Enterococci

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) ≥ 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 by the dideoxy termination method . The PFGE patterns revealed a high degree of clonal diversity . Structural analyses of the van gene detected IS1542 and IS1216V in the genomes of all 20 isolates, whereas it did not detect IS1476 or IS1251 in the genomes of any of the isolates . In addition, IS19 was detected in the vanS-vanH intergenic region of one isolate . These data indicate that identification of the IS within a vanA gene cluster could be a useful tool in epidemiological investigations . In addition, the distribution of ISs associated with Tn1546-like elements among the Korean isolates is therefore similar to that among European vancomycin-resistant enterococci.

J Antimicrob Chemother, 2004 Jun, 53(6), 1105 - 8 Epub 2004 May 05.
Relationship between myalgias/arthralgias occurring in patients receiving quinupristin/dalfopristin and biliary dysfunction; Raad I et al.; OBJECTIVES: To determine whether myalgias/arthralgias occurring in cancer patients who receive quinupristin/dalfopristin are associated with biliary tract dysfunction . METHODS: We studied 56 patients with vancomycin-resistant enterococcal infections who were treated with quinupristin/dalfopristin 7.5 mg/kg every 8 h for a mean duration of 12 days (range 2-52 days) . Liver function tests, including a test for alkaline phosphatase, were performed before, during and after the end of therapy . All patients were followed for 1 month after completion of therapy . RESULTS: Thirty-eight (68%) of the 56 patients responded . Myalgias/arthralgias were the leading adverse events occurring in 20 (36%) of the patients . Patients with myalgias/arthralgias had significantly higher levels of alkaline phosphatase (mean 318.7 IU/L) during the mid-term therapy cycle compared with patients without any joint or muscular pain (mean 216.3 IU/L, P = 0.05) . In addition, 3/18 (16.6%) patients with myalgias/arthralgias had more than five-fold the normal levels of alkaline phosphatase, which did not occur in any of the other patients who did not develop myalgias/arthralgias (P = 0.04) . All myalgias/arthralgias resolved after the discontinuation of quinupristin/dalfopristin . By univariate analysis, other factors associated with myalgias/arthralgias were relapse of haematological malignancy (P = 0.01), receiving tacrolimus within 1 month prior to treatment (P = 0.04) and receiving methotrexate during antimicrobial therapy (P = 0.05) . CONCLUSIONS: Myalgias/arthralgias occur frequently in cancer patients receiving quinupristin/dalfopristin and may be associated with biliary tract dysfunction, as measured by alkaline phosphatase or other factors that could lead to intra-hepatic cholestasis, such as relapse of haematological malignancy or treatment with tacrolimus or methotrexate.

Int J Antimicrob Agents, 2004 May, 23(5), 502 - 5
Rapid detection and differentiation method of VanA, VanB and VanC phenotypes in vancomycin-resistant enterococci; Hanaki H et al.; We developed a simple method that can replace the polymerase chain reaction (PCR) to distinguish between vancomycin-resistant enterococci (VRE) with the vanA, vanB and vanC genes . The method is based on induction of teicoplanin resistance by vancomycin in vanB-VRE, while the two compounds have a synergistic effect in vanC-VRE . In addition, vanA-VRE shows resistance to both vancomycin and teicoplanin, and both the compounds can induce resistance to vanA-VRE . Utilising these properties, we attempted to develop a simple method to distinguish between vanA, vanB and vanC . We compared our simple method with the PCR method in 43 strains of vanA-VRE, 35 strains of vanB-VRE and 37 strains of vanC-VRE . The results were 100% consistent with that obtained by PCR.

Clin Microbiol Infect, 2004 May, 10(5), 441 - 7
Microbiological and epidemiological studies of Enterococcus faecium resistant to amoxycillin in a university hospital in eastern France; Thouverez M et al.; Over 3 years, during six 1-month periods of surveillance, 69 cases of Enterococcus faecium colonisation or infection were detected in a university hospital in eastern France . Thirty-two cases involved strains resistant to amoxycillin (crude incidence of 0.21/1000 patient-days) . The risk of infection with E . faecium was higher if the patient was hospitalised in a haematology ward and/or treated with cephalosporins . Amoxycillin-resistant isolates (AmRE) were isolated from different wards and time periods, and none of the characteristics studied were associated significantly with amoxycillin resistance . Amoxycillin-sensitive and -resistant isolates were characterised by pulsed-field gel electrophoresis . Three epidemic patterns were identified which contained 87.5% (28/32) of the AmRE isolates, indicating that clonal spread was responsible, at least partially, for the high incidence of AmRE in this hospital.

Kansenshogaku Zasshi, 2004 Mar, 78(3), 283 - 7
{A case of probable bacterial translocation-associated sepsis in healthy adult}; Ohosone Y et al.; We report a patient with bacterial translocation-associated sepsis who was healthy and did not have any related-background . The 57-year-old male had been well until 16 hours before admission, when nausea and vomiting gradually developed and increased in intensity . In the morning of May 22, 2002, he had shaking chills, temperature of 38.6 degrees C and watery diarrhea, and was admitted to Kawasaki Municipal Hospital . On admission, temperature was 40.7 degrees C but otherwise physical examination revealed no particular abnormality . Laboratory data showed total white blood cells of 28,400/microliter, platelet count of 130,000/microliter, creatinine of 2.0 mg/dl and C-reactive protein of 7.5 mg/dl . 1 g of cefmetazole was administered every eight hours . In the early morning of May 23, he suddenly went into shock . At that time, laboratory findings revealed total white blood cells of 33,700/microliter, platelet count of 65,000/microliter, C-reactive protein of 24.9 mg/dl, creatinine of 5.6 mg/dl and serum potassium concentration of 5.7 mEq/l . Gram positive cocci and gram negative rods were isolated from blood culture obtained on admission . Cefmetazole was changed to 1.5 g/day of imipenem/cilastatin sodium and 600 mg/day of clindamycin . In addition, hemodialysis and endotoxin removal with an adsorbent column using polymyxin B were performed . Bacteria detected in the blood on admission were identified as Klebsiela oxytoca and Enterococcus faecium . Imipenem/cilastatin sodium and clindamycin were continued for 13 days . The patient recovered fully and was discharged on June 11 . This case suggests that bacterial translocation-associated sepsis might occur even in a hitherto healthy adult.

Clin Infect Dis, 2004 Apr 15, 38(8), 1108 - 15 Epub 2004 Apr 05.
Projected benefits of active surveillance for vancomycin-resistant enterococci in intensive care units; Perencevich EN et al.; Hospitals use many strategies to control nosocomial transmission of vancomycin-resistant enterococci (VRE) . Strategies include "passive surveillance," with isolation of patients with known previous or current VRE colonization or infection, and "active surveillance," which uses admission cultures, with subsequent isolation of patients who are found to be colonized with VRE . We created a mathematical model of VRE transmission in an intensive care unit (ICU) using data from an existing active surveillance program; we used the model to generate the estimated benefits associated with active surveillance . Simulations predicted that active surveillance in a 10-bed ICU would result in a 39% reduction in the annual incidence of VRE colonization when compared with no surveillance . Initial isolation of all patients, with withdrawal of isolation if the results of surveillance cultures are negative, was predicted to result in a 65% reduction . Passive surveillance was minimally effective . Using the best available data, active surveillance is projected to be effective for reducing VRE transmission in ICU settings.

J Agric Food Chem, 2004 Apr 21, 52(8), 2247 - 55
Performance and applications of polyclonal antipeptide antibodies specific for the enterococcal bacteriocin enterocin P; Gutierrez J et al.; Polyclonal antibodies with specificity for enterocin P (EntP) have been generated by immunization of rabbits with two chemically synthesized N-terminal peptides (P1 and P2) and a C-terminal peptide (P3) of this bacteriocin conjugated to the carrier protein KLH . The sensitivity and specificity of the peptide-KLH-generated antibodies were evaluated by a noncompetitive indirect enzyme-linked immunosorbent assay (NCI-ELISA) and a competitive indirect (CI)-ELISA . The NCI-ELISA but not the CI-ELISA was valuable for detecting the existence of EntP specific antibodies in the sera of the P2-KLH and P3-KLH immunized animals and to detect and quantify the EntP in the supernatant of producer strains . The anti-P2-KLH sera cross-reacted with the supernatant of a strain producer of sakacin A, a bacteriocin closely related to EntP . Immunoaffinity chromatography columns with anti-P2-KLH or anti-P3-KLH immunoglobulins retained the EntP from the supernatant of the producer strain . Western blotting of EntP with the anti-P2-KLH-generated antibodies suggests that purified EntP tends to the formation of aggregates with no antimicrobial activity . Monitoring the purification of EntP with antipeptide antibodies suggests that while the performance of the evaluated purification procedures would be reasonably acceptable in terms of recovery of the antimicrobial activity of the bacteriocin, their yield is far from attractive in terms of recovery of the initial concentration of enterocin P.

Indian J Pediatr, 2004 Mar, 71(3), 229 - 39
Antibacterial resistance; Ang JY et al.; Resistance to antimicrobial agents, which was recognized more than 50 years, continues to be a major cause of increased morbidity, mortality and health care cost . Overuse of antibiotics is considered the major contributing factor; however, poor implementation of infection control measures, prolonged hospitalization, admission to intensive care units and the use of invasive procedures are other contributing factors . The authors review the epidemiology, mechanism of resistance, treatment options and prevention measures of infections caused multi-drug resistant S . pneumoniae, methicillin-resistant S . aureus (MRSA), macrolide-resistant S . pyogenes and vancomycin-resistant enterococci (VRE) in pediatric patients . Antibacterial resistance among Gram-negative organisms, including extended spectrum beta-lactamase (ESBL) producing enteric bacteria, and the management and prevention of infections caused by these organisms are also discussed.

J Clin Microbiol, 2004 Apr, 42(4), 1785 - 6
Emergence of vanA Genotype Vancomycin-Resistant Enterococci with Low or Moderate Levels of Teicoplanin Resistance in Korea; Eom JS et al.; In Korea, vancomycin-resistant enterococci have become important nosocomial pathogens since the late 1990s, and most vancomycin-resistant enterococcal isolates have been VanA phenotype-vanA genotype strains . In 2001, we experienced an outbreak of VanB phenotype-vanA genotype vancomycin-resistant enterococci at a university hospital . This is the first report of VanB-vanA vancomycin-resistant enterococci from humans in Korea.

J Clin Microbiol, 2004 Apr, 42(4), 1751 - 2
Evaluation of the Velogene genomic assay for detection of vanA and vanB genes in vancomycin-resistant Enterococcus species; Appleman MD et al.; The Velogene (VEL) genomic assay is a qualitative DNA probe for vanA and vanB in enterococci . 150 clinical isolates were tested with the VEL assay and characterized by pigment production, catalase levels, motility, growth in 6 micro g of vancomycin/ml, vancomycin and teicoplanin susceptibility, API 20S assay, and genotyping by multiplex PCR . The VEL assay identified all enterococcal strains with vanA and vanB genes.

Diagn Microbiol Infect Dis, 2004 Apr, 48(4), 287 - 8
API 20 strep identification system may incorrectly speciate enterococci with low level resistance to vancomycin; Winston LG et al.; The API 20 Strep system was used to speciate 46 enterococcal isolates with vancomycin MICs between 16-32 microg/mL . All were identified as Enterococcus faecium . Further testing revealed that 42/46 isolates had been identified incorrectly . Enterococci with low-level vancomycin resistance should not be speciated solely with the API 20 Strep system.

Curr Microbiol, 2004 May, 48(5), 364 - 7
Inhibition of vancomycin-resistant enterococcus by continuous-flow cultures of human stool microflora with and without anaerobic gas supplementation; Hume ME et al.; A continuous-flow competitive exclusion (CFCE) culture model of human stool microflora was used to examine whether supplemental anaerobic gas is necessary for maintenance of anaerobes and inhibition of vancomycin-resistant Enterococcus (VRE) . CFCE cultures of human stool microflora were maintained with supplemental nitrogen, without supplemental nitrogen, or with percolated room air . Cultures with or without supplemental nitrogen maintained >9 log(10) CFU mL(-1) of obligate anaerobes and eliminated 10(6) CFU mL(-1) of VRE . When room air was percolated into the culture, anaerobes were detected at < or =2 log(10) CFU mL(-1), and the same VRE inoculum was not eliminated ( P < 0.001) . These data demonstrate that human stool CFCE cultures maintain high levels of obligate anaerobes and inhibit VRE without the addition of supplemental anaerobic gas.

J Biol Inorg Chem, 2004 Jun, 9(4), 396 - 402 Epub 2004 Apr 01.
Interaction kinetics of the copper-responsive CopY repressor with the cop promoter of Enterococcus hirae; Portmann R et al.; In Enterococcus hirae, copper homeostasis is controlled by the cop operon, which encodes the copper-responsive repressor CopY, the copper chaperone CopZ, and two copper ATPases, CopA and CopB . The four genes are under control of CopY, which is a homodimeric zinc protein, {Zn(II)CopY}2 . It acts as a copper-responsive repressor: when media copper is raised, CopY is released from the DNA, allowing transcription to proceed . This involves the conversion of {Zn(II)CopY}2 to {Cu(I)2CopY}2, which is no longer able to bind to the promoter . Binding analysis of {Zn(II)CopY}2 to orthologous promoters and to control DNA by surface plasmon resonance analysis defined the consensus sequence TACAnnTGTA as the repressor binding element, or " cop box", of Gram-positive bacteria . Association and dissociation rates for the CopY-DNA interaction in the absence and presence of added copper were determined . The dissociation rate of {Zn(II)CopY}2 from the promoter was 7.3 x 10(-6) s(-1) and was increased to 5 x 10(-5) s(-1) in the presence of copper . This copper-induced change may be the underlying mechanism of copper induction . Induction of the cop operon was also assessed in vivo with a biosensor containing a lux reporter system under the control of the E . hirae cop promoter . Half-maximal induction of this biosensor was observed at 5 microM media copper, which delineates the ambient copper concentration to which the cop operon responds in vivo .

Curr Microbiol, 2004 Mar, 48(3), 199 - 203
Characterization of the first VanB vancomycin-resistant Enterococcus faecium isolated in a Spanish hospital; Lorenzo-Diaz F et al.; We report the detection and characterization of the first vancomycin-resistant VanB-type Enterococcus faecium to be isolated in a Spanish hospital . Sequence analysis of the vanB gene showed that this isolate belonged to subtype vanB2 . Moreover, PCR amplification analysis indicated that the vanB gene cluster was linked to a Tn5382-like transposon.

Antimicrob Agents Chemother, 2004 Apr, 48(4), 1379 - 81
Reduction in glycopeptide resistance in vancomycin-resistant enterococci as a result of vanA cluster rearrangements; Lee WG et al.; The molecular characterization of five clinical isolates of vanA-containing vancomycin-resistant enterococci with altered resistance to glycopeptides was examined . One strain represented an IS1216V insertion accompanied by partial deletion of the reading frame of vanX following a transposition event . The other four strains represented IS1216V within the vanX-vanY intergenic region associated with deletion of vanY or vanZ.

S Afr Med J, 2004 Feb, 94(2), 121 - 4
An evaluation of nine probiotics available in South Africa, August 2003; Elliot E et al.; BACKGROUND AND OBJECTIVE: Although probiotics are not new, 5 new commercially available products have been launched onto the South African market in the last 2 years . Evaluations of products in the USA and Europe shown poor correlation between label claims and actual contents . We undertook an evaluation of 9 products currently available on the shelves in South Africa . METHODS AND ANALYSIS: An independent laboratory was used . A culture method involving serial dilutions on selective media was used to obtain a colony count per gram for indicated organisma . A non-culture method, denaturing gradient gel electrophoresis (DGGE), was used to determined the organisms present in the products . RESULTS: Disturbingly, we found a relatively poor correlation between the advertised and determined bacterial content . Only 3 of the 9 products tested contained the bacteria indicated on the label and 5 products contained sufficient bacteria for a probiotic effect . The Enterococcus faecium, a potential pathogen and Saccharomyces cerevisiae found in 2 of the products are of concern . CONCLUSION: This evaluation confirms that the contents of several probiotics available in South Africa do not correspond to the label claims . This is of concern as clinical efficacy is dependent on strain specificity and organism numbers . Current regulatory requirements do not address this discrepancy . As such, we recommend that commercially available probiotic products be screened annually and the results of such quality control measures be made available to the Medicines Control Council (MCC).

Bone Marrow Transplant, 2004 May, 33(9), 937 - 41
Incidence and clinical complications of vancomycin-resistant enterococcus in pediatric stem cell transplant patients; Tsiatis AC et al.; Vancomycin-resistant enterococcus (VRE) are increasingly important pathogens in stem cell transplant (SCT) . In all, 61 pediatric SCT patients had surveillance stool cultures for VRE between July 1999 and November 2002 . When VRE was identified, the patients were placed on strict contact isolation . VRE was detected in 15 patients (24.6%) . The median age was 3.6 years (range 0.6-18.5 years) . Of the 15, 13 (87%) received an allogeneic transplant (six unrelated and seven related) . Five of the 15 (33%) colonized patients developed VRE bacteremia . The bacteremia resolved in all five patients after therapy with quinupristin/dalfopristin; three patients required central line removal . Four patients died (38-153 days) post-SCT due to relapse or transplant complication not related to VRE . Of the 11 surviving patients, seven cleared the colonization at a median of 144 days (range 61-198 days) postcolonization . Four patients remain colonized at 68-702 days after the first positive culture . Intestinal colonization with VRE occurred commonly in pediatric SCT patients . Although the morbidity from VRE was not substantial, transplant patients were colonized for prolonged periods . Our results indicate that surveillance for VRE is an effective way to identify colonized patients and may lead to a decrease in transmission to other patients.

Infez Med, 1995 Mar, 3(1), 28 - 32
Enterococci: susceptibility patterns and therapeutic options; Stefani S et al.; Enterococcal isolates are increasingly responsible for nosocomial infections in the last decade and they are recently cited as being the second most common pathogen isolated from hospitalized patients . Enterococcal infections can be localized at different sites: endocarditis, CNS, intrabdominal, pelvic . The epidemiological trend may be a consequence of predisposing risk factors due to hospitalization such as instrumentation, immunosuppression, long-term hospitalization . Nevertheless the most alarming aspect of the problem is the increasing detection of antibiotic resistance Enterococcal strains, especially the high level resistance (HLR) to aminoglycosides, penicillin and glycopeptides must not be underestimated . Optimal antibiotic regimes for the treatment of multiple resistant strains in severe enterococcal infections are not been defined yet and antibiotic association such as aminoglycosides + glycopeptides or glycopeptides + fluoroquinolones could be suggested.

Risk Anal, 2004 Feb, 24(1), 271 - 88
Quantifying human health risks from virginiamycin used in chickens; Cox LA Jr et al.; The streptogramin antimicrobial combination Quinupristin-Dalfopristin (QD) has been used in the United States since late 1999 to treat patients with vancomycin-resistant Enterococcus faecium (VREF) infections . Another streptogramin, virginiamycin (VM), is used as a growth promoter and therapeutic agent in farm animals in the United States and other countries . Many chickens test positive for QD-resistant E . faecium, raising concern that VM use in chickens might compromise QD effectiveness against VREF infections by promoting development of QD-resistant strains that can be transferred to human patients . Despite the potential importance of this threat to human health, quantifying the risk via traditional farm-to-fork modeling has proved extremely difficult . Enough key data (mainly on microbial loads at each stage) are lacking so that such modeling amounts to little more than choosing a set of assumptions to determine the answer . Yet, regulators cannot keep waiting for more data . Patients prescribed QD are typically severely ill, immunocompromised people for whom other treatment options have not readily been available . Thus, there is a pressing need for sound risk assessment methods to inform risk management decisions for VM/QD using currently available data . This article takes a new approach to the QD-VM risk modeling challenge . Recognizing that the usual farm-to-fork ("forward chaining") approach commonly used in antimicrobial risk assessment for food animals is unlikely to produce reliable results soon enough to be useful, we instead draw on ideas from traditional fault tree analysis ("backward chaining") to reverse the farm-to-fork process and start with readily available human data on VREF case loads and QD resistance rates . Combining these data with recent genogroup frequency data for humans, chickens, and other sources (Willems et al., 2000, 2001) allows us to quantify potential human health risks from VM in chickens in both the United States and Australia, two countries where regulatory action for VM is being considered . We present a risk simulation model, thoroughly grounded in data, that incorporates recent nosocomial transmission and genetic typing data . The model is used to estimate human QD treatment failures over the next five years with and without continued VM use in chickens . The quantitative estimates and probability distributions were implemented in a Monte Carlo simulation model for a five-year horizon beginning in the first quarter of 2002 . In Australia, a Q1-2002 ban of virginiamycin would likely reduce average attributable treatment failures by 0.35 x 10(-3) cases, expected mortalities by 5.8 x 10(-5) deaths, and life years lost by 1.3 x 10(-3) for the entire population over five years . In the United States, where the number of cases of VRE is much higher, a 1Q-2002 ban on VM is predicted to reduce average attributable treatment failures by 1.8 cases in the entire population over five years; expected mortalities by 0.29 cases; and life years lost by 6.3 over a five-year period . The model shows that the theoretical statistical human health benefits of a VM ban range from zero to less than one statistical life saved in both Australia and the United States over the next five years and are rapidly decreasing . Sensitivity analyses indicate that this conclusion is robust to key data gaps and uncertainties, e.g., about the extent of resistance transfer from chickens to people.

Indian J Gastroenterol, 2003 Dec, 22 Suppl 2, S50 - 3
Role of gut bacteria in the therapy of hepatic encephalopathy with lactulose and antibiotics; Dasarathy S; The pathogenesis of hepatic encephalopathy is mediated to a large extent by ammonia, mainly derived from the gut . Both bacterial and nonbacterial mechanisms of ammoniagenesis have been shown, but ammoniagenesis mediated by colonic bacteria is probably of clinical significance . The therapy of hepatic encephalopathy is based on the putative etiological agents, including ammonia . Non-absorbable disaccharides and antibiotics have been shown to modify gut flora and decrease blood ammonia levels, but these are not necessarily related (indicating nonbacterial sources of ammonia, which may also be decreased by these compounds) . A combination of these has been suggested but not consistently demonstrated to be beneficial in hepatic encephalopathy . Sodium benzoate is an alternate method of nonbacterial, non-hepatic metabolic binding pathway for ammonia disposal . Other mechanisms of gut bacterial modification may be achieved albeit transiently by the use of resistant bacteria like Enterococcus faecium.

Int J Antimicrob Agents, 2004 Feb, 23(2), 200 - 3
Emergence of resistance to chloramphenicol among vancomycin-resistant enterococcal (VRE) bloodstream isolates; Lautenbach E et al.; Therapeutic options for vancomycin-resistant enterococcal (VRE) bloodstream infections are extremely limited . Chloramphenicol is effective when VRE isolates are susceptible to this agent . However, longitudinal trends in chloramphenicol-resistant VRE (CR-VRE) are unknown . The possible association between CR-VRE and antibiotic use has not been studied . We analyzed the antimicrobial susceptibility profiles of all VRE blood isolates from 1991-2000 at our institution . We performed a correlational study to examine the relationship between annual hospital-wide use of specific antibiotics and antibiotic classes and CR-VRE prevalence . During the 10-year study period, the prevalence of CR-VRE increased from 0 to 11% ( P< 0.001, trend) . CR-VRE prevalence was correlated only with chloramphenicol use (P=0.05 ) and quinolone use (P= 0.01 ) . If these trends continue, dependence on newer, more expensive agents will increase . The correlation between both chloramphenicol use and quinolone use and the prevalence of CR-VRE suggests that efforts to preserve the utility of chloramphenicol in VRE infections may depend on optimizing the use of these agents.

Eur J Biochem, 2004 Mar, 271(5), 993 - 1003
Metal-binding stoichiometry and selectivity of the copper chaperone CopZ from Enterococcus hirae; Urvoas A et al.; We studied the interaction of several metal ions with the copper chaperone from Enterococcus hirae (EhCopZ) . We show that the stoichiometry of the protein-metal complex varies with the experimental conditions used . At high concentration of the protein in a noncoordinating buffer, a dimer, (EhCopZ)2-metal, was formed . The presence of a potentially coordinating molecule L in the solution leads to the formation of a monomeric ternary complex, EhCopZ-Cu-L, where L can be a buffer or a coordinating molecule (glutathione, tris(2-carboxyethyl)phosphine) . This was demonstrated in the presence of glutathione by electrospray ionization MS . The presence of a tyrosine close to the metal-binding site allowed us to follow the binding of cadmium to EhCopZ by fluorescence spectroscopy and to determine the corresponding dissociation constant (Kd = 30 nm) . Competition experiments were performed with mercury, copper and cobalt, and the corresponding dissociation constants were calculated . A high preference for copper was found, with an upper limit for the dissociation constant of 10-12 m . These results confirm the capacity of EhCopZ to bind copper at very low concentrations in living cells and may provide new clues in the determination of the mechanism of the uptake and transport of copper by the chaperone EhCopZ.

Clin Infect Dis, 2004 Mar 15, 38(6), 780 - 6 Epub 2004 Feb 27.
Detection of vancomycin-resistant enterococci before and after antimicrobial therapy: use of conventional culture and polymerase chain reaction; Trick WE et al.; Antimicrobial therapy can increase the colonization density of gastrointestinal vancomycin-resistant enterococci (VRE) . Among previously VRE-colonized patients, we evaluated VRE colonization before and after initiation of antimicrobial therapy by means of polymerase chain reaction (PCR) and culture . Perianal swab samples were obtained at admission to the hospital and after receipt of antimicrobial therapy . At admission, 12 (21%) of 56 patients were culture positive, and 17 (30%) had vanA or vanB genes detected by PCR . Culture results showed that 25 (86%) of 29 culture-negative patients from whom a second swab sample was obtained remained culture negative, 2 (6.9%) had a relapse of colonization with a strain related to the previously colonizing strain type (2 and 6 days after admission), and 2 (6.9%) tested positive for a previously undetected strain type (16 and 19 days after admission) . PCR at admission detected VRE in 1 of the 2 patients who later relapsed . Patients with negative results of culture of the initial swab sample and of PCR were unlikely to relapse after receipt of antimicrobial therapy.

