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Oligonucleotide microarray for identification of Enterococcus species.

For detection of most members of the Enterococcaceae, the specificity of a novel oligonucleotide microarray (ECC-PhyloChip) consisting of 41 hierarchically nested 16S or 23S rRNA gene-targeted probes was evaluated with 23 pure cultures (including 19 Enterococcus species). Target nucleic acids were prepared by PCR amplification of a 4.5-kb DNA fragment containing large parts of the 16S and 23S rRNA genes and were subsequently labeled fluorescently by random priming. Each tested member of the Enterococcaceae was correctly identified on the basis of its unique microarray hybridization pattern. The evaluated ECC-PhyloChip was successfully applied for identification of Enterococcus faecium and Enterococcus faecalis in artificially contaminated milk samples demonstrating the utility of the ECC-PhyloChip for parallel identification and differentiation of Enterococcus species in food samples.

DNA, Bacterial↗

Bacterial flora of leg ulcers in patients admitted to Department of Dermatology, Poznań University of Medical Sciences, during the 1998-2002 period.

Venous leg ulcers are an important cause of morbidity in a significant percentage of the world population. The percentage of leg ulceration, either active or healed, in the European population is about 1%-2%. The aim of this study was to analyze the rate of colonization and qualitative composition of the bacterial flora isolated from leg ulcers in patients admitted to Dermatology Department, Poznań University of Medical Sciences, during the 1998-2002 period, with special reference to the infection risk factors. Bacteriological diagnosis of 175 wound swabs was performed in compliance with compulsory laboratory methods. In 173 positive results, the predominant culture composition included Staphylococcus aureus (56.57%), Pseudomonas aeruginosa (37.14%), Enterococcus faecalis (22.29%), Proteus mirabilis (13.71%) and Escherichia coli (12.57%). There was a significant increase in the incidence of Pseudomonas aeruginosa, Enterococcus faecalis and Escherichia coli, along with a decrease in the incidence of Staphylococcus aureus isolation during the study period. The rate of yeast-like fungi strains, mainly Candida albicans, recorded in culture composition showed a systematic increase. Changes in the qualitative and quantitative composition of bacterial flora, presence of multiple isolates, and concomitant diseases that may influence the characteristics of leg ulcer disease were closely monitored.

Academic Medical Centers↗

Sparfloxacin and clinafloxacin alone or in combination with gentamicin for therapy of experimental ampicillin-resistant enterococcal endocarditis in rabbits.

Sparfloxacin and clinafloxacin alone and in combination with gentamicin, were evaluated for the treatment of experimental endocarditis in rabbits caused by ampicillin-resistant enterococci. Clinafloxacin was tested against Enterococcus faecalis strain WH245, a beta-lactamase producer lacking high-level gentamicin resistance (MIC 12.5 mg/L). Sparfloxacin was tested against Enterococcus faecium strain SF2149 a non-producer of beta-lactamase (ampicillin MIC 400 mg/L, gentamicin MIC 12.5 mg/L). For strain WH245, clinafloxacin alone significantly reduced enterococcal counts in vegetations (7.7 log10 cfu/g) and for strain SF2149, sparfloxacin significantly reduced counts (7.0 log10 cfu/g) compared with untreated controls (WH245, 8.8 log10 cfu/g and SF2149, 9.3 log10 cfu) or treatment with ampicillin plus gentamicin (WH245, 9.7 log10 cfu/g). The addition of gentamicin resulted in no further reduction of bacterial counts in vegetations but resulted in an increase in sterilization of blood for strain SF2149. These results suggest that sparfloxacin and clinafloxacin and may prove useful in the therapy of infections due to ampicillin-resistant enterococci.

Ampicillin Resistance↗

[Frequency and antibiotic susceptibility of urinary pathogens isolated in the microbiology and virology laboratory of the Careggi General Hospital in the period July-December 1996].

