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PubMed1995

Enumeration of intestinal enterococci and interfering organisms with Slanetz-Bartley agar, KF streptococcus agar and the MUST method.

The recovery of intestinal species of enterococci and streptococci and potentially interfering nonfaecal species was measured on KF streptococcus agar, Slanetz-Bartley agar and in a medium based on 4-methylumbelliferyl-beta-D-glucoside (MUST method) using pure cultures. Both of the solid media yielded high recoveries of the target species. Their selectivity was better at elevated incubation temperature but nonfaecal Enterococcus and Staphylococcus species were not eliminated even at the elevated temperature. The MUST method tended to give slightly lower recoveries than the agar cultivation methods with some target species at 44 degrees C but recoveries were better at 41 degrees C.

Agar↗
PubMed2006

Identification and characterization of enterococci from bryndza cheese.

AIMS: To identify enterococci isolated from sheep milk cheese--bryndza, and to compare differences in the composition of enterococcal microflora affected by the season, and to evaluate the potential presence of vancomycin resistance and virulence determinants. METHODS AND RESULTS: Bacterial strains were isolated during analysis of bryndza cheese and identified on the genus and species level by phenotypic methods and with commercial biochemical sets. The identification of the species, Enterococcus faecium, Ent. durans and Ent. faecalis, was confirmed by PCR using species-specific primers for ddl genes. PCR was also used for assessment of presence of vanA and vanB genes and virulence determinants gelE, agg and cytolysin genes namely: cylL(L), cylL(S), cylM, cylB and cylA. Among 308 Enterococcus sp. strains, 177 isolates were proved to be Ent. faecium, 59 to be Ent. durans and 41 to be Ent. faecalis. Vancomycin resistance genes vanA and vanB were not detected. Agar plate testing confirmed their absence. Gene gelE, however, was found in 20 Ent. faecalis isolates, but only 13 of them showed gelatinase-positive phenotype. Seven isolates had five cytolysin genes, but none of the isolates exhibited a positive haemolytic phenotype. Four isolates possessed the agg gene. The prevalence of Ent. faecium species was highest in samples from the winter season harvest. CONCLUSIONS: Ent. faecium is the dominant enterococcal species in bryndza cheese and the most prevalent in the winter season product. None of the Enterococcus sp. strains was proved to have vanA or vanB genes and the vancomycin resistance. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first report of enterococcal microflora in bryndza cheese and its evaluation for the presence of vanA and vanB genes as well as virulence determinants.

Animals↗
PubMed2000

Preventing the spread of vancomycin-resistant enterococci in a long-term care facility.

OBJECTIVES: To test the hypothesis that infection control practices can prevent the spread of vancomycin-resistant enterococci (VRE) to residents of a long-term care facility (LCF) from an affiliated acute care facility with a high endemic rate of colonization. DESIGN: Point prevalence study of the rate of rectal colonization. SETTING: A state-supported veterans nursing home and an acute care veterans hospital. PARTICIPANTS: Residents in a state veterans home. INTERVENTIONS: Identification of patients with rectal colonization by VRE before transfer to the state veterans home, contact isolation for colonized veterans, use of oral bacitracin to eliminate colonization. MEASUREMENTS: Rectal swab and culture for VRE, review of clinical records and recording of presumptive risk factors for VRE colonization. The risk factors were age, gender, length of stay at nursing home, treatment with vancomycin or oral antibiotics, prior hospitalization at the acute care facility during the prior year, use of indwelling urethral catheters, presence of diarrhea, and fecal or urinary incontinence. RESULTS: Sixty-nine of 200 residents were cultured in the first study (1996) and 130 of 230 residents were cultured in the second study (1998). Residents who consented to culture differed from those who did not only with regards to gender (2 vs 7, P = .012). In neither study were any residents found to be colonized with VRE who had not already been identified as positive on admission. CONCLUSIONS: Adherence to infection control practices by the patient care staff of the LTCF was associated with the absence of transmission of VRE colonization among its residents. The presence of rectal colonization with VRE in an acute care patient should not be a barrier to acceptance in a nursing home.

