J Infect Dis, 2005 Feb 15, 191(4), 588 - 95 Epub 2005 Jan 17. Impact of Vancomycin Resistance on Mortality among Patients with Neutropenia and Enterococcal Bloodstream Infection; Diazgranados CA et al.; We performed a retrospective cohort study to measure the impact of vancomycin resistance on clinical outcome for 83 episodes of enterococcal bloodstream infection (BSI; 22 with vancomycin-resistant enterococci {VRE} and 61 with vancomycin-susceptible enterococci {VSE}) in 77 patients with neutropenia . Cox proportional hazards models showed that vancomycin resistance was an independent predictor of mortality, after controlling for severity of illness, enterococcal species, gram-negative copathogens, sex, race, duration of neutropenia before bacteremia, and early administration of active antibiotics . This effect was evident only >/=10 days after the onset of bacteremia (P=.0263; hazard ratio {HR}, 4.969) but not after adjustment for duration of bacteremia . The median duration of bacteremia was 4.5 days for VRE BSI and <1 day for VSE BSI (P = .0001) . The only independent predictor of bacteremia duration was vancomycin resistance (P=.0284; HR, 3.863) . Vancomycin resistance is associated with increased mortality in patients with neutropenia, possibly because of prolonged duration of bacteremia. Microb Drug Resist, 2004 Winter, 10(4), 341 - 5 Distribution of the erm (B) gene, tetracycline resistance genes, and Tn1545-like transposons in macrolide- and lincosamide-resistant enterococci from pigs and humans; De Leener E et al.; The distribution of the erm (B) and the tetracycline resistance genes tet(K), tet(L), tet(M), tet(O), and tet(S) was investigated among macrolide- and lincosamide-resistant enterococci originating from humans, pigs, and pork carcasses . The presence of transposons of the Tn916/Tn1545 family was also traced in these isolates . Furthermore, the porcine strains were tested for the presence of glycopeptide resistance genes vanA and vanB . The erm(B) gene was found in 85% of the porcine and in all human isolates . Ninety-eight percent of the porcine and 89% of the human erm(B)-positive enterococci carried the tet(M) gene . Seventy-seven percent and 70%, respectively, of these strains harbored a Tn1545-like element . Tet(L) was observed in 68% of the porcine and in 65% of the human enterococci . The other tetracycline resistance genes were very rare and the glycopeptide resistance genes vanA and vanB were not detected among the porcine isolates . The similar frequencies of resistance genes and the highly mobile Tn1545-like transposon among porcine and human enterococci might indicate exchange of resistant strains or their resistance genes between humans and pigs or the existence of a common reservoir. Can J Microbiol, 2004 Sep, 50(9), 767 - 770 Identification of an unusual VanA element in glycopeptide-resistant Enterococcus faecium in Brazil following international transfer of a bone marrow transplant patient; Camargo IL et al.; A vancomycin-resistant Enterococcus (VRE) was isolated from a blood culture of a patient in a Brazilian hospital who had a treatment history of a bone marrow transplant in the USA . The organism was identified as Enterococcus faecium, which exhibited an MIC (minimum inhibitory concentration) &ge; 256 microg/mL for vancomycin . This was confirmed by E-test and the vanA gene was detected by PCR . Overlapping PCR revealed a left IR deletion and an additional 1.5 kb fragment between vanSH genes . DdeI digestion of vanRSHAX genes showed the determinant to be a T type variant, and the element was cloned and sequenced . These results revealed an IS1251 downstream of nucleotide 5820 of the VanA element . Insertions like this have not been reported previously in Brazil, but have been detected in the USA . The genotype and association with a patient previously treated in the USA suggest that this VRE was introduced from abroad, probably through inter-hospital strain spread. Neoplasma, 2004, 51(5), 341 - 4 Influence of diet containing lyophilized Enterococcus faecium M-74 with organic selenium on tumor incidence in Apc1638N mice; Hlubinova K et al.; The aim of the present study was to test the effect of long-term application of diet containing Enterococcus faecium M-74 with organic selenium on tumor induction in transgenic mice carrying mutation in Apc gene . Heterozygosity for the Apc1638N mutation in mice causes development of small intestine and gastric tumors . Feeding of Apc1638N transgenic mice with enriched diet with probiotic components during 8 months have shown a minor therapeutic effect on the clinical manifestations in small intestine in comparison with control group. Bone Marrow Transplant . 2005 Jan 10; {Epub ahead of print} Early vancomycin-resistant enterococcus (VRE) bacteremia after allogeneic bone marrow transplantation is associated with a rapidly deteriorating clinical course; Avery R et al.; Summary:Vancomycin-resistant enterococcal (VRE) infection is a growing threat . We studied the incidence, risk factors, and clinical course of early-onset VRE bacteremia in allogeneic hematopoietic stem cell transplant recipients . We carried out a chart review of 281 allogeneic hematopoietic stem cell transplant recipients from 1997-2003, including preparative regimen, diagnosis, status of disease, graft-versus-host disease prophylaxis, antimicrobial therapy, and survival . VRE bacteremia developed in 12/281 (4.3%) recipients; 10 (3.6%) were within 21 days of transplant . Diagnoses were acute leukemia (7), NHL (2), and MDS (1) . In all, 70% had refractory/relapsed disease; 30% were in remission . In total, 50% had circulating blasts . Nine of 10 had matched unrelated donors (7/9 with CD8+ T-cell depletion) . The average time to positive VRE cultures was 15 days; average WBC was 0.05, and 80% had concomitant infections . Despite treatment, all patients died within 73 days of VRE bacteremia . Intra-abdominal complications were common . Causes of death included bacterial or fungal infection, multiorgan failure, VOD, ARDS, and relapse . A total of 60% of patients engrafted neutrophils, but none engrafted platelets . Early VRE bacteremia after allogeneic bone marrow transplant is associated with a rapidly deteriorating clinical course, although not always directly due to VRE . Early VRE may be a marker for the critical condition of these high-risk patients at the time of transplant.Bone Marrow Transplantation advance online publication, 10 January 2005; doi:10.1038/sj.bmt.1704821. Curr Opin Infect Dis, 2004 Dec, 17(6), 541 - 7 Approaches to vancomycin-resistant enterococci; Torres-Viera C et al.; PURPOSE OF REVIEW: This article reviews recent publications regarding new antimicrobial drugs for the treatment of vancomycin-resistant enterococci . RECENT FINDINGS: Newer drugs against vancomycin-resistant enterococci are now available or will soon be available . Quinupristin-dalfopristin, a streptogramin, and linezolid, an oxazolidinone, are effective and safe but only bacteriostatic against enterococi . Bacterial isolates resistant to either antibiotic have been described . Daptomycin, a lipopeptide antimicrobial, has good in-vitro bactericidal activity against enterococci, but very limited clinical data exist regarding the treatment of serious enterococcal infection with this compound . Ramoplanin, the first glycolipodepsipeptide antimicrobial in clinical trials, is not systemically absorbed after oral administration, and is being evaluated for the prevention of bloodstream infection in patients colonized with vancomycin-resistant enterococci . Oritavancin and dalbavancin (both glycopeptides) and tigecycline (a monocycline derivative) are being evaluated in phase II and III trials and are not yet commercially available . SUMMARY: Treatment of vancomycin-resistant enterococci continues to be problematical although these new drugs offer some hope . The rational use of antibiotics, strict guidelines for the use of new compounds, and adherence to infection control practices continue to be essential components of the management of vancomycin-resistant enterococci colonization and infection. Appl Environ Microbiol, 2005 Jan, 71(1), 512 - 8 Methods to increase fidelity of repetitive extragenic palindromic PCR fingerprint-based bacterial source tracking efforts; Hassan WM et al.; The goal of the study was to determine which similarity coefficient and statistical method to use to produce the highest rate of correct assignment (RCA) in repetitive extragenic palindromic PCR-based bacterial source tracking . In addition, the use of standards for deciding whether to accept or reject source assignments was investigated . The use of curve-based coefficients Cosine Coefficient and Pearson's Product Moment Correlation yielded higher RCAs than the use of band-based coefficients Jaccard, Dice, Jeffrey's x, and Ochiai . When enterococcal and Escherichia coli isolates from known sources were used in a blind test, the use of maximum similarity produced consistently higher RCAs than the use of average similarity . We also found that the use of a similarity value threshold and/or a quality factor threshold (the ratio of the average fingerprint similarity within a source to the average similarity of this source's isolates to an unknown) to decide whether to accept source assignments of unknowns increases the reliability of source assignments . Applying a similarity value threshold improved the overall RCA (ORCA) by 15 to 27% when enterococcal fingerprints were used and 8 to 29% when E . coli fingerprints were used . Applying the quality factor threshold resulted in a 22 to 32% improvement in the ORCA, depending on the fingerprinting technique used . This increase in reliability was, however, achieved at the expense of decreased numbers of isolates that were assigned a source. J Food Prot, 2004 Dec, 67(12), 2772 - 8 Cloning of the bile salt hydrolase (bsh) gene from Enterococcus faecium FAIR-E 345 and chromosomal location of bsh genes in food enterococci; Wijaya A et al.; Enterococcus faecium strain FAIR-E 345 isolated from food was shown to possess bile salt hydrolase (Bsh) activity in a plate screening assay and by high-performance liquid chromatography analysis . The bsh gene was cloned and sequenced . DNA sequence analysis revealed that it encoded a protein of 324 amino acids, with pI 4.877 . A bsh gene probe was prepared from the cloned bsh gene and was used for probing plasmid and total genomic DNA of Bsh-positive enterococci isolated from food to determine the genomic location of their bsh genes . This probe was able to detect the bsh gene among total genomic DNA preparations but not from plasmid preparations of 10 plasmid-bearing Enterococcus strains . However, the probe could detect the bsh gene from total genomic DNA preparations of 12 Enterococcus strains that did not contain detectable plasmid DNA . In no cases did the probe hybridize with plasmid DNA preparations, suggesting that the bsh gene among enterococci is probably generally chromosomally encoded . This presumptive chromosomal location of bsh genes among food enterococci suggests that transfer of this trait by conjugative plasmids is unlikely. Ann Agric Environ Med, 2004, 11(2), 355 - 8 Microbial quality of water in dental unit reservoirs; Szymanska J et al.; Microbial quality of water in a dental unit is of considerable importance since patients and dental staff are regularly exposed to water and aerosol generated by the unit . Water delivered to a dental unit by the so-called independent water system is the water coming from a reservoir which, at the same time, is an initial part of dental unit waterlines (DUWL) . Thus, microbiological quality of this water is extremely important for the quality of water flowing from dental handpieces . The aim of the study was to assess microbiologically the water contained in dental unit reservoirs . Water samples were collected aseptically from the water reservoirs of 19 dental units . Results concerning microbial contamination: potable water quality indices, and detection and isolation of Legionella species bacteria, were presented . Over a half of the samples did not comply with the norms for potable water . In 63.1% of the cases, the number of colony forming units (cfu/ml) and of coliform organisms significantly exceeded acceptable values . Enterococcus was not detected in the samples of examined water . Similarly, no Legionella were found in the samples of dental unit reservoirs water . Reservoirs as water supplies and initial segment of DUWL should be subject to protocol to eliminate microbial contamination and routine monitoring to guarantee an appropriate quality of water used in dental treatment. Int J Antimicrob Agents, 2005 Jan, 25(1), 51 - 6 First report of VanA Enterococcus gallinarum dissemination within an intensive care unit in Argentina; Corso A et al.; Enterococcusgallinarum is intrinsically resistant to low levels of vancomycin and has been described as a colonizing microorganism causing bacteraemia and infection among immunosupresed patients . Between August 2000 and February 2001, 15 highly glycopeptide-resistant E . gallinarum isolates, one from blood and the remaining from rectal swabs, were recovered in a general hospital of Buenos Aires Province, Argentina . All isolates were characterized by biochemical assays, and displayed MICs of vancomycin in the range 16-128mg/l and MICs of teicoplanin in the range 16-32mg/l . In all cases, PCR analysis yield positive results for both vanC1 and vanA genes . E . gallinarum isolates were classified as two clonal types by SmaI-PFGE: clone A (n = 8) and clone B (n = 7) and both harboured a transferable vanA