BACKGROUND: The aim of the study was to evaluate the isolation frequency and antibiotic susceptibility of pathogens isolated form urinocoltures. METHODS: 2192 bacterial strains obtained from urine cultures with bacterial count > 100,000 UFC/ml were examined in our Laboratory from July through December 1996. Inpatients as well as outpatients were considered. Five different ward typologies were taken into account (Surgery, Medicine, Obstetrics, Spinal Unit and High Risk Wards). Isolation frequencies were evaluated for each ward, but in order to get an adequate statistical sample they were divided into in two categories, e.g. strains isolated from inpatients and from outpatients. Antibiotic susceptibility was also evaluated dividing the data into the same two categories. For strains isolated with lower frequencies that was not possible, and data from inpatients were grouped with data from outpatients. RESULTS: The results obtained show that bacterial species most frequently isolated among inpatients are Escherichia coli (45.7%), Enterococcus faecalis (16.8%), Proteus mirabilis (9.1%), Pseudomonas aeruginosa (6.9%) and Klebsiella pneumoniae (6.4%). Those most frequently isolated among outpatients are Escherichia coli (58.1%), Enterococcus faecalis (12.3%), Proteus mirabilis (9.1%), Klebsiella pneumoniae (4.4%) and Coagulase negative staphylococci (4.0%). As far as antibiotic susceptibility is concerned, whenever a statistically significant difference in sensitivity could be observed between strains isolated from inpatients and strains isolated from outpatients, sensitivity was always higher for outpatients strains. CONCLUSIONS: Isolation frequency and antibiotic susceptibility are different depending on the place of origin of the patients (inpatients/outpatients). This implies a different approach to the empiric therapy in urinary tract infections.

Bacteria↗

Prevalence of high level aminoglycoside and vancomycin resistance among enterococci in Turkey.

Enterococcus spp. have become the third most common cause of nosocomial infections. High-level aminoglycoside resistance (HLAR), an important clinical concern, has been associated with some species of enterococci. The synergistic effect obtained by the combination of aminoglycosides with penicillin or vancomycin disappears in strains that show high-level resistance (HLR) to the aminoglycosides. We evaluated the agar dilution and high content disk diffusion tests for the detection of HLAR for 60 isolated enterococci. In our study 4 strains of Enterococcus faecalis (6.6%) and 8 strains of Enterococcus faecium (13.3%), totally 20% were found to be highly resistant to gentamicin; 2 strains of E. faecalis (13.3%), 8 strains of E. faecium (3.3%), totally 16.6% showed HLR to streptomycin by agar dilution method and the same rates were found by high-content disk diffusion method. 2 strains of E. faecalis (3.3%), 4 strains of E. faecium (6.6%), totally 10% showed vancomycin resistance by agar dilution method. The ratio for beta lactamase activity was found to be 26.6% for E. faecalis and 3.3% for E. faecium. Our results demonstrate that high-content disk diffusion test should be performed by using both gentamicin and streptomycin in routine screening of HLAR among enterococci and the determination of beta lactamase activity at the same time is also useful for detecting resistance to a beta lactam agent.

Anti-Bacterial Agents↗

Antimicrobial resistance of Enterococcus species isolated from produce.

The purpose of this study was to characterize the antibiotic resistance profiles of Enterococcus species isolated from fresh produce harvested in the southwestern United States. Among the 185 Enterococcus isolates obtained, 97 (52%) were Enterococcus faecium, 38 (21%) were Enterococcus faecalis, and 50 (27%) were other Enterococcus species. Of human clinical importance, E. faecium strains had a much higher prevalence of resistance to ciprofloxacin, tetracycline, and nitrofurantoin than E. faecalis. E. faecalis strains had a low prevalence of resistance to antibiotics used to treat E. faecalis infections of both clinical and of agricultural relevance, excluding its intrinsic resistance patterns. Thirty-four percent of the isolates had multiple-drug-resistance patterns, excluding intrinsic resistance. Data on the prevalence and types of antibiotic resistance in Enterococcus species isolated from fresh produce may be used to describe baseline antibiotic susceptibility profiles associated with Enterococcus spp. isolated from the environment. The data collected may also help elucidate the role of foods in the transmission of antibiotic-resistant strains to human populations.