Administration, Oral↗
PubMed1994

Conjugal transfer of the vancomycin resistance determinant vanB between enterococci involves the movement of large genetic elements from chromosome to chromosome.

Resistance to various levels of vancomycin and susceptibility to teicoplanin in enterococci (VanB phenotype) is mediated by the vanB gene cluster. VanB-type resistance was transferred by intra- and inter-specific conjugation between different strains of Enterococcus. Analysis of SfiI-digested genomic DNA by zero integrated-field electrophoresis followed by Southern hybridization revealed vanB-containing chromosomal insertions of approximately 90-250 kb in the transconjugants. Thus, transfer of VanB-type resistance is associated with the movement of large genetic elements from chromosome to chromosome.

Conjugation, Genetic↗
PubMed1997

Arrangement of the vanA gene cluster in enterococci of different ecological origin.

Glycopeptide-resistant enterococci (vanA) isolated from infections in humans, from non-hospitalized humans, from sewage, from animal feces and from meat products in Germany (20 Enterococcus faecium and one Enterococcus hirae) were investigated for the arrangement of the genes in the vanA gene cluster by means of overlapping PCR with five primer pairs. In 20 of these strains, the vanA gene clusters were uniform which suggests a horizontal spread among different ecosystems. In one clinical isolate a rearrangement was detected in the vanY-vanZ region.

Animals↗
PubMed1999

Strain typing among enterococci isolated from home-made Pecorino Sardo cheese.

Three molecular techniques (RAPD-polymerase chain reaction analysis, plasmid profile and pulsed-field gel electrophoresis) were used for a preliminary approach to type, at strain level, enterococci isolated from a 24-h-old home-made Pecorino Sardo (protected designation of origin) cheese. A high genetic polymorphism was found. Clusters obtained by the RAPD technique and plasmid profile analysis contained different strains. Pulsed-field gel electrophoresis proved to be an efficient and highly reproducible typing method. In addition, by combining the results from plasmid profile analysis and pulsed-field gel electrophoresis, it was possible to identify closely related strains probably belonging to the same clonal lineage.

Bacterial Typing Techniques↗
PubMed2000

Persistence of vancomycin-resistant enterococci (VRE) on Norwegian broiler farms.

Five Norwegian broiler farms previously identified as housing broilers carrying vancomycin-resistant enterococci (VRE) were examined for the presence of VRE 4 years after avoparcin was banned. Environmental samples were obtained from empty, cleaned broiler houses. Faecal samples were collected weekly from the flock housed after the environmental sampling. The hatchery from where the chicks originated was also sampled. VRE were found to be present in the farm environment after depopulation and cleanup of the broiler houses. Within 3 weeks after introduction to the farm, all broiler flocks tested positive for VRE. VRE were not isolated from the hatchery.

Animal Husbandry↗
PubMed2001

Antibiotic resistance and genotypic characterization by PFGE of clinical and environmental isolates of enterococci.

Fifty-four Enterococcus faecalis and 20 Enterococcus faecium isolates from clinical and non-human sources in Rome, Italy, were characterized by antibiotic resistance and pulsed field gel electrophoresis (PFGE). Resistance to vancomycin, teicoplanin, ampicillin, and ciprofloxacin was more frequent in E. faecium than in E. faecalis, whereas high-level resistance to aminoglycoside was found primarily in E. faecalis. Multi-resistance was found primarily among clinical isolates, but was also observed among environmental isolates. Common genotypes shared among clinical and environmental isolates were observed, however, the majority of isolates occurred as unique, source-specific clones. Several PFGE types were associated with shared features in their antibiotic resistance patterns; evidences of clonal spread between and within wards were also noted. This is the first report indicating clonal relatedness between human and environmental enterococci isolated in Italy.

Anti-Bacterial Agents↗
PubMed2006

Phenotypic and genetic characterization of vancomycin-resistant enterococci from hospitalized humans and from poultry in Korea.