element. Antimicrob Agents Chemother, 2005 Jan, 49(1), 454 - 6 Relationship between copper, glycopeptide, and macrolide resistance among Enterococcus faecium strains isolated from pigs in Denmark between 1997 and 2003; Hasman H et al.; A significant relationship between copper resistance (tcrB), glycopeptide resistance (Tn1546), and macrolide resistance {erm(B)} in Enterococcus faecium isolated from pigs was found . The tcrB gene was located closely upstream of the Tn1546 element . However, the continued use of copper sulfate has not been able to maintain high levels of macrolide and glycopeptide resistance. J Infect Chemother, 2004 Dec, 10(6), 331 - 4 No regional spread of vancomycin-resistant enterococci with vanA or vanB in Kitakyushu, Japan; Matsumoto T et al.; Outbreaks of vancomycin-resistant enterococci (VRE) infection occur sporadically in Japan, and their frequency has been gradually increasing . We experienced a nosocomial outbreak of VRE in two hospitals in the city of Kitakyushu, and the spread of VRE strains was suspected in this area . To examine the prevalence rate of infection and colonization of VRE in Kitakyushu, we screened a total of 24 297 clinical samples from patients in hospitals and clinics in Kitakyushu from October through December 2002 for VRE . The isolates screened as positive for VRE accounted for 2.3% (566/24 297) of the tested clinical samples . Polymerase chain reaction (PCR) analyses for vanA, vanB, vanC1, and vanC2/3 were performed to confirm the screening test results . Neither vanA nor vanB genes were detected in any isolates . The 265 vanC1-positive isolates were Enterococcus gallinarum, and the 150 vanC2/3-positive isolates were E . casseliflavus . Other Enterococcus species were negative in this PCR-detection test . In this study, the PCR procedure was considered reliable and successful because although neither vanA nor vanB was detected, vanC1 and vanC2/3 were completely detectable . Therefore, we concluded that the regional spread of VRE with vanA and vanB had not occurred in Kitakyushu in 2002 . In the near future, the prevalence of VRE with vanA or vanB is likely to increase in Japan, as it has in other countries . We should continue to find and prevent nosocomial outbreaks of infection and colonization by VRE. J Appl Microbiol, 2005, 98(1), 216 - 28 Molecular characterization, technological properties and safety aspects of enterococci from 'Hussuwa', an African fermented sorghum product; Yousif NM et al.; Abstract n.m.k . yousif, p . dawyndt, h . abriouel, a . wijaya, u . schillinger, m . vancanneyt, j . swings, h.a . dirar, w.h . holzapfel and c.m.a.p . franz . 2004.Aims: To identify enterococci from Hussuwa, a Sudanese fermented sorghum product, and determine their technological properties and safety for possible inclusion in a starter culture preparation . Methods and Results: Twenty-two Enterococcus isolates from Hussuwa were identified as Enterococcus faecium by using phenotypic and genotypic tests such as 16S rDNA gene sequencing, RAPD-PCR and restriction fragment length polymorphism of the 16S/23S intergenic spacer region fingerprinting . Genotyping revealed that strains were not clonally related and exhibited a considerable degree of genomic diversity . Some strains possessed useful technological properties such as production of bacteriocins and H(2)O(2) or utilization of raffinose and stachyose . None produced alpha-amylase or tannase . A safety investigation revealed that all strains were susceptible to the antibiotics ampicillin, gentamicin, chloramphenicol, tetracycline and streptomycin, but some were resistant to ciprofloxacin, erythromycin, penicillin and vancomycin . Production of biogenic amines or presence of genes encoding virulence determinants occurred in some strains . Conclusions: Enterococcus faecium strains are associated with fermentation of Sudanese Hussuwa . Some strains exhibited useful technological properties such as production of antimicrobial agents and fermentation of indigestible sugars, which may aid in stabilizing and improving the digestibility of the product respectively . Significance and Impact of the Study: Enterococci were shown to play a role in the fermentation of African foods . While beneficial properties of these bacteria are indicated, their presence in this food may also imply a hygienic risk as a result of antimicrobial resistances or presence of virulence determinants. J Org Chem, 2004 Dec 24, 69(26), 9025 - 9 A new structural theme in the imidazole-containing alkaloids from a calcareous Leucetta sponge; Ralifo P et al.; Further study of the Fijian sponge Leucetta sp., a source of (+)-calcaridine A (4) and (-)-spirocalcaridines A (5) and B (6), has yielded (-)-spiroleucettadine (8), the first natural product to contain a fused 2-aminoimidazole oxalane ring along with the known compounds N,N-dimethylnaamidine D (3) and isonaamine B (7) . NMR analysis allowed the unambiguous 2D structural assignment of 8, and its relative stereochemistry was determined by ROESY data . Good antibacterial activity was observed for 8 against Enterococcus durans with an MIC of less than 6.25 microg/mL. J Infect, 2005 Jan, 50(1), 84 - 8 A rare case of pleuropulmonary infection and septic shock associated with Enterococcus faecium endocarditis; Maceachern P et al.; Enterococci rarely cause pleuropulmonary infections . A novel case of definitively diagnosed nosocomial Enterococcus faecium pneumonia, empyema, and endocarditis associated with septic shock is reported. Ann Vasc Surg, 2004 Nov, 18(6), 755 - 7 A case of graft infection 10 years after ascending aorta replacement; Miyazaki S et al.; A 52-year-old man was admitted with anemia and slight fever, which he had for the last 2 months . He had undergone replacement of the ascending aorta for acute aortic dissection 10 years previously . Echocardiography demonstrated a flailing thin structure in the anterior wall of the ascending aorta corresponding to the proximal portion of the prosthetic graft . This abnormal echocardiographic finding led us to repeat blood cultivation . We finally detected Enterococcus facium and Staphilococcus epidermidis in his blood sample . We diagnosed this as a graft infection and prepared for surgical re-replacement of the infected graft . While he was waiting for the operation, an infectious aneurysm of a tibialis posterior artery ruptured and an emergency operation was done . Replacement of the infected ascending aorta graft was done thereafter . In surgery, 2-cm-long vegetation was found . It stuck to the graft wall near the former hole used for air removal in the first surgery . The patient recovered fully and left our hospital after 3 months of postoperative antibiotics therapy . This rare case of aortic graft infection long after the original replacement surgery suggests the importance of thorough echocardiographic investigation of prosthetic graft infection as a possible cause of fever of unknown origin. J Antimicrob Chemother, 2005 Jan, 55(1), 123 - 126 Epub 2004 Dec 1. Distribution of streptogramin resistance determinants among Enterococcus faecium from a poultry production environment of the USA; Hayes JR et al.; OBJECTIVES: The impact of agricultural use of antimicrobials on the present and future efficacy of therapeutic drugs in human medicine is a growing public concern . Quinupristin/dalfopristin has been approved to treat human disease caused by vancomycin-resistant Enterococcus faecium and is related to virginiamycin, a streptogramin complex that has long been used in USA agriculture poultry production . METHODS: Streptogramin-resistant isolates of E . faecium from poultry production environments on the eastern seaboard were recovered without selection for streptogramin resistance and examined using ribotyping to evaluate clonal bias . Colony PCR screening for the previously described streptogramin resistance determinants erm(A), erm(B), msr(C), vgb(A), vat(D) and vat(E) was performed to determine the prevalence of streptogramin resistance mechanisms from these environments . RESULTS: The collection of E . faecium isolates was unevenly distributed among 28 ribogroups and did not cluster geographically . The most prevalent ribogroups was composed of isolates that possessed diverse antimicrobial resistance profiles . Of the 127 isolates examined, 63% were resistant to quinupristin/dalfopristin . The resistance determinants erm(A) and erm(B) were observed among 6% and 10%, respectively, of streptogramin-resistant isolates . msr(C) was detected in a single isolate that was resistant to macrolide and lincosamide antimicrobials . The streptogramin B hydrolase vgb(A) and the streptogramin A acetyltransferases genes vat(D) and vat(E) were not detected in any of the E . faecium isolates . CONCLUSIONS: These results indicate that there is widespread resistance to streptogramin antimicrobials among E . faecium throughout the poultry production region in this study and that the mechanisms of resistance to streptogramin antimicrobials within this population remain largely uncharacterized. Am J Infect Control, 2004 Dec, 32(8), 462 - 9 Impact of a formulary switch from ticarcillin-clavulanate to piperacillin-tazobactam on colonization with vancomycin-resistant enterococci; Winston LG et al.; BACKGROUND: The prevalence of vancomycin-resistant enterococci (VRE) is increasing, despite infection control measures . Limited data link ticarcillin-clavulanate to higher VRE prevalence . METHODS: Active surveillance for VRE was conducted before and after a formulary switch from ticarcillin-clavulanate to piperacillin-tazobactam . Rectal swabs were obtained serially in 863 adult patients admitted to intensive care units (ICUs) between November 1, 2000 and September 30, 2004 . RESULTS: In the postswitch period, 38 of 497 (7.6%) patients acquired VRE versus 42 of 366 (11.5%) patients in the preswitch period . Survival analysis showed an overall hazard ratio (HR) of .68 postswitch versus preswitch ( P = .07), with the greatest change in the surgical ICU (HR = .17, P = .006) . Multivariate analysis showed an overall HR = .51 ( P = .004) . Hospital-wide, nonstool VRE clinical cultures fell from 39 (.58/1000 patient days) in the 10-month preswitch period to 27 (.33/1000 patient days) in the 12-month postswitch period . Infection control practices and use of other antibiotics remained stable . CONCLUSIONS: VRE acquisition appeared to decrease in association with a formulary change from ticarcillin-clavulanate to piperacillin-tazobactam. Transpl Infect Dis, 2004 Sep, 6(3), 117 - 9 Successful treatment of vancomycin-resistant Enterococcus faecium endocarditis with linezolid in a renal transplant recipient with human immunodeficiency virus infection; Archuleta S et al.; Infections with vancomycin-resistant Enterococci cause significant morbidity and mortality in hospitalized patients, including transplant recipients . We report the successful use of oral linezolid to treat a case of vancomycin-resistant Enterococcus faecium endocarditis in a renal transplant recipient with human immunodeficiency virus infection. Vet Res Commun, 2004 Oct, 28(7), 587 - 98 Mode of binding of fibrinogen, fibronectin and iron-binding proteins by animal enterococci; Styriak I et al.; Sixty-two animal enterococci were examined for their binding of bovine fibrinogen, porcine fibronectin, bovine lactoferrin, bovine apotransferrin and human holotransferrin in the particle agglutination assay (PAA) . Individual strains expressed binding of selected glycoproteins to various degrees (0, 1, 2, 3), whereas bovine fibrinogen binding of enterococci from goats, rabbits and rodents was the strongest (3) in general . Porcine fibronectin was bound weakly (1 or 2) by enterococci from horses, dogs, poultry, rabbits and rodents, while most of the goat isolates and half of the dog feed isolates did not bind fibronectin (0) . Bovine lactoferrin was bound especially by the isolates from rodents and rabbits . Bovine apotransferrin was bound very weakly (1) by only a few isolates . Human holotransferrin was bound to a greater extent than apotransferrin by some isolates from rabbits and rodents . Since multiresistant strains are preferred in our binding studies, enterococci were also examined for their antibiotic resistance pattern . Almost all investigated isolates were resistant at least to one antibiotic . However, some strains displayed resistance to five or six antibiotics of 10 antibiotics tested . In a study of the inhibitory effect of heparin, porcine mucin and hyaluronic acid, the greatest effect was observed after heparin treatment of bacterial cells . These observations, as well as the expression of heparin binding by most strains, may suggest that at least one mode of enterococcal attachment utilizes glycosaminoglycan chains present on the surface of adherent cells. Clin Dev Immunol, 2004 Sep-Dec, 11(3-4), 267 - 73 The effects of Enterococcus faecium and selenium on methotrexate treatment in rat adjuvant-induced arthritis; Rovensky J et al.; The effects of probiotic bacteria Enterococcus faecium (EF) and selenium were studied on methotrexate (MTX) treatment in rats with adjuvant arthritis (AA) . Arthritic rats were preventive treated orally with the following substances: lyophilized EF (15mg/kg/day, 5 days a week); sodium selenite pentahydrate (SSe, 0.050mg/kg containing 0.015 mg/kg selenium, 5 days a week); MTX (0.6 mg/kg/week), and their combinations for the period of 50 days from adjuvant application . Levels of serum albumin, serum nitrite/nitrate concentrations, hind paw swelling, arthrogram scores, whole body bone mineral density (BMD), and bone erosions were evaluated as markers of inflammation