Anti-Bacterial Agents↗

Species distribution and antibiotic resistance patterns of enterococci isolated from food of animal origin in Germany.

Presently, enterococci take the third place of bacterial pathogens associated with nosocomial infections, after staphylococci and Escherichia coli. Especially, the resistances of enterococci to several available antibiotics are threatening. We attempted to determine which species of enterococci could be found in food of animal origin and their significance according to their antibiotic resistances for human beings. From November 2000 to May 2002 we investigated 155 samples of food of animal origin bought in retail outlets in Germany: 27 samples of sausages, 19 of ham, 83 of minced meat, 26 of cheese. From these food samples we isolated 416 enterococcal strains. The most frequent species was Enterococcus faecalis (299 strains); furthermore, we found Enterococcus faecium (54 strains), Enterococcus durans together with Enterococcus hirae (24 strains), Enterococcus casseliflavus (22 strains), Enterococcus avium (9 strains) and Enterococcus gallinarum (8 strains). We focused on the resistance patterns of 118 selected E. faecium and E. faecalis strains to 13 antimicrobial active agents (ampicillin, amoxicillin/clavulanic acid, avilamycin, chloramphenicol, enrofloxacin, erythromycin, flavomycin, gentamicin, penicillin, quinupristin/dalfopristin, teicoplanin, tetracycline and vancomycin). From the clinical point of view, the situation of antibiotic resistance to the examined antimicrobial agents seemed to be favourable. The investigated strains were sensitive to ampicillin and amoxicillin/clavulanic acid. These antibiotics are, in combination with an aminoglycoside, for example gentamicin, agents of choice for the treatment of enterococcal infections in human medicine. Only one E. faecium strain was resistant to penicillin, while all strains were sensitive to the glycopeptide antibiotics, vancomycin and teicoplanin. Resistances found against the antibiotics, tetracycline, quinupristin/dalfopristin and erythromycin, are causes for concern.

Animals↗

Antibacterial activity of rhodium, iridium, and ruthenium tripodal phosphine complexes.

Twenty-eight rhodium, iridium, or ruthenium complexes were evaluated for their in vitro antibacterial activity against Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 25922 and Pseudomonaa aeruginosa ATCC 27853. Ten compounds showed an antibacterial activity against Enterococcus faecalis ATCC 29212 and Staphylococcus aureus ATCC 29213 with a range of the minimum inhibitory concentrations (MICs) between 16 and 125 micrograms/ml. None of the compounds exhibited antimicrobial activity against the gram-negative strains E. coli ATCC 25922 or Pseudomonas aeruginosa ATCC 29213 with a concentration range between 4 and 500 micrograms/ml.

Anti-Bacterial Agents↗

Phylogenetic analysis of Streptococcus saccharolyticus based on 16S rRNA sequencing.

Reverse transcriptase sequencing of 16S rRNA of Streptococcus saccharolyticus was performed in order to determine the phylogenetic position of this organism. On the basis of the sequence data Streptococcus saccharolyticus formed a distinct group with Enterococcus faecalis (type species of the genus Enterococcus) and other enterococcal species. Streptococcus saccharolyticus was found to be only distantly related to members of the genus Streptococcus sensu stricto. It is therefore proposed that Streptococcus saccharolyticus be reclassified in the genus Enterococcus, as Enterococcus saccharolyticus comb. nov.

Base Sequence↗

The killing activity of microwaves on some non-sporogenic and sporogenic medically important bacterial strains.

