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PubMed2000

Characteristics of polyarthritis in rabbits by hyperimmunization with attenuated Enterococcus faecalis.

To produce polyarthritis and rheumatoid factor like substance (RFLS), rabbits were hyperimmunized intravenously with 0.02% thimelosal (TMS)-treated Enterococcus faecalis (E. faecalis) as a persistent bacterial flora. Swelling of knee joints occurred at a rate of 41% (27/66), and of shoulder joints at a rate of 25% (17/66) while that of elbow joints occurred at a rate of 4.5% (3/66). On culturing of knee joint fluids, no colonies appeared while 2/4 fluid specimens from the shoulder joints gave positive colonies for 78 days after the first immunization; thereafter, no colonies appeared. On histological examination, in early stages, acute inflammatory reactions with degenerative changes of synovial tissue was observed. In later stages, chronic inflammatory changes, proliferation of synovial cells with pannus formation, destruction of articular cartilage and subchondral bone were observed. RFLS titer showed bi-phasic peaks at 11 days and 41 days after the first immunization. A high incidence of polyarthritis, particularly knee joints, occurred. Thus, hyperimmunization with attenuated E. faecalis as a normal intestinal flora may provide an animal model of chronic polyarthritis.

Agglutinins↗
PubMed2002

Composition of enterococcal and streptococcal flora from pigeon intestines.

AIMS: To identify the streptococcal and enterococcal components of the microflora from pigeon intestines to species level by tDNA-intergenic polymerase chain reaction. METHODS AND RESULTS: Gram-positive coccal strains were isolated from the intestines or faeces of 50 different pigeons, all from different lofts, on Columbia CNA blood agar. Sodium azide-sensitive enterococci were found to constitute the major coccal members of the intestinal flora of pigeons. Forty-two of 69 isolates were identified as Enterococcus columbae (84% of animals tested positive), while E. cecorum was encountered in 28% of all animals (14 isolates). Minor species (less than 10% of pigeons positive) were E. faecalis, E. faecium, E. gallinarum, E. casseliflavus, Streptococcus alactolyticus and Strep. gallolyticus. Contrary to the species description of E. columbae and E. cecorum, some of the strains were found to be motile. Certain other minor differences with the species description of E. columbae are also indicated. CONCLUSIONS: Enterococcus columbae is the major Gram-positive facultatively anaerobic component of the pigeon intestinal flora. Certain phenotypic characteristics differ from the original species description. The potentially pathogenic Strep. gallolyticus is a minor component of the microflora. Enterococcus faecium and E. faecalis are rare in pigeons. SIGNIFICANCE AND IMPACT OF THE STUDY: The enterococcal and streptococcal flora of pigeons is composed of mainly host-specific bacteria. These are not likely to act as zoonotic pathogens nor as carriers of antibiotic resistance determinants possibly spreading to humans.

Animals↗
PubMed1991

Antibiotic disk diffusion testing revisited. Single strain regression analysis. Review article.

The standardized (NCCLS, ICS, DIN etc.) disk diffusion method is the most widespread technique for antibiotic susceptibility testing. Interpretive zone breakpoints are calculated from the regular regression line between minimum inhibitory concentrations (MIC) of bacterial isolates and the corresponding inhibition zone diameters around the disk containing the antibiotic. Studies of the regression line has revealed marked differences between different bacterial species. A newly described equation, the single strain regression analysis (SRA) equation, can be used to determine the regression line constants for individual strains. This method was applied to ciprofloxacin and S. aureus, E. faecalis, E. coli, P. mirabilis, P. aeruginosa, and P. maltophilia. The slope and intercept constants were determined for all 40 strains and showed a strong similarity within each species. A close similarity was also observed between the two Pseudomonas species and between S. aureus and E. faecalis. When the regression lines calculated by SRA for individual strains were extrapolated towards higher MIC values, the lines obtained for the more susceptible strains predicted the zones of more resistant strains within the species. The applications of SRA to several other antibiotics and bacterial species in earlier studies were reviewed. One exception to the predictive power of SRA has been detected earlier, H. influenzae and erythromycin. This led to the formulation of the standard curve regression analysis (SCA) equation which requires the use of two or more strains. Methodological aspects of SRA/SCA applications were presented. Three areas are particularly well suited for the use of SRA/SCA: 1. Calculation of interpretive zone breakpoints corresponding to recommended MIC limits in the individual laboratory. 2. Analysis of the effects of various disk contents of antibiotic on the resulting inhibition zones for various bacteria when new antibiotics are introduced. 3. Analytical tool as part of external quality control programmes.