and destructive changes associated with arthritis . Long-term preventive treatment with low-dose MTX significantly inhibited the markers of both inflammation and arthritis . EF or SSe when administered singly or in combination had no significant effect on given parameters in arthritic rats . EF but not SSe potentiated the beneficial effects of MTX, which resulted in a more significant reduction of hind paw swelling, arthrogram scores and whole body BMD decrease . EF had a tendency to improve also the effect of MTX on serum albumin and nitrite/nitrate concentrations . Our results indicate that EF may increase the preventive effect of MTX treatment in rat AA by improving its anti-inflammatory and anti-arthritic effects. Int J Antimicrob Agents, 2004 Dec, 24(6), 613 - 5 In vitro antibacterial activity of echinomycin and a novel analogue, YK2000, against vancomycin-resistant enterococci; Kim JB et al.; The in vitro inhibitory and bactericidal activity of echinomycin and its the novel synthetic analogues of echinomycin,YK2000 and YK2005, were evaluated using 93 clinical isolates of vancomycin-resistant enterococci (VRE) . In agar dilution tests, the MIC(90) of echinomycin and YK2000 were 0.125 and 8 mg/l, respectively, using Mueller-Hinton II agar, while that of YK2005 was 32 mg/l . Bactericidal activity of echinomycin and YK2000 were two to four times higher than the MIC in time-kill assay experiments . These results suggest that echinomycin and its analogues might be useful as anti-VRE drugs. Rinsho Biseibutshu Jinsoku Shindan Kenkyukai Shi, 2003, 14(2), 151 - 8 {Detection of vancomycin-resistant enterococci by a fully automated microbiology system, RAISUS}; Nakasone I et al.; A fully automated microbiology system, RAISUS recently developed (Nissui Pharmaceuticals Co., Ltd., Tokyo) was evaluated for identification of enterococci and for detection of vancomycin-resistant enterococci (VRE) . When a total of 124 enterococcal isolates were tested, RAISUS correctly identified 122 (98.4%) isolates . Two isolates resulted in species-identifications disagreed with the reference but agreed as belonging to the genus of Enterococcus . When a total of fifty-seven VRE isolates confirmed to be positive for vanA and/or vanB genes were tested against vancomycin, the current RAISUS susceptibility program version 1.76 could detect 41 (71.9%) isolates of VRE as having > or = 32 microg/ml MIC for vancomycin, but one was intermediate (MIC, 8.0 microg/ml) and the remaining 15 vanB-type isolates were incorrectly interpreted as vancomycin-susceptible (MIC, < or = 4.0 microg/ml) . The test program based on the algorism to determine bacterial growth in the presence of vancomycin was developed and evaluated . With this test program, all the VRE isolates positive for vanA and/or vanB genes were identified as being vancomycin-resistant or intermediate interpretation . However, eight of 19 clinical isolates of E . casseliflavus and E . gallinarum intrinsically possessing vanC gene were determined as being < or = 4.0 microg/ml MIC for vancomycin . With the influence of program revision, RAISUS became to incubate the test plate longer than with the current program, but 50% of enterococcal isolates including vancomycin-resistant and vancomycin-susceptible isolates were determined within 5 hour-incubations and 90% were within 9 to 10 hour-incubations . With these results, we can conclude that the revised test program for enterococcal isolates could rapidly and correctly identify vancomycin-resistance, and will be applicable to the routine susceptibility test in clinical laboratories. J Infect Dis, 2004 Dec 15, 190(12), 2162 - 6 Epub 2004 Dec 15. In vitro antienterococcal activity explains associations between exposures to antimicrobial agents and risk of colonization by multiresistant enterococci; Rice LB et al.; We compared ceftriaxone and piperacillin-tazobactam at doses ranging from 0.1 to 2 times the human equivalent daily dose (HEDD), to determine their impact on gastrointestinal colonization by ampicillin- and vancomycin-resistant Enterococcus faecium C68 in a mouse model . Ceftriaxone failed to promote colonization at doses up to 0.25 times the HEDD, whereas piperacillin-tazobactam promoted colonization at doses up to 0.5 times the HEDD . Ceftriaxone promoted colonization at doses at least 0.5 times the HEDD, whereas piperacillin-tazobactam inhibited colonization at doses at least 0.75 times the HEDD . Both piperacillin-tazobactam and ceftriaxone inhibited colonization by an enterococcal strain devoid of low-affinity penicillin-binding protein-5 (significantly increasing its susceptibility to these agents), at doses that promoted colonization by E . faecium C68 . These results support a model in which the impact that different beta -lactam agents have on colonization by VRE is related to the level of the beta -lactam agent's intrinsic antienterococcal activity against the colonizing strain. Int J Syst Evol Microbiol, 2004 Nov, 54(Pt 6), 2175 - 9 Enterococcus saccharominimus sp . nov., from dairy products; Vancanneyt M et al.; Four isolates, which were obtained from Belgian, Moroccan and Romanian dairy products, constituted a homogeneous but unidentified taxon after screening with whole-cell protein fingerprinting . Complete 16S rRNA gene sequence analysis classified representative strains in the genus Enterococcus . Highest sequence similarities of 98.6 and 98.0 % were obtained with the species Enterococcus sulfureus and Enterococcus saccharolyticus, respectively . Growth characteristics, biochemical features, tRNA intergenic length polymorphism analysis, DNA-DNA hybridization and DNA G+C contents of selected strains demonstrated that they represent a single, novel Enterococcus species . It differs phenotypically from other enterococci in characteristics commonly considered as typical of this genus: no growth in 6.5 % NaCl or 0.4 % sodium azide, and no acid production from a wide range of carbohydrates . The name Enterococcus saccharominimus sp . nov . is proposed for this novel species; the type strain (LMG 21727(T)=CCM 7220(T)) was isolated from contaminated pasteurized cow's milk. Skinmed, 2004 Nov-Dec, 3(6), 336 - 8 Cutaneous zygomycosis following attempted radial artery cannulation; Kapadia S et al.; A 70-year-old man was seen in a hospital consultation for evaluation of cellulitis of the left arm . The patient had multiple medical problems, including advanced liver disease due to alcohol, diabetes mellitus, congestive heart failure, atrial fibrillation, chronic renal in sufficiency, and hypopituitarism requiring steroid replacement . Most recently, he was admitted to the intensive care unit, where he required intubation and mechanical ventilation support following respiratory failure secondary to pneumonia . At that time, an attempt was also made to place an arterial line in the left radial artery . The patient had multiple areas of ecchymosis on both arms . A large bulla was found on the lateral aspect of the left wrist several days after the attempted arterial line placement . Subsequently, the lesion drained serosanguineous fluid, and, during the next 2 days, it ulcerated with necrosis extending around the wrist and to the elbow . He was started on ampicillin/sulbactam and clindamycin for presumed necrotizing fasciitis . The surgical service performed a very limited debridement,which was partially limited by his coagulopathy from liver disease . The initial tissue culture was positive only for Enterococcus faecium.At the time of the consultation, his temperature was 95' F (35 degrees C), pulse 82 bpm, respirations 16 BPM, and blood pressure 101/56 mmHg . He was awake but not oriented or responsive . His cardiopulmonary exam was unremarkable . Abdominal exam disclosed ascites . His extremities were all grossly edematous with multiple ecchymoses . His left forearm had a circumferential area of ecchymosis and necrosis with macerated margins, sparing only the lateral ulnar epicondyle, and involving deeper structures of subcutaneous fat and muscle(Figures 1-2 showing evolution of the lesion in a period of 1 week) . Small tissue clippings were taken from the edge of the lesion and placed on culture plates . By the next morning, the patient's tissue culture grew a mold, later identified as Rhizopus . Amphotericin B was initiated . Surgical intervention (wide debridement with potential conversion to amputation of the left arm) was considered to offer little benefit in view of the patient's multiple and severe comorbidities and his poor prognosis . Amphotericin B was then stopped; the patient died within a week from his multiple medical complications . The family refused an autopsy. Chemotherapy, 2004 Nov, 50(5), 245 - 9 Epub 2004 Nov. The emergence of glycopeptide-resistant Enterococcus faecium in a university hospital in southwestern Greece; Kolonitsiou F et al.; BACKGROUND: Enterococci and especially glycopeptide-resistant strains (GRE) are widely distributed in the hospital environment, by acquiring resistance determinants and virulence factors . METHODS: The study included 48 GRE isolated during a 1-year period from different inpatients in a tertiary hospital in southwestern Greece . Antibiotic susceptibility was determined by the Etest, and the presence of resistance and virulence genes was shown by PCR . Clonal types were identified by pulsed-field gel electrophoresis of SmaI DNA digests . RESULTS: All GRE were multi-resistant of the VanA phenotype, verified by the detection of the gene by PCR . Two major clones were distributed in all hospital wards . The majority of the strains (46 of 48) harbored the esp gene, while 27 GRE expressed also the gelE and/or as genes . CONCLUSIONS: The spread of two clones expressing the vanA gene and virulence factors were responsible for the emergence of GRE in the University Hospital of Patras. Int J Food Microbiol, 2004 Dec 1, 97(1), 85 - 91 Growth inhibition of heat-injured Enterococcus faecium by oligophosphates in a cured meat model; Houben JH et al.; Cells of two heat-resistant strains of Enterococcus faecium were heated and incubated in meat suspensions containing curing ingredients . The concentrations of the curing ingredients were those frequently used for pasteurized ham-type products, except that the concentrations of the oligophosphates (triphosphate and diphosphate) varied . Heating tests at 69 degrees C were performed with inoculated meat suspensions in heat-sealed plastic pouches . Numbers of bacteria were counted immediately after heating and in parallel series of heated pouches incubated at 37 degrees C . Plating was performed in Tryptone Dextrose Yeast Meat Peptonised Milk Agar (TDYMP); in TDYMP Agar to which the curing ingredients were added; and in TDYMP Agar to which the curing ingredients except oligophosphates were added . The inclusion of oligophosphates in the heating medium increased the heat-injury sustained by the E . faecium cells, and in combination with rather severe heat treatment even completely blocked the growth of surviving organisms in the meat suspension incubated at 37 degrees C . The presence of oligophosphates in the culture medium TDYMP Agar severely reduced the counts of freshly heated cells; however, this effect disappeared after repair and growth of the surviving organisms in the meat suspension. Clin Microbiol Infect, 2004 Nov, 10(11), 1009 - 11 Influence of rifaximin treatment on the susceptibility of intestinal Gram-negative flora and enterococci; DuPont HL et al.; The development of rifaximin- and rifampicin-resistant intestinal coliforms was studied in 27 subjects receiving rifaximin for 3 days by plating stool samples on media containing rifaximin 200 mg/L or rifampicin 64 mg/L before treatment (day 0), after treatment was completed (day 3), and after a further 2 days (day 5) . The susceptibility of enterococci grown on day 0 and day 3 was also studied in 71 subjects . Significant increases in antimicrobial-resistant coliform flora were not seen in either the rifaximin-treated or the placebo-treated subjects . Enterococci recovered pre- and post-treatment showed similar susceptibilities . Rifaximin did not select for significant resistance in the Gram-negative and Gram-positive intestinal flora during therapy. Scand J Infect Dis, 2004, 36(10), 771 - 2 Impact of prolonged treatment with trimethoprim-sulfamethoxazole on the human gut flora; Kofteridis DP et al.; The case of a mentally ill man inadvertently treated with trimethoprim-sulfamethoxazole (TMP-SMX) for 2 y is presented . Quantitative stool cultures revealed a substantially suppressed Gram-negative aerobic flora, while Enterococcus spp . and anaerobes were not affected . Yeasts were moderately increased . TMP-SMX represents an attractive antimicrobial for immunocompromized patients who need the integrity of their intestinal anaerobic flora for colonization resistance. J Food Prot, 2004 Oct, 67(10), 2292 - 5 Resistance to gentamicin and vancomycin in enterococcal strains isolated from retail broiler chickens in Japan; Harada T et al.; A total of 137 Enterococcus strains isolated from chicken meat were subjected to antimicrobial susceptibility tests . Strains with the vanCl gene were isolated from seven of nine samples of chicken meat processed in Japan and from all chickens from China and Brazil between July 2001 and April 2002 . The pulsed-field gel electrophoresis (PFGE) patterns of the isolates were distinguishable from each other, suggesting that VanCl-type vancomycin-resistant Enterococcus is preferentially colonized in broiler chickens in these countries . The incidence of high-level gentamicin resistant (HLGR) enterococci that harbored the aac(6')-le-aph(2")-la or