The killing activity of microwaves of 2450 MHz frequency and 325 W, 650 W and 1400 W power on some bacterial strains was investigated. Vegetative strains of Staphylococcus aureus, Streptococcus pyogenes Group A, Escherichia coli, Pseudomonas aeruginosa and Enterococcus faecalis and spores of Bacillus subtilis and Bacillis stearothermophilus in aqueous suspensions were exposed to 325 W and 650 W waves for different lengths of time. Enterococcus faecalis and spores of B. subtilis and B. stearothermophilus were exposed additionally to 1400 W waves in aqueous and 'dried' suspensions. Vegetative bacteria were promptly killed in 5 min or less, E. faecalis being slightly more resistant. Bacterial spores were only killed in aqueous suspension when a 1400 W setting was used for 10 to 20 min. Bacterial spores adhering to the tube walls after the aqueous suspension was poured out were reduced in number. We assume that the conventional microwave ovens available on the market may be used for a high level of disinfection but not for sterilization, and only then if sufficient water is present.

Bacteria↗

[Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (1995). I. Susceptibility distribution].

The frequencies of isolation and susceptibilities to antimicrobial agents were investigated on 704 bacterial strains isolated from patients with urinary tract infections (UTIs) in 11 hospitals during the period of June 1995 to May 1996. Of the above bacterial isolates, Gram-positive bacteria accounted for 29.8% and a majority of them were Enterococcus faecalis. Gram-negative bacteria accounted for 70.2% and most of them were Escherichia coli. Susceptibilities of several isolated bacteria to antimicrobial agents were as followed; 1. Enterococcus faecalis Ampicillin (ABPC) and imipenem (IPM) showed the highest activities against E. faecalis isolated from patients with UTIs. The MIC90S of them were 1 microgram/ml. Vancomycin (VCM) and piperacillin (PIPC) were also active with the MIC90S of 2 micrograms/ml and 4 micrograms/ml, respectively. The others had low activities with the MIC90S of 16 micrograms/ml or above. 2. Staphylococcus aureus including MRSA VCM showed the highest activities against S. aureus isolated from patients with UTIs. Its MIC90 was 1 microgram/ml against both S. aureus and MRSA. Arbekacin (ABK) was also active with the MIC90 of 2 micrograms/ml. The other except minocycline (MINO) had very low activities with the MIC90S of 64 micrograms/ml or above. 3. Staphylococcus epidermidis ABK and MINO showed the strongest activities against S. epidermidis isolated from patients with UTIs. The MIC90S of them were 0.25 microgram/ml. VCM was also active with the MIC90 of 1 microgram/ml. The MIC90S of cephems ranged from 2 micrograms/ml to 16 micrograms/ml in 1994, but they ranged from 8 micrograms/ml to 128 micrograms/ml in 1995. These results indicated that some resistances existed among S. epidermidis to cephems. 4. Streptococcus agalactiae All drugs except gentamicin (GM) were active against S. agalactiae. ABPC, cefmenoxime (CMX), IPM, erythromycin (EM), clindamycin (CLDM) and clarithromycin (CAM) showed the highest activities. The MICs for all strains were lower than 0.125 microgram/ml. The MIC90S of the others were 2 micrograms/ml or below. 5. Citrobacter freundii IPM showed the highest activity against C. freundii isolated from patients UTIs. Its MIC90 was 1 microgram/ml. GM was also active with the MIC90 of 2 micrograms/ml. Cefpirome (CPR), cefozopran (CZOP) and amikacin (AMK) were also active with the MIC90S of 4 micrograms/ml. Penicillins and cephems except CMX, CPR and CZOP showed low activities with MIC90S of 256 micrograms/ml or above. 6. Enterobacter cloacae IPM showed the highest activity against E. cloacae. The MICs for all strains were equal to or lower than 1 microgram/ml. MINO and tosufloxacin (TFLX) were also active with the MIC90S of 8 micrograms/ml. Penicillins and cephems except CPR and CZOP showed lower activities with the MIC90S of 256 micrograms/ml or above. 7. Escherichia coli. Most of the antimicrobial agents were active against E. coli. Particularly CPR, CZOP and IPM showed the highest activities against E. coli. The MICs for all strains were equal to or lower than 0.5 microgram/ml. CMX and TFLX were also active with the MIC90S of 0.125 microgram/ml or below. Penicillins were slightly active with MIC90S of 128 micrograms/ml or above. 8. Klebsiella pneumoniae K. pneumoniae was susceptible to all drugs except penicillins, with MIC90S of 2 micrograms/ml or below. Carumonam (CRMN) had the strongest activity against K. pneumoniae, the MICs for all strains were equal to or lower than 0.125 microgram/ml. Comparing with the result of 1994, the sensitivities of K. pneumoniae against all drugs had obviously changed into a better state. For example, the MIC90S of cephems ranged from 0.25 microgram/ml to 16 micrograms/ml in 1994, but they were all lower than 2 micrograms/ml in 1995. 9. Proteus mirabilis P. mirabilis was susceptible to a majority of drugs. CMX, ceftazidime (CAZ), cefixime (CFIX), and CRMN showed the highest activities against P. mirabilis isolated from patients with UTIs. MICs of CRMN for all