J Infect Dis, 2004 Mar 15, 189(6), 1113 - 8 Epub 2004 Mar 01.
Beta-lactam antibiotics and gastrointestinal colonization with vancomycin-resistant enterococci; Rice LB et al.; We studied the effect of different subcutaneously administered beta-lactam antibiotics on the establishment of gastrointestinal colonization by vancomycin-resistant Enterococcus faecium C68 in a mouse model . Aztreonam, cefazolin, cefepime, and, to a lesser extent, ceftazidime, which neither have significant antienterococcal activity nor are secreted into human bile at high concentrations, did not promote significant vancomycin-resistant enterococci (VRE) colonization . Piperacillin-tazobactam, which has antienterococcal activity and is secreted in human bile at high concentrations, inhibited colonization after limited exposure to the inoculum but was associated with progressively increased VRE colony counts in stool samples after repeated exposure to the VRE inoculum . Ceftriaxone and cefotetan, which lack antienterococcal activity but are secreted into human bile at high concentrations, were associated with rapid and high-level colonization . These data suggest that the risk of VRE colonization varies during exposure to different beta-lactam antimicrobial agents and that the risk is related to biliary concentration and antienterococcal activity of the specific beta-lactam.

J Antimicrob Chemother, 2004 Apr, 53(4), 646 - 9 Epub 2004 Mar 03.
Prospective, randomized study comparing quinupristin-dalfopristin with linezolid in the treatment of vancomycin-resistant Enterococcus faecium infections; Raad I et al.; OBJECTIVES: Quinupristin-dalfopristin and linezolid have been shown to be efficacious in the treatment of vancomycin-resistant Enterococcus faecium (VREF) infections . However, the two antibiotics have not been compared in terms of safety and efficacy in a prospective randomized study . The objective of this study was to compare the safety and efficacy of the two drugs in the treatment of VREF infections in cancer patients . PATIENTS AND METHODS: Forty cancer patients with VREF infection were randomized to receive linezolid 600 mg every 12 h or quinupristin-dalfopristin 7.5 mg/kg every 8 h . All patients were followed up for 30 days after discontinuation of study drugs . RESULTS: Linezolid and quinupristin-dalfopristin had comparable clinical responses (58% and 43%, respectively, P = 0.6) . Myalgias and/or arthralgias occurred at a frequency of 33% in patients who received quinupristin-dalfopristin, but were not observed in the linezolid group (P = 0.03) . In contrast, drug-related thrombocytopenia occurred in 11% of patients who received linezolid, but was not observed in the quinupristin-dalfopristin group (P = 0.2) . CONCLUSION: In cancer patients, quinupristin-dalfopristin treatment is associated with a relatively high frequency of myalgias/arthralgias; however, profound thrombocytopenia might limit the choice of linezolid in a subpopulation of cancer patients.

J Int Med Res, 2004 Jan-Feb, 32(1), 84 - 93
Cefepime versus ceftazidime for treatment of pneumonia; Konstantinou K et al.; Consecutive patients with pneumonia, treated with cefepime (n = 66) or ceftazidime (n = 132), were evaluated in a retrospective, observational study . There was no significant difference between the two treatment groups with respect to age, underlying diseases, acute physical and chronic health evaluation score, intensive care unit admission, presence of sepsis, community or hospital acquisition, causative organism, duration of therapy, death, cure or improvement in infection, adverse events, superinfections, presence of vancomycin-resistant enterococcus (VRE) and resistance to therapy . Post-therapy hospitalization (days) and vancomycin co-administration were significantly lower, and time to vancomycin initiation significantly higher, in the cefepime compared with the ceftazidime group . The results suggest a trend towards less resistance on therapy, less VRE, reduced vancomycin use and shorter post-therapy hospitalization in patients treated with cefepime compared with ceftazidime . The clinical outcomes for hospitalized patients treated for serious pneumonia were similar between the two groups.

Infect Control Hosp Epidemiol, 2004 Feb, 25(2), 138 - 45
Chloramphenicol resistance in vancomycin-resistant enterococcal bacteremia: impact of prior fluoroquinolone use?
Gould CV, Fishman NO, Nachamkin I, Lautenbach E.
OBJECTIVE: The prevalence of vancomycin-resistant enterococci (VRE) has increased markedly during the past decade . Few data exist regarding the epidemiology of resistance of VRE to chloramphenicol, one of the few therapeutic options . DESIGN: Survey and case-control study . SETTING: A 725-bed, tertiary-care academic medical center and a 344-bed urban community hospital . PATIENTS: Hospitalized patients with blood cultures demonstrating VRE . METHODS: We examined the trends in the prevalence of chloramphenicol resistance in VRE blood isolates at our institution from 1991 through 2002 and conducted a case-control study to identify risk factors for chloramphenicol resistance among these isolates . RESULTS: From 1991 through 2002, the annual prevalence of chloramphenicol-resistant VRE increased from 0% to 12% (P < .001, chi-square test for trend) . Twenty-two case-patients with chloramphenicol-resistant VRE bloodstream isolates were compared with 79 randomly selected control-patients with chloramphenicol-susceptible VRE . Independent risk factors for chloramphenicol-resistant VRE were prior chloramphenicol use (odds ratio {OR}, 10.9; 95% confidence interval {CI95}, 1.72-68.91; P = .01) and prior fluoroquinolone use (OR, 4.74; CI95, 1.15-19.42; P = .03) . Chloramphenicol-resistant VRE isolates were more likely to be susceptible to beta-lactams and resistant to tetracycline than were chloramphenicol-susceptible VRE isolates . CONCLUSIONS: Significant increases in the prevalence of chloramphenicol-resistant VRE may limit the future utility of chloramphenicol in the treatment of VRE infections, and close monitoring of susceptibility trends should continue . The association between fluoroquinolone use and chloramphenicol-resistant VRE, reflecting possible co-selection of resistance, suggests that recent dramatic increases in fluoroquinolone use may have broader implications than previously recognized.

Infect Control Hosp Epidemiol, 2004 Feb, 25(2), 105 - 8
Co-carriage rates of vancomycin-resistant Enterococcus and extended-spectrum beta-lactamase-producing bacteria among a cohort of intensive care unit patients: implications for an active surveillance program; Harris AD et al.; OBJECTIVE: To assess the co-colonization rates of extended-spectrum beta-lactamase (ESBL)-producing bacteria and vancomycin-resistant Enterococcus (VRE) obtained on active surveillance cultures . DESIGN: Prospective cohort study . SETTING: Medical and surgical intensive care units (ICUs) of a tertiary-care hospital . PATIENTS: Patients admitted between September 2001 and November 2002 to the medical and surgical ICUs at the University of Maryland Medical System had active surveillance perirectal cultures performed . Samples were concurrently processed for VRE and ESBL-producing bacteria . RESULTS: Of 1,362 patients who had active surveillance cultures on admission, 136 (10%) were colonized with VRE . Among these, 15 (positive predictive value, 11%) were co-colonized with ESBL . Among the 1,226 who were VRE negative, 1,209 were also ESBL negative (negative predictive value, 99%) . Among the 1,362 who had active surveillance cultures on admission, 32 (2%) were colonized with ESBL . Among these, 15 (47%) were co-colonized with VRE . Of the 32 patients colonized with ESBL, 10 (31%) had positive clinical cultures for ESBL on the same hospital admission . For these 10 patients, the surveillance cultures were positive an average of 2.7 days earlier than the clinical cultures . CONCLUSIONS: Patients who are colonized with VRE can also be co-colonized with other antibiotic-resistant bacteria such as ESBL-producing bacteria . Our study is the first to measure co-colonization rates of VRE and ESBL-producing bacteria . Isolating VRE-colonized patients would isolate 47% of the ESBL-colonized patients without the need for further testing . Hence, active surveillance for VRE should also theoretically diminish the amount of patient-to-patient transmission of ESBL-producing bacteria.

Biosci Biotechnol Biochem, 2004 Feb, 68(2), 293 - 9
Identification of nucleotide binding sites in V-type Na+-ATPase from Enterococcus hirae; Hosaka T et al.; A and B subunits of the V-type Na+-ATPase from Enterococcus hirae were suggested to possess nucleotide binding sites (Murata, T . et al., J . Biochem., 132, 789-794 (2002)), although the B subunit did not have the consensus sequence for nucleotide binding . To further characterize the binding sites in the V-ATPase, we did the photoaffinity labeling study using 8-azido-{alpha-32P}ATP . A and B subunits were labeled with 8-azido-{alpha-32P}ATP when analysed with SDS polyacrylamide gel electrophoresis . The peptide fragment of A subunit obtained by lysyl endopeptidase digestion after labeling showed a molecular size of 9 kDa and its amino acid sequencing revealed that it corresponded to residues Arg423-Lys494 . The peptide fragment from B subunit after photoaffinity labeling and lysyl endopeptidase digestion showed the size of 5 kDa and corresponded to residues Phe404-Lys443 . In our structure model, these peptides were close to the adenine ring of ATP . We suggest that non-catalytic B subunit of E . hirae V-ATPase has a nucleotide binding site, similarly to eukaryotic V-ATPases and F-ATPases.

Obes Surg, 2004 Jan, 14(1), 16 - 22
Analysis of the incidence and risk factors for wound infections in open bariatric surgery; Christou NV et al.; BACKGROUND: Our clinical impression derived from >95% follow-up of patients was that our wound infection rate was higher than the 1-10% reported in the literature . The purpose of this study was to determine the incidence and risk factors for wound infection in open bariatric surgery . METHODS: We queried our prospectively acquired bariatric surgery outcomes database, and retrospectively audited the charts of patients operated from April 1 to March 31, 2003 . Risk categories were obtained using the National Nosocomial Infection Surveillance (NNIS) definitions and stratification . Expected site-specific rates were adjusted for duration of operation, degree of wound contamination, and underlying disease condition . RESULTS: 269 patients undergoing a standardized open Roux-en-Y gastric bypass were studied . The mean age (SD) was 39.5 (10.5) years and the mean BMI was 54.3 (9.9) . Operating time averaged 63 (17) minutes, and length of stay was 4.1 (1.3) days . Based on NNIS categories 10.9 wound infections were expected, but 54 were observed, for a rate of 20% . Bacterial isolates included S . aureus (39%), alpha-hemolytic strep (26%), Enterococcus (16%), P.mirabilis (9%), and multiple other bacteria at 10% . Epidural analgesia and delayed antibiotic prophylaxis administration (after the incision was made) increased the odds of developing a wound infection, whereas gender, age, BMI, duration of surgery, and incidence of diabetes had no effect.There was a high correlation between wound infection and subsequent incisional hernia formation . CONCLUSION: The incidence of wound infections following open bariatric surgery is high, and the current recommendations for antibiotic prophylaxis are ineffective . As these infections carry significant morbidity, effective methods to prevent them are needed.

Epidemiol Infect, 2004 Jan, 132(1), 95 - 105
Relations between the consumption of antimicrobial growth promoters and the occurrence of resistance among Enterococcus faecium isolated from broilers; Emborg HD et al.; The present study investigates, at farm level, the effect of the time-span between sampling and the last time a particular antimicrobial growth promoter (AGP) was included in the feed on the probability of selecting an AGP-resistant Enterococcus faecium isolate from a broiler flock . The probability that a randomly selected E . faecium isolate was resistant to avilamycin, erythromycin or virginiamycin was 0.91, 0.92 and 0.84, respectively if the isolate originated from a broiler flock fed either avilamycin- or virginiamycin-supplemented feed . As the time-span between sampling and the last AGP consumption increased, the probability of isolating an E . faecium isolate resistant to a particular AGP decreased (probability <0.2 within 3-5 years after last exposure to AGPs) . The decrease in probability over time showed little farm-to-farm variation . The number of times a particular AGP was given to previous flocks reared in the same house had no effect on the probability of isolating a resistant isolate.

J Antimicrob Chemother, 2004 Mar, 53(3), 530 - 2 Epub 2004 Feb 12.
Daptomycin synergy with rifampicin and ampicillin against vancomycin-resistant enterococci; Rand KH et al.; We used a novel screening method to look for synergy between daptomycin and 18 other antibiotics against 19 strains of high-level vancomycin-resistant enterococci (VRE) (vancomycin MIC > or = 256 mg/L) . In this approach, daptomycin was incorporated into Ca(2+)-supplemented Mueller-Hinton agar at subinhibitory concentrations, and synergy was screened by comparing test antibiotic Etest MICs on agar with and without daptomycin . A striking reduction in the rifampicin MIC was seen in 11/15 (73.3%) VRE that were resistant to rifampicin, from > or =12 mg/L to a mean +/- s.d . of 0.22 +/- 0.21 mg/L at daptomycin 0.25 x MIC and 0.85 +/- 0.90 mg/L at daptomycin 0.125 x MIC . Synergy was also observed for 13/19 (68%) isolates with ampicillin (MIC > or = 128 mg/L) . There was no significant synergy between daptomycin and any other antibiotic by this screening method . If confirmed by further studies, daptomycin with either rifampicin or ampicillin may be useful in the management of infections caused by VRE.

Klin Padiatr, 2004 Jan-Feb, 216(1), 21 - 3
{Linezolid in two premature babies with necrotizing enterocolitis and infection with vancomycin-resistant enterococcus}; Klingkowski U et al.; The use of the antibiotic Linezolid in two premature babies with necrotizing enterocolitis and infection with vancomycin-resistant enterococcus is described . Therapy with Linezolid in cause of necrotizing enterocolitis is an exception . Actually there are no proven recommendations for the use in this age group . Despite the successfully results more investigations for the use of linezolid in newborns and prematures are necessary.

J Microbiol Methods, 2004 Feb, 56(2), 143 - 9
Autoclave method for rapid preparation of bacterial PCR-template DNA; Simmon KE et al.; An autoclave method for preparing bacterial DNA for PCR template is presented, it eliminates the use of detergents, organic solvents, and mechanical cellular disruption approaches, thereby significantly reducing processing time and costs while increasing reproducibility . Bacteria are lysed by rapid heating and depressurization in an autoclave . The lysate, cleared by microcentrifugation, was either used directly in the PCR reaction, or concentrated by ultrafiltration . This approach was compared with seven established methods of DNA template preparation from four bacterial sources which included boiling Triton X-100 and SDS, bead beating, lysozyme/proteinase K, and CTAB lysis method components . Bacteria examined were Enterococcus and Escherichia coli, a natural marine bacterial community and an Antarctic cyanobacterial-mat . DNAs were tested for their suitability as PCR templates by repetitive element random amplified polymorphic DNA (RAPD) and denaturing gradient gel electrophoresis (DGGE) analysis . The autoclave method produced PCR amplifiable template comparable or superior to the other methods, with greater reproducibility, much shorter processing time, and at a significantly lower cost.

Bone Marrow Transplant, 2004 Mar, 33(5), 519 - 29
Outbreaks of infectious diseases in stem cell transplant units: a silent cause of death for patients and transplant programmes; McCann S et al.; Following the closure of the National Blood and Bone Marrow Transplant Unit in Dublin, because of an outbreak of vancomycin-resistant enterococcal infection, a survey was carried out by the EBMT to investigate the occurrence of outbreaks of infection in SCT units and the impact on patient morbidity, mortality and the administration of the transplant programme over a 10-year period from 1991 to 2001 . A total of 13 centres reported 23 outbreaks of infection involving 231 patients: 10 bacterial, eight viral and five fungal outbreaks were reported and 56 deaths were attributed to infection . All fungal and bacterial deaths and the majority of viral deaths occurred in allograft recipients . In all outbreaks, the infection was reported to be hospital acquired and in all the viral, and half the bacterial infections, cross-infection was a major factor . All viral, four of 10 bacterial and three of five fungal outbreaks occurred in HEPA filtered rooms . A total of 12 SCT units reported a partial or total closure . The introduction of mandatory quality management systems such as JACIE should result in a change in attitude to 'incident reporting' and together with future surveys should reduce the incidence of infectious outbreaks in SCT units.

Antimicrob Agents Chemother, 2004 Feb, 48(2), 556 - 60
Duplex real-time PCR assay for rapid detection of ampicillin-resistant Enterococcus faecium; Mohn SC et al.; Rapid and accurate identification of carriers of resistant microorganisms is an important aspect of efficient infection control in hospitals . Traditional identification methods of antibiotic-resistant bacteria usually take at least 3 to 4 days after sampling . A duplex real-time PCR assay was developed for rapid detection of ampicillin-resistant Enterococcus faecium (ARE) . Primers and probes that are used in this assay specifically detected the D-Ala-D-Ala ligase gene of E . faecium and the modified penicillin-binding protein 5 gene (pbp5) carrying the Glu-to-Val substitution at position 629 (Val-629) in a set of 129 tested E . faecium strains with known pbp5 sequence . Presence of the Val-629 in the strain set from 11 different countries was highly correlated with ampicillin resistance . In a screening of hospitalized patients, the real-time PCR assay yielded a sensitivity and a specificity for the detection of ARE colonization of 95% and 100%, respectively . The results were obtained 4 h after samples were harvested from overnight broth of rectal swab samples, identifying both species and the resistance marker mutation in pbp5 . This novel assay reliably identifies ARE 2 to 3 days more quickly than traditional culture methods, thereby increasing laboratory throughput, making it useful for rectal screening of ARE . The assay demonstrates the advantages of real-time PCR for detection of nosocomial pathogens.

Eur J Clin Microbiol Infect Dis, 2004 Feb, 23(2), 73 - 7 Epub 2004 Jan 21.
Are there patients with peritonitis who require empiric therapy for enterococcus?
Harbarth S, Uckay I.
Enterococci are an increasingly important cause of nosocomial infections . While the clinical impact of enterococci in cases of bacteremia and super-infections in selected patient populations has been well-established, their role as primary pathogens in polymicrobial intra-abdominal infections remains controversial . While it has been suggested that the presence of enterococci increases the rate of infectious post-operative complication, it has also been demonstrated that polymicrobial intra-abdominal infections involving enterococci can be treated successfully with appropriate surgical drainage and antibiotics, such as cephalosporins, that are not active against enterococci . Therefore, the question arises of whether or not antibiotic coverage against enterococci should be included in the empirical treatment of peritonitis in certain high-risk patient populations . An extensive literature review revealed some evidence arguing in favour of using empirical therapy with enterococcal coverage for intra-abdominal infections in the following cases: (i) immunocompromised patients with nosocomial, post-operative peritonitis; (ii) patients with severe sepsis of abdominal origin who have previously received cephalosporins and other broad-spectrum antibiotics selecting for Enterococcus spp.; (iii) patients with peritonitis and valvular heart disease or prosthetic intravascular material, which place them at high risk of endocarditis . The ideal therapeutic regimen for these high-risk patients remains to be determined, but empirical therapy directed against enterococci should be considered.

Shokuhin Eiseigaku Zasshi, 2003 Oct, 44(5), 220 - 6
{Production of tyramine in "moromi" mash during soy sauce fermentation}; Ibe A et al.; The concentrations of 7 non-volatile amines, tyramine (Tym), histamine (Him), phenethylamine (Phm), putrescine (Put), cadaverine (Cad), spermidine (Spd) and spermine (Spm) in the liquid part of "moromi" mash during soy sauce fermentation were studied . These amines, except for him and Cad, were detected during fermentation by the conventional production method in the laboratory . Put and Spd were detected at the beginning, and Tym, Phm and Spm appeared later; these 5 amines increased gradually during the fermentation . Put, Spd, Spm and Cad were present in the raw starting material for soy sauce; thus, Tym and Phm were produced by the fermentation . When "moromi" mash was added to liquid medium and cultivated, Tym was detected in some "moromi" mash and the other amines were not detected . Tym-producing bacterial strains were isolated from the liquid culture media of Tym-positive "moromi" mash . The Tym-producing strain was a gram-positive coccus . The conditions for production of amines by Tym-producing bacterial strains were examined . These strains grew and produced tyramine under various conditions, which may occur during soy sauce fermentation . Namely, Tym was produced at pH 5-10, at salt concentrations of less than 8%, under either aerobic or anaerobic conditions . During soy sauce fermentation, it is assumed that Tym would be produced by these strains during the early stages of soy sauce aging within a short period when the salt concentration and pH conditions are optimal for growth . Based on the bacteriological properties, the strains were identified as Enterococcus faecium . With the exception of Phm and Him, which did not exist in the starting raw material, non-volatile amines (including Put, Cad, Spd and Spm) were not produced and microorganisms producing them are not believed to be present during "moromi" fermentation.

Biochemistry, 2004 Jan 20, 43(2), 446 - 54
Molecular mechanism of the enterococcal aminoglycoside 6'-N-acetyltransferase': role of GNAT-conserved residues in the chemistry of antibiotic inactivation; Draker KA et al.; The Gram-positive pathogen Enterococcus faecium is intrinsically resistant to aminoglycoside antibiotics due to the presence of a chromosomally encoded aminoglycoside 6'-N-acetyltransferase {AAC(6')-Ii} . This enzyme is a member of the GCN5-related N-acetyltransferase (GNAT) superfamily and is therefore structurally homologous to proteins that catalyze acetyl transfer to diverse acyl-accepting substrates . This study reports the investigation of several potential catalytic residues that are present in the AAC(6')-Ii active site and also conserved in many GNAT enzymes . Site-directed mutagenesis of Glu72, His74, Leu76, and Tyr147 with characterization of the purified site mutants gave valuable information about the roles of these amino acids in acetyl transfer chemistry . More specifically, steady-state kinetic analysis of protein activity, solvent viscosity effects, pH studies, and antibiotic resistance profiles were all used to assess the roles of Glu72 and His74 as potential active site bases, Tyr147 as a general acid, and the importance of the amide NH group of Leu76 in transition-state stabilization . Taken together, our results indicate that Glu72 is not involved in general base catalysis, but is instead critical for the proper positioning and orientation of aminoglycoside substrates in the active site . Similarly, His74 is also not acting as the active site base, with pH studies revealing that this residue must be protonated for optimal AAC(6')-Ii activity . Mutation of Tyr147 was found not to affect the chemical step of catalysis, and our results were not consistent with this residue acting as a general acid . Last, the amide NH group of Leu76 is implicated in important interactions with acetyl-CoA and transition-state stabilization . In summary, the work described here provides important information regarding the molecular mechanism of AAC(6')-Ii catalysis that allows us to contrast our findings with those of other GNAT proteins and to demonstrate that these enzymes use a variety of chemical mechanisms to accelerate acyl transfer.

Biochemistry, 2004 Jan 20, 43(2), 362 - 73
Structures of enterococcal glycerol kinase in the absence and presence of glycerol: correlation of conformation to substrate binding and a mechanism of activation by phosphorylation; Yeh JI et al.; The first structure of a glycerol kinase from a Gram-positive organism, Enterococcus casseliflavus, has been determined to 2.8 A resolution in the presence of glycerol and to 2.5 A resolution in the absence of substrate . The substrate-induced closure of 7 degrees is significantly smaller than that reported for hexokinase, a model for substrate-mediated domain closure that has been proposed for glycerol kinase . Despite the 78% level of sequence identity and conformational similarity in the catalytic cleft regions of the En . casseliflavus and Escherichia coli glycerol kinases, remarkable structural differences have now been identified . These differences correlate well with their divergent regulatory schemes of activation by phosphorylation in En . casseliflavus and allosteric inhibition in E . coli . On the basis of our structural results, we propose a mechanism by which the phosphorylation of a histidyl residue located 25 A from the active site results in a 10-15-fold increase in the activity of the enterococcal glycerol kinase.

J Clin Microbiol, 2004 Jan, 42(1), 487 - 9
Disseminated mucormycosis in a patient with acute myeloblastic leukemia misdiagnosed as infection by Enterococcus faecium; Sammassimo S et al.; Mucormycosis is a rare complication in cancer patients . This report presents the case of a acute myeloblastic leukemia patient who developed an ascending paralysis due to disseminated mucormycosis . The presentation was unusual because the early symptoms were fever and pain, and the disease was misdiagnosed because of a concomitant infection by Enterococcus faecium.

Appl Environ Microbiol, 2004 Jan, 70(1), 293 - 300
Census of the bacterial community of the gypsy moth larval midgut by using culturing and culture-independent methods; Broderick NA et al.; Little is known about bacteria associated with Lepidoptera, the large group of mostly phytophagous insects comprising the moths and butterflies . We inventoried the larval midgut bacteria of a polyphagous foliivore, the gypsy moth (Lymantria dispar L.), whose gut is highly alkaline, by using traditional culturing and culture-independent methods . We also examined the effects of diet on microbial composition . Analysis of individual third-instar larvae revealed a high degree of similarity of microbial composition among insects fed on the same diet . DNA sequence analysis indicated that most of the PCR-amplified 16S rRNA genes belong to the gamma-Proteobacteria and low G+C gram-positive divisions and that the cultured members represented more than half of the phylotypes identified . Less frequently detected taxa included members of the alpha-Proteobacterium, Actinobacterium, and Cytophaga/Flexibacter/Bacteroides divisions . The 16S rRNA gene sequences from 7 of the 15 cultured organisms and 8 of the 9 sequences identified by PCR amplification diverged from previously reported bacterial sequences . The microbial composition of midguts differed substantially among larvae feeding on a sterilized artificial diet, aspen, larch, white oak, or willow . 16S rRNA analysis of cultured isolates indicated that an Enterococcus species and culture-independent analysis indicated that an Entbacter sp . were both present in all larvae, regardless of the feeding substrate; the sequences of these two phylotypes varied less than 1% among individual insects . These results provide the first comprehensive description of the microbial diversity of a lepidopteran midgut and demonstrate that the plant species in the diet influences the composition of the gut bacterial community.