aph(2')-Id gene varied among the countries from which the chickens originated (Japan, 2 of 65; China, 11 of 43; Brazil, 6 of 29) . Moreover, the PFGE patterns of the HLGR strains were distinguishable from each other, except for two strains obtained from chickens from Brazil . The results suggest that HLGR Enterococcus is highly prevalent in broiler chickens. Antimicrob Agents Chemother, 2004 Nov, 48(11), 4427 - 9 Evidence for biliary excretion of vancomycin into stool during intravenous therapy: potential implications for rectal colonization with vancomycin-resistant enterococci; Currie BP et al.; Sixty-three stool samples and five bile samples were prospectively collected from 33 patients receiving intravenous vancomycin therapy and were quantitatively analyzed for vancomycin by a competitive immunoassay . Vancomycin was excreted via bile into the stools of almost all patients at concentrations of 3.3 to 94.8 microg/ml after >/=5 days of a therapy of 1 g every 12 h. FEMS Microbiol Lett, 2004 Nov 1, 240(1), 69 - 74 Glycopeptide resistance determinants from the teicoplanin producer Actinoplanes teichomyceticus; Serina S et al.; In enterococci and other pathogenic bacteria, high-level resistance to vancomycin and other glycopeptide antibiotics requires the action of the van genes, which direct the synthesis of peptidoglycan terminating in the depsipeptide D-alanyl-D-lactate, in place of the usual D-Ala-D-Ala . The Actinoplanes teichomyceticus tcp cluster, devoted to the biosynthesis of the glycopeptide antibiotic teicoplanin, contains van genes associated to a murF-like sequence (murF2) . We show that A . teichomyceticus contains also a house-keeping murF1 gene, capable of complementing a temperature sensitive Escherichia coli murF mutant . MurF1, expressed in Streptomyces lividans, can catalyze the addition of either D-Ala-D-Ala or D-Ala-D-Lac to the UDP-N-acetyl-muramyl-L-Ala-D-Glu-d-Lys . However, similarly expressed MurF2 shows a small enzymatic activity only with D-Ala-D-lactate . Introduction of a single copy of the entire set of van genes confers resistance to teicoplanin-type glycopeptides to S . coelicolor. Mar Pollut Bull, 2004 Oct, 49(7-8), 624 - 9 Assessing Oregon's twenty-six coastal beach areas for recreational water quality standards; Benedict RT et al.; Water samples from 26 Oregon beaches were analyzed for Escherichia coli (E . coli) and enterococci concentrations by the Oregon Department of Human Services (ODHS) Public Health Laboratory . Nine Oregon beaches exceeded US Environmental Protection Agency's (USEPA) single sample maximum density of 104 enterococci colony forming units (cfu) per 100 mL with levels ranging from 121 to 4325 most probable number (MPN)/100 milliliters (mL) . Otter Rock at South Cove had the highest enterococci concentration at 4352 MPN/100 mL . The results from two Oregon beaches exceeded Oregon Department of Environmental Quality's (ODEQ) estuarine E . coli standard of 406 organisms/100 mL . Otter Rock at South Cove had the highest E . coli concentration at 1850 MPN/100 mL while Road's End had an E . coli density of 771 MPN/100 mL . Results of this study suggest that adopting USEPA's marine enterococci standard in lieu of the ODEQ's estuarine standard will lead to increased Oregon beach water standard failures. J Hosp Infect, 2004 Oct, 58(2), 97 - 103 Control of vancomycin-resistant enterococci in a hospital: a five-year experience in a Taiwanese teaching hospital; Wang JT et al.; In order to prevent transmission of hospital-acquired vancomycin-resistant enterococci (VRE), the infection control team (ICT) of the National Taiwan University Hospital (NTUH) introduced practical guidelines from January 1997 to June 2000 . All patients at NTUH found to be infected or colonized with VRE were placed in strict contact and cohort isolation . Surveillance cultures were obtained from other patients in close proximity in order to determine any spread of VRE . If identified, these patients were also placed in contact and cohort isolation, and their isolates were subjected to antimicrobial susceptibility testing and molecular typing by pulsed-field gel electrophoresis . During this period, 20 patients were found to have VRE . Based on typing results, there were three occasions where the same VRE strain had spread between index patients and roommates or patients staying in neighbouring rooms . No further spread occurred after applying strict contact isolation for these patients . The hospital-acquired VRE infection rate was around 0.03 to 0.09 per 1000 discharges during the intervention period . After July 2000, however, members of the ICT did not actively monitor or implement any interventions to control VRE . The rate then increased to 0.20 per 1000 discharges in 2001 . This study suggests that interventions for the control of VRE, based on the guidelines from the Hospital Infection Control Practice Advisory Committee, are effective for control of VRE spread . Failure to adhere to these guidelines may result in an increase in hospital-acquired VRE. J Clin Microbiol, 2004 Oct, 42(10), 4503 - 11 Multiple-locus variable-number tandem repeat analysis, a novel typing scheme to study the genetic relatedness and epidemiology of Enterococcus faecium isolates; Top J et al.; Multiresistant Enterococcus faecium is a major cause of hospital acquired infections and outbreaks . Here, we describe the development of multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA) as a novel typing method to assess the genetic relatedness of E . faecium isolates . Six VNTR loci were used to genotype 392 isolates recovered from different animals and human community, hospital survey, and clinical isolates . From 3 to 13 alleles were found per locus, resulting in 127 different MLVA profiles . Clustering of MLVA profiles confirmed the host-specific genogroups found by multilocus sequence typing (MLST) and showed the grouping of clinical and epidemic isolates that belonged to the MLST-C1 cluster in a distinct MLVA-C1 cluster (sensitivity of 97% and specificity of 90%) . Furthermore, the discriminatory power of MLVA is comparable to MLST . MLVA profiles appeared to be relatively stable, since isolates from a single outbreak shared the same MLVA profile, which is a prerequisite when MLVA is used to study hospital outbreaks . Our data show that MLVA is a highly reproducible and portable typing method; in contrast to MLST, it is fast, relatively cheap, and easy to perform . Furthermore, it has the abilities of MLST to recognize genetically related and potential epidemic isolates . Submission of MLVA profiles is possible via a Web-based database for international comparison. Int J Hyg Environ Health, 2004 Sep, 207(4), 386 - 9 Two episodes of vancomycin-resistant Enterococcus faecium outbreaks caused by two genetically different clones in a newborn intensive care unit; Borgmann S et al.; In 2001 two outbreak episodes (January-March and June-July) caused by vancomycin-resistant E . faecium (VRE) of the VanA-type were observed at a neonatal intensive care unit (NICU) of a university hospital in south-west Germany . To identify the initial source and the route of transmission environmental samples were examined as well as stool samples from patients and the staff . VRE was not found in environmental samples . However, stool samples from 24 hospitalised children tested positive and bacterial clonality was assessed by Sma1-based macro restriction analysis . Furthermore, esp gene and vancomycin resistance gene carriage were examined as well as bacteriocin production . PCR analysis showed that all 24 isolates carried vanA gene cluster, encoding resistance to vancomycin and teicoplanin . However, five of the vanA-positive isolates were resistant to vancomycin but not to teicoplanin . Only these five isolates produced bacteriocin, but in none of the isolates esp gene was detected . PFGE revealed that both outbreaks were caused by two different clones . The patient initiating the first episode, was identified whereas the origin of the second episode remained unknown . From one of the 40 staff stool samples VRE was isolated . This strain was related to the clone of the summer outbreak . In conclusion there were two independent episodes of self limiting VRE outbreaks and transmission on the ward is highly probable. J Food Prot, 2004 Sep, 67(9), 1948 - 52 Antibiotic resistance and virulence traits of enterococci isolated from Baylough, an Irish artisanal cheese; Gelsomino R et al.; Eight representative Enterococcus strains from a collection of over 600 previously isolated from an Irish artisanal cheese were subjected to phenotypic and genotypic analysis of antibiotic resistance and virulence properties . Genes encoding resistance to tetracycline (tet(M) and tet(L)) and/or erythromycin (erm(B)) were detected in five strains . In addition, all strains contained two or more of the virulence genes tested (agg, gel, cyl, esp, ace, efaAfs, and efaAfm) . Further investigation into the transferability and environmental dissemination of these resistance and virulence traits will allow risk assessment and safety evaluation of artisanal cheeses. Int J Syst Evol Microbiol, 2004 Sep, 54(Pt 5), 1823 - 7 Enterococcus hermanniensis sp . nov., from modified-atmosphere-packaged broiler meat and canine tonsils; Koort J et al.; Isolates 302, 334, 356, 377 and 379, detected in modified-atmosphere-packaged broiler meat, together with strains LMG 12317T and LMG 13617, detected in dog tonsils, were analysed in a polyphasic taxonomy study, including numerical analysis of ribopatterns and whole-cell protein patterns, 16S rRNA gene sequence analysis, DNA-DNA hybridization and determination of some phenotypic properties . The results indicated that these isolates represent a novel species in the genus Enterococcus . The isolates showed classical phenotypic reactions for the genus Enterococcus with the exception of not possessing the Lancefield group D antigen . Isolates 334, LMG 12317T and LMG 13617 showed the highest 16S rRNA gene sequence similarity (98.3-99.0 %) to the Enterococcus pallens type strain . In the distance matrix tree based on 16S rRNA gene sequences, the three isolates were located in the Enterococcus avium group with E . pallens as their closest phylogenetic neighbour . Numerical analyses of whole-cell protein patterns and HindIII/EcoRI ribotypes placed all seven isolates together in a single cluster separated from the E . avium group reference strains . The DNA-DNA hybridization level between strains 334 and LMG 12317T was 93.5 %, confirming that they represent the same species . Low hybridization levels (12-30 %) were, by contrast, obtained with the E . pallens and Enterococcus raffinosus type strains . The name Enterococcus hermanniensis sp . nov . is proposed, with strain LMG 12317T (= CCUG 48100T) as the type strain. Int J Syst Evol Microbiol, 2004 Sep, 54(Pt 5), 1717 - 21 Molecular analysis of artisanal Italian cheeses reveals Enterococcus italicus sp . nov; Fortina MG et al.; The taxonomic positions of seven atypical Enterococcus strains, isolated from artisanal Italian cheeses, were investigated in a polyphasic study . By using 16S rRNA gene sequencing, DNA-DNA hybridization and intergenic transcribed spacer analysis, as well as by examining the phenotypic properties, the novel isolates were shown to constitute a novel enterococcal species . Their closest relatives are Enterococcus sulfureus and Enterococcus saccharolyticus, having a 16S rRNA gene sequence similarity of 96.7 % . This group of strains can be easily differentiated from the other Enterococcus species by DNA-DNA hybridization and by their phenotypic characteristics: the strains do not grow in 6.5 % NaCl, and they do not produce acid from L-arabinose, melezitose, melibiose, raffinose or ribose . The name Enterococcus italicus sp . nov . is proposed for this species, with strain DSM 15952T (= LMG 22039T) as the type strain. Peptides, 2004 Sep, 25(9), 1377 - 88 Enterococcal peptide sex pheromones: synthesis and control of biological activity; Chandler JR et al.; The enterococcal pheromone-inducible plasmids such as pCF10 represent a unique class of mobile genetic elements whose transfer functions are induced by peptide sex pheromones . These pheromones are excreted by potential recipient cells and detected by plasmid-containing donor cells at the cell surface, where the pheromone is imported and signals induction of the plasmid transfer system . Pheromone is processed from a chromosomally encoded lipoprotein and excreted by both the donor and recipient cells, but a carefully controlled detection system prevents a response to self-pheromone while still allowing an extremely sensitive response to exogenous pheromone. J Infect Chemother, 2004 Aug, 10(4), 220 - 6 A rapid antimicrobial susceptibility test based on chemiluminescence assay and its application to screening of genotypes in vancomycin-resistant enterococci; Nagasawa Z et al.; Minimum inhibitory concentrations (MICs) of vancomycin (VCM) and teicoplanin (TEIC) were measured using a novel susceptibility test based on the chemiluminescence assay (CA) method (Rapid-Lumi Eiken; Eiken Chemicals, Tokyo, Japan) against 33 strains in total: 7, 5, and 10 strains of which are VCM-resistant enterococci (VRE) with vanA, vanB, and vanC genes, respectively, and the other 11 strains are vancomycin-susceptible enterococci (VSE) . The results were in good accordance with the values determined by the standard broth dilution method approved by the National Committee for Clinical Laboratory Standards (NCCLS): i.e., 88% (29/33) of consistency for VCM and 97% (32/33) for TEIC, respectively . In addition, genotypes