Vancomycin resistant enterococci (VRE) isolates from humans (23 isolates) and poultry (20 isolates) were characterized by antibiotic susceptibility, vancomycin resistance transferability, pulsed-field gel electrophoresis (PFGE), and structural analysis of Tn1546-like elements. VRE isolates from humans and poultry showed different resistance patterns, transferability, and transfer rate. In addition to these phenotypic differences between humans and poultry VRE, PFGE and the structure of Tn1546-like elements were also distinct. Most poultry isolates (16/20) were identical to the prototype vanA transposon, Tn1546, while most human isolates (21/23) had multiple integrations of insertion sequence. The transmission of VRE and vancomycin resistance determinant between humans and poultry could not be demonstrated in this study.

Animals↗
PubMed2006

Comparison of workflow and accuracy of identification and antimicrobial susceptibility testing of clinical isolates of Enterobacteriaceae, Pseudomonas aeruginosa and enterococci by Vitek 2 and routine methods.

Three hundred and fifty-three consecutive urine cultures growing Enterobacteriaceae, Pseudomonas aeruginosa or enterococci were subjected to parallel identification (ID) and antimicrobial susceptibility testing (AST) by Vitek 2 and routine methods, including simple screening tests or API 20 E for ID and standardized disc diffusion for AST. Accuracy of results, technician hands-on time required by both methods and time to results were compared. Vitek 2 correctly identified 322 (94.7%) of the 340 gram-negative isolates and 17 (81%) of the 21 Enterococcus faecalis strains. AST by Vitek 2 and disc diffusion gave category agreement for 4,058 (95.5%) of 4,248 organism-antimicrobial agent combinations. With MIC determination by E-test as reference, AST by Vitek 2 and disc diffusion produced 15 and 3 very major errors, respectively. Six (40%) of the fifteen very major errors by Vitek 2 were associated with trimethoprim-sulfamethoxazole. With an average of 22 specimens processed per day, use of Vitek 2 saved 80 min per day of technician hands-on time as compared to routine methods. Regarding the cost of hands-on worktime and consumables, use of Vitek 2 for identification of Escherichia coli-screened Enterobacteriaceae saved 0.70 p per sample in comparison to API 20 E. More than 80% of Enterobacteriaceae introduced to Vitek 2 in the morning could be reported by 16:00.

Anti-Bacterial Agents↗
PubMed2007

In vitro activity of the oxazolidinone RWJ-416457 against linezolid-resistant and -susceptible staphylococci and enterococci.

RWJ-416457, a novel oxazolidinone, had modal MICs of 0.5 to 1 microg/ml for linezolid-susceptible staphylococci and enterococci, versus linezolid MICs for these organisms of 1 or 2 microg/ml. RWJ-416457 MICs for mutants with 23S rRNA mutations were 2 to 32 microg/ml, versus linezolid MICs of 8 to 64 microg/ml; actual values reflected the proportion of gene copies mutated.

Acetamides↗
PubMed1976

Synergism of cefazolin-gentamicin against enterococci.

A miniaturized technique for detecting antibiotic synergy by using microliter volumes was perfected. With this microtiter serial dilution method, the effect of the previously untried combination of cefazolin and gentamicin on 10 strains of enterococci was evaluated. At clinically achievable concentrations, the combination of cefazolin and gentamicin was found to be synergistic against all 10 strains when the data were plotted in the form of an isobologram. These results were compared with the conventional method of detecting synergy in vitro, namely, the determination of the rate of killing of the microorganisms with the antibiotics, singly and in combination. Similar results were also observed. These findings indicate that the microtiter serial dilution technique is a useful method for the routine determination of drug synergy. Moreover, in a patient with penicillin hypersensitivity and enterococcal infection, the combination of cefazolin and gentamicin should be considered for possible therapy.

Cefazolin↗
PubMed1977

Synergy of penicillin-netilmicin combinations against enterococci including strains highly resistant to streptomycin or kanamycin.