Pol J Vet Sci, 2003, 6(4), 267 - 9
Survival of Enterococcus hirae in ripened cheese subjected to ultra high pressure; Szczawinski J et al.; The samples of a sliced, hard cheese were placed in commercially used polyamide-polyethylene bags, inoculated with E . hirae and sealed under vacuum . The samples were exposed to high hydrostatic pressure of 300, 400 and 500 MPa for 5, 10 and 15 min . The number of surviving E . hirae and Aerobic Plate Counts (APC) were determined . The bacterial counts were transformed to logarithms and D-values (time required for decimal reduction of E . hirae at given pressure) were calculated using linear regression method . It was found that numbers of E . hirae decreased with the increase of pressure and prolongation of treatment time . D-values amounted to: D300MPa - 33.67 min, D400MPa - 17.83 min and D500MPa - 16.03 min . The obtained results indicate that E . hirae is one of the most resistant vegetative bacteria to ultra high pressure treatment.

Antimicrob Agents Chemother, 2004 Jan, 48(1), 326 - 8
Effect of parenteral fluoroquinolone administration on persistence of vancomycin-resistant Enterococcus faecium in the mouse gastrointestinal tract; Donskey CJ et al.; We examined the effect of subcutaneous fluoroquinolone antibiotic administration on persistence and density of vancomycin-resistant Enterococcus faecium stool colonization in mice . Levofloxacin and ciprofloxacin did not promote colonization in comparison to saline controls, whereas moxifloxacin and gatifloxacin promoted persistent overgrowth in a dose-dependent fashion.

J Infect Chemother, 2003 Dec, 9(4), 348 - 50
Meningitis caused by Enterococcus gallinarum in patients with ventriculoperitoneal shunts; Takayama Y et al.; Vancomycin-resistant enterococci have recently emerged as significant nosocomial pathogens . Here we describe two Japanese patients, a 57-year-old man and a 12-year-old boy, with ventriculoperitoneal shunts for hydrocephalus who developed meningitis caused by vancomycin-resistant Enterococcus gallinarum . The infection of the central nervous system in these two patients may have been associated with E . gallinarum derived from the gut . Removal of the shunts and antimicrobial treatment promoted apparent improvement in these patients . The risk factors and management of vancomycin-resistant enterococcal infections in the central nervous system are discussed.

Pediatr Infect Dis J, 2003 Dec, 22(12), 1101 - 3
Vancomycin-resistant Enterococcus faecium endocarditis in a premature infant successfully treated with linezolid; Ang JY et al.; A 4 1/2-month-old, 26-week premature infant with multiple complications of prematurity required a central venous catheter for venous access and antibiotic treatment of bacterial nosocomial infections . He developed tricuspid valve endocarditis with vegetation caused by Enterococcus faecium resistant to ampicillin, vancomycin and quinupristin-dalfopristin but susceptible to linezolid . He was successfully treated with linezolid intravenously (7 weeks) and then orally (2 weeks).

Scand J Gastroenterol, 2003 Nov, 38(11), 1162 - 8
Bacteriobilia in percutaneous transhepatic biliary drainage: occurrence over time and clinical sequelae . A prospective observational study; Rosch T et al.; BACKGROUND: In the diagnosis and treatment of biliary disorders, establishing percutaneous transhepatic biliary drainage (PTBD) is an invasive procedure that can potentially lead to infectious complications in both the short and long-term . We therefore prospectively analysed the time course and spectrum of biliary bacteria in patients undergoing PTBD . METHODS: Forty-nine patients (19 F, 30 M; mean age 64 years) with malignant (65%) or benign (35%) biliary disorders were included, 20 of whom had a newly established PTBD (group A), while the remaining 29 had already had their PTBD in situ (group B) for a mean of 8 months . Bacteriological analyses of bile and blood were carried out, and clinical symptoms and laboratory values were obtained . RESULTS: Biliary bacteria were found in 60% of cases during the initial PTBD placement, and 24 h later this rate had already increased to 85%; two or more microorganisms were found in 40% initially and in 70% after a few days . At later PTBD exchanges, bacteriobilia was found in 100%, with all patients harbouring multiple organisms . Whereas the initial spectrum was mixed, Escherichia coli and enterococci (97% each), Klebsiella (73%) and Bacteroides species (37%) later predominated; Candida increased initially from 15% to 80%, but later decreased to 30% . Clinical signs of cholangitis were observed in 30% initially (no sepsis), but decreased to 6% at later exchanges . CONCLUSIONS: Bacteriobilia is initially a frequent, and later a regular, event in PTBD; however, clinically significant complications are rare during the long-term course and limited to the initial, more invasive, phase of PTBD . A knowledge of the bacterial spectrum is important for selecting appropriate antibiotic coverage if complications arise and/or major interventions such as surgery are planned.

Clin Infect Dis, 2004 Jan 1, 38(1), 53 - 61 Epub 2003 Dec 05.
Clinical features and outcomes of bacteremia caused by Enterococcus casseliflavus and Enterococcus gallinarum: analysis of 56 cases; Choi SH et al.; The clinical significance and virulence potential of Enterococcus casseliflavus/flavescens and Enterococcus gallinarum are still uncertain . We retrospectively analyzed 56 cases of significant bacteremia caused by E . casseliflavus or E . gallinarum . Of these cases, 25 (44.6%) were associated with polymicrobial bacteremia, and 43 (76.8%) were associated with entry via the biliary tract . Resistance to vancomycin was observed in 17 (30.4%) of these 56 patients, and this resistance was significantly associated with E . gallinarum bacteremia (adjusted odds ratio {AOR}, 10.56; 95% confidence interval {CI}, 2.41-46.27) and bacteremia without biliary tract origin (AOR, 6.74; 95% CI, 1.44-31.67) . The crude mortality rate was 13%, and the bacteremia-related mortality rate was 1.9% . In conclusion, bacteremia due to E . casseliflavus and E . gallinarum is commonly associated with biliary tract disease and may be associated with a low risk of mortality.

J Am Chem Soc, 2003 Dec 24, 125(51), 15702 - 3
Using biofunctional magnetic nanoparticles to capture vancomycin-resistant enterococci and other gram-positive bacteria at ultralow concentration; Gu H et al.; Covalently linked to vancomycin (Van), chemically stable and highly magnetic anisotropic FePt magnetic nanoparticles (3-4 nm) become water-soluble and capture vancomycin-resistant enterococci (VRE) and other Gram-positive bacteria at concentrations approximately 10(1) cfu/mL via polyvalent ligand-receptor interactions . When a pyramidal end of a magnet "focuses" the nanoparticles into approximately 1 mm(2) area, the bacteria can be observed by an optical microscope and further identified by electron micrograph (EM) . Compared to the conventional use of magnetic particles (with the sizes of 1-5 microm) in biological separation or drug delivery, magnetic nanoparticles, combined with specific receptor-ligand interactions, promise a sensitive and rapid protocol to detect pathogens.

J Clin Microbiol, 2003 Dec, 41(12), 5742 - 6
Detection of enterococcal surface protein gene (esp) and amplified fragment length polymorphism typing of glycopeptide-resistant Enterococcus faecium during its emergence in a Greek intensive care unit; Routsi C et al.; The emergence of glycopeptide-resistant Enterococcus faecium (GREF) in a Greek intensive care unit was studied by amplified fragment length polymorphism analysis and esp gene detection . Three GREF clones harboring the esp gene were recovered from 17 out of 21 patients, indicating the dissemination of genetically homogenous and virulent strains of GREF.

J Clin Microbiol, 2003 Dec, 41(12), 5377 - 83
Acquisition and duration of vancomycin-resistant enterococcal carriage in relation to strain type; Mascini EM et al.; In May 2000, the first outbreak of vancomycin-resistant Enterococcus faecium (VREF) was detected in the University Medical Center Utrecht in the nephrology ward . The question arose why some VREF strains spread among hospitalized patients, whereas other strains do not . Thirty patients who were found to be colonized with VREF between May and November 2000 were included in the study . Molecular typing confirmed that 19 of them carried an identical epidemic strain which harbored the esp gene while 11 were colonized by nonepidemic strains that were all esp negative . Acquisition of the outbreak strain was significantly associated with diabetes mellitus, renal transplantation, and extensive use of antibiotics, especially cephalosporins, in the 2-month period before the first isolation of VREF . To establish the duration of colonization, prospective surveillance of VREF carriage for a 6-month period starting from the first isolation of VREF was realized for 20 patients . After 6 months, VREF was still recovered from 60% of carriers of the outbreak strain versus 20% of carriers of nonepidemic strains (P < 0.01) . However, antibiotic use during the follow-up period was significantly higher by carriers of the outbreak strain than by carriers of nonepidemic strains . The fact that the outbreak strain was recovered for a longer period of time than nonepidemic strains may facilitate dissemination of the strain . The results support a careful restrictive antibiotic policy for wards at risk for spread of VREF and implementation of isolation precautions for patients who are colonized with esp-positive outbreak strains.

Berl Munch Tierarztl Wochenschr, 2003 Nov-Dec, 116(11-12), 506 - 9
{Multiplex PCR in the framework of an enterococci monitoring project}; Mac K et al.; A multiplex-PCR (MPCR) assay was optimised in the framework of an enterococci monitoring project in order to accelerate the previous time-consuming biochemical testing of enterococcal strains isolated from animal materials and food . The MPCR was optimised by variation of annealing temperature, MgCl2-concentration but more over by primer combinations and primer concentrations . Reference strains were used during the optimization . Afterwards 522 enterococcal field strains were examined by the MPCR . Ambiguous results occurred for 20 strains (17 with false-positive and 3 with false-negative results) which could be explained by the minor specificity ("EM"-primers) and sensitivity (ddlE.faecium-primers) respectively . Differences also existed between the genus-specific primers "E" and "Ent" . The first mentioned primers showed weaknesses for the identification of enterococci whereas the use of the "Ent" primers inhibited other primers in the MPCR . However, the majority of the examined strains could be detected unambiguously, so that a differentiation of enterococcal isolates can be recommended by MPCR in contrast to the high time-, material- and work-consumption of conventional biochemical testings.

Mol Microbiol, 2004 Jan, 51(1), 271 - 81
Characterization of cis-acting prgQ mutants: evidence for two distinct repression mechanisms by Qa RNA and PrgX protein in pheromone-inducible enterococcal plasmid pCF10; Bae T et al.; The pCF10-encoded negative regulators PrgX and Qa (prgQ antisense) RNA inhibit pCF10 transfer by blocking prgQ transcription extension past a potential transcription terminator sequence IRS1 . To identify potential target sites for negative regulation, we isolated and analysed 13 cis-acting mutations in the prgXQ region . Determination of the 3' end of Qa RNA showed that eight mutations mapped in the region encoding Qa RNA . Four mutations were in the Qa promoter region and one was in IRS1 . Three mutations in Qa greatly reduced the intracellular level of this RNA but did not affect that of PrgX . However, both Qa RNA and PrgX protein were reduced in three Qa promoter region mutants and the expression of prgQ transcripts extending 3' from IRS1 became constitutive . Qa RNA could mediate its negative regulatory activity in the absence of PrgX, and this activity was not abolished by cCF10, the peptide pheromone that induces pCF10 transfer . RNA analysis showed that Qa RNA abolished transcription readthrough . Based on the experimental data as well as computer analysis of predicted secondary structures of prgQ mRNA in the presence or absence of Qa, we concluded that Qa RNA is a pheromone-insensitive effector of prgQ mRNA termination or degradation at IRS1 . In cells lacking a Qa target sequence, expression of PrgX repressed transcription from the prgQ promoter, and this repression was relieved by addition of exogenous cCF10 . Thus, even though the synthesis of these negative regulators is coupled, they each act independently on separate targets to regulate expression of conjugation functions.

Microb Drug Resist, 2003, 9 Suppl 1, S69 - 77
Association between antecedent intravenous antimicrobial exposure and isolation of vancomycin-resistant enterococci; Chavers LS et al.; Vancomycin-resistant enterococci (VRE) have become important causes of nosocomial infections . This study evaluated the association between a variety of intravenous antimicrobial exposures and the isolation of VRE using two control groups: (1) a vancomycin-susceptible enterococci (VSE) group, to assess factors associated with development of VRE, and (2) a nonenterococci control group, to assess factors associated with positive cultures for enterococci without regard to vancomycin resistance . After adjusting for the effect of other antimicrobials, time at risk, and patient morbidity, compared to vancomycin-susceptible enterococci controls, exposures to imipenem (OR = 4.9, 95% CI = 1.6-14.1) and ceftazidime (OR = 2.6, 95% CI = 1.1-6.1) were significant predictors of VRE . When compared to nonenterococci controls, exposures to ampicillin (OR = 20.1, 95% CI = 1.5-263.1) and imipenem (OR = 5.1, 95% CI = 1.5-17.1) were significantly associated with VRE . Neither piperacillin nor vancomycin was associated with VRE compared to either control group . This study offers further evidence that the replacement of broad-spectrum cephalosporins by extended-spectrum penicillins, specifically piperacillin, may be effective in reducing VRE.

Lett Appl Microbiol, 2003, 37(6), 475 - 81
Thermal inactivation of Enterococcus faecium: effect of growth temperature and physiological state of microbial cells; Martinez S et al.; AIMS: To provide data on the effects on culture temperature and physiological state of cells on heat resistance of Enterococcus faecium, which may be useful in establishing pasteurization procedures . METHODS AND RESULTS: The heat resistance of this Ent . faecium (ATCC 49624 strain) grown at different temperatures was monitored at various stages of growth . In all cases, the bacterial cells in the logarithmic phase of growth were more heat sensitive . For cells which had entered in the stationary phase, D70 values of 0.53 min at 5 degrees C, 0.74 min at 10 degrees C, 0.83 min at 20 degrees C, 0.79 min at 30 degrees C, 0.63 min at 37 degrees C, 0.48 min at 40 degrees C and 0.41 min at 45 degrees C were found . By extending the incubation times cells were more heat resistant as stationary phase progressed, although a different pattern was observed for cells grown at different temperatures . At the lower temperatures heat resistance increased progressively, reaching D70 values of 1.73 min for cells incubated at 5 degrees C for 50 days and 1.04 min for those grown at 10 degrees C for 16 days . At other temperatures assayed heat resistance became stable for late stationary phase cells, reaching D70 values of 1.05, 1.08 and 1.01 min for cultures incubated at 20, 30 and 37 degrees C . Heat resistance of cells obtained at higher temperatures, 40 and 45 degrees C, was significantly lower, with D70 values of 0.76 and 0.67 min, respectively . Neither the growth temperature nor the growth phase modified the z-values significantly . CONCLUSIONS: D70 values obtained for Ent . faecium (ATCC 49624) varies from 0.33 to 1.73 min as a function of culture temperature and physiological state of cells . However, z values calculated were not significantly influenced by these factors . A mean value of 4.50 +/- 0.39 degrees C was found . SIGNIFICANCE AND IMPACT OF THE STUDY: Overall results strongly suggest that, to establish heat processing conditions of pasteurized foods ensuring elimination of Ent . faecium, it is advisable to take into account the complex interaction of growth temperature and growth phase of cells acting on bacterial thermal resistance.

J Altern Complement Med, 2003 Oct, 9(5), 631 - 40
Enteric disposition and recycling of flavonoids and ginkgo flavonoids; Liu Y et al.; OBJECTIVE: The objective of this study was to determine the intestinal and microbial disposition of flavonoids and how these disposition processes affect their enteric recycling . DESIGN: Studies were performed using a perfused rat intestinal model or using enrichment cultures and a pure isolate of Enterococcus avium (LY1) . RESULTS: In the rat intestine, aglycones, such as quercetin and apigenin, were as permeable (P*(eff) > or = 2) as compounds such as propranolol (100% absorption) . However, a significant portion of the absorbed aglycones was conjugated and the metabolites were excreted into the lumen . Flavonoid glycosides, such as isoquercitrin and apigenin-7-O-glucoside, also had high apparent P*(eff) values (> or = 2) in the upper small intestine because of rapid hydrolysis . However, isoquercitrin was absorbed much slower (P*(eff) < or = 0.7, p < 0.05) when hydrolysis was absent or inhibited by 20 mmol gluconolactone . Absorption of other intact glycosides was similar to intact isoquercitrin and was much slower than the corresponding aglycones (P*(eff) < or = 0.7, p < 0.05) . Intestinal bacteria, such as LY1, hydrolyzed the flavonoid glycosides used in the study . Excreted glycosidases were involved in the hydrolysis of glycosides because glycosides were poorly taken up by LY1 . In conclusion, glycosidase-catalyzed hydrolysis is a critical first step in the intestinal and microbial disposition of flavonoid glycosides . Aglycones were not only rapidly absorbed, but also rapidly metabolized into phase II conjugates, which were then excreted back into the lumen . Therefore, intestinal and microbial glycosidases and intestinal phase II enzymes make a significant contribution to the disposition of flavonoids via the proposed enteric and enterohepatic recycling scheme.

Korean J Intern Med, 2003 Sep, 18(3), 187 - 90
Quinupristin/dalfopristin-induced Sweet's syndrome; Choi HS et al.; Quinupristin/Dalfopristin is a new combination of streptogramin antibiotics designed specifically to treat clinically significant infections due to Vancomycin-resistant Enterococcus Faecium . Sweet's syndrome is characterized by painful skin plaques, which is associated with dermal neutrophilic infiltration, fever and peripheral blood leukocytosis . Drug-induced Sweet's syndrome has a temporal relationship between drug ingestion, clinical presentation and the temporally-related resolution of lesions following drug withdrawal or on treatment with systemic corticosteroids . A 63-year-old woman received Quinupristin/Dalfopristin for acute pyelonephritis developed fever, arthralgia, vomiting, and painful erythematous skin plaques . A skin biopsy showed neutrophilic dermatosis, and there was rapid resolution of the symptoms and cutaneous lesions after discontinuation of Quinupristin/Dalfopristin, consistent with drug-induced Sweet's syndrome . To date, there has been no reported case of Sweet's syndrome associated with the use of Quinupristin/Dalfopristin.

J Bacteriol, 2003 Dec, 185(23), 7024 - 8
Highly conjugative pMG1-like plasmids carrying Tn1546-like transposons that encode vancomycin resistance in Enterococcus faecium; Tomita H et al.; A total of 12 VanA-type vancomycin-resistant enterococci, consisting of 10 Enterococcus faecium isolates and two Enterococcus avium isolates, were examined in detail . The vancomycin resistance conjugative plasmids pHTalpha (65.9 kbp), pHTbeta (63.7 kbp), and pHTgamma (66.5 kbp) were isolated from each of three different E . faecium strains . The plasmids transferred highly efficiently between enterococcus strains during broth mating and were homologous with pMG1 (Gm(r); 65.1 kb).

Transpl Infect Dis, 2003 Sep, 5(3), 121 - 5
Linezolid in the treatment of vancomycin-resistant Enterococcus faecium in solid organ transplant recipients: report of a multicenter compassionate-use trial; El-Khoury J et al.; Vancomycin-resistant Enterococcus faecium (VRE) is increasing in incidence in solid organ transplant recipients and has a high (up to 83%) associated mortality rate . Until recently, there have been no consistently effective antimicrobial therapies for VRE infection . Linezolid is a new antibiotic that belongs to the class of oxazolidinones approved by the FDA for the treatment of VRE infections, including those with bacteremia . Here, we report the experience with linezolid in an open-label, compassionate-use trial at 53 US centers for the treatment of documented VRE infections in patients with solid organ transplants . Eighty-five patients with solid organ transplants and documented VRE infections were studied . Blood cultures were positive for VRE in 43 patients, while 42 patients had other, non-rectal, sites of infection . Fifty-three patients responded well to treatment, with clinical resolution of the infection (62.4% survival rate) . Of these, 47 had documented negative cultures post therapy . The mean duration of therapy for cured patients was 23.5 days . Thirty-two (37.6%) patients died, 28 due to sepsis and organ failure (32.9% failure rate), and 4 due to unrelated causes . Mortality rates for patients with bacteremia were comparable to mortality rates observed with patients who had positive cultures from other sites . Adverse reactions to linezolid included thrombocytopenia (4.7%), decreased leukocyte count (3.5%), and an increase in blood pressure (1.2%), none of which led to discontinuation of therapy . Linezolid appears to be a safe and effective treatment option for VRE, even in the presence of bacteremia, and may lead to decreased mortality in solid organ transplant recipients with VRE infection.

Mol Microbiol, 2003 Nov, 50(3), 1069 - 83
A six amino acid deletion, partially overlapping the VanSB G2 ATP-binding motif, leads to constitutive glycopeptide resistance in VanB-type Enterococcus faecium; Depardieu F et al.; Enterococcus faecium clinical isolate BM4524, resistant to vancomycin and susceptible to teicoplanin, harboured a chromosomal vanB cluster, including the vanSB/vanRB two-component system regulatory genes . Enterococcus faecium strain BM4525, isolated two weeks later from the same patient, was resistant to high levels of both glycopeptides . The ddl gene of BM4525 had a 2 bp insertion leading to an impaired d-alanine:d-alanine ligase . Sequencing of the vanB operon in BM4525 also revealed an 18 bp deletion in the vanSB gene designated vanSBDelta . The resulting six amino acid deletion partially overlapped the G2 ATP-binding domain of the VanSBDelta histidine kinase leading to constitutive expression of the resistance genes . Sequence analysis indicated that the deletion occurred between two tandemly arranged heptanucleotide direct repeats, separated by 11 base-pairs . The VanSB, VanSBDelta and VanRB proteins were overproduced in Escherichia coli and purified . In vitro autophosphorylation of the VanSB and VanSBDelta histidine kinases and phosphotransfer to the VanRB response regulator did not differ significantly . However, VanSBDelta was deficient in VanRB phosphatase activity leading to accumulation of phosphorylated VanRB . Increased glycopeptide resistance in E . faecium BM4525 was therefore a result of the lack of production of d-alanyl-d-alanine ending pentapeptide and to constitutive synthesis of d-alanyl-d-lactate terminating peptidoglycan precursors, following loss of d-alanine:d-alanine ligase and of VanSB phosphatase activity respectively . We suggest that the heptanucleotide direct repeat in vanSB may favour the appearance of high level constitutively expressed vancomycin resistance through a 'slippage' type of genetic rearrangement in VanB-type strains.

J Microbiol Methods, 2003 Dec, 55(3), 739 - 43
Antimicrobial resistance in swine and chickens fed virginiamycin for growth promotion; Donabedian S et al.; In a prospective controlled study, we evaluated pigs (5-month period) and chickens (11-week period) fed subtherapeutic levels of virginiamycin . A total of 13 Enterococcus faecium were isolated from 10 pigs and 17 from 8 chickens . There were 8 pulsed-field gel electrophoresis (PFGE) patterns in E . faecium isolates from pigs and 17 from chickens . Resistance to quinupristin/dalfopristin resistance occurred in 2 of 13 E . faecium from pigs and 2 of 17 E . faecium from chickens . There were no strains exhibiting high-level gentamicin (MIC> or =2000 microg/ml) or vancomycin resistance . There was no relative weight gain in animals that received virginiamycin . The mean weight increase for the pigs in the group fed virginiamycin was 107.6 lb vs . 126.4 lb in the group that did not receive virginiamycin (P=n.s.) . Chickens fed virginiamycin had a mean weight increase of 1672 g vs . 1886 g in the group that did not receive virginiamycin (P=n.s.) . There was no correlation between receipt of virginiamycin or weight gain and presence of quinupristin/dalfopristin-resistant strains.

Appl Environ Microbiol, 2003 Nov, 69(11), 6405 - 11
Receiver operating characteristic curve analysis of beach water quality indicator variables; Morrison AM et al.; Receiver operating characteristic (ROC) curve analysis is a simple and effective means to compare the accuracies of indicator variables of bacterial beach water quality . The indicator variables examined in this study were previous day's Enterococcus density and antecedent rainfall at 24, 48, and 96 h . Daily Enterococcus densities and 15-min rainfall values were collected during a 5-year (1996 to 2000) study of four Boston Harbor beaches . The indicator variables were assessed for their ability to correctly classify water as suitable or unsuitable for swimming at a maximum threshold Enterococcus density of 104 CFU/100 ml . Sensitivity and specificity values were determined for each unique previous day's Enterococcus density and antecedent rainfall volume and used to construct ROC curves . The area under the ROC curve was used to compare the accuracies of the indicator variables . Twenty-four-hour antecedent rainfall classified elevated Enterococcus densities more accurately than previous day's Enterococcus density (P = 0.079) . An empirically derived threshold for 48-h antecedent rainfall, corresponding to a sensitivity of 0.75, was determined from the 1996 to 2000 data and evaluated to ascertain if the threshold would produce a 0.75 sensitivity with independent water quality data collected in 2001 from the same beaches.

Ann Thorac Surg, 2003 Nov, 76(5), 1719 - 20
Successful treatment of vancomycin resistant Enterococcus faecium mediastinitis associated with left ventricular assist devices; Malani PN et al.; We present 2 cases of vancomycin-resistant Enterococcus faecium mediastinitis associated with left ventricular assist devices in the setting of heart transplantation . Despite complicated operative courses and deep infection secondary to antimicrobial resistant organisms, both patients were successfully treated and have remained infection free in the long term.