in VRE strains (vanA, vanB, vanC-1, and vanC-2/3 genes) were properly estimated from the results of the CA method and the NCCLS interpretive categories, even though the incubation time was very short (2-4 h) . In conclusion, it was found that the new method is reliable and rapid to detect VRE strains in clinical laboratories. J Microbiol, 2004 Jun, 42(2), 143 - 6 DD1.5k, the gene preferentially expressed in bloodstream isolates of vancomycin-resistant Enterococcus faecium; Kim SH et al.; Vancomycin-resistant Enterococcus faecium (VREFM) is becoming a threatening pathogen . We identified a gene called DD1.5K by differential display-PCR, which was preferentially expressed in the bloodstream isolates of VREFM . Due to its amino acid similarity to transfer complex protein, trsE, and tissue-specific expression, this gene may be involved in virulence of VREFM. Chemistry, 2004 Sep 6, 10(17), 4334 - 40 Solid-phase synthesis of dihydrovirginiamycin S1, a streptogramin B antibiotic; Shaginian A et al.; We describe the first solid-phase synthesis of dihydrovirginiamycin S(1), a member of the streptogramin B family of antibiotics, which are nonribosomal-peptide natural products produced by Streptomyces . These compounds, along with the synergistic group A components, are "last line of defense" antimicrobial agents for the treatment of life-threatening infections such as vancomycin-resistant enterococci . The synthesis features an on-resin cyclization and is designed to allow production of streptogramin B analogues with diversification at positions 1', 1, 2, 3, 4, and 6 . Several synthetic challenges known to hinder the synthesis of this class of compounds were solved, including sensitivity to acids and bases, and epimerization and rearrangements, through the judicious choice of deprotection conditions, coupling conditions, and synthetic strategy . This work should enable a better understanding of structure-activity relationships in the streptogramin B compounds, possible identification of analogues that bypass known resistance mechanisms, and perhaps the identification of analogues with novel biological activities. J Hosp Infect, 2004 Sep, 58(1), 28 - 33 Implications of colonization of vancomycin-resistant enterococci (VRE) in renal dialysis patients . Learning to live with it? Humphreys H, Dolan V, Sexton T, Conlon P, Rajan L, Creamer E, Walshe J, Donohoe J, Smyth EG. Vancomycin-resistant enterococci (VRE) commonly colonize, but less frequently infect, debilitated patients, such as those on chronic renal dialysis . The emergence of VRE amongst our cohort of renal replacement therapy patients posed considerable challenges in our attempts to prevent spread . Although 60 of 451 (13%) patients became colonized, only two patients required systemic antibiotics for confirmed or suspected invasive infection . Mortality and inpatient stay was greater in VRE-positive compared with VRE-negative patients (50% versus 10%) and patients who were screened on three or more occasions were likely to remain positive (e.g . 56% of patients screened on six occasions were positive) . The application of recommended guidelines for the control of VRE, however, severely disrupted our renal dialysis programme and therefore had to be abandoned . As patients on renal dialysis are more likely to acquire VRE, remain colonized, require antibiotics and require regular inpatient or outpatient care more frequently than other patients, control measures should be adapted to minimize spread but not disrupt important and essential medical services. Antimicrob Agents Chemother, 2004 Sep, 48(9), 3583 - 5 Clinical-use-associated decrease in susceptibility of vancomycin-resistant Enterococcus faecium to linezolid: a comparison with quinupristin-dalfopristin; Raad II et al.; The susceptibility of 135 vancomycin-resistant Enterococcus faecium bacteremic isolates to linezolid and quinupristin-dalfopristin was determined . All were susceptible to linezolid, while 88% were susceptible to quinupristin-dalfopristin prior to the clinical use of the drugs at our hospital . More than 6 months after their clinical use, a decrease in susceptibility was noted for only linezolid at 83% . This was related in part to a single G2576U gene mutation in domain V of the 23S rRNA gene. Emerg Infect Dis, 2004 Jul, 10(7), 1277 - 81 Nosocomial infection with vancomycin-dependent enterococci; Tambyah PA et al.; We report three patients infected with unique strains of vancomycin-dependent enterococci . Two were first infected by genetically identical strains of vancomycin-resistant enterococci (VRE) . All three patients had much greater exposure to vancomycin and third-generation cephalosporins than did two control groups (patients infected with VRE and hospitalized patients without enterococcal infections) . While antimicrobial pressure promotes nosocomial colonization by VRE, prolonged exposure to vancomycin may foster the transition from vancomycin resistance to dependence. J Hosp Infect, 2004 Aug, 57(4), 281 - 4 Debate-guidelines for control of glycopeptide-resistant enterococci (GRE) have not yet worked; McGowan JE; Glycopeptide-resistant enterococci (GRE) have become a focus of concern in many countries because options for antimicrobial therapy of GRE infection are limited . Several guidelines for the control and prevention of GRE colonization and infection have been developed for healthcare settings, and occasional journal articles now report "control" (usually relative reduction of incidence or prevalence rate rather than elimination) of GRE infections . Yet, rates of infection and colonization with GREcontinue to climb in many parts of the world, showing that true control has not been achieved . Programmes to control GRE will be effective only when they (1) are less expensive to implement; (2) are shown to be cost-effective despite the fact that they merely reduce prevalence levels rather than eradicating the problem; (3) do not require almost perfect implementation to be effective; (4) are shown to be sustainable; (5) are shown to work in acute-care settings other than selected academic centres; and (6) are shown to work in non-acute care settings . Until then, it is clear that guidelines for control of GRE have not worked . New guidelines that truly control GRE must be developed, and this must be done quickly. Intern Med J, 2004 Aug, 34(8), 510 - 2 Rate of nosocomial transmission of vancomycin-resistant enterococci from isolated patients; Cheng AC et al.; To evaluate an isolation policy for patients colonised with vancomycin-resistant enterococci (VRE), we instituted active surveillance for transmission to uncolonised patients . Surveillance rectal swabs were taken and pulsed-field gel electrophoresis was performed on positive isolates . VRE transmission with an identical genotype occurred in 5 patients, giving a transmission rate of 3.7 per 1000 patient days, or 1 patient per ward each week . The present study provides a baseline for -assessment of VRE transmission and will be useful in evaluation of the effectiveness of infection control interventions. J Antimicrob Chemother, 2004 Sep, 54(3), 680 - 3 Epub 2004 Aug 12. Enterococcus faecium N03-0072 carries a new VanD-type vancomycin resistance determinant: characterization of the VanD5 operon; Boyd DA et al.; OBJECTIVES: To genotypically characterize the vancomycin resistance mechanism of Enterococcus faecium N03-0072, which was negative by PCR for the currently known van genotypes . METHODS: PCR was used to amplify the entire vancomycin resistance operon and the complete nucleotide sequence was determined by dideoxy cycle sequencing . RESULTS: Analysis revealed a VanD-type operon with 94% nucleotide identity to the VanD4 operon and 90% nucleotide identity to the VanD1/D3 operons . A set of universal primers was designed in order to identify all current vanD variants by PCR . CONCLUSIONS: E . faecium N03-0072 carries a new VanD-type operon, designated VanD5. Cas Lek Cesk, 2004, 143(5), 324 - 8 {Complex therapy of chronic hepatic encephalopathy completed with probiotic: comparison of two studies}; Boca M et al.; BACKGROUND: Several reports about usefulness of probiotics, including Enterococcus faecium in the treatment of chronic hepatic encephalopathy in patients with liver cirrhosis have been recently published . The results obtained by the administration of Enterococcus faecium M-74 + selenium will be evaluated and compared with those published by Loguercio et al . for Enterococcus faecium SF 68 . METHODS AND RESULTS: Fifteen patients with liver cirrhosis, portal hypertension and chronic hepatic encephalopathy were treated, beside the standard therapy, also with one capsule of probiotic "Enterococcus faecium M-74 + Selenium" per day in three four-week periods, separated by two fortnight pauses . During the treatment, every patient was examined 9-times . The severity of chronic hepatic encephalopathy was evaluated with the aid of the portal systemic encephalopathy index, calculated from 5 parameters (mental stage, asterixis, number connecting test, blood ammonia, EEG) . The indexes of initial and final values were compared . The time course of blood ammonia levels and that of results from number connecting test in either study were mutually compared on the basis of interval estimates of quadratic regression function . Significant improvement of the portal systemic encephalopathy index after the treatment was found: it decreased in average by 70% (55 to 85%, interval 95% confidence interval) . The mental stage improved and the asterixis disappeared . The blood ammonia levels as well as the results from the number-connecting test after 8-9 weeks significantly approached the normal pattern . EEG findings improved and they were often normalised . Our starting blood ammonia levels (increased by 31% above norm) and results from the connecting test (increased by 60%) were significantly lower accordingly the Loguercio et al . (increased by 243 and 238%, respectively) . During the treatment, the values in our study decreased by 25% and 30%, in the compared study by 50% and 70% . CONCLUSIONS: Our results proved the hypothetical favourable effect of probioticum Enterococcus faecium M-74 + Selenium on chronic hepatic encephalopathy . Interestingly, a markedly higher relative therapeutic effect has been achieved in more serious disorder in comparison with the less severe disorders in the present study. J Clin Microbiol, 2004 Aug, 42(8), 3558 - 65 Use of a genus- and species-specific multiplex PCR for identification of enterococci; Jackson CR et al.; The 16S rRNA gene has previously been used to develop genus-specific PCR primers for identification of enterococci . In addition, the superoxide dismutase gene (sodA) has been identified as a potential target for species differentiation of enterococci . In this study, Enterococcus genus-specific primers developed by Deasy et al . (E1/E2) were incorporated with species-specific primers based upon the superoxide dismutase (sodA) gene for development of a multiplex PCR . This assay provides simultaneous genus and species identification of 23 species of enterococci using seven different reaction mixtures . Accuracy of identification of the multiplex PCR was determined by comparisons to standard biochemical testing, the BBL Crystal kit, VITEK, and API Rapid ID 32 Strep . Isolates from swine feces, poultry carcasses, environmental sources, and retail food were evaluated and, overall, results for 90% of the isolates tested by PCR agreed with results obtained using standard biochemical testing and VITEK . Eighty-five percent and 82% of PCR results agreed with results from the API Rapid ID 32 Strep and BBL Crystal tests, respectively . With the exception of concurrence between identification using standard biochemical testing and VITEK (85%) and between BBL Crystal and VITEK (83%), the percent agreement for PCR was higher than or equal to any other pairwise comparison . Multiplex PCR for genus and species determination of enterococci provides an improved, rapid method for identification of this group of bacteria. Poult Sci, 2004 Jul, 83(7), 1099 - 105 Competitive exclusion of a glycopeptide-resistant Enterococcus faecium in the presence of vancomycin but not equivalent concentrations of tylosin or gentamicin; Poole TL et al.; The effect of subtherapeutic concentrations of antibiotics (10.0 and 40.0 microg/mL of vancomycin, gentamicin, and tylosin) on the efficacy of a mixed anaerobe culture of chicken microflora (CCF) was studied in a continuous-flow fermentation system . Efficacy of CCF posttreatment was assessed by challenge with glycopeptide-resistant Enterococcus faecium (GRE) at 6.0 log10 cfu/mL . Bacterial enumeration of endogenous CCF isolates, volatile fatty acid (VFA) analysis, and challenge with GRE indicated that CCF efficacy was affected by all antibiotic treatments . Although CCF treated with 10.0 microg/mL of vancomycin eliminated GRE13 at a rate of 0.61 log10 cfu/ mL per day, it was unable to eliminate E . coli, a gram-negative challenge organism . All other antibiotic treatments allowed GRE persistence at approximately 2.0 to 6.5 log10 cfu/mL . All antibiotic-treated cultures had decreased concentrations of acetic and propionic acids . Our data suggest that low concentrations of antimicrobials may adversely affect the microbial ecology of gut microflora with respect to its ability to exclude exogenous bacteria . Moreover, gentamicin had an adverse effect on the inhibitory stringency of CCF even though it showed little anti-anaerobic activity against CCF strict anaerobes in pure culture . Verification of the results in live animals will be necessary to determine if