Anti-Bacterial Agents↗

Comparison of the effect of delayed entry into 2 different blood culture systems (BACTEC 9240 and BacT/ALERT 3D) on culture positivity.

The effect of delayed entry (2-48 h) into BacTAlert 3D and BACTEC 9240 and the effect of 2 storage temperatures (22 degrees C vs 35 degrees C) on bacterial growth was evaluated. The delay in transportation of blood culture bottles stored at room temperature had no effect on the recovery rate for the first 12 h. Culture positivity was between 74.4% and 100% for different microorganisms at less than 24 h preincubation time. The positivity rate decreased significantly for Acinetobacter baumannii, Bacteroides fragilis, Escherichia coli, Enterococcus faecalis, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Streptococcus pneumoniae for more than 24 h of delay. Culture positivity was higher at 22 degrees C for all microorganisms especially for Enterococcus faecalis and P. aeruginosa. Effects of instrument, preincubation time, and temperature showed that the risk of culture negativity increased 1.5 times for BACTEC compared with BacTAlert 3D and increased 2.5 times for 35 degrees C compared with 22 degrees C. The negativity increased 5.5 times and 8.5 times at 24 and 48 h of delay respectively, compared with no delay.

Bacteria↗

Efficacy of daptomycin in the treatment of experimental endocarditis due to susceptible and multidrug-resistant enterococci.

OBJECTIVES: Daptomycin was tested in vitro and in rats with experimental endocarditis against the ampicillin-susceptible and vancomycin-susceptible Enterococcus faecalis JH2-2, the vancomycin-resistant (VanA type) mutant of strain JH2-2 (strain JH2-2/pIP819), and the ampicillin-resistant and vancomycin-resistant (VanB type) Enterococcus faecium D366. METHODS: Rats with catheter-induced aortic vegetations were treated with doses simulating intravenously kinetics in humans of daptomycin (6 mg/kg every 24 h), amoxicillin (2 g every 6 h), vancomycin (1 g every 12 h) or teicoplanin (12 mg/kg every 12 h). Treatment was started 16 h post-inoculation and continued for 2 days. RESULTS: MICs of daptomycin were 1, 1 and 2 mg/L, respectively, for strains JH2-2, JH2-2/pIP819 and D366. In time-kill studies, daptomycin showed rapid (within 2 h) bactericidal activity against all strains. Daptomycin was highly bound to rat serum proteins (89%). In the presence of 50% rat serum, simulating free concentrations, daptomycin killing was maintained but delayed (6-24 h). In vivo, daptomycin treatment resulted in 10 of 12 (83%), 9 of 11 (82%) and 11 of 12 (91%) culture-negative vegetations in rats infected with strains JH2-2, JH2-2/pIP819 and D366, respectively (P < 0.001 compared to controls). Daptomycin efficacy was comparable to that of amoxicillin and vancomycin for susceptible isolates. Daptomycin, however, was significantly (P < 0.05) more effective than teicoplanin against the glycopeptide-susceptible strain JH2-2 and superior to all comparators against resistant isolates. CONCLUSIONS: These results support the use of the newly proposed daptomycin dose of 6 mg/kg every 24 h for treatment of enterococcal infections in humans.