Int J Food Microbiol, 2003 Dec 1, 88(2-3), 247 - 54
EU assessment of enterococci as feed additives; Becquet P; Particular micro-organisms such as enterococcal strains are used as probiotics in feed . Observations indicate a positive effect of such strains on the gut flora, especially for young animals or during feed transition phases . This favourable effect is perceived by the farmers as a means of maintenance of the health status of the animals (e.g . less diarrhoea) and results in significant improvement of animal performance . Micro-organisms have been used since the end of the 1980s in animal feeds and were strictly regulated in 1993, when they were introduced under the scope of Council Directive 70/524/EEC of 23 November 1970 on additives in animal nutrition on feed additives . After a transition period, which ended in the year 2000, every microbial strain must now be assessed by the EU bodies and authorised by a Commission Regulation, before it can be placed on the market for use in feedingstuffs . Council Directive 70/524/EEC on feed additives is based on three main principles: (1) pre-market authorisation, (2) positive list principle, and (3) thorough risk assessment of the effect of a particular strain on human and animal health as well as on the environment . Therefore, before introducing a new enterococcal strain preparation or promote a new use of an approved product, a dossier has to be submitted to the authorities, following the guidelines, as published in Commission Directive 94/40/EEC {Commission Directive 94/40/EC of 22 July 1994 amending Council Directive 87/153/EEC fixing guidelines for the assessment of additives in animal nutrition} . These guidelines contain detailed evaluation methods . The safety requirements refer to (1) the target animal categories, (2) the consumer and the environment (presence of toxins and virulence factors as well as antibiotic resistance and transferability are assessed), and (3) the workers, based on requirements of Council Directive 89/391/EEC.

Int J Food Microbiol, 2003 Dec 1, 88(2-3), 165 - 88
Methods used for the isolation, enumeration, characterisation and identification of Enterococcus spp . 2 . Pheno- and genotypic criteria; Domig KJ et al.; This paper reviews the methodology applied for the identification and characterisation of enterococci and covers phenotypic, genotypic and phylogenetic techniques . Although conventional phenotypic typing schemes are useful for rapid and simple identification of enterococcal species for routine applications, other methods like standardised sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), multilocus enzyme electrophoresis (MLEE), antimicrobial susceptibility testing, serotyping, pyrolysis mass spectrometry (pyMS) and vibrational spectroscopic methods allow a more in-depth characterisation of enterococci . Many of the recently described enterococcal species exhibit deviations from hitherto so-called classical enterococci with regard to their phenotypical properties . Therefore, genotypic methods have to be used to clarify their possible assignment to the genus Enterococcus . In this review, special emphasis is given on recently developed polymerase chain reaction (PCR)-based typing methods such as random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), specific and random amplification (SARA) and modifications of PCR-ribotyping as well as pulsed-field gel electrophoresis (PFGE) and partial sequence analysis . The use of PCR and probes for genus and species identification of enterococci is also considered like the application of sequence data of conserved DNA regions (e.g., ribosomal ribonucleic acid (rRNA) genes) in the case of species identification.

Int J Food Microbiol, 2003 Dec 1, 88(2-3), 147 - 64
Methods used for the isolation, enumeration, characterisation and identification of Enterococcus spp . 1 . Media for isolation and enumeration; Domig KJ et al.; Due to their significance in food, feed, environmental and clinical samples, the detection and enumeration of enterococci have become an important issue not only in daily routine but also in current research activities . Several media and protocols have been published for diverse purposes, but there is no single method, which universally meets all requirements . Depending on the nature of the accompanying microflora and its level, certain substrates and modifications thereof have to be used, taking into account various drawbacks and advantages . In addition to the historical applications (examination of water, different kinds of foods, intestinal and other clinical specimen), the detection of vancomycin-resistant enterococci (VRE) has become an important task, since VRE have found to be frequently involved in nosocomial infections . Moreover, contradictory methodological recommendations can be found in the literature . This paper will give a systematic survey of the different media and methods proposed during the last two decades . Emphasis is placed on compositional details and on specific applications of the media described.

Pharmacotherapy, 2003 Oct, 23(10), 1322 - 6
Peritoneal dialysis fluid concentrations of linezolid in the treatment of vancomycin-resistant Enterococcus faecium peritonitis; DePestel DD et al.; OBJECTIVE: To determine linezolid concentrations in peritoneal dialysis fluid after multiple oral doses of the drug in a 46-year-old man with vancomycin-resistant Enterococcus faecium peritonitis who was undergoing peritoneal dialysis . METHODS: After administration of oral linezolid 600 mg twice/day was started, peritoneal dialysis fluid was collected at the end of several 4- and 8-hour dwell times and submitted for analysis of linezolid concentration . Before linezolid therapy was begun, and immediately after several peritoneal dialysis exchanges, 30 ml of expended peritoneal dialysis fluid was collected in a sterile container and immediately frozen at -70 degrees C until analysis by high-performance liquid chromatography . RESULTS: Peritoneal dialysis concentrations of linezolid greater than 4 microg/ml were achieved after the first dose of linezolid and maintained after repeated doses . During the course of therapy, mean linezolid concentrations in peritoneal dialysis fluid tended to increase (mean 7.60 pg/ml, range 3.54-16.2 microg/ml) . All assayed peritoneal dialysis samples demonstrated linezolid concentrations greater than 4 microg/ml at the end of 4- or 8-hour dwell times, except for one level after a missed dose on linezolid treatment day 3 . Duration of dwell times did not appear to correlate with linezolid concentrations . CONCLUSION: In this patient, linezolid 600 mg twice/day penetrated into peritoneal dialysis fluid at or above the concentrations necessary to treat common gram-positive bacteria . Linezolid therapy is likely to have a role in peritoneal dialysis-associated peritonitis based on its antimicrobial activity, pharmacokinetic properties, ease of administration, and tolerability.

Bioorg Med Chem Lett, 2003 Nov 17, 13(22), 4117 - 20
Synthesis and in vitro activity of novel isoxazolyl tetrahydropyridinyl oxazolidinone antibacterial agents; Lee JS et al.; A series of isoxazolyl tetrahydropyridinyl oxazolidinones with various substituents at the 3-position of the isoxazole ring have been synthesized and their in vitro antibacterial activities (MIC) were evaluated against several Gram-positive strains including the resistant strains of Staphyloccus and Enterococcus, such as MRSA and VRE . One of the most potent compounds synthesized, 4f, showed comparable or better activity against selected bacterial strains than those of linezolid and vancomycin.

J Med Chem, 2003 Nov 6, 46(23), 4904 - 9
Multivalent antibiotics via metal complexes: potent divalent vancomycins against vancomycin-resistant enterococci; Xing B et al.; Dimers of vancomycin (Van), linked by a rigid metal complex, {Pt(en)(H(2)O)(2)}(2+), exhibit potent activities (MIC approximately 0.8 mug/mL, approximately 720 times more potent than that of Van itself) against vancomycin-resistant enterococci (VRE) . The result suggests that combining metal complexation and receptor/ligand interaction offers a useful method to construct multivalent inhibitors.

Antimicrob Agents Chemother, 2003 Nov, 47(11), 3620 - 2
Rapid detection and estimation by pyrosequencing of 23S rRNA genes with a single nucleotide polymorphism conferring linezolid resistance in Enterococci; Sinclair A et al.; Pyrosequencing was used to detect rapidly and estimate the number of 23S rRNA genes with a G2576T mutation in 43 linezolid-resistant and -susceptible clinical isolates of enterococci . The method showed 100% concordance with PCR-restriction fragment length polymorphism for detecting isolates homozygous for either G2576 or T2576 or heterozygous for this mutation . A good correlation was found between linezolid MICs and the number of 23S rRNA gene copies carrying the mutation.

Antimicrob Agents Chemother, 2003 Nov, 47(11), 3515 - 8
Influence of VanD type resistance on activities of glycopeptides in vitro and in experimental endocarditis due to Enterococcus faecium; Lefort A et al.; The consequences of VanD type glycopeptide resistance on the activity of vancomycin and teicoplanin were evaluated in vitro and in a rabbit model of aortic endocarditis with VanD type clinical isolate Enterococcus faecium BM4339 (MICs: vancomycin, 64 microg/ml; teicoplanin, 4 microg/ml) and its susceptible derivative BM4459 (MICs: vancomycin, 1 microg/ml; teicoplanin, 1 microg/ml) . The two antibiotics were inactive against BM4339 in vivo, in terms both of reduction of bacterial counts and of prevention of emergence of glycopeptide-resistant subpopulations, despite using teicoplanin at concentrations greater than the MIC for VanD strains . This could be due to the high inoculum effect also observed in vitro with BM4339 and two other VanD type isolates against both antibiotics . These results suggest that detection of VanD type resistance is of major importance because it abolishes in vivo glycopeptide activity and allows the emergence of mutants highly resistant to glycopeptides.

Curr Opin Microbiol, 2003 Oct, 6(5), 439 - 45
Antibiotics in animal feed and their role in resistance development; Wegener HC; Animals and humans constitute overlapping reservoirs of resistance, and consequently use of antimicrobials in animals can impact on public health . For example, the occurrence of vancomycin-resistant enterococci in food-animals is associated with the use of avoparcin, a glycopeptide antibiotic used as a feed additive for the growth promotion of animals . Vancomycin-resistant enterococci and vancomycin resistance determinants can therefore spread from animals to humans . The bans on avoparcin and other antibiotics as growth promoters in the EU have provided scientists with a unique opportunity to investigate the effects of the withdrawal of a major antimicrobial selective pressure on the occurrence and spread of antimicrobial resistance . The data shows that although the levels of resistance in animals and food, and consequently in humans, has been markedly reduced after the termination of use, the effects on animal health and productivity have been very minor.

Hum Pathol, 2003 Sep, 34(9), 944 - 5
Enterococcus gastritis; El-Zimaity HM et al.; Helicobacter pylori infection is the most common cause of gastritis with its associated sequelae . Gastritis secondary to other bacteria is rare . This report describes Enterococcus-associated gastritis in a 59-year-old diabetic man . Nine months after receiving treatment for H . pylori-associated gastritis, he underwent endoscopy to confirm H . pylori eradication and to evaluate the status of previously seen ulcers . Mucosal biopsy specimens revealed severe active but focal gastritis adjacent to gram-positive coccobacilli in short to long chains with no H . pylori . Culture grew an Enterococcus similar to E . hirae and E . durans . No treatment was given, and endoscopy done 2 months later showed complete resolution of the gastritis and absence of H . pylori or enterococci . Our patient's gastritis represents a previously undescribed manifestation of Enterococcus infection . It is possible that the presence of NSAID gastric mucosal injury and diabetes predisposed this individual to the development of transient Enterococcus gastritis.

J Infect, 2003 Nov, 47(4), 344 - 7
Polymerase chain reaction aids in the diagnosis of an unusual case of Aspergillus niger endocarditis in a patient with acute myeloid leukaemia; McCracken D et al.; Endocarditis secondary to Aspergillus niger has not been described in a leukaemic patient . We describe a case of A . niger endocarditis in a patient with acute myeloid leukaemia and refractory fever . The microbiological cause of his endocarditis was initially misdiagnosed because he fulfilled the Duke criteria for enterococcal endocarditis . A polymerase chain reaction test utilizing pan-fungal primers detected a product from an Aspergillus sp . The DNA was subsequently sequenced and was found to have 100% homology with A . niger . A postmortem revealed fungal endocarditis secondary to disseminated aspergillosis, without evidence of bacterial endocarditis . The patient was found to have a lung aspergilloma that was possibly occupationally acquired, and may have been long standing.

Vet Q, 2003 Sep, 25(3), 124 - 30
First cases of animal diseases published since 2000 . 2 . Cats; Elsinghorst TA; In this second article of a series of papers listing first case reports of animal diseases published since 2000, the following nine cases of cat diseases are discussed: Congenital spongiform change in the brain stem nuclei . Enterococcus hirae enteropathy . Focal cerebral angiomatosis . Glomus tumor . Intraocular extramedullary plasmacytoma . Lens epithelial neoplasias . Phaeohyphomycosis due to Fonsecaea pedrosoi . Pulmonary lymphomatoid granulomatosis . Systemic amyloidosis in a Devon rex . After a short introduction, the bibliographical data, the abstract of the author(s) and some additional information derived from the article are given . The article will be regularly updated adding overlooked as well as new first reports.

J Clin Microbiol, 2003 Oct, 41(10), 4805 - 7
Comparison of PCR assay to culture for surveillance detection of vancomycin-resistant enterococci; Paule SM et al.; Direct multiplex PCR assay using vanA and vanB primers, which provides rapid results, was more sensitive than culture on selective media for samples collected by rectal swab (20 of 46 versus 8 of 46; P < 0.001) or perianal swab (17 of 58 versus 12 of 58; P = 0.059) for the detection of gastrointestinal colonization by vancomycin-resistant enterococci.

Int J Food Microbiol, 2003 Nov 15, 88(1), 11 - 8
The potential of vancomycin-resistant enterococci to persist in fermented and pasteurised meat products; Houben JH; Experiments with 148 isolates of vancomycin-resistant enterococci (VRE) were performed to assess their potential to persist and grow in fermented sausages and pasteurised meat products . All strains were meat isolates and Van-type A, except a single VanC1 strain . In total, 143 strains of Enterococcus faecium were involved . Eight selected strains were examined for their potential to grow at high salt and nitrite levels and at reduced pH . The same isolates were used in experiments with fermented sausages . All available strains were subjected to heating tests in meat suspensions with added curing ingredients . All but one of the eight tested isolates grew at pH 4.0 in tryptone soya broth (TSB) . With the combination of 8% w/w NaCl, 400 ppm NaNO2 and 0.5% w/w glucose in the meat suspension, all isolates grew at 37 degrees C, whereas none grew at 7 degrees C even after 56 days . With the addition of 10% w/w NaCl, 200 ppm NaNO2 and 0.5% w/w glucose, still one E . faecium isolate grew at 37 degrees C, although very slowly . Overall, the strains tolerated high salt and nitrite concentrations and reduced pH very well, even beyond levels applied in the regular production of fermented and/or pasteurised meat products . The tested strains could be isolated after the fermentation and further ripening of "boerenmetworst" and "snijworst" . Overall, their colony counts decreased on average about 1 log-unit over a period of 60 days after batter manufacture . All 148 isolates demonstrated a relatively weak thermal resistance compared to results for selected vancomycin-sensitive enterococci strains reported in the literature and to results collected under identical experimental conditions in this laboratory . None of the strains (log inoculation level about 5-6 ml(-1) for each isolate) could be cultured after heating at 70 degrees C for 10 min.

J Bacteriol, 2003 Oct, 185(20), 5925 - 35
Redefining the role of psr in beta-lactam resistance and cell autolysis of Enterococcus hirae; Sapunaric F et al.; The contribution of penicillin-binding protein 5 (PBP5) and the PBP5 synthesis repressor (Psr) to the beta-lactam resistance, growth, and cell autolysis of wild-type strain ATCC 9790 and resistant strain R40 of Enterococcus hirae was investigated by disruption or substitution of the corresponding pbp5 and psr genes by Campbell-type recombination . The resulting modifications were confirmed by hybridization and PCR . The low susceptibility of E . hirae to beta-lactams was confirmed to be largely dependent on the presence of PBP5 . However, against all expectations, inactivation of psr in ATCC 9790 or complementation of R40 cells with psr did not modify the susceptibility to benzylpenicillin or the growth and cell autolysis rates . These results indicated that the psr gene does not seem to be involved in the regulation of PBP5 synthesis and consequently in beta-lactam resistance or in the regulation of cell autolysis in E . hirae.

Microbiology, 2003 Oct, 149(Pt 10), 2901 - 8
Characterization of Enterococcus faecium mutants resistant to mundticin KS, a class IIa bacteriocin; Sakayori Y et al.; The emergence and spread of mutants resistant to bacteriocins would threaten the safety of using bacteriocins as food preservatives . To determine the physiological characteristics of resistant mutants, mutants of Enterococcus faecium resistant to mundticin KS, a class IIa bacteriocin, were isolated . Two types of mutant were found that had different sensitivities to other antimicrobial agents such as nisin (class I) and kanamycin . Both mutants were resistant to mundticin KS even in the absence of Mg(2+) ions . The composition of unsaturated fatty acids in the resistant mutants was significantly increased in the presence of mundticin KS . The composition of the phospholipids in the two resistant mutants also differed from those in the wild-type strain . The putative zwitterionic amino-containing phospholipid in both mutants significantly increased, whereas amounts of phosphatidylglycerol and cardiolipin decreased . These changes in membrane structure may influence resistance of enterococci to class IIa and class I bacteriocins.

J Antimicrob Chemother, 2003 Nov, 52(5), 772 - 5 Epub 2003 Sep 30.
Molecular analysis of Tn1546-like elements mediating high-level vancomycin resistance in Enterococcus gallinarum; Foglia G et al.; OBJECTIVES: Four Enterococcus gallinarum isolates, all highly resistant to vancomycin, were studied in order to investigate their relationship and to gain insight into the molecular events responsible for their acquired resistance . METHODS: Extensive molecular analysis was performed to compare the four E . gallinarum isolates and their Tn1546-like elements . RESULTS: The four strains had very similar random amplified polymorphic DNA (RAPD) patterns and different but related PFGE profiles . Genotypic analysis demonstrated that all carried both vanC-1 and vanA genes . Using a vanA probe, no hybridization was detected to plasmid DNA, whereas hybridization to different SmaI fragments of the four strains was obtained with total DNA . Amplification and sequencing experiments showed that all four strains carried a Tn1546-like element that contained the orf2, vanR, vanS, vanH, vanA and vanX genes and was flanked at both ends by oppositely oriented IS1216V sequences . On the left side of the vanA cluster, all lacked IRL, and all had, upstream from orf2, 1029 bp of the 3' end of orf1 . On the right side, one of the strains lacked vanY, vanZ and IRR, whereas in one of the other three there was an IS1542 element inserted within the vanZ gene . In one strain, an additional IS1216V element was inserted in the intergenic region vanX-vanY . CONCLUSIONS: This is the first study providing a molecular analysis of chromosomal Tn1546-like elements (possibly composite transposons) associated with high-level vancomycin resistance in human and animal strains of E . gallinarum . These molecular findings, together with those from PFGE and RAPD, suggest that the four E . gallinarum isolates are related and might have a common ancestor.

Emerg Infect Dis, 2003 Sep, 9(9), 1108 - 15
Epidemic and nonepidemic multidrug-resistant Enterococcus faecium; Leavis HL et al.; The epidemiology of vancomycin-resistant Entero- coccus faecium (VREF) in Europe is characterized by a large community reservoir . In contrast, nosocomial outbreaks and infections (without a community reservoir) characterize VREF in the United States . Previous studies demonstrated host-specific genogroups and a distinct genetic lineage of VREF associated with hospital outbreaks, characterized by the variant esp-gene and a specific allele-type of the purK housekeeping gene (purK1) . We investigated the genetic relatedness of vanA VREF (n=108) and vancomycin-susceptible E . faecium (VSEF) (n=92) from different epidemiologic sources by genotyping, susceptibility testing for ampicillin, sequencing of purK1, and testing for presence of esp . Clusters of VSEF fit well into previously described VREF genogroups, and strong associations were found between VSEF and VREF isolates with resistance to ampicillin, presence of esp, and purK1 . Genotypes characterized by presence of esp, purK1, and ampicillin resistance were most frequent among outbreak-associated isolates and almost absent among community surveillance isolates . Vancomycin-resistance was not specifically linked to genogroups . VREF and VSEF from different epidemiologic sources are genetically related; evidence exists for nosocomial selection of a subtype of E . faecium, which has acquired vancomycin-resistance through horizontal transfer.

South Med J, 2003 Aug, 96(8), 818 - 20
Endocarditis due to vancomycin-resistant Enterococcus faecium in an immunocompromised patient: cure by administering combination therapy with quinupristin/dalfopristin and high-dose ampicillin; Thompson RL et al.; A 56-year-old man with diabetes mellitus and cadaveric renal transplantation had vancomycin-resistant Enterococcus faecium tricuspid valve endocarditis . Relapse followed 6 weeks of treatment with intravenous gentamicin and high-dose ampicillin . On the basis of previous data suggesting the potential for synergistic activity of quinupristin/dalfopristin plus high-dose ampicillin, therapy with this combination was administered for 63 days . Cure was achieved and later confirmed at 2-year follow-up.

Infect Control Hosp Epidemiol, 2003 Sep, 24(9), 690 - 8
Outcomes associated with vancomycin-resistant enterococci: a meta-analysis; Salgado CD et al.; BACKGROUND: Because patients with vancomycin-resistant Enterococcus bacteremia (VREB) usually have a higher severity of illness, it has been unclear whether VREB is worse than vancomycin-susceptible Enterococcus bacteremia (VSEB) . METHODS: Data on morbidity and case fatality rates and costs were pooled from studies comparing VREB and VSEB, identified by Medline January 1986 to April 2002) and meeting abstracts . Heterogeneity across studies was assessed with contingency table chi-square . Multivariate analyses (MVAs) controlling for other predictors were evaluated . RESULTS: Thirteen studies compared case-fatality rates of VREB and VSEB . VREB case fatality was significantly higher (48.9% vs 19%; RR, 2.57; CI95, 2.27 to 2.91; attributable mortality = 30%) . Five studies compared VREB with VSEB when bacteremia was the direct cause of death; VREB case fatality was significantly higher (39.1% vs 21.8%; RR, 1.79; CI95, 1.28 to 2.5; attributable mortality = 17%) . Four MVAs found significant increases in case-fatality rates (OR, 2.10 to 4.0), 3 showed trends toward increase (OR, 1.74 to 3.34 with wide confidence intervals), and 3 with low statistical power found no difference . VREB recurred in 16.9% versus 3.7% with VSEB (P < .0001) . Three studies reported significant increases in LOS, costs, or both with VREB . CONCLUSION: Most studies have had inadequate sample size, inadequate adjustment for other predictors of adverse outcomes, or both, but available data suggest that VREB is associated with higher recurrence, mortality, and excess costs than VSEB including multiple studies adjusting for severity of illness.

Antimicrob Agents Chemother, 2003 Oct, 47(10), 3061 - 6
Mutations in the DNA mismatch repair proteins MutS and MutL of oxazolidinone-resistant or -susceptible Enterococcus faecium; Willems RJ et al.; Mutations in mutS and mutL, which encode DNA mismatch repair (MMR) proteins, can confer hypermutator phenotypes and may facilitate the emergence of mutational antibiotic resistance in bacteria . Linezolid-resistant enterococci (LRE) rarely emerge during therapy and contain mutations in 23S rRNA genes . As enterococci with defective MMR could be prone to the development of oxazolidinone resistance mutations, we investigated 13 clinical isolates of Enterococcus faecium, including 2 LRE, for mutations in mutSL . A 4,944-bp fragment spanning mutSL was sequenced from two pairs of linezolid-resistant (MICs, 64 micro g/ml) and linezolid-susceptible (MICs, 2 micro g/ml) E . faecium isolates (one pair from Austria and one pair from the United Kingdom) identical by pulsed-field gel electrophoresis . The pairs represented distinct strains in which linezolid resistance had emerged during therapy . The MutSL peptides of all four isolates had amino acid substitutions compared with the sequence of E . faecium strain DO (used for genome sequencing) . These were Val352Ile (one pair of isolates only) and Met628Leu in MutS and Leu387Pro, Tyr406Phe, Thr415Ser, Phe427Leu, and Phe565Ile in MutL . The significance of these changes remains unknown; these isolates did not show a demonstrable hypermutator phenotype . The same substitutions were found in two of nine geographically diverse linezolid-susceptible enterococcal isolates; the other seven isolates had MutSL sequences identical to that of strain DO . Multilocus sequence typing revealed that all isolates with alternate MutSL peptides belonged to a distinct lineage of a prevalent E . faecium clonal complex, designated CC17 . Further studies are needed to investigate the prevalence of these MutSL mutations and their possible roles in the emergence of E . faecium strains resistant to oxazolidinones and other antibiotic classes.

Curr Infect Dis Rep, 2003 Oct, 5(5), 372 - 378
Can Enterococcal Infections Initiate Sepsis Syndrome?
Linden P.
Enterococci are the third most common nosocomial bloodstream pathogen and frequently are the causative pathogen(s) of intra-abdominal, genitourinary, surgical wound, endovascular, or other serious infections . In addition to a diverse spectrum of intrinsic and acquired antimicrobial resistance mechanisms, some strains of enterococci exhibit numerous virulence factors that facilitate mucosal adherence, tissue invasiveness, endovascular attachment, and inflammation production . However, the opportunistic nature of enterococcal infection coupled with animal models of experimental enterococcal and mixed infection have caused many to question the independent capacity of this organism to incite severe sepsis or septic shock . Despite evidence suggesting that this organism possesses low intrinsic virulence, observational and interventional trials of enterococcal bacteremia and sepsis strongly support the notion that Enterococcus is capable of promulgating sepsis as a solitary or copathogen with more serious sequelae in the immunocompromised hosts . Monomicrobial enterococcal bacteremia appears to be increasing during the glycopeptide-resistant era, which bolsters the notion that pure enterococcal sepsis is a real entity.

Chem Commun (Camb), 2003 Sep 7, (17), 2224 - 5
Self-assembled multivalent vancomycin on cell surfaces against vancomycin-resistant enterococci (VRE); Xing B et al.; A vancomycin (Van) derivative self-assembles in a phosphate buffer as a divalent Van and on cell surfaces as a multivalent Van, which offers potent activity against VRE.