antimicrobial treatment could compromise the effectiveness of normal microflora to serve as a natural host defense against infection. Antimicrob Agents Chemother, 2004 Aug, 48(8), 3028 - 32 Impact of specific pbp5 mutations on expression of beta-lactam resistance in Enterococcus faecium; Rice LB et al.; We tested the impact of individual PBP 5 mutations on expression of ampicillin resistance in Enterococcus faecium using a shuttle plasmid designed to facilitate expression of cloned pbp5 in ampicillin-susceptible E . faecium D344SRF . Substitutions that had been implicated in contributing to the resistance of clinical strains conferred only modest levels of resistance when they were present as single point mutations . The levels of resistance were amplified when some mutations were present in combination . In particular, a methionine-to-alanine change at position 485 (in close proximity to the active site) combined with the insertion of a serine at position 466 (located in a loop that forms the outer edge of the active site) was associated with the highest levels of resistance to all beta-lactams . Affinity for penicillin generally correlated with beta-lactam MICs for the mutants, but these associations were not strictly proportional. Infect Immun, 2004 Aug, 72(8), 4512 - 20 Cytotoxicity of hydrogen peroxide produced by Enterococcus faecium; Moy TI et al.; Although the opportunistic bacterial pathogen Enterococcus faecium is a leading source of nosocomial infections, it appears to lack many of the overt virulence factors produced by other bacterial pathogens, and the underlying mechanism of pathogenesis is not clear . Using E . faecium-mediated killing of the nematode worm Caenorhabditis elegans as an indicator of toxicity, we determined that E . faecium produces hydrogen peroxide at levels that cause cellular damage . We identified E . faecium transposon insertion mutants with altered C . elegans killing activity, and these mutants were altered in hydrogen peroxide production . Mutation of an NADH oxidase-encoding gene eliminated nearly all NADH oxidase activity and reduced hydrogen peroxide production . Mutation of an NADH peroxidase-encoding gene resulted in the enhanced accumulation of hydrogen peroxide . E . faecium is able to produce hydrogen peroxide by using glycerol-3-phosphate oxidase, and addition of glycerol to the culture medium enhanced the killing of C . elegans . Conversely, addition of glucose, which leads to the down-regulation of glycerol metabolism, prevented both C . elegans killing and hydrogen peroxide production . Lastly, detoxification of hydrogen peroxide either by exogenously added catalase or by a C . elegans transgenic strain overproducing catalase prevented E . faecium-mediated killing . These results suggest that hydrogen peroxide produced by E . faecium has cytotoxic effects and highlight the utility of C . elegans pathogenicity models for identifying bacterial virulence factors. Microb Drug Resist, 2004 Summer, 10(2), 177 - 83 Vancomycin-resistant enterococcus (VRE) carriage and infection in intensive care units; Yeh KM et al.; From July, 1997, through December, 2001, patients who were admitted to intensive care units (ICUs) were enrolled in the study of vancomycin resistance enterococcus (VRE) colonization . Among 4,538 patients admitted to the ICUs, 363 (8.0%) patients were found to have positive culture of VRE at the day of admission to the ICUs and 453 (10.0%) of patients were negative to the first day of admission but became colonized with VRE during the stay in ICU . Among 816 patients, 9 (1.1%) with VRE isolated from sterile sites were selected for further analysis . Pulsed-field gel electrophoresis (PFGE) revealed a total of four PFGE banding patterns in the colonized and infected Enterococcus faecium isolates . Six of nine 9 were found to have an identical PFGE type Ia, suggesting the circulation of an endemic strain . All of these type Ia isolates also contained two potential virulence genes, the esp and hly genes and were first identified in Asia . After the further typing of 540 isolates that were randomly selected from each month, the endemic strain was not identified before the first patient was colonized and infected with this strain in November, 1998, but was isolated from other ICU patients during each month thereafter throughout the remainder of the study period . Although colonization of VRE is the first step toward infection, a low infection rate was observed, except in patients with prolonged hospitalization and severe illness . Use of the isolation room and reminders regarding hand hygiene failed to prevent the circulation of endemic strain . Thus, the SHEA guideline (Muto et al., Infect . Control Hosp . Epidemiol . 2003;24:362-386) for preventing nosocomial transmission of VRE should be enforced. Microb Drug Resist, 2004 Summer, 10(2), 114 - 23 Vancomycin-resistant Enterococcus faecium isolates causing hospital outbreaks in northern Italy belong to the multilocus sequence typing C1 lineage; Bonora MG et al.; Multilocus sequence typing (MLST) was used to obtain insights into the genetic relationships between 14 vancomycin-resistant Enterococcus faecium (VREF) isolates from humans (hospitalized patients, 5 strains) and nonhuman sources (meat and poultry, 9 strains) in northern Italy over the period 1993-2001 . The typing scheme (Homan et al., 2002, J . Clin . Microb., 40:1963-1971) based on seven housekeeping genes--adk (adenylate kinase), atpA (ATP synthase, alpha subunit), ddl (D-alanine-D-alanine ligase), gyd (glyceraldehyde-3-phosphate dehydrogenase), gdh (glucose-6-phosphate dehydrogenase), purK (phosphoribosylaminoimidazole carboxylase ATPase subunit), and pstS (phosphate ATP-binding cassette transporter)--was used . In the 14 VREF analyzed, the number of unique alleles ranged from 1 (gyd) to 8 (atpA) . Isolates from hospitalized patients were defined by the unique allele purK 1 . Nine sequence types (STs) were identified . All of the epidemic strains isolated over the period 2000-2001 showed identical or closely related pulsed-field gel electrophoresis (PFGE) patterns and clustered in the same ST78 . These strains shared six of the seven alleles with the strain CA20 representative of the 1993-1999 outbreaks, which PFGE indicated as being unrelated to those of the recent outbreaks . MLST confirmed the unrelatedness of human and nonhuman strains already detected by PFGE . All isolates clustered in three main genetic lineages: group A comprised two of the three isolates from meat; group C the human strains of all outbreaks and one poultry strain; and group B four of the five poultry strains and one meat strain . All human strains carried the esp gene and clustered in the C1 sublineage that has been described as having emerged recently worldwide. Semin Dial, 2004 Jul-Aug, 17(4), 310 - 9 National surveillance of dialysis-associated diseases in the United States, 2001; Tokars JI et al.; In December 2001, all U.S . chronic hemodialysis (HD) centers were surveyed regarding selected patient care practices and dialysis-associated diseases . The results were compared with similar surveys conducted in previous years . During 1997-2001, the percentage of patients vaccinated against hepatitis B virus (HBV) infection increased from 47% to 60% and the percentage of staff vaccinated increased from 87% to 89% . In 2001, an estimated 65% of patients had been vaccinated for influenza and 26% for pneumococcal pneumonia . In 2001, routine testing for antibody to hepatitis C virus (anti-HCV) was performed on staff at 42% of centers and on patients at 62% of centers; anti-HCV was found in 1.5% of staff and 8.6% of patients . In 2001, the incidence of HBV infection was higher among patients in centers where injectable medications were prepared at the dialysis station, and both HCV prevalence and incidence were higher among patients in centers where injectable medications were prepared at the dialysis station compared to a dedicated medication room . During 1995-2001, the percentage of patients who received dialysis through central catheters increased from 13% to 25%; this trend is worrisome, as infections and antimicrobial use are higher among patients receiving dialysis through catheters . However, during the same period, the percentage of patients receiving dialysis through fistulas increased from 22% to 30% . In 2001, 25% of catheters were used for new patients awaiting an arteriovenous (AV) access, 28% for established patients with a failed access awaiting new AV access, 40% 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 12% in 1995 to 31% in 2001. Appl Environ Microbiol, 2004 Jul, 70(7), 4205 - 10 Effects of tylosin use on erythromycin resistance in enterococci isolated from swine; Jackson CR et al.; The effect of tylosin on erythromycin-resistant enterococci was examined on three farms; farm A used tylosin for growth promotion, farm B used tylosin for treatment of disease, and farm C did not use tylosin for either growth promotion or disease treatment . A total of 1,187 enterococci were isolated from gestation, farrowing, suckling, nursery, and finishing swine from the farms . From a subset of those isolates (n = 662), 59% (124 out of 208), 28% (80 out of 281), and 2% (4 out of 170) were resistant to erythromycin (MIC >/= 8 microg/ml) from farms A, B, and C, respectively . PCR analysis and Southern blotting revealed that 95% (65 out of 68) of isolates chosen from all three farms for further study were positive for ermB, but all were negative for ermA and ermC . By using Southern blotting, ermB was localized to the chromosome in 56 of the isolates while 9 isolates from farms A and B contained ermB on two similar-sized plasmid bands (12 to 16 kb) . Pulsed-field gel electrophoresis revealed that the isolates were genetically diverse and represented a heterogeneous population of enterococci . This study suggests that although there was resistance to a greater number of enterococcal isolates on a farm where tylosin was used as a growth promotant, resistant enterococci also existed on a farm where no antimicrobial agents were used. APMIS, 2004 Apr-May, 112(4-5), 291 - 8 Role of penicillin-binding protein 5 C-terminal amino acid substitutions in conferring ampicillin resistance in Norwegian clinical strains of Enterococcus faecium; Jureen R et al.; The importance of amino acid sequence differences in the C-terminal part and levels of mRNA expression of penicillin-binding protein 5 (PBP5) for ampicillin resistance in Enterococcus faecium was investigated . Seventeen isolates from Norwegian hospitalized patients (ampicillin MIC 0.064->256 mg/L) with different C-terminal pbp5 DNA sequences encoding 11 different amino acid sequences were analyzed with a 14C-radiolabeled penicillin- binding assay to PBP5 and with real-time PCR quantification of pbp5 mRNA expression . Using multiple logistic regression analysis the amino acid substitution Met 485 was linked to ampicillin MIC and levels of 14C-radiolabeled penicillin bound to PBP5; however, there were isolates with identical PBP5 alleles and different ampicillin MICs . There was no relation between the quantity of pbp5 mRNA transcripts and ampicillin resistance . The results cannot explain ampicillin resistance in Norwegian clinical strains of E . faecium and indicate that other factors besides the properties of the C-terminal part of PBP5 are most likely involved. Dtsch Tierarztl Wochenschr, 2004 May, 111(5), 209 - 12 {Discussion of a biometrical model for the evaluation of feeding, age, and animal effects on transport properties of small intestinal mucosa}; Lodemann U et al.; This paper is concerned with the question in which ways study results can depend on the choice of the statistical model and factors included in this model . This is shown using example data of a study dealing with the effects of an Enterococcus faecium as probiotic in the diet of pigs . We focused on the effects on transport properties of pig jejunum . The experimental design was the following: the sows and piglets were randomly assigned to two different feeding groups . The control group was fed a conventional diet and the experimental group was additionally supplemented with a probiotic preparation of Enterococcus faecium NCIMB 10415 . The animals were divided into four age groups . Two samples of five animals of each feeding and age group were taken and mounted into conventional Ussing chambers . Glucose transport rates were measured by changes in short-circuit current (Isc) of the pig jejunum epithelium . The appropriate reference base for evaluation of effects of feeding or age on Isc is the variation between animals which are submitted to identical conditions relating to these factors . To refer explicitly to this variation a random animal effect has to be included in the statistical model of variance analysis . Otherwise the variation between animals could be underestimated . With the example data set conclusions for the factor "feeding" would be different depending on whether a random animal effect is included in the model or not. Indian J Med Res, 2004 May, 119 Suppl, 77 - 9 Aminoglycoside resistance in enterococci isolated from paediatric septicaemia in a tertiary care hospital in north India; Randhawa VS et al.; BACKGROUND & OBJECTIVES: Enterococci are important nosocomial agents and serious infections caused by them are often treated with a combination of cell wall inhibitor and aminoglycoside . However, the presence of high level aminoglycoside resistance in these isolates makes this treatment combination ineffective . The prevalence of such isolates in a tertiary care set up has important diagnostic and therapeutic