial effects of individual fatty acids released from hydrolysed milkfat were at least additive and suggest that hydrolysis of milkfat may be a significant factor in controlling growth of organisms imbibed with food in pre-weaned animals . The amount of pregastric catalyzed triglyceride hydrolysis in the digestive tract is sufficient to produce an antibacterial concentration of fatty acids and monoglycerides. Int J Food Microbiol, 2002 Jul 25, 77(1-2), 109 - 15 Gastrointestinal transit survival of an Enterococcus faecium probiotic strain administered with or without vancomycin; Lund B et al.; The primary aim of this study was to evaluate if an ingested probiotic, containing viable Enterococcus faecium could survive gastrointestinal transit and if so, correlate the amount of the recovered probiotic strain with the host's own enterococci . The second aim was to investigate if simultaneous vancomycin intake influenced the survival and persistence of the probiotic strain and the stability of endogenous enterococci strains . Twenty healthy volunteers were given the probiotic product once daily for 10 days . Half of the subjects were simultaneously given vancomycin . Isolates of E . faecium strains were genotypically or phenotypically analysed with pulsed-field gel electrophoresis (PFGE) and the PhenePlate system, respectively . In eight of the ten volunteers given only the probiotic, the ingested E . faecium could be detected on day 10, while in none on day 31 . From subjects given both probiotic and vancomycin no ingested E . faecium could be detected on day 10 or day 31 . The estimated amount of ingested E . faecium recovered from faeces on day 10 ranged from 1.2 x 10(3) to 4.2 x 10(6) colony forming units per gram faeces, which in several cases were a substantial part of the total amount of E . faecium . The E . faecium isolated before probiotic plus vancomycin administration showed no close relationship to the ones isolated 3 weeks after ceased intake in any subjects . In conclusion, the ingested E . faecium strain can survive gastrointestinal transit . After intake, the E . faecium probiotic strain might become a large part of the total E . faecium population . The occurrence of the probiotic strain in the human gut seems to be transient after intake stop . Re-colonization of E . faecium after simultaneous probiotic plus vancomycin intake occurs mainly with strains without close genetic relationship to the strains harboured before treatment or to the ingested E . faecium strain. FEMS Microbiol Rev, 2002 Jun, 26(2), 163 - 71 Enterococci from foods; Giraffa G; Enterococci have recently emerged as nosocomial pathogens . Their ubiquitous nature determines their frequent finding in foods as contaminants . In addition, the notable resistance of enterococci to adverse environmental conditions explains their ability to colonise different ecological niches and their spreading within the food chain through contaminated animals and foods . Enterococci can also contaminate finished products, such as fermented foods and, for this reason, their presence in many foods (such as cheeses and fermented sausages) can only be limited but not completely eliminated using traditional processing technologies . Enterococci are low grade pathogens but their intrinsic resistance to many antibiotics and their acquisition of resistance to the few antibiotics available for treatment in clinical therapy, such as the glycopeptides, have led to difficulties and a search for new drugs and therapeutic options . Enterococci can cause food intoxication through production of biogenic amines and can be a reservoir for worrisome opportunistic infections and for virulence traits . Clearly, there is no consensus on the acceptance of their presence in foodstuffs and their role as primary pathogens is still a question mark . In this review, the following topics will be covered: (i) emergence of the enterococci as human pathogens due to the presence of virulence factors such as the production of adhesins and aggregation substances, or the production of biogenic amines in fermented foods; (ii) their presence in foods; (iii) their involvement in food-borne illnesses; (iv) the presence