Clin Infect Dis, 2003 Oct 1, 37(7), 921 - 8 Epub 2003 Sep 12.
Active surveillance reduces the incidence of vancomycin-resistant enterococcal bacteremia; Price CS et al.; The impact of active surveillance of patients at risk for infection with vancomycin-resistant enterococci (VRE) was examined, and VRE bacteremia rates and the degree of VRE clonality in 2 similar neighboring hospitals were compared . Hospital A did not routinely screen patients for VRE rectal colonization; hospital B actively screened high-risk patients . Retrospective observations were made over the course of 6 years, beginning when initial VRE bloodstream isolates were recovered at each institution . The rate of VRE bacteremia was 2.1-fold higher at hospital A, and the majority of hospital A isolates were clonally related: 4 clones were responsible for infection in >75% of patients with VRE bacteremia, and isolates from 30% of patients were from the most common clone . The 4 most common clones at hospital B were responsible for infection in 37% of patients, and isolates from 14.5% of patients were from the most common clone . Lower VRE bacteremia rates and a more polyclonal population, representing less horizontal transmission, may result from routine screening of patients who are at risk for VRE and prompt contact isolation of colonized individuals.

Chest, 2003 Sep, 124(3), 1030 - 8
Antibiotic-impregnated catheters associated with significant decrease in nosocomial and multidrug-resistant bacteremias in critically ill patients; Hanna HA et al.; OBJECTIVE: To evaluate the impact of using central venous catheters (CVCs) impregnated with the combination of minocycline and rifampin on nosocomial bloodstream infections (BSIs), morbidity, and mortality in cancer patients in the ICU . DESIGN: Prospective surveillance study consisting of the following two time periods: September 1997 through August 1998 (ie, fiscal year {FY} 1998); and from September 1998 through August 1999 (ie, FY 1999) . SETTING: ICUs of a tertiary care hospital in Houston, TX . PATIENTS: Cancer patients in the medical ICU (MICU) and surgical ICU (SICU) . INTERVENTIONS: ICUs started using CVCs impregnated with the minocycline-rifampin combination at the beginning of FY 1999 . Measurements and main results:The rates of nosocomial BSIs and other patients' characteristics were compared for the two study periods to determine the impact of using the impregnated catheters in the ICU . Patients' characteristics, including antibiotic use, were comparable for the two study periods in both the MICU and the SICU . The rate of nosocomial BSIs in the MICU unit decreased from 8.3 to 3.5 per 1,000 patient-days (p < 0.01), and decreased in the SICU from 4.8 to 1.3 per 1,000 patient-days (p < 0.01) in FY 1999 . Nosocomial vancomycin-resistant enterococcus (VRE) bacteremia also decreased significantly (p = 0.004) . Length of stay in the MICU and SICU significantly decreased in FY 1999 (p < 0.01 and p = 0.03, respectively) . The duration of hospitalization decreased for MICU and SICU patients (p = 0.06 and p < 0.01, respectively) . The rate of catheter-related infections decreased from 3.1 to 0.7 per 1,000 patient-days in FY 1999 (p = 0.02) . The decrease in infections resulted in net savings of at least $1,450,000 for FY 1999 . CONCLUSIONS: The use of antibiotic-impregnated CVCs in the MICU and SICU was associated with a significant decrease in nosocomial BSIs, including VRE bacteremia, catheter-related infections, and lengths of hospital and ICU stays.

Chest, 2003 Sep, 124(3), 1021 - 9
The influence of infection on hospital mortality for patients requiring > 48 h of intensive care; Osmon S et al.; OBJECTIVE: To determine the influence of microbiologically confirmed infection on hospital mortality among patients requiring intensive care for > 48 h . DESIGN: Prospective cohort study . SETTING: Medical ICU of the Barnes-Jewish Hospital, an urban teaching hospital . PATIENTS: A total of 893 patients requiring intensive care for > 48 h . INTERVENTIONS: Prospective patient surveillance and data collection . MEASUREMENTS AND MAIN RESULTS: Three hundred seventy-two patients (41.7%) requiring intensive care for > 48 h had a microbiologically confirmed infection . Only six patients (0.7% {1.6% of patients with microbiologically confirmed infections}) received inadequate antimicrobial therapy during the first 24 h of treatment, and 248 patients (27.8%) died during hospitalization . Compared to hospital survivors, hospital nonsurvivors were significantly more likely to have a microbiologically confirmed infection (53.2% vs 37.2%, respectively; p < 0.001) and to develop severe sepsis (45.6% vs 28.7%, respectively; p < 0.001) . Cirrhosis and the requirement for vasopressors were the only variables identified by multiple logistic regression analysis as independent risk factors for hospital mortality in all patient groupings of severity of illness . Multiple logistic regression analysis also demonstrated that underlying malignancy (adjusted odds ratio {AOR}, 1.98; 95% CI, 1.55 to 2.53), chronic renal insufficiency (AOR, 1.57; 95% CI, 1.31 to 1.87), cirrhosis (AOR, 1.94; 95% CI, 1.48 to 2.53), temperature > 38.3 degrees C (AOR, 1.72; 95% CI, 1.47 to 2.02), severe sepsis (AOR, 2.78; 95% CI, 1.94 to 3.98), positive culture for vancomycin-resistant enterococci (AOR, 1.78; 95% CI, 1.51 to 2.09), and the presence of multiple infections (AOR, 1.65; 95% CI, 1.28 to 2.14) were independently associated with the requirement for therapy with vasopressors . CONCLUSIONS: Microbiologically confirmed infections are common among patients requiring medical intensive care for > 48 h . Despite the administration of adequate antimicrobial therapy, microbiologically confirmed infections appear to be an important cause of hemodynamic instability and increased hospital mortality . These data suggest that clinical efforts aimed at the prevention of infections and improvements in the medical management of patients with severe infections, especially those associated with hemodynamic instability and the need for vasopressors, are required to achieve further improvements in patient outcomes.

APMIS, 2003 Jun, 111(6), 669 - 72
Genetic characterization of glycopeptide-resistant enterococci of human and animal origin from mixed pig and poultry farms; Bogo Jensen L et al.; Glycopeptide resistant enterococci (GRE) isolated from animals and humans were characterised using both AFPL typing and genetic characterisation of the glycopeptide resistance transposon Tn1546 . All isolates were collected in 1997 when the glycopeptide avoparcin was still being used as growth promoter . All investigated animal isolates were from mixed pig and poultry farms in the Netherlands and the human isolated from the farmers of these farms . A total of 24 isolates were investigated . AFLP and Tn1546 typing revealed that both pig and poultry related enterococcal and vanA transposon genotypes were found among the human isolates indicating spread of glycopeptide resistance from both pig and poultry to the farmers . These findings contradict previous finding that showed that GRE recovered from the general population were genotypically undistinguishable from GRE isolated from pigs but are in line with other studies that demonstrated spread of GRE from poultry to farmers in poultry farms.

Environ Sci Technol, 2003 Aug 1, 37(15), 3275 - 82
Escherichia coli and enterococci at beaches in the Grand Traverse Bay, Lake Michigan: sources, characteristics, and environmental pathways; Haack SK et al.; This study quantified Escherichia coli (EC) and enterococci (ENT) in beach waters and dominant source materials, correlated these with ambient conditions, and determined selected EC genotypes and ENT phenotypes . Bathing-water ENT criteria were exceeded more frequently than EC criteria, providing conflicting interpretations of water quality . Dominant sources of EC and ENT were bird feces (10(8)/d/bird), storm drains (10(7)/d), and river water (10(11)/d); beach sands, shallow groundwater and detritus were additional sources . Beach-water EC genotypes and ENT phenotypes formed clusters with those from all source types, reflecting diffuse inputs . Some ENT isolates had phenotypes similar to those of human pathogens and/or exhibited high-level resistance to human-use antibiotics . EC and ENT concentrations were influenced by collection time and wind direction . There was a 48-72-h lag between rainfall and elevated EC concentrations at three southern shoreline beaches, but no such lag at western and eastern shoreline beaches, reflecting the influence of beach orientation with respect to cyclic (3-5 d) summer weather patterns . In addition to local contamination sources and processes, conceptual or predictive models of Great Lakes beach water quality should consider regional weather patterns, lake hydrodynamics, and the influence of monitoring method variables (time of day, frequency).

Arch Intern Med, 2003 Sep 8, 163(16), 1905 - 12
Role of environmental contamination as a risk factor for acquisition of vancomycin-resistant enterococci in patients treated in a medical intensive care unit; Martinez JA et al.; BACKGROUND: Colonization pressure, proximity to another case, exposure to a nurse who cares for another case, enteral feeding, and the use of sucralfate, vancomycin hydrochloride, cephalosporins, or antibiotics are among the defined risk factors for acquisition of vancomycin-resistant enterococci (VRE) in the intensive care unit (ICU) setting . However, the role of rooms with contaminated environmental surfaces has not been well delineated . METHODS: Retrospective case-control study conducted on patients admitted to the medical ICU (MICU) of a tertiary-care, university-affiliated medical center during a 9-month period . Patients who acquired VRE (cases) were matched with 2 randomly selected control subjects who did not acquire VRE and had been in the MICU for at least the same number of days . RESULTS: Thirty cases were matched with 60 appropriate controls . Cases were more likely to have been in the hospital for longer than 7 days before MICU admission (P =.009); to have occupied a specific room with persisting contaminated surfaces (P =.06); to have had a central venous catheter (P =.05); to have received vancomycin (P =.02), cephalosporins (P =.03), and quinolones (P =.006) before MICU admission; and to have received vancomycin (P =.02) and metronidazole sodium phosphate (P =.03) after MICU admission . Multivariate analysis showed that a hospital stay of longer than 1 week before MICU admission (P =.04), use of vancomycin before or after MICU admission (P =.03), use of quinolones before MICU admission (P =.03), and placement in a contaminated room (P =.02) were the best predictors of VRE acquisition . CONCLUSIONS: Among all other factors associated with VRE transmission, VRE acquisition may depend on room contamination, even after extensive cleaning . This study underscores the need for better cleaning and the role of the environment in transmission of VRE.

Microb Drug Resist, 2003 Fall, 9(3), 307 - 12
Recurrent episodes of shock-like syndrome caused by the same strain of vancomycin-resistant Enterococcus faecium in a pediatric patient; Papp L et al.; We present the case of a hospitalized pediatric patient with short bowel syndrome who was dependent upon total parenteral nutrition for 17 months . Shortly after admission she became colonized with vancomycin-resistant Enterococcus faecium (VRE) and developed 12 distinct episodes of serious infection associated with it . The course of VRE colonization and infections in this patient was studied through analysis of 40 representative isolates obtained from different sites during distinct episodes of infection . Standard microbiological techniques, automated ribosomal DNA typing, polymerase chain reaction, and pulsed-field gel electrophoresis (PFGE) were used . All isolates except for the one associated with the initial episode of bacteremia were VRE, and were multidrug resistant . The last four episodes of infection were caused by isolates resistant to all tested antibiotics except for intermediate susceptibility to chloramphenicol . The vanA genotype was a source of vancomycin resistance in all VRE isolates . Both ribotyping and PFGE showed two distinct clones of VRE in clinical and stool surveillance isolates: one was associated with clinical illness and the other was not associated with infection . Recurrent VRE infections occur as a consequence of prolonged gastrointestinal colonization . Morbidity is associated with host factors, the presence of co-pathogens, and possibly intrinsically more virulent VRE strain.

Front Biosci, 2003 Sep 01, 8, s1229 - 39
Acute cholangitis; Lipsett PA et al.; Acute cholangitis is an infectious disease of the biliary tract with a wide spectrum of presentation ranging in severity from a mild form with fever and jaundice, to a severe form with septic shock . Supportive care with hydration, antibiotics, and biliary decompression remain the cornerstones of care . Broad-spectrum antibiotics should include coverage of E.coli, Klebsiella sp., Enterococcus sp., and in severely critically ill patients, coverage of additional pathogens such as Bacteriodes sp., Pseudomonas, and yeast should be considered . Biliary decompression should be performed early in the course of the illness when the patient has not improved or if they worsen with hydration and antibiotics . Stable patients should have biliary decompression usually within 72 hours when the fever has resolved . Urgent decompression with a percutaneous or endoscopic stent is preferred over an operative decompression in most institutions . Outcome is dependent on the etiology of the obstruction (benign versus malignant) and the ability to achieve biliary decompression.

Bioorg Med Chem Lett, 2003 Oct 6, 13(19), 3253 - 6
2-piperidin-4-yl-benzimidazoles with broad spectrum antibacterial activities; He Y et al.; A series of 2-piperidin-4-yl-benzimidazoles were synthesized and evaluated for antibacterial activities . Certain compounds inhibit bacterial growth with low micromolar minimal inhibitory concentration (MIC) . These benzimidazoles are effective against both Gram-positive and Gram-negative bacteria of clinical importance, particularly enterococci, and represent a new class of potential antibacterial agents.

Antimicrob Agents Chemother, 2003 Sep, 47(9), 2993 - 6
Multiple antibiotic resistance gene transfer from animal to human enterococci in the digestive tract of gnotobiotic mice; Moubareck C et al.; It has been proposed that food animals represent the source of glycopeptide resistance genes present in enterococci from humans . We demonstrated the transfer of vanA and of other resistance genes from porcine to human Enterococcus faecium at high frequency in the digestive tract of gnotobiotic mice . Tylosin in the drinking water favored colonization by transconjugants.

Rev Argent Microbiol, 2003 Apr-Jun, 35(2), 96 - 9
{Bacteremia caused by Enterococcus gallinarum with a high level of glycopeptide resistance: 1st documented cases in Argentina}; Togneri A et al.; A case of bacteremia due to high-level-vancomycin- (MIC = 64 micrograms/ml) and high-level-teicoplanin- (MIC = 32 micrograms/ml) resistant Enterococcus gallinarum is described . Both genes, van C1 and van A, respectively conferring natural low-level resistance and acquired high-level resistance to vancomycin, were found in the enterococcal genoma . The present is the first report of an E . gallinarum isolate showing the van A genotype in Argentina.

Dis Aquat Organ, 2003 Jul 8, 55(2), 161 - 7
Metschnikowia bicuspidata and Enterococcus faecium co-infection in the giant freshwater prawn Macrobrachium rosenbergii; Chen SC et al.; In May 2001, an epizootic yeast and bacterial co-infection in the giant freshwater prawn Macrobrachium rosenbergii occurred in Taiwan causing a cumulative mortality of 25% . The diseased prawns had a yellowish-brown body color, milky hemolymph, opaque, whitish muscles, and were approximately 7 mo old with total lengths ranging from 8 to 10 cm . Histopathological examination showed marked edema, yeast infiltration, and necrotic lesions with inflammation in the muscles, hepatopancreas and other internal organs . We isolated 2 pathogens from the diseased prawns, one was a yeast (AOD081MB) and the other a gram-positive coccus (AOD081EF) . The gram-positive coccus was identified as Enterococcus faecium by the API 20 Strepsystem, conventional biochemical tests, and it had 99% 16S rDNA sequence identity (GenBank Accession Number AJ276355) to E . faecium (GenBank Accession Number AF529204) . The sequence of a PCR product from the D1/D2 domain of 26S rDNA (GenBank Accession Number AF529297) from the yeast gave 99% sequence identity to Metschnikowia bicuspidata (GenBank Accession Number U44822) . Experimental infections with these isolates produced gross signs and histopathological changes similar to those observed in the naturally infected prawns . The lethal doses (LD50) for isolate E . faecium AOD081EF, M . bicuspidata AOD081MB and the co-infection were 4.7 x 10(4), 2.6 x 10(2), and 2.4 x 10(2) colony-forming units prawn(-1), respectively . This is the first report of a confirmed co-infection of M . bicuspidata and E . faecium in prawn aquaculture.

Med Dosw Mikrobiol, 2003, 55(1), 25 - 32
{Expression of proteins regulated by iron levels in enterococci}; Lisiecki P et al.; Enterococci respond to iron deprivation in vitro by increasing the expression of a number of iron-regulated proteins . They detected in whole protoplasts lysates and corresponded to proteins with apparent molecular masses of the region 14.4-43 Kda . Their occurrence were correlated to siderophore production.

Am J Clin Oncol, 2003 Aug, 26(4), e86 - 91
A simple method of producing low oxygen conditions with oxyrase for cultured cells exposed to radiation and tirapazamine; Ho KC et al.; Several methods of establishing low O(2) conditions have been used in studies on the response of cultured cells to radiation and other agents . These methods, eg, gassing culture vessels with O(2)-free nitrogen with or without carbon dioxide or placing high cell-density suspensions in sealed glass ampoules to consume O(2) in the ampules, can be technically demanding and have experimental limitations . We introduce a simple, versatile, and reliable method of producing low O(2) conditions without special equipment or changes in culture conditions unrelated to hypoxia . The method is based on the ability of Oxyrase (Oxyrase, Inc., Mansfield, OH), membrane fragments prepared from Enterococcus coli, to consume O(2) in solution and is confirmed in the present study by 2 analytical methods . The effects of low O(2) conditions induced by Oxyrase on cellular responses to radiation and treatment with the bioreductive agent tirapazamine (TPZ) were examined with Chinese hamster V79 and human glioma U373 cells . Measured by clonogenic and MTT assays, these cells were less sensitive to radiation but more sensitive to TPZ in treatment media containing native Oxyrase than in media containing heat-inactivated Oxyrase . In addition, Oxyrase treatment increased the basal activity of mitogen-activated protein kinase (ERK1/2) but suppressed its activation induced by radiation . The results suggest that this method might also be useful for other in vitro cancer biologic investigations requiring a low O(2) condition.

Zhonghua Wai Ke Za Zhi, 2003 Jun, 41(6), 420 - 2
{Integrative therapy for postoperative intractable enterococcus faecium infections}; Tang Z et al.; OBJECTIVE: To assess the treatment of patients with infection of enterococcus faecium after surgery who failed to respond to antibiotics . METHODS: Five patients after surgery were proved to have Enterococcus faecium infection by bacterial culture . They were treated by sensitive antibiotics but failed . Comprehensive treatment prescribed included immunoenhancements, enteral nutrition, and traditional Chinese medicines . RESULTS: Four patients were discharged from the hospital after recovery, and was cared else where after 1 month treatment . CONCLUSION: Comprehensive treatment is a better way to treat patients with refractory enterococcus faecium infection after surgery.

Int J Syst Evol Microbiol, 2003 Jul, 53(Pt 4), 1069 - 74
Description of Enterococcus canis sp . nov . from dogs and reclassification of Enterococcus porcinus Teixeira et al . 2001 as a junior synonym of Enterococcus villorum Vancanneyt et al 2001; De Graef EM et al.; Strains from anal swabs and chronic otitis externa in dogs were shown to be phylogenetically related to the Enterococcus faecium species group . They shared a number of phenotypic characteristics with these species, but they could be easily differentiated by biochemical reactions . In addition, the canine strains were unusual in their nearly complete failure to grow on sodium azide-containing enterococci-selective media and in their Voges-Proskauer reactions (usually negative) . By using 16S rRNA sequencing and DNA-DNA hybridization of representative strains, as well as tDNA interspacer gene PCR and SDS-PAGE of whole-cell proteins, the group of canine strains was shown to constitute a novel enterococcal species . The name Enterococcus canis sp . nov . is proposed for this species, with LMG 12316T (= CCUG 46666T) as the type strain . Concurrently, the taxonomic situation and nomenclatural position of Enterococcus porcinus were investigated . As no phenotypic or genotypic differences were found between this species and Enterococcus villorum, the name E . porcinus is considered to be a junior synonym of E . villorum.

Clin Infect Dis, 2003 Aug 1, 37(3), 326 - 32 Epub 2003 Jul 14.
Control of endemic vancomycin-resistant Enterococcus among inpatients at a university hospital; Calfee DP et al.; We sought to determine the ability of surveillance cultures and isolation of vancomycin-resistant Enterococcus (VRE)-colonized patients to control nosocomial VRE infection and colonization during a 5-year period (November 1994 through October 1999) . During this period, VRE colonization was limited to 0.82% of admissions . The incidence of VRE infection was 0.12 cases per 1000 patient-days (attack rate, 0.07%) . Colonized patients were first identified by surveillance (95%) or routine clinical cultures (5%); 14% of colonized patients had a positive clinical culture a median of 15 days after a positive surveillance culture . Ten percent of colonized patients were identified by surveillance at the time of transfer from another health care facility . Identification of these colonized patients was associated with reduction from a peak incidence rate of 2.07% to a rate of 1.25% and stabilization at this lower level . The use of surveillance cultures to identify and isolate patients with asymptomatic colonization can provide sustained control of the spread of VRE within a health care facility.

Rapid Commun Mass Spectrom, 2003, 17(16), 1889 - 96
Analysis of the metal-binding selectivity of the metallochaperone CopZ from Enterococcus hirae by electrospray ionization mass spectrometry; Urvoas A et al.; Metallochaperones are soluble proteins involved in metal transport and regulation in vivo . Copper metallochaperones belong to a structural family of metal binding domains displaying a ferredoxin-like fold (betaalphabetabetaalphabeta) and a consensus metal-binding motif MXCXXC . The metal-binding selectivities for this class of proteins are poorly documented so far . The present study focuses on the measurement of the selectivity of the copper metallochaperone CopZ from Enterococcus hirae for different metal ions using an experimental approach based on electrospray ionization mass spectrometry (ESI-MS) . All the metal cations tested, i.e . Cu(I), Cu(II), Hg(II), Cd(II) and Co(II), form specific metal complexes with CopZ . The study of a chemically modified CopZ as well as variants of CopZ in the active site demonstrated that the complexes observed by ESI-MS, i.e . in the gas phase, corresponded to the complexes previously observed by other analytical methods in solution . Competition experiments allowed the classification of the metal ions by increasing affinities for CopZ as follows: Co << Cd < Hg < Cu . A dissociation constant in the range of 20 microM was determined for cobalt . The affinity of CopZ for the other metals tested was found to be higher, with dissociation constants smaller than micromolar .

Vet Microbiol, 2003 Aug 29, 95(1-2), 91 - 101
Susceptibility of Escherichia coli and Enterococcus faecium isolated from pigs and broiler chickens to tetracycline degradation products and distribution of tetracycline resistance determinants in E . coli from food animals; Sengelov G et al.; One hundred Escherichia coli isolates from diseased and healthy pigs, cattle and broiler chickens were screened for the presence of tetracycline resistance genes tet(A), (B), (C), (D) or (E) . The tet(A) gene was the most abundant (71% of the 100 isolates) followed by tet(B) (25%) . The predominance of tet(A) and tet(B) applied to all three animal species, and there was no difference between the distribution of tet(A) and tet(B) genes among non-pathogenic and pathogenic E . coli in any of the animal species . The susceptibility of 20 of these isolates together with 10 tetracycline sensitive E . coli and 18 tetracycline resistant and 10 sensitive Enterococcus faecium to tetracyclines and tetracycline degradation products was determined . The resistant isolates showed reduced resistance to anhydrotetracycline, 4-epi-anhydrotetracycline, anhydrochlortetracycline and 4-epi-anhydrochlortetracycline . In general both the tetracycline resistant and susceptible E . faecium were more susceptible to the compounds tested than E . coli.

J Appl Microbiol, 2003, 95(2), 390 - 7
Potential of the enterocin regulatory system to control expression of heterologous genes in Enterococcus; Hickey RM et al.; AIMS: To exploit the enterocin regulatory system for regulated expression of genes in Enterococcus . METHODS AND RESULTS: Production of some pediocin-like bacteriocins such as enterocin A in Enterococcus is regulated by a three-component system comprising a histidine kinase (entK ), a response regulator (entR) and an induction factor (entF ) . Exposure to the induction factor results in the transcription of gene(s) under the control of the enterocin A promoter, including entA which encodes the bacteriocin . In an effort to exploit this system for expression of genes in Enterococcus, a number of vectors were constructed which contain the entA promoter followed by convenient cloning sites to introduce gene(s) of interest . These vectors were used in an enterococcal background which does not produce induction factor but does produce both the kinase and regulator proteins . The system was tested using the reporter genes ltnI (lacticin 3147 immunity) and gusA (beta-glucuronidase) under the control of the entA promoter . CONCLUSIONS: Upon addition of the induction factor, the beta-glucuronidase activity increased 20-fold when compared with uninduced cells . In addition, concentrations of as little as 0.2 nm synthetic EntF were sufficient to give maximal expression . SIGNIFICANCE AND IMPACT OF THE STUDY: The potential benefit of having an expression system based on EntF is that gene expression can be finely controlled upon addition of low concentrations of a peptide that can easily be artificially synthesized.

J Hosp Infect, 2003 Jul, 54(3), 188 - 95
Clustering of enterococcal infections in a general intensive care unit; Samuelsson A et al.; This is a retrospective study comparing patients' characteristics, antibiotic consumption and environmental contamination before the impact of a new regimen of intensified infection control measures in a general intensive care unit (ICU) at a university-affiliated tertiary-care teaching hospital . The new regimen consisted of (1) reorganization of patient rooms (2) improved hygienic measures including strict hygiene barrier nursing (3) more isolated patient care and (4) more restrictive use of antibiotics . The regimen was introduced after a cluster of enterococcal infections . All patients admitted to the ICU from 1 March 1995 to 28 february 1997 were included . A study period of 12 months after reorganization of the ward was compared with the 12 months immediately before it . The antibiotic consumption, the individual patient's severity of disease (APACHE score), and the extent of therapeutic interventions (TISS score) were recorded . Enterococci were typed biochemically, antibiograms were established and the relation between the isolates was investigated with pulsed-field gel electrophoresis . The bacteriological results and the patient data suggested a hospital-acquired spread as the cause of the ICU enterococcal outbreak . After implementation of the new regimen, we observed a reduction in the rate of enterococcal bloodstream infections from 3.1 to 1.8% . The consumption of antibiotics fell from 6.11 to 4.24 defined daily doses per patient . The introduction of strict hygiene and barrier nursing, more restrictive use of antibiotics, isolation of infected patients, thorough cleaning and disinfection of the unit was followed by an absence of enterococcal infection clustering and reduction in incidence of enterococcal bacteraemia . We were not able to determine whether the reduction in antibiotic consumption was due to the intervention programme.