implications . The present study was carried out to find out the occurrence of high level aminoglycoside resistant isolates of enterococci in paediatric septicaemia cases in a tertiary care set up in north India . METHODS: Blood of paediatric cases with a clinical diagnosis of septicaemia was cultured to isolate and identify enterococci . Agar screen method was used to detect high level streptomycin and gentamicin resistance in these isolates . Vancomycin susceptibility of these isolates were determined as per the National Committee for Clinical Laboratory Standards (NCCLS) recommendations . RESULTS: Fifty one enterococcal strains were isolated from 21 neonates, nine infants and 21 children with a clinical diagnosis of septicaemia . Sixty eight per cent of these isolates had high level gentamicin resistance and forty three per cent had high level streptomycin resistance . All the isolates with high level streptomycin resistance also had high level gentamicin resistance . More than ninety five per cent of these isolates were sensitive to vancomycin . INTERPRETATION & CONCLUSION: The occurrence of high level gentamicin and high level streptomycin resistance in enterococcal isolates in our set up was high . This would require routine testing of the enterococcal isolates for high level aminoglycoside resistance . Alternative treatment regimes need to be sought, besides prudent use of antibiotics. Folia Microbiol (Praha), 2004, 49(2), 179 - 82 Anaerobic bacteria in the gut of terrestrial isopod Crustacean Porcellio scaber; Kostanjsek R et al.; Anaerobic bacteria from Porcellio scaber hindgut were identified and, subsequently, isolated using molecular approach . Phylogenetic affiliation of bacteria associated with the hindgut wall was determined by analysis of bacterial 16S rRNA gene sequences which were retrieved directly from washed hindguts of P . scaber . Sequences from bacteria related to obligate anaerobic bacteria from genera Bacteroides and Enterococcus were retrieved, as well as sequences from 'A1 subcluster' of the wall-less mollicutes . Bacteria from the genus Desulfotomaculum were isolated from gut wall and cultivated under anaerobic conditions . In contrast to previous reports which suggested the absence of anaerobic bacteria in the isopod digestive system due to short retention time of the food in the tube-like hindgut, frequent renewal of the gut cuticle during the moulting process, and unsuccessful attempts to isolate anaerobic bacteria from this environment our results indicate the presence of resident anaerobic bacteria in the gut of P . scaber, in spite of apparently unsuitable, i.e . predominantly oxic, conditions. Emerg Infect Dis, 2004 Apr, 10(4), 679 - 83 Antimicrobial resistance gene delivery in animal feeds; Lu K et al.; Avoparcin, a glycopeptide antimicrobial agent related to vancomycin, has been used extensively as a growth promoter in animal feeds for more than 2 decades, and evidence has shown that such use contributed to the development of vancomycin-resistant enterococci . A cluster that includes three genes, vanH, vanA, and vanX, is required for high-level resistance to glycopeptides . In the vancomycin producer Amycolatopsis orientalis C329.2, homologs of these genes are present, suggesting an origin for the cluster . We found substantial bacterial DNA contamination in animal feed-grade avoparcin . Furthermore, nucleotide sequences related to the cluster vanHAX are present in this DNA, suggesting that the prolonged use of avoparcin in agriculture led to the uptake of glycopeptide resistance genes by animal commensal bacteria, which were subsequently transferred to humans. Infect Control Hosp Epidemiol, 2004 May, 25(5), 436 - 8 Control of nosocomial acquisition of vancomycin-resistant Enterococcus through active surveillance of hemodialysis patients; Axon RN et al.; Hemodialysis-dependent patients are an important VRE source . After implementation of active surveillance for VRE targeting hemodialysis patients, the hospital-wide nosocomial VRE rate increased by 41%, but decreased by 41% among non-hemodialysis patients (P = .05) . To assess the effectiveness of active surveillance, patients undergoing active surveillance should be analyzed separately from other patients. Infect Control Hosp Epidemiol, 2004 May, 25(5), 418 - 24 A cost-benefit analysis of gown use in controlling vancomycin-resistant Enterococcus transmission: is it worth the price? Puzniak LA, Gillespie KN, Leet T, Kollef M, Mundy LM. OBJECTIVE: To determine the net benefit and costs associated with gown use in preventing transmission of vancomycin-resistant Enterococcus (VRE) . DESIGN: A cost-benefit analysis measuring the net benefit of gowns was performed . Benefits, defined as averted costs from reduced VRE colonization and infection, were estimated using a matched cohort study . Data sources included a step-down cost allocation system, hospital informatics, and microbiology databases . SETTING: The medical intensive care unit (MICU) at Barnes-Jewish Hospital, St . Louis, Missouri . PATIENTS: Patients admitted to the MICU for more than 24 hours from July 1, 1997, to December 31, 1999 . INTERVENTIONS: Alternating periods when all healthcare workers and visitors were required to wear gowns and gloves versus gloves alone on entry to the rooms of patients colonized or infected with VRE . RESULTS: On base-case analysis, 58 VRE cases were averted with gown use during 18 months . The annual net benefit of the gown policy was dollar 419,346 and the cost per case averted of VRE was dollar 1,897 . The analysis was most sensitive to the level of VRE transmission . CONCLUSIONS: Infection control policies (eg, gown use) initially increase the cost of health services delivery . However, such policies can be cost saving by averting nosocomial infections and the associated costs of treatment . The cost savings to the hospital plus the benefits to patients and their families of avoiding nosocomial infections make effective infection control policies a good investment. Infect Control Hosp Epidemiol, 2004 May, 25(5), 391 - 4 Impact of surveillance for vancomycin-resistant enterococci on controlling a bloodstream outbreak among patients with hematologic malignancy; Hachem R et al.; OBJECTIVE: To determine the impact of stool surveillance cultures of critically ill patients on controlling vancomycin-resistant enterococci (VRE) outbreak bacteremia . DESIGN: Stool surveillance cultures were performed on patients who had hematologic malignancy or were critically ill at the time of hospital admission to identify those colonized with VRE . Hence, contact isolation was initiated . SETTING: A tertiary-care cancer center with a high prevalence of VRE . PARTICIPANTS: All patients with hematologic malignancy who were admitted to the hospital as well as all of those admitted to the intensive care unit were eligible . RESULTS: Active stool surveillance cultures performed between 1997 and 2001 decreased the incidence density of VRE bacteremias eightfold while vancomycin use remained constant . In fiscal year (FY) 1997 and FY 1998, there were five and three VRE outbreak bacteremias, respectively . The outbreak clones were responsible for infection in 69% of those patients with VRE bacteremia . However, the stool surveillance program resulted in the complete control of VRE bacteremia by FY 1999 until the end of the study . CONCLUSION: Despite the steady use of vancomycin, the active surveillance program among high-risk patients with hematologic malignancy and those who were critically ill resulted in the complete control of VRE outbreak bacteremia at our institution. Infect Control Hosp Epidemiol, 2004 May, 25(5), 384 - 90 Eradication of a large outbreak of a single strain of vanB vancomycin-resistant Enterococcus faecium at a major Australian teaching hospital; Christiansen KJ et al.; OBJECTIVE: To demonstrate that nosocomial transmission of vancomycin-resistant enterococci (VRE) can be terminated and endemicity prevented despite widespread dissemination of an epidemic strain in a large tertiary-care referral hospital . INTERVENTIONS: Two months after the index case was detected in the intensive care unit, 68 patients became either infected or colonized with an epidemic strain of vanB vancomycin-resistant Enterococcus faecium despite standard infection control procedures . The following additional interventions were then introduced to control the outbreak: (1) formation of a VRE executive group; (2) rapid laboratory identification (30 to 48 hours) using culture and polymerase chain reaction detection of vanA and vanB resistance genes; (3) mass screening of all hospitalized patients with isolation of carriers and cohorting of contacts; (4) environmental screening and increased cleaning; (5) electronic flagging of medical records of contacts; and (6) antibiotic restrictions (third-generation cephalosporins and vancomycin) . RESULTS: A total of 19,658 patient and 24,396 environmental swabs were processed between July and December 2001 . One hundred sixty-nine patients in 23 wards were colonized with a single strain of vanB vancomycin-resistant E . faecium . Introducing additional control measures rapidly brought the outbreak under control . Hospital-wide screening found 39 previously unidentified colonized patients, with only 7 more nonsegregated patients being detected in the next 2 months . The outbreak was terminated within 3 months at a cost of dollar 2.7 million (Australian dollars) . CONCLUSION: Despite widespread dissemination of VRE in a large acute care facility, eradication was achievable by a well-resourced, coordinated, multifaceted approach and was in accordance with good clinical governance. Infect Control Hosp Epidemiol, 2004 May, 25(5), 380 - 3 Effect of the increasing use of piperacillin/tazobactam on the incidence of vancomycin-resistant enterococci in four academic medical centers; Stiefel U et al.; BACKGROUND: The substitution of piperacillin/tazobactam, ampicillin/sulbactam, or both for third-generation cephalosporins has been associated with reduced vancomycin-resistant enterococci (VRE) . However, piperacillin/tazobactam came into widespread use during a period in which the prevalence of VRE increased . We hypothesized that the increasing use of piperacillin/tazobactam and other agents with relatively enhanced anti-enterococcal activity (ie, piperacillin, ampicillin/sulbactam, and ampicillin) has been associated with increased or unchanged rates of VRE in some hospitals . DESIGN: We retrospectively evaluated the correlation between hospital antibiotic use (defined daily doses per 10,000 patient-days of care) and incidence of stool or non-stool VRE isolation . We assessed whether a high or increasing proportion of use of beta-lactam agents with relatively enhanced versus minimal (ie, third-generation cephalosporins and ticarcillin/clavulanate) anti-enterococcal activity would prevent increased VRE . SETTING: Four academic medical centers . RESULTS: With the increasing use of piperacillin/tazobactam, the use of beta-lactam agents with enhanced activity against enterococci surpassed the combined use of third-generation cephalosporins and ticarcillin/clavulanate in each hospital . In one hospital, the incidence of VRE was positively correlated with the use of piperacillin/tazobactam or beta-lactam agents with enhanced anti-enterococcal activity (P < .0001) . The incidence of VRE rose steadily in another hospital despite relatively high use of beta-lactam agents with enhanced versus minimal anti-enterococcal activity . A negative correlation between VRE and piperacillin/tazobactam or beta-lactam agents with enhanced anti-enterococcal activity was observed in one hospital, but this correlation was not statistically significant . CONCLUSION: Increasing the hospital use of piperacillin/tazobactam and other beta-lactams with relatively enhanced anti-enterococcal activity may not be an effective control measure for VRE. Infect Control Hosp Epidemiol, 2004 May, 25(5), 373 - 9 Increased susceptibility to vancomycin-resistant Enterococcus intestinal colonization persists after completion of anti-anaerobic antibiotic treatment in mice; Stiefel U et al.; BACKGROUND: Antibiotic-associated disruption of the indigenous intestinal microflora may persist beyond the treatment period . Although piperacillin/tazobactam inhibits the establishment of vancomycin-resistant Enterococcus (VRE) stool colonization in mice during treatment, we hypothesized that this agent and other anti-anaerobic antibiotics would increase susceptibility to colonization during the period of recovery of the intestinal microflora . DESIGN: Mice received 10(4) colony-forming units of vancomycin-resistant E . faecium by orogastric inoculation 2, 5, or 10 days after completing 5 days of subcutaneous antibiotic treatment, or both during and 2 days after the completion of treatment . Denaturing gradient gel electrophoresis (DGGE) was performed to assess changes in the intestinal microflora . RESULTS: Anti-anaerobic antibiotics (ie, piperacillin/ tazobactam, cefoxitin, and clindamycin) caused significant disruption of the indigenous microflora (mean DGGE similarity indices < or = 27% in comparison with saline controls) and promoted the establishment of high-density colonization when VRE was inoculated 2 or 5, but not 10, days following treatment (P < .001) . Piperacillin/tazobactam exhibited a biphasic effect on the establishment of colonization (ie, inhibition when exposed to VRE during treatment and promotion when exposed to VRE after discontinuation of treatment), resulting in greater overall promotion of colonization than did agents with minimal anti-anaerobic