The in vitro activity of combinations of penicillin and netilimicin was determined against 20 clinical isolates of enterococci and compared with that obtained in simultaneous tests with penicillin/sisomicin, penicillin/streptomycin, and penicillin/kanamycin. Synergy between the two drugs in each combination was determined by the use of quantitative kill curves and was defined as a killing by the combination at least 100-fold greater than that produced by the most effective drug alone. Penicillin/netilmicin and penicillin/sisomicin combinations were found to be synergistic against the majority of isolates tested, including strains resistant to penicillin/streptomycin or penicillin/kanamycin combinations. This synergy with penicillin could be demonstrated at a concentration of </=7 mug/ml for either netilmicin or sisomicin. Studies on the kinetics of killing produced by these combinations showed the rate and extent of killing to be directly dependent upon the organism's relative susceptibility to the aminoglycoside alone and the aminoglycoside concentration in the combination. Results also indicated that the interaction between penicillin and netilmicin was true synergy; i.e., rapid and complete killing was produced by combinations containing each drug at concentrations insufficient to produce any killing alone, and the killing observed could not be produced by either drug alone at a concentration equivalent to the total drug concentration in the combination. The potential clinical application of this synergistic interaction should be investigated further, especially in view of recent reports showing netilmicin to be considerably less toxic than gentamicin in experimental animals.

Anti-Bacterial Agents↗
PubMed1980

Incidence of thymidine-dependent enterococci detected on Mueller-Hinton agar with low thymidine content.

It was observed that 10% of urine culture isolates of enterococci tested for antimicrobial susceptibility failed to grow on commercially prepared Mueller-Hinton agar with low levels of thymidine and thymine. All strains could utilize exogenous thymidine and thymine and required only low levels (0.4-microgram disk) to support growth. All thymidine-thymine-requiring strains were resistant to trimethoprim-sulfamethoxazole.

Adult↗
PubMed1981

Susceptibility of enterococci and Listeria monocytogenes to N-Formimidoyl thienamycin alone and in combination with an aminoglycoside.

The susceptibilities of 12 strains of enterococci and 10 stains of Listeria monocytogenes to N-formimidoyl thienamycin were determined by a standard broth dilution method. Minimal inhibitory concentrations for L. monocytogenes strains were less than 0.25 micrograms/ml. As a group, strains of Streptococcus faecium were less susceptible than Streptococcus faecalis strains, but even for the latter, the minimal inhibitory concentrations were slightly greater than those previously reported. Minimal bactericidal concentrations against all organisms were many-fold higher than the corresponding inhibitory concentrations. In time-kill studies, combinations of N-formimidoyl thienamycin with gentamicin were synergistic against (or they completely sterilized cultures of) all the enterococcal strains and nine to 10 strains of L. monocytogenes. The magnitude of killing by the combinations was comparable to that previously observed with penicillin-gentamicin combinations. N-Formimidoyl thienamycin-tobramycin combinations were synergistic against those strains of S. faecalis and L. monocytogenes tested, but not against those of S. faecium.

Aminoglycosides↗
PubMed1983

Enterococci from Bangkok, Thailand, with high-level resistance to currently available aminoglycosides.

Enterococcal endocarditis is usually treated with a combination of a penicillin and an aminoglycoside. Recent reports have documented the emergence of enterococci in France with high-level resistance to gentamicin, tobramycin, and kanamycin and the emergence of strains in Houston, Tex. with high-level resistance to all of these drugs and streptomycin. In this study, we examined strains from a geographic area where newer aminoglycosides have been less commonly used. Of 125 distinct patient isolates, 18 (14%) were resistant to greater than 2,000 micrograms of gentamicin and most other aminoglycosides per ml. Four of these strains transferred gentamicin resistance to a laboratory recipient. One strain, chosen for further study, was resistant to synergism between penicillin and gentamicin, tobramycin, kanamycin, streptomycin, and amikacin and demonstrated the following enzymatic activities: 3'- and 2"-aminoglycoside phosphotransferases, 6'-aminoglycoside acetyltransferase, and adenylylation of streptomycin. Optimal therapy for endocarditis caused by such highly resistant strains is currently unknown.

Aminoglycosides↗