J Antimicrob Chemother, 2003 Aug, 52(2), 214 - 9 Epub 2003 Jul 01.
Gentamicin resistance in dairy and clinical enterococcal isolates and in reference strains; Lopes Mde F et al.; Enterococci isolated from Portuguese dairy products (milk and cheese) and clinical settings (hospitals and veterinary clinics), together with reference strains from the genus Enterococcus, were screened for low- and high-level gentamicin resistance using the standard disc diffusion method (10 and 120 microg gentamicin discs) . MICs were also determined using both the macrodilution method and the Etest . Four genes {aac(6')-Ie-aph(2")-Ia, aph(2")-Ib, aph(2")-Ic and aph(2")-Id} responsible for high- and mid-level gentamicin resistance were sought using PCR . Although enterococci generally are regarded as being intrinsically resistant to low levels of gentamicin, results revealed that many dairy enterococci (around 30% of the isolates used) are not intrinsically resistant to gentamicin, showing MICs of < or = 4 mg/l . High-level gentamicin resistance was not detected in any of the dairy isolates studied, except for aph(2")-Ib, which was found in one . Therefore, gentamicin resistance should be monitored in dairy enterococci, although it does not seem to be a problem at present . In contrast, all clinical isolates studied were, as expected, intrinsically resistant to low levels of gentamicin, presenting MICs > 8 mg/l . Fifteen percent of these clinical isolates showed high-level gentamicin resistance (MICs > 512 mg/l), with the bifunctional gene aac(6')-aph(2") being detected in four of them . However, discs with gentamicin 120 microg failed to detect some isolates with high-level gentamicin resistance.

Rev Argent Microbiol, 2003 Jan-Mar, 35(1), 41 - 4
{1st isolation of vancomycin-resistant Enterococcus faecium with the vanB genotype in Argentina: presentation of 2 cases}; Miranda G et al.; We describe the first isolate of van B vancomycin-resistant Enterococcus faecium in Argentina . The strains were recovered from ambulatory patients admitted to a hospital of Buenos Aires city in July 2000 . They were not high-risk patients, they had not received previous antibiotic therapy, and they were assisted in different services . MICs for vancomycin were 32 micrograms/ml for both strains, whereas MICs for teicoplanin were 0.12 microgram/ml in case 1 and 0.25 microgram/ml in case 2 . PCR was performed to confirm the vanB genotype . The molecular fingerprints of the isolations by PFGE revealed that they were identical . No further VanB strains were isolated in the hospital.

FEMS Microbiol Rev, 2003 Jun, 27(2-3), 183 - 95
Copper homeostasis in Enterococcus hirae; Solioz M et al.; Copper is an essential component of life because of its convenient redox potential of 200-800 mV when bound to protein . Extensive insight into copper homeostasis has only emerged in the last decade and Enterococcus hirae has served as a paradigm for many aspects of the process . The cop operon of E . hirae regulates copper uptake, availability, and export . It consists of four genes that encode a repressor, CopY, a copper chaperone, CopZ, and two CPx-type copper ATPases, CopA and CopB . Most of these components have been conserved across the three evolutionary kingdoms . The four Cop proteins have been studied in vivo as well as in vitro and their function is understood in some detail.

Infect Control Hosp Epidemiol, 2003 Jun, 24(6), 461 - 4
A pseudo-outbreak of vancomycin-resistant Enterococcus faecium; Grinbaum RS et al.; Five VRE isolates were reported from sterile samples . No infections were diagnosed among the patients, who were on different wards . PFGE showed that all five isolates were clonal . All samples were manipulated by the same worker, but the source could not be found . Contamination was probably related to manipulation of a source specimen in the laboratory.

Environ Health Perspect, 2003 Jun, 111(8), 1102 - 9
Do U.S . Environmental Protection Agency water quality guidelines for recreational waters prevent gastrointestinal illness? A systematic review and meta-analysis; Wade TJ et al.; Despite numerous studies, uncertainty remains about how water quality indicators can best be used in the regulation of recreational water . We conducted a systematic review of this topic with the goal of quantifying the association between microbial indicators of recreational water quality and gastrointestinal (GI) illness . A secondary goal was to evaluate the potential for GI illness below current guidelines . We screened 976 potentially relevant studies and from these identified 27 studies . From the latter, we determined summary relative risks for GI illness in relation to water quality indicator density . Our results support the use of enterococci in marine water at U.S . Environmental Protection Agency guideline levels . In fresh water, (Italic)Escherichia(/Italic) coli was a more consistent predictor of GI illness than are enterococci and other bacterial indicators . A log (base 10) unit increase in enterococci was associated with a 1.34 {95% confidence intervals (CI), 1.00-1.75} increase in relative risk in marine waters, and a log (base 10) unit increase in E . coli was associated with a 2.12 (95% CI, 0.925-4.85) increase in relative risk in fresh water . Indicators of viral contamination were strong predictors of GI illness in both fresh and marine environments . Significant heterogeneity was noted among the studies . In our analysis of heterogeneity, studies that used a nonswimming control group, studies that focused on children, and studies of athletic or other recreational events found elevated relative risks . Future studies should focus on the ability of new, more rapid and specific microbial methods to predict health effects, and estimating the risks of recreational water exposure among susceptible persons.

Am J Transplant, 2003 Jul, 3(7), 902 - 5
Low prevalence of colonization with vancomycin-resistant Enterococcus in patients awaiting liver transplantation; Hagen EA et al.; The orthotopic liver transplant (OLT) population has been particularly affected by the increase in vancomycin-resistant enterococcus (VRE) infections in recent years . Pre-transplant colonization prevalence, the role of spontaneous bacterial peritonitis (SBP) antimicrobial prophylaxis as a risk factor, and the risk of post-OLT infection in colonized patients are all unknowns . We prospectively evaluated OLT candidates at our center with the aim of answering these questions . Vancomycin-resistant enterococcus colonization status was determined by rectal culture . Data collected included illness severity, antibiotic use (including SBP prophylaxis), waiting time, previous hospitalizations, and invasive procedures . Eighty-eight patients (31 female, 57 male, median age 52 years) were enrolled . The most common diagnoses were hepatitis C (49%), primary sclerosing cholangitis (13.6%), and alcoholic liver disease . Median MELD score was 11.5 (range 7-24), and median waiting time was 551 days (range 1-2224) . Vancomycin-resistant enterococcus risk factors were common in our patients: recent hospitalization in 16%, recent antibiotic exposure in 39%, and renal insufficiency in 7% . Seventeen percent were receiving SBP prophylaxis . Despite the presence of established risk factors, VRE colonization prevalence was 3.4% . Preliminary limited data showed poor correlation between screening rectal cultures and operative/peri-operative cultures . Vancomycin-resistant enterococcus colonization prevalence in an OLT candidate population with mid-level MELD scores was low, and SBP prophylaxis was not a significant risk factor.

Acta Pharm Hung, 2002, 72(4), 245 - 51
{Analysis of antibiotic consumption of five different clinical departments, especially considering the features of hematology departments}; Benko R et al.; The importance of antibiotic consumption and bacterial resistance analysis is unquestionable . Although the WHO has been recommending the use of ATC/DDD method for the analysis of drug utilization for about 20 years, only limited numbers of publications are available considering the antibiotic consumption of in-patients . In the present study--with the help of the mentioned method--we analysed the utilization of antibiotics in 5 clinical departments . We also investigated the connection between antibiotic consumption, bacterial resistance and enterococcal sepsis at the haematologic department . We could follow the successful impact of the implementation of the new guideline.

Int J Food Microbiol, 2003 Aug 1, 84(3), 273 - 84
Relations between the occurrence of resistance to antimicrobial growth promoters among Enterococcus faecium isolated from broilers and broiler meat; Emborg HD et al.; From 1995 to 2001, Enterococcus faecium isolates were collected from broiler flocks at slaughter and broiler meat products at retail outlets and were tested for susceptibility to classes of antimicrobials used for growth promotion in broilers in Denmark, namely: evernimicin, glycopeptide, macrolide and streptogramin . By February 1998, all antimicrobial growth promoters (AGPs) were withdrawn from the Danish broiler production . The present study investigates, by logistic regression analyses, the (1) changes in the occurrence of AGP resistance among E . faecium from broilers and broiler meat from the fourth quarter of 1995 to the fourth quarter of 2001 and (2) relations between the occurrence of AGP resistance among E . faecium isolates from Danish broilers and AGP resistance among E . faecium isolates from the broiler meat of Danish and unknown origin collected in the same quarter within the year . In the present study, we showed that after the AGP withdrawal, a significant decline in resistance to avilamycin, erythromycin, vancomycin and virginiamycin was observed among E . faecium from broilers and broiler meat . In addition, a decline in the occurrence of AGP resistance among E . faecium from Danish broilers was associated with a decrease in the predicted probability of isolating an AGP-resistant E . faecium isolate from a randomly selected broiler meat product.In the analyses "relations between the occurrence of AGP resistance among E . faecium isolated from broilers and broiler meat collected in the same quarter" errors in the explanatory variable were expected . Therefore, a simulation study was performed to validate the results from logistic regression analyses . The results obtained by the two methods were similar.

Commun Dis Intell, 2003, 27 Suppl, S97 - 102
Screening and electronic labelling of ward contacts of vancomycin-resistant Enterococcus faecium vanB carriers during a single-strain hospital outbreak and after discharge from hospital; Pearman JW et al.; A large single-strain outbreak of vancomycin-resistant Enterococcus faecium (VREF) vanB occurred in Royal Perth Hospital from July to December 2001 . When a VREF-carrying patient was discovered on a ward, all patients on the ward were screened with rectal swabs . A total of 172 patients were colonised, four with infections, but no deaths were attributable to VREF . The number of rectal swabs required to detect each carrier was recorded . On average four rectal swabs, each collected on separate days, were needed to detect more than 90 per cent of the 172 VREF carriers who were epidemiologically linked to the Royal Perth Hospital outbreak . An electronic alert system (Micro-Alert) was used to identify ward contacts of VREF carriers and enabled those who had not been screened before discharge to be followed-up and screened . Ninety-six contacts were actively followed-up in October 2001 and 32 (33.3%) were found to be VREF carriers . From 28 September 2001 to 30 April 2002, a total of 1,977 ward contacts were screened after discharge from hospital and 54 (2.73%) were found to be carrying VREF . We conclude that during single-strain outbreaks of vancomycin-resistant enterococci in hospitals, patient contacts need to be screened on more than three occasions in order to detect most of the carriers and control the outbreak . Secondly, electronic labelling and active follow-up of ward contacts of VREF carriers resulted in a significant number of carriers being detected who otherwise posed a risk of initiating further outbreaks in hospitals if they were readmitted.

J Antimicrob Chemother, 2003 Jul, 52(1), 113 - 5 Epub 2003 Jun 12.
Intra-hospital dissemination of quinupristin/dalfopristin- and vancomycin-resistant Enterococcus faecium in a paediatric ward of a German hospital; Werner G et al.; OBJECTIVES: To demonstrate nosocomial transmission of Enterococcus faecium resistant to quinupristin/dalfopristin and vancomycin/teicoplanin among paediatric patients in a German hospital ward . MATERIALS AND METHODS: Multiply-resistant E . faecium were isolated from three female patients aged 9 months, 2 and 15 years during a 10 day time span . Antibiotic susceptibilities were determined by microbroth dilution . Clonal relatedness among the isolates was investigated via SmaI-macrorestriction analysis by PFGE, multilocus sequence typing (MLST), and plasmid profiling . Presence of virulence and resistance determinants was tested by polymerase chain reaction (PCR) . Selected resistance genes were localized by Southern hybridizations . RESULTS: A single E . faecium isolate per patient was investigated . All exhibited resistances to quinupristin/dalfopristin, vancomycin/teicoplanin, streptomycin (high-level), penicillin/ampicillin, erythromycin, oxytetracycline, chloramphenicol, rifampicin and fusidic acid . The isolates were susceptible to linezolid only and intermediately resistant to fluoroquinolones including moxifloxacin . PFGE revealed identical patterns for all three isolates . PCRs for virulence determinants hyaluronidase and enterococcal surface protein, esp, were negative, whereas PCR for the enterocin A gene was positive . MLST identified clonal type {8-5-1-1-1-1-1} belonging to a clonal subgroup C1 of hospital- and outbreak-related E . faecium . Southern hybridizations located several resistance genes (erm(B), vat(D), vanA) on a large plasmid, which was transferable in mating experiments with an E . faecium recipient . CONCLUSIONS: These data show routes of dissemination of resistance to multiple antibiotics including streptogramins and glycopeptides in E . faecium via vertical and/or horizontal gene transfer . The isolates spread in the absence of a direct selective pressure, as none of the patients had received earlier streptogramin or glycopeptide therapy.

J Antimicrob Chemother, 2003 Jun, 51 Suppl 3, iii23 - 30
Therapeutic and preventative options for the management of vancomycin-resistant enterococcal infections; Kauffman CA; Enterococci are naturally resistant to a wide range of antimicrobial agents . In addition, some enterococci, known as vancomycin-resistant enterococci (VRE) have become resistant to glycopeptide antibiotics . The therapeutic options for VRE infections are therefore very limited . New antimicrobials have been developed that are active against VRE, such as linezolid and quinupristin/dalfopristin . Others, e.g . tigecycline, daptomycin and oritavancin, are in the later stages of development . However, resistance has already been detected to some of these agents . Some success has been enjoyed through the application of older antibiotics against VRE . The lack of therapeutic options has led to the consideration of measures to prevent infection with VRE . In addition to standard infection control procedures such as isolation and hand washing, decolonization of the gastrointestinal tract has been investigated as a method for the prevention of VRE infection in vulnerable patient groups . Several decolonization regimens have been investigated . These include the use of ramoplanin, a new glycolipodepsipeptide antibiotic that has features that particularly suit it for decolonization . Ramoplanin is not absorbed from the gastrointestinal tract, has potent bactericidal activity against Gram-positive organisms and limited side effects . These features and current clinical evidence suggest that ramoplanin may have a role in future gastrointestinal decolonization regimens.

J Antimicrob Chemother, 2003 Jun, 51 Suppl 3, iii13 - 21
Clinical impact of vancomycin-resistant enterococci; Patel R; In humans, vancomycin-resistant enterococci (VRE) most commonly result in intestinal colonization, which does not result in symptoms, may persist for a long time and serves as a reservoir for transmission of VRE to other patients . Certain VRE-colonized patients are at risk of infection, including haematology and oncology patients, patients in intensive care units and recipients of solid (especially abdominal) organ transplants . Controlling the spread of VRE colonization and preventing colonized patients from becoming infected are important aims . Ramoplanin is a member of a new class of antimicrobial agents; it may have a role in preventing infection in patients colonized with VRE.

Bioorg Med Chem Lett, 2003 Jul 7, 13(13), 2227 - 30
Synthesis and in vitro activity of new methylenepiperidinyl and methylenepyrrolidinyl oxazolidinone antibacterial agents; Kim HY et al.; We have prepared and evaluated the antibacterial activities of a series of substituted methylenepiperidinyl and methylenepyrrolidinyl oxazolidinones against several gram-positive strains including the resistant strains of Staphyloccus and Enterococcus, such as MRSA, CRSA, MSSA and VRE . Some of them showed comparable or superior in vitro activities (MIC) to vancomycin.

J Clin Microbiol, 2003 Jun, 41(6), 2703 - 5
Comparison of microscan broth microdilution, synergy quad plate agar dilution, and disk diffusion screening methods for detection of high-level aminoglycoside resistance in enterococcus species; Murdoch DR et al.; We compared the dried MicroScan microdilution panel, Synergy Quad plate agar dilution, and high-potency disk diffusion screening methods for the detection of high-level aminoglycoside resistance in 815 enterococcal bloodstream isolates . Agreement between the three methods was 99% when testing for high-level gentamicin resistance and 96% when testing for high-level streptomycin resistance.

J Clin Microbiol, 2003 Jun, 41(6), 2644 - 6
Use of colony morphology to distinguish different enterococcal strains and species in mixed culture from clinical specimens; Qamer S et al.; Colony morphology on kanamycin esculin azide agar was investigated as a means of selecting different species and strains of enterococci from clinical specimens . Four representative colonies of each morphotype were indistinguishable by pulsed-field gel electrophoresis, biotype, and antibiogram analysis . The optimum time for identification of different colony morphotypes was 72 h.

J Clin Microbiol, 2003 Jun, 41(6), 2641 - 3
Diversity of tn1546-like elements in vancomycin-resistant enterococci isolated from humans and poultry in Korea; Yu HS et al.; To investigate the possible spread of vancomycin resistance among enterococci through horizontal gene transfer between two sources of vancomycin-resistant enterococci (VRE) isolated from farm animals and humans in Korea, molecular characterization of VRE isolated from poultry (20 isolates) and humans (35 isolates) was performed by PCR-restriction fragment length polymorphism typing of the vanA gene cluster (Tn1546) . PCR mapping of Tn1546 finally distinguished seven different transposon types (types A to G) . Type A was observed only in the poultry isolates, while the other six types were present only in the human isolates . An insertion sequence and deleted sequence were detected in most of the human isolates in the orf2-vanR and vanX-vanY regions in the Tn1546-like element, but not in the poultry isolates . Tn1546-like elements were found in conjugal plasmids of most human VRE, whereas they were detected in the chromosomes of all poultry VRE . Accordingly, no evidence was found of any recent transmission of vancomycin resistance genes between poultry and humans in Korea.

J Clin Microbiol, 2003 Jun, 41(6), 2569 - 76
Comparative study using type strains and clinical and food isolates to examine hemolytic activity and occurrence of the cyl operon in enterococci; Semedo T et al.; The hemolytic ability, the presence of cyl genes, and the diagnostic accuracy of cytolysin molecular detection were investigated in the genus Enterococcus by using 164 strains from 20 different species (26 reference strains, 42 clinical isolates from human and veterinary origin, and 96 isolates from ewe cheese and milk) . Hemolysis was assayed with sheep and horse erythrocytes and under aerobic or anaerobic conditions . Screening of cytolysin genes (cylL(L), cylL(S), cylM, cylB, and cylA) was performed with new specific primers and the anaerobic assay of beta-hemolysis was used as the "gold standard" for the evaluation of cyl gene-based PCRs . Since beta-hemolysis and cyl genes were found in 10 and 14 species, respectively, the hemolytic ability seems to be spread throughout the genus ENTEROCOCCUS: Beta-hemolysis was observed in 6 of 26 (23%) reference strains, 14 of 42 (33%) clinical isolates, and 6 of 96 (6%) food isolates . The presence of cyl genes was detected in 15 of 26 (58%) reference strains, 37 of 42 (88%) clinical isolates, and 67 of 96 (70%) food isolates . These data indicate a virulence potential in food isolates, reinforcing the need of their safety assessment . Analysis of phenotypic-genotypic congruence suggests a divergent sequence evolution of cyl genes and the effect of environmental factors in the regulation of cytolysin expression . Evaluation of the diagnostic accuracy of cytolysin molecular detection points to cylL(L)-based PCR and cylL(L)L(S)MBA-based PCR as the most reliable approaches . Nevertheless, the low sensitivity (46%) and gene variability indicated by our study strongly recommend the phenotypic assay for the assessment of hemolytic ability in enterococci, followed by the molecular screening of cyl genes in nonhemolytic strains to evaluate their virulence potential.

J Clin Microbiol, 2003 Jun, 41(6), 2330 - 6
Molecular characterization of ampicillin-resistant Enterococcus faecium isolates from hospitalized patients in Norway; Jureen R et al.; The genetic relationship of 81 ampicillin-resistant and 21 ampicillin-susceptible Enterococcus faecium isolates from clinical infections and rectal screening in hospitalized patients in Norway was studied by pulsed-field gel electrophoresis (PFGE) and amplified fragment length polymorphism (AFLP) . PFGE showed 55 different banding patterns, and 65 of the isolates could be grouped into one large group . With AFLP, 46 patterns were discerned, and 74 isolates clustered in one group . In general, the isolates had a higher degree of similarity than with PFGE . The purK gene, which is one of the targets of the E . faecium multilocus sequence typing scheme, was sequenced . Eleven different purK alleles could be discerned, with the majority of isolates (n = 80) harboring allele 1 . With only two exceptions, all strains carrying purK-1 clustered in the same PFGE and AFLP groups, indicating a good correlation between PFGE type, AFLP type, and purK allele . Genetic polymorphism of a 571-bp PCR fragment of the C-terminal domain of the penicillin-binding protein 5 gene (pbp5) was determined, and sequence differences were associated with the level of ampicillin resistance . This study indicates that the majority of ampicillin-resistant E . faecium strains in Norway belong to a distinct genetic lineage of closely related genotypes . Rectal and clinical isolates were generally indistinguishable, and differences in clonal distribution and allele polymorphism were found mainly between ampicillin-resistant and -susceptible isolates.

Int J Food Microbiol, 2003 Jul 25, 84(2), 197 - 206
Growth and energy generation by Enterococcus faecium FAIR-E 198 during citrate metabolism; Sarantinopoulos P et al.; Citrate metabolism by Enterococcus faecium FAIR-E 198, isolated from Greek Feta cheese, was studied in various growth media containing citrate either in the presence of glucose, or as the sole carbon source, both under aerobic and anaerobic conditions . In de Man-Rogosa-Sharpe (MRS) broth with increasing citrate concentrations, cometabolism of citrate and glucose took place . Glucose was stoichiometrically converted into lactate, while citrate into acetate . Glucose consumption and biomass yield were enhanced with increasing initial citrate concentrations, even though maximum specific growth rate was not . When citrate was used as the sole carbon source in increasing initial concentrations, the main end product was acetate . Small amounts of lactate, formate, ethanol, and acetoin were also produced . In all cases, no significant differences were observed between aerobic and anaerobic conditions . However, when citrate was used as sole carbon source, formate production was favored in the absence of oxygen . The present work shows that E . faecium is able to utilize citrate in synthetic media, either in the presence of glucose or as the sole carbon source, resulting in energy production and the formation of aroma compounds.

J Biol Chem, 2003 Aug 8, 278(32), 29963 - 70 Epub 2003 May 27.
Structural basis of Synercid (quinupristin-dalfopristin) resistance in Gram-positive bacterial pathogens; Kehoe LE et al.; Synercid, a new semisynthetic streptogramin-derived antibiotic containing dalfopristin and quinupristin, is used in treatment of life-threatening infections caused by glycopeptide-resistant Enterococcus faecium and other bacterial pathogens . However, dissemination of genes encoding virginiamycin acetyltransferases, enzymes that confer resistance to streptogramins, threatens to limit the medical utility of the quinupristin-dalfopristin combination . Here we present structures of virginiamycin acetyltransferase D (VatD) determined at 1.8 A resolution in the absence of ligands, at 2.8 A resolution bound to dalfopristin, and at 3.0 A resolution in the presence of acetyl-coenzyme A . Dalfopristin is bound by VatD in a similar conformation to that described previously for the streptogramin virginiamycin M1 . However, specific interactions with the substrate are altered as a consequence of a conformational change in the pyrollidine ring that is propagated to adjacent constituents of the dalfopristin macrocycle . Inactivation of dalfopristin involves acetyl transfer from acetyl-coenzyme A to the sole (O-18) hydroxy group of the antibiotic that lies close to the side chain of the strictly conserved residue, His-82 . Replacement of residue 82 by alanine is accompanied by a fall in specific activity of >105-fold, indicating that the imidazole moiety of His-82 is a major determinant of catalytic rate enhancement by VatD . The structure of the VatD-dalfopristin complex can be used to predict positions where further structural modification of the drug might preclude enzyme binding and thereby circumvent Synercid resistance.

Biochemistry, 2003 Jun 3, 42(21), 6565 - 74
Kinetic mechanism of the GCN5-related chromosomal aminoglycoside acetyltransferase AAC(6')-Ii from Enterococcus faecium: evidence of dimer subunit cooperativity; Draker KA et al.; The aminoglycoside 6'-N-acetyltransferase AAC(6')-Ii from Enterococcus faecium is an important microbial resistance determinant and a member of the GCN5-related N-acetyltransferase (GNAT) superfamily . We report here the further characterization of this enzyme in terms of the kinetic mechanism of acetyl transfer and identification of rate-contributing step(s) in catalysis, as well as investigations into the binding of both acetyl-CoA and aminoglycoside substrates to the AAC(6')-Ii dimer . Product and dead-end inhibition studies revealed that AAC(6')-Ii follows an ordered bi-bi ternary complex mechanism with acetyl-CoA binding first followed by antibiotic . Solvent viscosity studies demonstrated that aminoglycoside binding and product release govern the rate of acetyl transfer, as evidenced by changes in both the k(cat)/K(b) for aminoglycoside and k(cat), respectively, with increasing solvent viscosity . Solvent isotope effects were consistent with our viscosity studies that diffusion-controlled processes and not the chemical step were rate-limiting in drug modification . The patterns of partial and mixed inhibition observed during our mechanistic studies were followed up by investigating the possibility of subunit cooperativity in the AAC(6')-Ii dimer . Through the use of AAC-Trp(164) --> Ala, an active mutant which exists as a monomer in solution, the partial nature of the competitive inhibition observed in wild-type dead-end inhibition studies was alleviated . Isothermal titration calorimetry studies also indicated two nonequivalent antibiotic binding sites for the AAC(6')-Ii dimer but only one binding site for the Trp(164) --> Ala mutant . Taken together, these results demonstrate subunit cooperativity in the AAC(6')-Ii dimer, with possible relevance to other oligomeric members of the GNAT superfamily.