activity (ie, levofloxacin, cefepime, and aztreonam) when VRE was inoculated both during and 2 days after treatment (P < .001) . CONCLUSION: Patients receiving anti-anaerobic antibiotics, including piperacillin/tazobactam, may be susceptible to the establishment of high-density VRE colonization during the period of recovery of the anaerobic microflora. J Clin Microbiol, 2004 Jun, 42(6), 2636 - 43 Comparison of the Roche LightCycler vanA/vanB detection assay and culture for detection of vancomycin-resistant enterococci from perianal swabs; Sloan LM et al.; We compared the performance characteristics of a real-time PCR method, the LightCycler vanA/vanB detection assay (Roche Diagnostics Corporation, Indianapolis, Ind.) to that of Enterococcosel agar (BBL, Sparks, Md.) for direct detection of vancomycin-resistant enterococci (VRE) from 894 perianal stool swabs . For 421 of 894 swabs, the result for LightCycler PCR was compared to an Enterococcosel plate containing vancomycin at 6 microg/ml; for the remaining 473 swabs, the result for LightCycler PCR was compared to an Enterococcosel plate containing 8 microg/ml vancomycin . The LightCycler method produced considerably more positive results than either the Enterococcosel plate containing vancomycin at 6 microg/ml (n = 25 versus n = 11; sensitivity, 100%; specificity, 97%; positive predictive value {PPV}, 42%; negative predictive value {NPV}, 100%) or the Enterococcosel plate containing vancomycin at 8 microg/ml (n = 31 versus n = 10; sensitivity, 100%; specificity, 95%; PPV, 32%; NPV, 100%) . When possible, additional testing, including culture, LightCycler PCR, and/or a conventional PCR method (PCR-restriction fragment length polymorphism assay), were performed on either the original specimens or original cultures or subsequent specimens for cases in which the original specimen was positive by LightCycler PCR but the Enterococcosel plate was negative . This additional testing demonstrated positive results for 7 of 14 (50%) evaluable discordant specimens which initially tested as LightCycler PCR positive but culture negative using the Enterococcosel plate containing vancomycin at 6 microg/ml and 12 of 17 (71%) evaluable discordant specimens which initially tested as LightCycler positive but culture negative using the Enterococcosel plate containing vancomycin at (8 microg/ml) . These results demonstrate that the LightCycler VRE detection assay is considerably more sensitive than the standard culture method for detecting VRE directly from perianal swab specimens . The LightCycler assay also provides results much faster than culture (approximately 3.5 versus > or =72 h) . The use of this test could have important implications for the effective control and prevention of nosocomial outbreaks of VRE. FEMS Microbiol Lett, 2004 Jun 15, 235(2), 385 - 91 Conjugal transfer of aminoglycoside and macrolide resistance between Enterococcus faecium isolates in the intestine of streptomycin-treated mice; Lester CH et al.; The purpose was to study conjugal transfer of resistance genes between a multi-resistant Enterococcus faecium isolate and a sensitive E . faecium isolate . Co-transfer of erm(B)-Tn5405-like element and aac(6')-Ie-aph(2'')-Ia was obtained in both in vivo and in vitro . Plasmid profiles and Southern blots showed that both the erm(B)-Tn5405-like element and aac(6')-Ie-aph(2'')-Ia were placed on the same large plasmid (>147 kb) . These data show to our knowledge the first co-transfer of the erm(B)-Tn5405-like element and aac(6')-Ie-aph(2'')-Ia . The in vivo study also indicates that transfer of resistance genes between enterococci might occur under natural conditions in the gut of animals. J Antimicrob Chemother, 2004 Jul, 54(1), 236 - 9 Epub 2004 Jun 02. Molecular study of vancomycin-resistant enterococci isolated from humans and from food in a cattle-rearing area of France; Garnier F et al.; OBJECTIVES: Study possible links between vancomycin-resistant enterococci strains isolated from human stool samples and from pork or poultry food products . METHODS: One hundred and eleven vancomycin-resistant enterococci strains (15 VanA and 96 VanC) were isolated from human stool samples and from pork or poultry food products . Characterization of the Tn1546-like element of the 15 VanA strains was realized by restriction analysis of PCR products and polymorphism study . The 96 strains of VanC phenotype (75 Enterococcus gallinarum and 21 Enterococcus casseliflavus) were analysed by pulsed-field gel electrophoresis (PFGE) . RESULTS: In the study described here, polymorphism of the Tn1546-like element enabled the establishment of five groups . Groups III, IV and V were found only in human strains . Groups I and II were found to occur in strains isolated from humans and from food, suggesting a possible transfer of the Tn1546-like element . The isolates from Group I harboured the whole Tn1546 element . In Group II, the five strains possessed a novel Tn1546-like element, characterized by a single-nucleotide difference in the vanX gene and a deletion upstream from the nucleotide 164 . Analysis by PFGE of the 75 E . gallinarum strains revealed 20 different patterns . One pattern was shared by isolates from pork food and human samples . None of the 21 E . casseliflavus strains tested was found to share similar PFGE patterns . CONCLUSIONS: Results tend to support the possible transfer of the Tn1546-like element between strains of VanA phenotype . Concerning VanC phenotype strains, the transfer was not demonstrated. Int J Food Microbiol, 2004 Jul 1, 94(1), 23 - 31 The presence of Enterococcus, coliforms and E . coli in a commercial yeast manufacturing process; O'Brien SS et al.; This study evaluated a typical commercial yeast manufacturing process for bacterial contamination . Product line samples of a commercial yeast manufacturing process and the corresponding seed yeast manufacturing process were obtained upstream from the final compressed and dry yeast products . All samples were analysed before (non-PI) and after preliminary incubation (PI) at 37 degrees C for 24 h . The PI procedure was incorporated for amplification of bacterial counts below the lower detection limit . Enterococcus, coliform and Escherichia coli counts were quantified by standard pour-plate techniques using selective media . Presence at all stages and progressive increases in counts of Enterococcus, coliforms and E . coli during processing in the commercial manufacturing operation suggested that the primary source of contamination of both compressed and dry yeast with these bacteria was the seed yeast manufacturing process and that contamination was amplified throughout the commercial yeast manufacturing process . This was confirmed by surveys of the seed yeast manufacturing process which indicated that contamination of the seed yeast with Enterococcus, coliforms and E . coli occurred during scale up of seed yeast biomass destined as inoculum for the commercial fermentation. Yao Xue Xue Bao, 2004 Mar, 39(3), 184 - 9 {Synthesis and antibacterial activity of 7-(7-aminomethyl-5-azaspiro {2,4} hept-5-yl)-1-cyclopropyl-6-fluoro-8-methoxy-1,4-dihydro-4-oxo-3-quinolinecarboxylic acid and its analogues}; Qi JJ et al.; AIM: To find new antibacterial agents of quinolone with high activity and low toxicity . METHODS: To design and synthesize 7-(7-aminomethyl-5-azaspiro {2,4} hept-5-yl)-1-cyclopropyl-6-fluoro-8-methoxy-1,4-dihydro-4-oxo-3-quinolinecarboxylic acid and its analogues, and to study their antibacterial activity in vitro and in vivo . RESULTS: Twenty new compounds (2 - 11, 17 - 26) were obtained including five targeted compounds (22 - 26) . The structures of the compounds were confirmed by 1HNMR, MS and HRMS . Compounds 22 - 26 showed broad spectrum of antibacterial activity against Gram-positive and Gram-negative organisms . Especially for compound 24, the relevant MIC values for 13 strains of Gram-positive organisms were < 0.001 - 0.03 mg(-1), including 4 strains of S . pneumoniae, 2 strains of S . pyogenes, 3 strains of S . aureus and 2 strains of Enterococci which exhibited more potent activity than contrast agents (clinafloxacin and gatifloxacin) . The MIC values of 24 for 6 strains Gram-positive organisms were 0.01 - 1 mg x L(-1), which exhibited equal or lower activity than contrast agents . They were more effective than ciprofloxacin and gatifloxacin against intraperitoneal infections caused by S . pneumoniae and S . aureus in mice . CONCLUSION: Compounds (23, 24 and 26) showed excellent antibacterial activity in vitro and in vivo and should be worth further investigation. Curr Opin Infect Dis, 2004 Jun, 17(3), 231 - 6 Late-onset infection and the role of antibiotic prescribing policies; Gordon A et al.; PURPOSE OF REVIEW: Late-onset infection is a significant cause of morbidity and mortality in low-birth-weight and premature infants . Empirical antibiotic treatment is used as infants can deteriorate rapidly without treatment . Current data on the epidemiology of late-onset infection, the types of antibiotics used, duration of antibiotic use, and antibiotic prescribing policies are reviewed . RECENT FINDINGS: Epidemiological data on late-onset sepsis is dominated by information concerning developed countries; large prospective data collections have been set up in many such countries . Recent data indicate that late-onset sepsis occurs in one-fifth of very-low-birth-weight infants . There are increasing concerns regarding antibiotic resistance . Antibiotic regimens that do not include third-generation cephalosporins produce less resistance . Strategies of antibiotic rotation have not been documented as producing a marked effect on the development of resistant micro-organisms, but there is a lack of randomized trials . Recommendations for preventing the spread of vancomycin-resistant enterococci, produced by the Hospital Infection Control Practices Advisory Committee, have been shown to be effective in a number of situations . Recent reports have documented the success of multidisciplinary, systems-orientated approaches for reducing neonatal nosocomial infection . SUMMARY: Antibiotic prescribing policies have an important role to play in the treatment of late-onset neonatal infection . There is enough evidence to state that narrow-spectrum antibiotics should be used wherever possible and that potent broad-spectrum antibiotics should be kept in reserve . Ongoing prospective surveillance of infection rates, micro-organisms, resistance and antibiotic use is essential. J Heart Lung Transplant, 2004 May, 23(5), 564 - 9 Management of aortopulmonary collaterals in children following cardiac transplantation for complex congenital heart disease; Krishnan US et al.; BACKGROUND: Heart transplantation (HTx) is increasingly utilized as therapy for end-stage cyanotic congenital heart disease . This study investigates the presence and impact of aortopulmonary collaterals (APCs) associated with cyanotic heart disease on the early post-operative course of patients undergoing transplantation . High output cardiac failure due to residual aortopulmonary collaterals can affect outcome following heart transplantation . METHODS: Seven patients with hemodynamically significant APCs post-transplant were identified among 40 patients with cyanotic congenital heart disease undergoing HTx . The peri- and intra-operative courses of these patients were reviewed . All 7 patients required prolonged inotropic support despite normal ventricular function and no allograft rejection; 5 were ventilator-dependent due to significant pulmonary vascular congestion . Selective angiography demonstrated the presence of significant aortopulmonary collaterals at 7 to 19 days post-transplant . Coil embolization of aortopulmonary collaterals was performed in all patients; a mean of 6 (2 to 16) vessels/patient were embolized . RESULTS: After embolization, pulmonary edema resolved and heart size normalized in all patients; inotropic support was weaned within 2 to 10 days in 5 patients . One patient developed transient renal failure secondary to excessive contrast load and another had enterococcal sepsis within 24 hours after the procedure . All patients were asymptomatic from 4 to 10 years of follow-up post-HTx . CONCLUSIONS: Aortopulmonary collaterals should be considered a cause of early donor heart failure in children following HTx for cyanotic congenital heart disease . Early detection and treatment of aortopulmonary collaterals by coil embolization is necessary to improve the post-transplant course in these complex patients. J Clin Microbiol, 2004 May, 42(5), 1897 - 902 Distribution of insertion sequences associated with Tn1546-like elements among Enterococcus faecium isolates from patients in Korea; Huh JY et al.; The vanA gene cluster is carried as a part of Tn1546-like elements . The genetic diversity in Tn1546-like elements has been documented previously . The differences described thus far have included the integration of insertion sequence (IS) elements IS1216V, IS1251, IS1476, and IS1542 . Among these, IS1216V has been reported to be widespread in VanA enterococci of diverse geographic areas, whereas IS1542 and IS1476 have been reported only in the United Kingdom and Canada, respectively . We investigated the distribution of ISs among 20 vanA-containing Enterococcus faecium isolates from human patients in nine different university hospitals in Korea . Pulsed-field gel electrophoresis (PFGE) was performed to identify the clonality of the isolates . Moreover, PCR amplification of the internal regions of Tn1546 was performed for structural analysis of the van gene, and both DNA strands of the PCR amplicons were directly sequenced