Amoxicillin↗

Species identification and determination of high-level aminoglycoside resistance among enterococci. Comparison study of sterile body fluid isolates, 1985-1991.

Enterococcus spp. have become the third most common cause of nosocomial infections. High-level aminoglycoside resistance (HLR), an important clinical concern, has been associated with some species of the enterococci. We evaluated the Vitek and API 20S systems for species identification and the Vitek for the detection of HLR. Enterococci from nosocomial infections (208 strains) at the University of Iowa Hospital (1985-1991) were tested by Vitek, API 20S, and reference methods. The error rate for species identification was 6.7% for the API 20S and 5.8% for the Vitek Gram-positive identification (GPI) cards. Both systems tended to incorrectly identify other enterococcal species as Enterococcus faecium. HLR was found in Enterococcus faecalis and E. faecium isolates only. The highest rates of HLR to streptomycin alone (17.9%) and with gentamicin (13.5%) was observed among E. faecalis strains, and to gentamicin alone (7.3%) was found among E. faecium isolates. No apparent differences in HLR rates were found from year-to-year over the 7-year enterococcus sample interval. Susceptibility errors for Vitek were among the streptomycin tests only. Our results demonstrated acceptable performance by the Vitek cards for enterococcal species identification and the detection of HLR. API 20S also provided an acceptable ability to speciate the enterococci within its data base, however, both systems must be improved by adding other clinical important Enterococcus species.

Aminoglycosides↗

[Bacterial pathogens and resistance patterns in community acquired pediatric urinary tract infection: experience of 152 cases].

OBJECTIVE: This study investigated the pathogen distribution and resistance patterns in childhood urinary tract infection in order to provide references for optimal use of antibiotics in the treatment of this disorder. METHODS: The clinical data of 152 children with community acquired urinary tract infection (urinary culture positive) between December 2001 and December 2004 were studied retrospectively. The bacterial pathogens of urinary tract infection and antimicrobial resistance were analyzed. RESULTS: Gram-negative bacilli was predominant pathogenic bacteria, accounting for 79.0% of the cases, and Escherichia coli (E. coli) was most commonly found (56.2%). Gram-positive cocci accounted for 18.4%, including 15.1% of Enterococcus faecalis. Fungi was rarely seen, accounting for only 2.6%. E. coli had a resistance rate of more than 50% to ampicillin, amoxicillin/clavulate, co-trimoxazole, cefradine, and fosomycin, but a very low resistance rate (< 4%) to 3rd generation cefalosporin, nitrofurantoi, azactom and amikacin. Enterococcus faecalis had a low resistance rate (< 20%) to ampicillin, vancomycin, penicillin, and nitrofurantoin. CONCLUSIONS: E. coli is the major pathogen in community acquired pediatric urinary tract infection, and Enterococcus has been become another important pathogen. Selection of antibiotics for the treatment of this disorder should base on drug-sensitive test results.

Adolescent↗

Effect of growth promotant usage on enterococci species on a poultry farm.

The species population of enterococci isolated from four poultry houses for six grow-outs on one farm was determined. Two houses on the farm were control houses and did not use any antimicrobials, while two other houses on the farm used flavomycin, virginiamycin, and bacitracin during different poultry grow-outs. Litter, chick boxliners, feed, and poultry carcass rinses were obtained from each house and cultured for the presence of enterococci. Nine species of enterococci (Enterococcusfaecalis, E. faecium, E. avium, E. casselifiavus, E. cecorum, E. durans, E. gallinarum, E. hirae, and E. malodoratus) were identified from the study. Enterococcus faecalis was isolated more frequently from chick boxliners (n=176; 92%) and carcass rinses (n=491; 69%), whereas E. faecium was found more frequently in litter (n=361; 77%) and feed (n=67; 64%). Enterococcus faecalis (n=763; 52%) and E. faecium (n=578; 39%) were isolated most often from the farm and houses, regardless of antimicrobial treatment. Fifty-two percent of E. faecalis and 39% of E. faecium were isolated from both control (n=389 and 295, respectively) and treatment (n=374 and 283, respectively) samples. This study indicates that antimicrobial usage on this farm did not alter the resident population of enterococci.