Clin Infect Dis, 2003 Jun 1, 36(11), E146 - 8 Epub 2003 May 20.
Linezolid-resistant, vancomycin-resistant Enterococcus faecium infection in patients without prior exposure to linezolid; Rahim S et al.; We describe 2 patients without prior exposure to linezolid who were infected with closely related strains of linezolid- and vancomycin-resistant Enterococcus faecium (LRVREF) that may have been hospital acquired . Polymerase chain reaction amplification of the domain V region of the 23S ribosomal RNA gene demonstrated the presence of the G2576U mutation previously reported to be associated with linezolid resistance . Nosocomial transmission of LRVREF is an ominous sign and underscores the importance of meticulous infection-control measures.

Saudi Med J, 2003 Apr, 24(4), 331 - 6
Febrile neutropenia . Etiology of infection, empirical treatment and prophylaxis; Eltahawy AT; Much has changed in the treatment of patients with fever and neutropenia, including the patterns of microbial flora and drug resistance, and the drugs used . Gram-positive organisms have overshadowed the gram-negative ones as causes of bacteremia . Changes in therapy may include antimicrobials directed against gram-positive bacteria, resistant gram-negative bacteria, or fungi . Due to the high risk for colonization by vancomycin resistant Enterococci, vancomycin use is restricted as first line empiric therapy unless the patient is at high-risk for serious gram-positive infection . Prophylactic antibiotic therapy may increase the selection of resistant strains and should be avoided . Therapy with colony stimulating factor is only considered for patients who remain severely neutropenic and have documented infections that do not respond to appropriate antibacterial therapy . Patients stratification for risk of infection-associated morbidity and mortality is essential to facilitate treatment decision.

Syst Appl Microbiol, 2003 Mar, 26(1), 13 - 22
Virulence factors in food, clinical and reference Enterococci: A common trait in the genus?
Semedo T, Santos MA, Lopes MF, Figueiredo Marques JJ, Barreto Crespo MT, Tenreiro R.
The occurrence of several virulence traits (cytolysin, adhesins and hydrolytic enzymes) was investigated in a collection of 164 enterococci, including food and clinical isolates (from human and veterinary origin), as well as type and reference strains from 20 enterococcal species . Up to fifteen different cyl genotypes were found, as well as silent cyl genes . The occurrence of the cyl operon and haemolytic potential seems to be widespread in the genus . A significant association of this virulent trait with clinical isolates was found (p < 0.05) . High levels of incidence were also observed for genes encoding surface adhesins (esp, efaA(fs), efaA(fm)), agg and gelE, irrespectively of species allocation and origin of strains . Although gelE behaves as silent in the majority of the strains, gelatinase activity predominates in clinical isolates, whereas lipase and DNase were mainly detected in food isolates pointing to their minor role as virulence determinants . No hyaluronidase activity was detected for all strains . Numerical hierarchic data analysis grouped the strains in three main clusters, two of them including a total of 50 strains with low number of virulence determinants (from 2 to 7) and the other with 114 strains with a high virulence potential (up to 12 determinants) . No statistical association was found between virulence clusters and species allocation (p > 0.10), strongly suggesting that virulence determinants are a common trait in the genus Enterococcus . Clinical strains seem to be significantly associated with high virulence potential, whereas food, commensal and environmental strains harbour fewer virulence determinants (p < 0.01) . A high level of relative diversity in virulence patterns was observed (Shannon's index varies from 0.95 to 1.0 among clusters), reinforcing the strain-specific nature of the association of virulence factors . Although a low risk seems to be associated with the use of enterococci in long-established artisanal cheeses, screening of virulence traits and their cross-synergies must be performed, particularly for commercial starters, probiotic strains and products to be used by high risk population groups.

Chest, 2003 May, 123(5 Suppl), 504S - 18S
Vancomycin-resistant enterococci: a road map on how to prevent the emergence and transmission of antimicrobial resistance; DeLisle S et al.; Nosocomial acquisition of microorganisms resistant to multiple antibiotics represents a threat to patient safety . Here we review the mechanisms that have allowed highly resistant strains belonging to the Enterococcus genus to proliferate within our health-care institutions . These mechanisms indicate that decreasing the prevalence of resistant organisms requires active surveillance, adherence to vigorous isolation, hand hygiene and environmental decontamination measures, and effective antibiotic stewardship . We suggest how to tailor such a complex, multidisciplinary program to the needs of a particular health-care setting so as to maximize cost-effectiveness.

Infez Med, 1999, 7(3), 142 - 155
{Bacterial colangitis: therapeutic features}; Russo M et al.; Cholangitis results from the combination of bactibilia and biliary tract obstruction . In recent years considerable progress has been made in the diagnosis and treatment of cholangitis; advances in endoscopic techniques and antibiotic therapy have ameliorated the prognosis of cholangitis . The choice of an antimicrobial regimen for cholangitis should take into account the antibiotic sensitivities of bacteria colonizing biliary tree, the antibiotic excretion into bile and whether biliary obstruction or bacteremia is present . Successful treatment depends on relieving biliary obstruction and administering antibiotics effective against bacteria implicated . The initial therapy should be active against E . coli and Klebsiella spp., while it is controversial whether the empirical antibiotic regimen should also include coverage against Enterococcus, Pseudomonas and anaerobes . The ureidopenicillins are the preferred initial treatment; the combination piperacillin-tazobactam may be active against the resistant species . Second generation cephalosporins like cefamandole and cefoxitin are still useful, cefoperazone gives excellent coverage against gram-negative bacteria, while cefepime may be suitable as treatment for acute cholangitis . In severe cholangitis an aminoglycoside can be added to the beta-lactamin; once-daily aminoglycoside administration is associated with a reduced incidence of nephrotoxicity also in patients with cholestasis . Whether the fluoroquinolones are effective in treatment for cholangitis has not been fully evaluated . In patients with suppurative cholangitis prompt endoscopic drainage is mandatory, since antibiotics alone will not sterilize the biliary tract in the face of obstruction . Antibiotic prophylaxis to prevent cholangitis after ERCP should be administered particularly to patients in whom biliary drainage is expected to be difficult; antimicrobial prophylaxis with piperacillin effectively prevents ERCPinduced cholangitis . Antibiotic maintenance therapy can be highly successful in the treatment of recurrent cholangitis in patients with a compromised biliary tract.

Curr Microbiol, 2003 May, 46(5), 313 - 7
Heat-resistance and heat-shock response in the nosocomial pathogen Enterococcus faecium; Silva Laport M et al.; We have characterized the heat-shock response of the nosocomial pathogen Enterococcus faecium . The growth of E . faecium cells was analyzed at different temperatures; little growth was observed at 50 degrees C, and no growth at 52 degrees C or 55 degrees C . In agreement, a marked decrease of general protein synthesis was observed at 52 degrees C, and very light synthesis was detected at 55 degrees C . The heat resistance of E . faecium cells was analyzed by measuring the survival at temperatures higher than 52 degrees C and, after 2 h of incubation, viable cells were still observed at 70 degrees C . By Western blot analysis, two heat-induced proteins were identified as GroEL (65 kDa) and DnaK (75 kDa) . Only one isoform for either GroEL or DnaK was found . The gene expression of these heat-shock proteins was also analyzed by pulsed-labeled experiments . The heat-induced proteins showed an increased rate of synthesis during the first 5 min, reaching the highest level of induction after 10 min and returning to the steady-state level after 20 min of heat treatment.

Am J Geriatr Cardiol, 2003 May-Jun, 12(3), 183 - 9
Infective endocarditis in the elderly: diagnosis and management; Gregoratos G; Infective endocarditis is seen with increasing frequency in older patients . This increase is due to the general aging of the population, improved survival of patients with congenital and valvular heart disease, and the increasing use of catheters and other prosthetic devices with resulting higher incidence of nosocomial endocarditis . In older patients, infective endocarditis frequently develops in the absence of underlying structural heart disease; atheromatous deposits and mitral annular calcification are two important risk factors in this population . Infective organisms in older patients are frequently enterococci and other gastrointestinal tract bacteria . A marked febrile response is uncommon whereas central nervous system symptoms are more common in older patients . Transesophageal echocardiography can be performed safely and is a major diagnostic tool with sensitivity of more than 90% in detecting vegetations as small as 2-5 mm . Appropriately drawn blood cultures provide bacteriologic diagnosis in 80%-99% of patients . Prolonged antibiotic therapy may be required in many instances depending on the infective organism . Early surgical therapy is advisable for patients who develop heart failure as a result of severe acute aortic valvular regurgitation . Valve replacement surgery can be performed with acceptable mortality and morbidity even in very elderly patients .

Bioorg Med Chem Lett, 2003 May 19, 13(10), 1683 - 6
Identification of potent and broad-spectrum antibiotics from SAR studies of a synthetic vancomycin analogue; Ahrendt KA et al.; Dimeric vancomycin analogues based on a lead compound identified from a library of synthetic analogues of vancomycin have up to 60-fold greater activity than vancomycin against vancomycin-resistant Enterococcus faecium (VRE, VanA phenotype) . Simplified analogues have also been prepared and found to maintain activity against VRE and have broad-spectrum antibiotic activity.

Infez Med, 1999, 7(1), 33 - 38
{Survey on ciprofloxacin resistance: the epidemiological situation in Pordenone (Italy)}; Camporese A et al.; An epidemiological survey on 2859 bacterial strains was performed in the first nine months of 1998, to verify the predominant susceptibility to ciprofloxacin of the main microrganisms isolated in this period . Our results showed that the use of ciprofloxacin would probably decrease in future because of increasing resistance with respect to previous period . An increase in antimicrobial resistance occurred in some gram positive (as Enterococci) and especially gram negative strains such as Escherichia coli, that are common and frequent bacterial causes of infections in hospital and in the community . The study demonstrated that there are also interesting and important variations in resistance patterns with respect to previous studies

Infect Control Hosp Epidemiol, 2003 Apr, 24(4), 264 - 8
The changing epidemiology of vancomycin-resistant enterococci; Lai KK et al.; OBJECTIVE: To determine the distribution of vancomycin-resistant enterococci (VRE) cases in our hospital and those from outside of our hospital from 1993 through 1998 . METHODS: Weekly rectal surveillance was instituted whenever there were two or more cases present in the units . Cases were divided into acquired in our hospital, acquired outside of our hospital (VRE positive after and within 72 hours of admission, respectively), and indeterminate . Hospital cases were attributed to the originating ward or intensive care unit (ICU) if patients were noted to be positive within 72 hours of transfer . RESULTS: From 1993 to 1998, the rate of VRE per 1,000 admissions increased threefold, from 3.2 to 9.8, for the hospital . VRE cases acquired outside of the hospital increased by approximately 5% per year (r = 0.87; P = .03) . The rate of VRE per 1,000 admissions increased 1.7-fold in the ICUs and 3.6-fold in the wards . The ICUs had an average of 75.3 cases per year, with the number of new cases per year increasing by approximately 9 (r = 0.80; P = .028) . In the wards, there were an average of 22.0 new cases per year, with a slight upward trend of 3 additional new cases per year (r = 0.69; P = .64) . There was a highly significant increasing linear trend (P = .0007) for VRE colonization and infection . CONCLUSION: Although VRE still predominate in the ICUs, cases originating from outside of our hospital and the wards are becoming more common . VRE colonization remained more frequent than infection.

Infect Control Hosp Epidemiol, 2003 Apr, 24(4), 257 - 63
The epidemiology of vancomycin-resistant Enterococcus colonization in a medical intensive care unit; Warren DK et al.; OBJECTIVE: To determine the epidemiology of colonization with vancomycin-resistant Enterococcus (VRE) among intensive care unit (ICU) patients . DESIGN: Ten-month prospective cohort study . SETTING: A 19-bed medical ICU of a 1,440-bed teaching hospital . METHODS: Patients admitted to the ICU had rectal swab cultures for VRE on admission and weekly thereafter . VRE-positive patients were cared for using contact precautions . Clinical data, including microbiology reports, were collected prospectively during the ICU stay . RESULTS: Of 519 patients who had admission stool cultures, 127 (25%) had cultures that were positive for VRE . Risk factors for VRE colonization identified by multiple logistic regression analysis were hospital stay greater than 3 days prior to ICU admission (adjusted odds ratio {AOR}, 3.6; 95% confidence interval {CI95}, 2.3 to 5.7), chronic dialysis (AOR, 2.4; CI95, 1.2 to 4.5), and having been admitted to the study hospital one to two times (AOR, 2.3; CI95, 1.4 to 3.8) or more than two times (AOR, 6.5; CI95, 3.7 to 11.6) within the past 12 months . Of the 352 VRE-negative patients who had one or more follow-up cultures, 74 (21%) became VRE positive during their ICU stay (27 cases per 1,000 patient-ICU days) . CONCLUSION: The prevalence of VRE culture positivity on ICU admission was high and a sizable fraction of ICU patients became VRE positive during their ICU stay despite contact precautions for VRE-positive patients . This was likely due in large part to prior VRE exposures in the rest of the hospital where these control measures were not being used.

Infect Control Hosp Epidemiol, 2003 Apr, 24(4), 251 - 6
Effect of nosocomial vancomycin-resistant enterococcal bacteremia on mortality, length of stay, and costs; Song X et al.; OBJECTIVE: To determine the impact of vancomycin-resistant enterococcal bacteremia on patient outcomes and costs by assessing mortality, excess length of stay, and charges attributable to it . DESIGN: A population-based, matched, historical cohort study . SETTING: A 1,025-bed, university-based teaching facility and referral hospital . PATIENTS: Two hundred seventy-seven vancomycin-resistant enterococcal bacteremia case-patients and 277 matched control-patients identified between 1993 and 2000 . RESULTS: The crude mortality rate was 50.2% and 19.9% for case-patients and control-patients, respectively, yielding a mortality rate of 30.3% attributable to vancomycin-resistant enterococcal bacteremia . The excess length of hospital stay attributable to vancomycin-resistant enterococcal bacteremia was 17 days, of which 12 days were spent in intensive care units . On average, dollars 77,558 in extra charges was attributable to each vancomycin-resistant enterococcal bacteremia . To adjust for severity of illness, 159 pairs of case-patients and control-patients, who had the same severity of illness (All Patient Refined-Diagnosis Related Group complexity level), were further analyzed . When patients were stratified by severity of illness, the crude mortality rate was 50.3% among case-patients compared with 27.7% among control-patients, accounting for an attributable mortality rate of 22.6% . Attributable excess length of stay and charges were 17 days and dollars 81,208, respectively . CONCLUSION: Vancomycin-resistant enterococcal bacteremia contributes significantly to excess mortality and economic loss, once severity of illness is considered . Efforts to prevent these infections will likely be cost-effective.

Antiviral Res, 2003 Mar, 58(1), 17 - 24
Enterocin CRL35 inhibits late stages of HSV-1 and HSV-2 replication in vitro; Wachsman MB et al.; The replication of herpes simplex virus (HSV) type 1 and 2 in Vero cells is inhibited in the presence of enterocin CRL35 (ECRL), a bacteriocin produced by Enterococcus faecium CRL35 . Attempts to resolve the mode of action of ECRL indicate that virus adsorption and penetration are not affected . Instead, a late step of virus multiplication is hindered since the addition of 100 microg/ml of ECRL at 8h post infection still causes a 90% inhibition of virus release . The effect of ECRL on HSV antigen expression was studied by immunofluorescence using a polyclonal serum and a monoclonal antibody against glycoprotein D (gamma protein) . These studies indicated that ECRL impeded the second round of infection, apparently as a consequence of the inhibition of glycoprotein D expression . The replication of syncytial mutants of HSV-1 was significantly inhibited at a ECRL concentration of 25 microg/ml . Both the percentage of fused cells and the polykaryocyte size were affected . Studies on the effect of ECRL on viral protein synthesis showed that in the presence of ECRL, HSV late gamma proteins were not synthesized . From these findings, it is concluded that inhibition of HSV spreading by ECRL is due to the prevention of mainly late glycoprotein synthesis.

Zh Mikrobiol Epidemiol Immunobiol, 2000 Jul-Aug, (4 Suppl), 101 - 4
{Characterization of the biological properties of enterococci of different origin}; Cherdantseva GA et al.; The species composition of enterococci isolated from different ecological niches in newborn infants was determined . Pathogenicity and persistence markers were unified into a single complex which integrally determined the pathogenic potential of enterococci . The mathematical model suitable for differentiation pathogenic and nonpatogenic enterococci was developed.

Ann Oncol, 2003 May, 14(5), 795 - 801
Safety, efficacy and pharmacokinetics of linezolid for treatment of resistant Gram-positive infections in cancer patients with neutropenia; Smith PF et al.; BACKGROUND: Linezolid is a recently approved oxazalidinone with extended activity against Gram-positive bacteria . We evaluated the results of linezolid therapy in neutropenic cancer patients with Gram-positive bacterial infections from a compassionate-use program . PATIENTS AND METHODS: This was a prospective, multicenter, open-label, non-comparative, non-randomized compassionate-use treatment program in patients with serious Gram-positive infections . To qualify for enrollment patients were required to have an infection resistant to available antimicrobial agents, or in whom available agents had failed or to which they were intolerant . Patients with absolute neutrophil counts (ANC) <500 cells/mm(3) or <1000 cells/mm(3) and expected to decrease to <500 cells/mm(3), and who received linezolid 600 mg twice daily were included . Plasma samples for population pharmacokinetic analysis were collected . Clinical and microbiological assessments of outcomes were made at the end of therapy and at short-term follow-up . RESULTS: Of the patients in the compassionate-use trial, 103 were neutropenic . The mean {standard deviation (SD)} age was 50.1 (17.5) years, 47% were female, and 47.6% had a baseline ANC </=100 cells/mm(3) . The mean (SD) duration of linezolid therapy was 14.6 (11.4) days . The most common site of infection was the bloodstream (90.3%), and the most commonly identified pathogen was vancomycin-resistant Enterococcus faecium (83%) . A total of 83 (80.5%) and 52 (50.4%) patients were evaluable for clinical and microbiological outcomes at the end of therapy, respectively . Clinical and microbiological cure rates in the evaluable patients were 79% and 86%, respectively . Linezolid was well-tolerated in this patient population, with an overall adverse event rate of 17.5%; 5% of patients required discontinuation of the drug due to side-effects . The pharmacokinetics of linezolid in patients with neutropenia did not differ from the overall compassionate-use population . CONCLUSIONS: Linezolid was safe and effective in treating resistant Gram-positive infections in neutropenic cancer patients . Comparative clinical trials to evaluate further the effectiveness and safety of linezolid in this patient population are warranted.

J Endod, 2003 Apr, 29(4), 261 - 4
The comparative sealing ability of hydroxyapatite cement, mineral trioxide aggregate, and super ethoxybenzoic acid as root-end filling materials; Mangin C et al.; A comparison was made of the ability of hydroxyapatite cement, mineral trioxide aggregate, and super ethoxybenzoic acid to prevent the leakage of bacteria from root canals, when used as root-end filling materials . The materials were tested in a double-chamber device in which a root segment connects the upper (delivery) chamber and the lower (receiving) chamber . The root segment was prepared by having the root canal instrumented to a #45 file, and a 3-mm-deep, root-end preparation placed at the apical foramen . The canal of each root segment was filled with gutta-percha, and the root-end preparation was filled with one of three test materials, mixed according to the manufacturer's directions . Negative controls were constructed with sticky wax sealing the apical foramen . A titered suspension of radioactively (3H-thymidine)-labeled bacteria (Enterococcus fecalis) was placed into the delivery chamber, and sterile saline was placed into the receiving chamber such that the apical third of each root section was immersed . At various time points, samples were taken from the receiving chamber, and measured for 3H activity . The results indicated that (a) all the test materials leaked significantly compared with the negative controls; and (b) there was no significant difference found between the leakage rates of the three materials tested.

J Clin Microbiol, 2003 Apr, 41(4), 1499 - 506
Influence of transferable genetic determinants on the outcome of typing methods commonly used for Enterococcus faecium; Werner G et al.; A variety of methods is used for a molecular typing of Enterococcus spp . and related gram-positive bacteria including macrorestriction analysis using pulsed-field gel electrophoresis (PFGE), ribotyping, rapid amplification of polymorphic DNA (RAPD), and amplified fragment length polymorphism (AFLP) . To test the influence of transferable determinants on the outcome of different typing methods commonly used for enterococci, we established a homogenous strain collection of 24 transconjugants resulting from filter matings with antibiotic-resistant Enterococcus faecium . As expected, AFLP, RAPD, and PFGE all identified our model bacteria as strongly related . However, distinct differences in the resolving and discriminatory power of the tested methods could be clearly addressed . In PFGE, 22 of 24 transconjugants possessed less than a three-band difference to the recipient pattern and would be regarded as strongly related . Three different RAPD PCRs were tested; in two reactions, identical patterns for all transconjugants and the recipient were produced . One RAPD PCR produced an identical pattern for 18 transconjugants and the recipient and a clearly different pattern for the remaining 6 transconjugants due to a newly appearing fragment resulting from acquisition of the tetL gene . AFLP clusters all transconjugants into a group of major relatedness . Percent similarities were highly dependent on the method used for calculating the similarity coefficient (curve-based versus band-based similarity coefficient) . Fragment patterns of digested plasmids showed the possession of nonidentical plasmids in most transconjugants . PFGE still could be recommended as the method of choice . Nevertheless, the more-modern AFLP approach produces patterns of comparable discriminatory power while possessing some advantages over PFGE (less-time-consuming internal standards) . Plasmid fingerprints can be included to subdifferentiate enterococcal isolates possessing identical macrorestriction and PCR typing patterns.

Lancet Infect Dis, 2003 Apr, 3(4), 241 - 9
Assessing risks for a pre-emergent pathogen: virginiamycin use and the emergence of streptogramin resistance in Enterococcus faecium; Smith DL et al.; Vancomycin-resistant enterococci (VRE) are an important cause of hospital-acquired infections and an emerging infectious disease . VRE infections were resistant to standard antibiotics until quinupristin/dalfopristin (QD), a streptogramin antibiotic, was approved in 1999 for the treatment of vancomycin-resistant Enterococcus faecium infections in people . After that decision, the practice of using virginiamycin in agriculture for animal growth promotion came under intense scrutiny . Virginiamycin, another streptogramin, threatens the efficacy of QD in medicine because streptogramin resistance in enterococci associated with food animals may be transferred to E faecium in hospitalised patients . Policy makers face an unavoidable conundrum when assessing risks for pre-emergent pathogens; good policies that prevent or delay adverse outcomes may leave little evidence that they had an effect . To provide a sound basis for policy, we have reviewed the epidemiology of E faecium and streptogramin resistance and present qualitative results from mathematical models . These models are based on simple assumptions consistent with evidence, and they establish reasonable expectations about the population-genetic and population-dynamic processes underlying the emergence of streptogramin-resistant E faecium (SREF) . Using the model, we have identified critical aspects of SREF emergence . We conclude that the emergence of SREF is likely to be the result of an interaction between QD use in medicine and the long-term use of virginiamycin for animal growth promotion . Virginiamycin use has created a credible threat to the efficacy of QD by increasing the mobility and frequency of high-level resistance genes . The potential effects are greatest for intermediate rates of human-to-human transmission (R0 approximately equal 1).

Environ Health Perspect, 2003 Apr, 111(4), 598 - 603
Evaluation of recreational health risk in coastal waters based on enterococcus densities and bathing patterns; Turbow DJ et al.; We constructed a simulation model to compute the incidences of highly credible gastrointestinal illness (HCGI) in recreational bathers at two intermittently contaminated beaches of Orange County, California . Assumptions regarding spatial and temporal bathing patterns were used to determine exposure levels over a 31-month study period . Illness rates were calculated by applying previously reported relationships between enterococcus density and HCGI risk to the exposure data . Peak enterococcus concentrations occurred in late winter and early spring, but model results showed that most HCGI cases occurred during summer, attributable to elevated number of exposures . Approximately 99% of the 95,010 illness cases occurred when beaches were open . Model runs were insensitive to 0-10% swimming activity assumed during beach closure days . Comparable illness rates resulted under clustered and uniform bather distribution scenarios . HCGI attack rates were within federal guidelines of tolerable risk when averaged over the study period . However, tolerable risk thresholds were exceeded for 27 total days and periods of at least 6 consecutive days . Illness estimates were sensitive to the functional form and magnitude of the enterococcus density-HCGI relationships . The results of this study contribute to an understanding of recreational health risk in coastal waters.

Int J Food Microbiol, 2003 May 25, 83(1), 27 - 38
The stimulating effect of a harsh environment on the bacteriocin activity by Enterococcus faecium RZS C5 and dependency on the environmental stress factor used; Leroy F et al.; Bacteriocin production by Enterococcus faecium RZS C5 occurs in a growth-associated way but is generally switched off in the very early growth phase . The influence of environmental stress on the bacteriocin production kinetics by E . faecium RZS C5 was analysed at a controlled temperature of 35 degrees C and constant pH 6.5 . The effect of environmental stress on bacteriocin production was depending on the type of stress applied . Oxidative stress did not interfere with cell growth or bacteriocin activity . In contrast, salt stress decreased both the cell growth and the specific bacteriocin production . Nevertheless, moderate levels of sodium chloride improved bacteriocin activity because they increased the biomass concentration at which bacteriocin production was switched off . Environmental stress due to limitations in sugar or complex nutrients did not affect the early shut-off mechanism or the specific bacteriocin production . However, bacteriocin stability decreased or increased at low levels of sugar or complex nutrients, respectively.