Agriculture↗

Synthesis and antimicrobial activity of electron rich olefin derived cyclic ureas.

Seventeen cyclic ureas containing imidazolidine and benzimidazoline nuclei were synthesised by the reaction of electron-rich olefins with appropriate group 16 elements. (O, S, Se, Te). The compounds synthesised were identified by 1H, 13C-NMR, FT-IR and mass spectroscopic techniques and micro analysis. All compounds studied in this work were screened for their in vitro antimicrobial activity against standard strains: Enterococcus faecalis (ATCC 29212), Staphylococcus aureus (ATCC 29213), Escherichia coli (ATCC 25922) and Pseudomonas aeruginosa (ATCC 27853). Eight of the compounds were found effective to inhibit the growth of Gram-positive bacteria (Enterococcus faecalis and Staphylococcus aureus) at MIC values between 25-400 micrograms/ml. None of the compounds exhibit antimicrobial activity against gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) at the concentrations studied (6.25-800 micrograms/ml).

Alkenes↗

[Epidemiological study on vancomycin-resistant enterococci from fecal samples in the east area of Japan].

Recently, Vancomycin-resistant enterococci (VRE) have become important nosocomial pathogens in the world. In Japan, the VRE-infection was first reported in 1996. However, an epidemiological study on VRE has not been aggressively done in Japan. We conducted a survey study to explore the incidence and antimicrobial susceptibility of vancomycin-resistant enterococci isolated from fecal samples at 45 hospitals in the east area of Japan (Kanto, Koshinetsu, Tohoku, and Hokkaido) during June 1998 to March 1999. The Enterococcosel agar containing vanocomycin (BBL) was used for screening VRE from fecal samples in each hospital. The susceptibilities of the isolates to 8 antimicrobials were determined by the broth microdilution method and the definitions of resistance were based on current standards of the NCCLS standards. The VRE genotypes (vanA, vanB, vanC1, and van C2/3) were confirmed by amplifying the respective genes by PCR. Eight hundred and ninety four strains of enterococci were tested by the microtiter plates hybridization method (WAKUNAGA SEIYAKU, Japan). One thousand five hundred eighty three strains of enterococci were collected from 6,914 patients in 45 hospitals. These strains included 72 (4.5%) strains Enterococcus faecalis, 33 (2.1%) strains Enterococcus faecium, 17 (1.1%) strains Enterococcus avium, 1,040 (65.7%) strains Enterococcus gallinarum, 386 (24.4%) strains Enterococcus cassliflavus, and 35 (2.2%) strains Enterococcus flavescens. These strains of vancomycin-resistant E. faecalis were isolated from 3 patients, two of these 3 strains had van A gene and other one had van B gene. Those 3 strains were in the Kanto area, and 2 of 3 strains were in Tokyo, Generally, though van A type VRE was highly resistaant to both vancomycin and teicoplanin. In our study, two strains of van A type E. faecalis were highly resistant to vancomycin (MICs > 128 micrograms/ml) and susceptible to teicoplanin with MICs 4 micrograms/ml. Those two strains were different in susceptibilities of minocycline and ofloxacin. The result of the analysis of PFGE had also different patterns. VanB type E. fecalis was highly resistant to vancomycin and susceptible to teicoplanin (MICs 0.25 microgram/ml). For ampicillin and imipenem, 3 strains of E. faecalis were susceptible (MIC < or = 1 microgram/ml). One of 562 strains of E. gallinarum had vanB and vanC1 genes and was moderately resistant to vancomycin and susceptible to teicoplanin. All strains of E. casseliflavus and E. flavescens had vanC2/C3 gene only. All strains of E. faecium and E. avium did not detect van genes. From this result, it was supposed that VRE were very rare in the east of Japan.

Enterococcus↗