Diagn Microbiol Infect Dis, 2003 Mar, 45(3), 213 - 5
Treatment of central nervous system infection by vancomycin-resistant enterococcus faecium; Kanchanapoom T et al.; Enterococci are uncommon causes of CNS infection . We describe a case of ventriculitis and Ommaya reservoir infection due to vancomycin-resistant Enterococcus faecium successfully treated with the combination of i.v . quinupristin/dalfopristin and i.v . linezolid . The patient deteriorated after receiving three dosages of intraventricular quinupristin/dalfopristin . He recovered after discontinuation of intraventricular quinupristin/dalfopristin.

Antimicrob Agents Chemother, 2003 Apr, 47(4), 1423 - 6
Multiplex PCR for detection of aminoglycoside resistance genes in enterococci; Vakulenko SB et al.; A multiplex PCR procedure for detecting the aminoglycoside resistance genes aac(6')-Ie-aph(2")-Ia, aph(2")-Ib, aph(2")-Ic, aph(2")-Id, aph(3')-IIIa, and ant(4')-Ia was evaluated and found to determine accurately the presence of these genes in enterococci.

Rinsho Byori, 2002 Nov, Suppl 123, 62 - 70
{Vancomycin resistant enterococcal infections and detection of their resistance genes}; Fujita N; Vancomycin-resistant enterococci(VRE) is a troublesome pathogens causing nosocomial infections worldwide . VRE infections or colonization occur mainly in immuno-compromised patients and in the course of antimicrobial treatment for other nosocomial infectious diseases . VRE are classified into 6 types by their resistance genes and of these types, both VanA and VanB type VRE are significantly important in clinical situations . Susceptibility tests for VCM and TEIC are often useful in the phenotypic screening for differentiating VanA from VanB type VRE . However, detection of the resistance genes using PCR methods is easy and essential for the conclusive identification of VRE.

J Biol Chem, 2003 Jun 6, 278(23), 21162 - 7 Epub 2003 Mar 21.
The membrane domain of the Na+-motive V-ATPase from Enterococcus hirae contains a heptameric rotor; Murata T et al.; In F-ATPases, ATP hydrolysis is coupled to translocation of ions through membranes by rotation of a ring of c subunits in the membrane . The ring is attached to a central shaft that penetrates the catalytic domain, which has pseudo-3-fold symmetry . The ion translocation pathway lies between the external circumference of the ring and another hydrophobic protein . The H+ or Na+:ATP ratio depends upon the number of ring protomers, each of which has an essential carboxylate involved directly in ion translocation . This number and the ratio differ according to the source, and 10, 11, and 14 protomers have been found in various enzymes, with corresponding calculated H+ or Na+:ATP ratios of 3.3, 3.7, and 4.7 . V-ATPases are related in structure and function to F-ATPases . Oligomers of subunit K from the Na+-motive V-ATPase of Enterococcus hirae also form membrane rings but, as reported here, with 7-fold symmetry . Each protomer has one essential carboxylate . Thus, hydrolysis of one ATP provides energy to extrude 2.3 sodium ions . Symmetry mismatch between the catalytic and membrane domains appears to be an intrinsic feature of both V- and F-ATPases.

Med Dosw Mikrobiol, 2002, 54(4), 317 - 24
{Susceptibility of enterococci to natural and synthetic iron chelators}; Lisiecki P et al.; A total of 79 strains of enterococci belonging to 10 species were tested for susceptibility to natural and synthetic iron chelators . All strains produced siderophores . These enterococci were susceptible to three synthetic iron chelators only: 8-hydroxyquinoline, disodium versenate (EDTA) and o-phenanthroline . They were resistant to all other synthetic chelators: ethylenediamine-di(o-hydroxyphenylacetic acid) (EDDHA), nitrilotriacetate, 2,2'-bipiridyl, salicylic acid, 8-hydroxy-5-sulphonic acid and to all natural chelators: ovotransferrine, human apotransferrine, horse apoferritine, desferrioxamine B, ferrichrome and rhodotorulic acid . The relations between susceptibility/resistance, iron assimilation and structure and stability constants of iron chelators were discussed.

J Med Chem, 2003 Mar 27, 46(7), 1204 - 9
Role of the glycopeptide framework in the antibacterial activity of hydrophobic derivatives of glycopeptide antibiotics; Printsevskaya SS et al.; The antibacterial properties of glycopeptide antibiotics are based on their interaction with the d-Ala-d-Ala containing pentapeptide of bacterial peptidoglycan . The hydrophobic amides of vancomycin (1), teicoplanin (2), teicoplanin aglycon (3), and eremomycin (4) were compared with similar amides of minimally or low active des-(N-methyl-d-leucyl)eremomycin (5), eremomycin aglycon (6), des-(N-methyl-d-leucyl)eremomycin aglycon (7), and a teicoplanin degradation product TB-TPA (8) . All hydrophobic amides of 1, 3, 4, and 6 were almost equally active against glycopeptide-resistant enterococci (GRE) {minimum inhibitory concentrations (MIC) <or= 4 microg/mL} and had better activity against Gram-positive strains sensitive to glycopeptides than against GRE . Extensive degradation of the glycopeptide framework in amides of 7 and 8 led to a decrease of anti-GRE activity (MIC = 16-64 microg/mL), and for these derivatives MIC values for bacterial strains sensitive and resistant to glycopeptides were very close . These results suggest that in sensitive bacteria two mechanisms of action are operating for the hydrophobic derivatives of glycopeptide antibiotics with the nondamaged peptide core-interaction with the d-Ala-d-Ala moiety and the inhibition of bacterial membrane bound enzymatic reactions, whereas for GRE lacking the d-Ala-d-Ala fragment, only the second mechanism is operating . It appears that a minimal glycopeptide core is required for activity, and that more extensive degradation results in a serious decrease of antibacterial activity.

APMIS, 2002 Dec, 110(12), 869 - 74
Modified pulsed-field gel electrophoresis protocol for typing of enterococci; Saeedi B et al.; Controlling the spread of vancomycin-resistant enterococci (VRE) is an important task in hospital epidemiology . Pulsed-field gel electrophoresis (PFGE) has become the golden standard for molecular epidemiological characterisation of enterococcal isolates . For separation of DNA fragments by PFGE, different electrophoresis conditions have been recommended, but none of these protocols allows a satisfactory separation of both small and large DNA fragments of enterococci simultaneously . In this study we have speeded up the preparation of chromosomal DNA and defined new electrophoresis conditions that enhance separation of small and large DNA fragments for subtyping of enterococci with a 24 h PFGE.

Euro Surveill, 1997 Dec, 2(12), 97 - 98
Increase of the number of vancomycin resistant enterococci (VRE) isolated in Helsinki, Finland; Vuopio-Varkila J et al.; The emergence of hospital acquired infections with bacteria resistant to antimicrobials such as vancomycin resistant enterococci (VRE) has become a worldwide concern . In hospitals in the United States, VRE have spread quickly and currently account for eve

Folia Microbiol (Praha), 2002, 47(6), 649 - 53
PCR-based methods for identification of Enterococcus species; Drahovska H et al.; Two DNA-based techniques were used for species identification of enterococci . PvuII digestion of the genus-specific PCR product yielded four different restriction profiles among 20 enterococcal species; one of them was species-specific for E . faecium . In the second case, 32 reference strains belonging to 20 enterococcal species were divided to 12 groups by amplification of internal transcribed spacer of rRNA operon . Interspecies and some intraspecies profile variability was determined . Both methods gave similar results.

J Hosp Infect, 2003 Mar, 53(3), 159 - 71
Vancomycin-resistant enterococci: 15 years and counting; Chavers LS et al.; We review the history of vancomycin-resistant enterococci (VRE) and propose a causal model illustrating the roles of exposure to VRE reservoirs, patient characteristics, antimicrobial exposure, and prevalence of VRE in the progression from potential VRE reservoirs to active disease in hospitalized patients . Differences in VRE colonization and VRE infection are discussed with respect to hospital surveillance methodology and implications for interventions . We further document clonal transmission of VRE in a large, urban, teaching hospital and demonstrate VRE susceptibility to a wide array of antimicrobial agents . This model can guide the identification of mutable factors that are focal points for intervention.

J Antimicrob Chemother, 2003 Mar, 51(3), 703 - 6
The vanC-3 vancomycin resistance gene cluster of Enterococcus flavescens CCM 439; Dutta I et al.; Enterococcus flavescens CCM 439 is phenotypically similar to Enterococcus casseliflavus; it possesses intrinsic low-level resistance to vancomycin and has the VanC phenotype . The complete vanC-3 vancomycin resistance gene cluster was cloned and sequenced, and found to contain five open reading frames . These encoded five proteins that displayed a high degree of amino acid identity to the proteins of the vanC-2 cluster of E . casseliflavus . The serine racemases displayed the lowest degree of identity (97%), whereas the response regulators VanR(C-2) and VanR(C-3) were 100% identical . Long-PCR-RFLP analysis of the vanC-3 and vanC-2 gene clusters distinguished E . flavescens CCM 439 from E . casseliflavus ATCC 25788 due to the absence of a single EcoRV restriction endonuclease site from the E . flavescens gene cluster . However, the lack of nucleotide divergence between the sequences of the vanC-2 and vanC-3 clusters casts doubt on the validity of E . flavescens and E . casseliflavus being classed as distinct species.

J Dairy Sci, 2003 Jan, 86(1), 331 - 5
Direct-fed microbial supplementation on the performance of dairy cattle during the transition period; Nocek JE et al.; The influence of a direct-fed microbial (DFM) on the prepartum period and the effects on production performance during the postpartum period was investigated using 64 multiparous Holstein cows . Two close-up dry cow diets were fed to two groups of 32 cows each starting 21 d precalving as follows: 1) no DFM and 2) DFM . Post-calving cows were fed a lactation ration with or without DFM supplementation . Direct-fed microbial supplementation contained approximately 5 x 10(9) cfu of yeast and 5 x 10(9) cfu of bacteria (Enterococcus faecium, two strains) topdressed in a 90-g supplement/cow per day in both pre- and postpartum periods . The DMI during the prepartum period was not affected by DFM supplementation . During the postpartum period, DMI, milk yield, and milk protein content were higher for cows receiving DFM supplementation compared with no DFM . Blood glucose and insulin levels were higher and NEFA levels were lower for cows receiving DFM during the postpartum period . These data suggest that targeted DFM supplementation increased DMI and milk production postpartum . Blood metabolite information would suggest this response was associated with more glucose being made available and less fatty acids being mobilized from lipid stores.

Kansenshogaku Zasshi, 2002 Dec, 76(12), 1035 - 9
{A critical pneumonia by multidrug-resistant Enterococcus faecium in a chronic hemodialysis patient . A case report}; Morii K et al.; Enterococcus faecium is usually known as a pathogen of nosocomial infections in compromised patients . Recently it is a problem that some of the E . faecium become resistant to many antibiotics . It is not usual that this pathogen causes pneumonia . We report a community-acquired pneumonia by multidrug-resistant E . faecium in an outpatient who had been receiving hemodialysis for the past year . The patient recovered from pneumonia with vancomycin, but had nosocomial pneumonia caused by MRSA later but recovered.

Antimicrob Agents Chemother, 2003 Mar, 47(3), 1154 - 6
Vancomycin resistance is maintained in enterococci in the viable but nonculturable state and after division is resumed; Lleo MM et al.; Stressed vancomycin-resistant enterococci (VRE) can activate a survival strategy known as the viable but nonculturable (VBNC) state and are able to maintain vancomycin resistance . During restoration of division they continue to express the vancomycin resistance trait . We suggest that VBNC enterococci may constitute further reservoirs of VRE and therefore represent an additional risk for human health.

Cir Pediatr, 2002 Apr, 15(2), 79 - 81
{Decrease in bacterial translocation in burned children treated with controlled nutritional support}; Rivas S et al.; INTRODUCTION: Alteration in epithelial enteric cells are the main cause of bacteria translocation and local and systemic infections . Our aim is to evaluate these infections in burned children treated with enteral nutrition and compare the results with our previous observations . Material and methods . We designed a prospective study including 50 patients (0-14 years) with TBSA > 10% (total body surface area) . Bacteriological cultures were performed at their admittance and weekly until the discharge . Nutritional necessities were evaluated using calorimetry . Nutritional support was supplied by nasoduodenal tube at continuous rates . RESULTS: Three (6%) patients with enteral nutrition (Group 1) showed positive blood cultures (two of them by enteric bacteriae) . 14 patients (28%) showed local infections (five by enterococci) . We performed a retrospective study in a homogenous group of 44 patients (Group 2), previously treated in our Department without controlling nutrition . We found five (11.3%) positive blood cultures (four by enteric bacteriae) and 22 (50%) local infections (eleven by enteric bacteriae) . The improvement of systemic and local infection rates and the decrease in infections caused by enteric bacteriae were statistically significative (p < 0.05) . CONCLUSIONS: A controlled nutritional support is a main cause of the improvement in infection rates and decrease in bacterial translocation in pediatric burned patients.

Rev Argent Microbiol, 2002 Oct-Dec, 34(4), 222 - 5
{Detection of vancomycin-resistant Enterococcus faecium: 2 years' experience in a high complexity hospital }; Lopreto CR et al.; The presence of vancomycin-resistant enterococci (VRE) in our hospital prompted us to apply an appropriate method for assessing its rectal carriage . A screening method with bile-esculin azide agar plus different concentrations of vancomycin was used . The antimicrobial susceptibility study of enterococci isolated from clinical samples was also emphasized . The present study includes the surveillance and detection of VRE in our hospital during two years . A total of 260 samples corresponding to 138 patients were studied, 158 of them resulting positive . All EVR were Van A Enterococcus faecium, with MICs of vancomycin > or = 256 micrograms/ml . The analysis of susceptibility patterns shows variations with chloramphenicol, tetracycline and high level gentamicin concentrations . This method was easily applied because materials could be available in any clinical microbiology laboratory, and in our hands it has demonstrated to be useful for epidemiological surveillance for EVR.

J Mol Biol, 2003 Mar 7, 326(5), 1437 - 48
A comprehensive view on proteasomal sequences: implications for the evolution of the proteasome; Gille C et al.; Proteasomes are large multimeric self-compartmentizing proteases, which play a crucial role in the clearance of misfolded proteins, breakdown of regulatory proteins, processing of proteins by specific partial proteolysis, cell cycle control as well as preparation of peptides for immune presentation . Two main types can be distinguished by their different tertiary structure: the 20S proteasome and the proteasome-like heat shock protein encoded by heat shock locus V, hslV . Usually, each biological kingdom is characterized by its specific type of proteasome . The 20S proteasomes occur in eukarya and archaea whereas hslV protease is prevalent in bacteria . To verify this rule we applied a genome-wide sequence search to identify proteasomal sequences in data of finished and yet unfinished genome projects . We found several exceptions to this paradigm: (1) Protista: in addition to the 20S proteasome, Leishmania, Trypanosoma and Plasmodium contained hslV, which may have been acquired from an alpha-proteobacterial progenitor of mitochondria . (2) Bacteria: for Magnetospirillum magnetotacticum and Enterococcus faecium we found that each contained two distinct hslVs due to gene duplication or horizontal transfer . Including unassembled data into the analyses we confirmed that a number of bacterial genomes do not contain any proteasomal sequence due to gene loss . (3) High G+C Gram-positives: we confirmed that high G+C Gram-positives possess 20S proteasomes rather than hslV proteases . The core of the 20S proteasome consists of two distinct main types of homologous monomers, alpha and beta, which differentiated into seven subtypes by further gene duplications . By looking at the genome of the intracellular pathogen Encephalitozoon cuniculi we were able to show that differentiation of beta-type subunits into different subtypes occurred earlier than that of alpha-subunits . Additionally, our search strategy had an important methodological consequence: a comprehensive sequence search for a particular protein should also include the raw sequence data when possible because proteins might be missed in the completed assembled genome . The structure-based multiple proteasomal alignment of 433 sequences from 143 organisms can be downloaded from the URL dagger and will be updated regularly.

Surg Infect (Larchmt), 2000, 1(1), 57 - 63
The role of beta-lactam antimicrobials as single agents in treatment of intra-abdominal infection; Powell LL et al.; Broad-spectrum beta-lactam antibiotics have several advantages in the treatment of intra-abdominal infections . These agents are effective against gram-negative rods and anaerobes, reach therapeutic levels rapidly after parenteral administration, and, in the absence of penicillin allergy, generally exhibit low toxicity . The second-generation cephalosporins (e.g., cefoxitin, cefotetan) are used widely in surgical prophylaxis, trauma, and treatment of mild-to-moderate community-acquired infections, but limitations in their spectra and microbial resistance restrict their utility in more serious infections . Extended-spectrum penicillin/beta-lactamase-inhibitor combinations are effective in the treatment of intra-abdominal infections and include enterococci in their spectrum . Gram-negative aerobe resistance has developed to ampicillin/sulbactam . Piperacillin/tazobactam, a ureidopenicillin with increased gram-negative coverage and enhanced antipseudomonal activity, has proved to be effective in clinical trial therapy for intra-abdominal infections . The very broad spectrum carbapenems--imipenem/cilastatin and meropenem--are effective for serious infections or resistant organisms and are often used in the intensive care unit or for nosocomial intra-abdominal infection . These classes of beta-lactams comprise a range of antimicrobials that can be targeted effectively as single agents to both prevention and treatment of intra-abdominal infection.

Surg Infect (Larchmt), 2000 Summer, 1(2), 95 - 107
Coverage of enterococci in community acquired secondary peritonitis: results of a randomized trial; Rohrborn A et al.; INTRODUCTION: The increasing number of enterococcal infections in hospitals and reports about the development of resistance of these bacteria make it necessary to review their importance as co-pathogens in secondary peritonitis . MATERIALS AND METHODS: A prospective randomized controlled trial on primary therapy of secondary peritonitis was carried out in six centers comparing cephalosporin-based antibiotic therapy to acylaminopenicillin-based therapy . RESULTS: Enterococci were only cultured in 6 of 110 cases from the abdomen and were found in only 5 cases of postoperative complications . No differences were found between penicillin-based vs . cephalosporin-based therapy . CONCLUSION: The study supports the view that these bacteria continue to play a minor role in secondary peritonitis . The point has to be emphasized, however, that the patients under study were in relatively good condition (APACHE II median 9 for cephalosporins and 10 for penicillins) and that postoperative cases of peritonitis were excluded.

Surg Infect (Larchmt), 2000 Autumn, 1(3), 187 - 96
Emerging bacterial pathogens: a consensus of the scientific data and the risk for development of multiple organ dysfunction syndrome; Rapp RP; Antibiotic resistance in the hospital setting is continuing to increase, particularly in intensive care units (ICUs) and other areas of the hospital such as oncology units, where the use of empiric broad-spectrum antibiotics is common . The problem of antibiotic resistance is also compounded in the immunocompromised patient . Multi-drug resistance is common among both Gram-positive and -negative bacteria, and becoming more prevalent among fungi (yeast) . Two major antibiotic-resistant pathogens include extended-spectrum beta-lactamase producing Klebsiella pneumoniae (ESBL-KP) and vancomycin-resistant enterococci (VRE) . When infections occur with ESBL-KP, a carbapenem antibiotic is usually the drug of choice . When infection occurs with VRE, specific therapy is bacteriostatic, and the clinician may have to rely on empirically selected antibiotics or combinations of antibiotics to achieve a positive outcome . Two newly-approved agents, linezolid and quinupristin/dalfopristin can be used to treat infections caused by resistant gram-positive cocci, but the latter is approved for use against VR-E . faecium . Risk factors for the development of ESBL-KP include the use of extended-spectrum cephalosporins such as ceftazidime . Risk factors for the development of VRE include inappropriate use of vancomycin, extended-spectrum cephalosporins, and antianaerobic drug therapy such as clindamycin . Several institutions have documented a reduction in one or both of these resistant pathogens following a decrease in the use of extended-spectrum cephalosporins combined with the increased use of extended-spectrum penicillins/beta-lactamase inhibitor combinations, such as piperacillin/tazobactam, for the empiric therapy of infections . For VRE, a reduction in the inappropriate use of vancomycin is also an important interventional strategy along with improved infection control practice.

Infection, 2003 Jan, 31(1), 9 - 14
Antibiotic use and the prevalence of antibiotic resistance in bacteria from healthy volunteers in the dutch community; Bruinsma N et al.; BACKGROUND: Although most antibiotics are consumed in the community (80%), the relation of antibiotic consumption and resistance in the community has not been thoroughly studied . MATERIALS AND METHODS: In the present study, antibiotic use was related to the prevalence of antibiotic resistance of Escherichia coli and enterococci isolated from healthy volunteers living in the southern part of the Netherlands in 1996 and 1999 . RESULTS: No change in the total antibiotic use in the Dutch community was observed between 1996 and 1999 (3542 and 3598 defined daily doses {DDD} per 1000 inhabitants/year) . However, the increased fluoroquinolone use (+ 18%) and the increased prevalence of ciprofloxacin resistance from 0 to 2% is a point of concern, especially since this was accompanied by a significant shift towards higher minimum inhibitory concentration (MIC) values (p < 0.05) . A significant decrease in the prevalence of vancomycin-resistant enterococci (p < 0.05) was found in addition to a significant shift towards lower MIC valued for avoparcin, a glycopeptide previously used as growth promoter in animal husbandry (p < 0.05) . This was very likely due to the banning of avoparcin for this purpose from April 1997 onwards . CONCLUSION: In order to maintain the low level of antibiotic use and resistance in The Netherlands, surveillance of antibiotic resistance in nonclinical isolates in relation to antibiotic use is very important.

FEMS Microbiol Rev, 2003 Jan, 26(5), 511 - 32
Mechanism of action of oritavancin and related glycopeptide antibiotics; Allen NE et al.; Oritavancin (LY333328) is a semisynthetic glycopeptide antibiotic having excellent bactericidal activity against glycopeptide-susceptible and -resistant Gram-positive bacteria . Oritavancin is the N-alkyl-p-chlorophenylbenzyl derivative of chloroeremomycin (LY264826) and is currently in phase III clinical trials for use in Gram-positive infections . Studies show that oritavancin and related alkyl glycopeptides inhibit bacterial cell wall formation by blocking the transglycosylation step in peptidoglycan biosynthesis in a substrate-dependent manner . As with other glycopeptide antibiotics, including vancomycin, the effects of oritavancin on cell wall synthesis are attributable to interactions with dipeptidyl residues of peptidoglycan precursors . Unlike vancomycin, however, oritavancin is strongly dimerized and can anchor to the cytoplasmic membrane, the latter facilitated by its alkyl side chain . Cooperative interactions derived from dimerization and membrane anchoring in situ can be of sufficient strength to enable binding to either dipeptidyl or didepsipeptidyl peptidoglycan residues of vancomycin-susceptible and -resistant enterococci, respectively . This review describes the antibacterial activity of oritavancin, and examines the evidence supporting the proposed mechanism of action for this agent and related analogs.

Enferm Infecc Microbiol Clin, 2003 Mar, 21(3), 157 - 64; quiz 165, 169
{Oxazolidinones and glycopeptides}; Pigrau C; We review the chemical structure, pharmacokinetics, antimicrobial spectrum, mechanisms of action and resistance, clinical use and adverse effects of oxazolidinones (linezolid) and glycopeptides . Oxazolidinones inhibit protein synthesis and glycopeptides cell wall synthesis . Antibacterial spectra cover grampositive pathogens, including multiresistant organisms . Linezolid has a 100% absorption and can be administered orally or intravenously; it is cleared by renal and hepatic routes, thus dose adjustments are not needed in moderate renal or hepatic failure . Oral absorption of glycopeptides is minimal . They are excreted by the kidneys (dose adjustments are needed in renal failure) . The main clinical indications are the treatment of infections caused by grampositive pathogens; linezolid is also useful for infections caused by glycopeptide-resistant enterococcal infections . Linezolid can cause trombocytopenia when treatment lasted longer than two weeks . The main side effect of vancomycin is its nefrotoxicity and teicoplanin can cause fever.

Lett Appl Microbiol, 2003, 36(3), 186 - 90
Identification of a novel insertion sequence in vanB2-containing Enterococcus faecium; Lee WG et al.; AIMS: The characterization of a novel insertion sequence (IS) in vanB2-containing Enterococcus faecium was conducted . METHODS AND RESULTS: Direct PCR amplification of ORFC region of Tn5382 from DNA extracted from vanB2-containing E . faecium, and sequence analysis were performed . A novel IS was identified . It is 1418 bp in length and contains one putative open reading frame that is similar to transposase . There exists inverted terminal repeats of 12 bp, but direct repeats are not present . According to high similarity to putative transposases of IS3 members, such as, IS150, IS861, IS1077 and IS911, we designated it ISEnfa3 . SIGNIFICANCE AND IMPACT OF STUDY: Since ISEnfa3 was detected in all vanB2-containing strains examined so far, it could be used as a tool for epidemiological study.

What is Food Microbiology?, What Is Yeast?, What Is Antibiotic?, What Is Molecular Biology?, What Is Biofilter?, o, Microorganisms, e, Microbe, r, Bacteriology, s, Microbiology, i, Bacteria, s, Streptococcal, c, Escherichia coli, n, Penicillin, e, Microorganisms, i, Shigella, i, Microorganism, n, Escherichia coli, a, Yeasts, s, Denitrifying, i, Microbiological, i, Micrococci, c, Corynebacterium, n, Corynebacterium, o, Cell cultures, a, Escherichia coli, i, Microbiological, s, S. cerevisiae, n, Staphylococcus aureus, s, Lactococci, e, Yeasts, i, S. cerevisiae

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005