Diffusion↗
PubMed2006

Genetic analysis of bacteriocin 43 of vancomycin-resistant Enterococcus faecium.

A total of 636 vancomycin-resistant Enterococcus faecium (VRE) isolates obtained between 1994 and 1999 from the Medical School Hospital of the University of Michigan were tested for bacteriocin production. Of the 277 (44%) bacteriocinogenic strains, 21 were active against E. faecalis, E. faecium, E. hirae, E. durans, and Listeria monocytogenes. Of those 21 strains, a representative bacteriocin of strain VRE82, designated bacteriocin 43, was found to be encoded on mobilizable plasmid pDT1 (6.2 kbp). Nine open reading frames (ORFs), ORF1 to ORF9, were presented on pDT1 and were oriented in the same direction. The bacteriocin 43 locus (bac43) consists of the bacteriocin gene bacA (ORF1) and the immunity gene bacB (ORF2). The deduced bacA product is 74 amino acids in length with a putative signal peptide of 30 amino acids at the N terminus. The bacB gene encodes a deduced 95-amino-acid protein without a signal sequence. The predicted mature BacA protein (44 amino acids) showed sequence homology with the membrane-active class IIa bacteriocins of lactic acid bacteria and showed 86% homology with bacteriocin 31 from E. faecalis YI717 and 98% homology with bacteriocin RC714. Southern analysis with a bac43 probe of each plasmid DNA from the 21 strains showed hybridization to a specific fragment corresponding to the 6.2-kbp EcoRI fragment, suggesting that the strains harbored the pDT1-like plasmid (6.2 kb) which encoded the bacteriocin 43-type bacteriocin. The bac43 determinant was not identified among non-VRE clinical isolates.

Amino Acid Sequence↗
PubMed2002

Enterococcus faecalis induces inflammatory bowel disease in interleukin-10 knockout mice.

Germ-free interleukin-10 knockout (IL-10 KO) mice developed inflammatory bowel disease (IBD) after they were colonized with a pure culture of Enterococcus faecalis. E. faecalis not only induced IBD (primarily in colon and rectum) but rectal dysplasia and adenocarcinoma was also found in the IL-10 KO mice. Conventional (complex-intestinal flora) IL-10 KO mice developed IBD within 10 to 15 weeks of age and showed more pathology in the cecum (typhlitis) than we observed with E. faecalis-induced IBD in gnotobiotic IL-10 KO mice. Conversely, neither germ-free IL-10 mice nor IL-10 KO mice colonized as adults, with a pure culture of Candida albicans, Escherichia coli, Lactobacillus casei, L. reuteri, L. acidophilus, a Bifidobacterium sp., Lactococcus lactis, or a Bacillus sp. developed IBD during the 25- to 30-week study. E. faecalis is a common intestinal microbe of man and animals that can trigger IBD, dysplasia, and carcinoma in a genetically susceptible murine host.

Animals↗
PubMed2006

Antibiotic resistance and mechanisms implicated in clinical enterococci in a Tunisian hospital.

Susceptibility testing for 15 antibiotics was performed in a series of 191 clinical enterococci recovered in a Tunisian Hospital during 2000-2003. Species detected were the following ones (number of isolates): E. faecalis (139), E. faecium (41), E. casseliflavus (5), E. gallinarum (3), E. avium (2) and E. hirae (1). The percentages of antibiotic resistance detected were as follows (E. faecalis/ E. faecium/ other species) : penicillin (0/ 73/ 9%), tetracycline (78/ 44/ 54%), chloramphenicol (52/ 29/ 27%), erythromycin (66/ 100/ 82%), spiramycin (84/ 83/ 64%), pristinamycin (100/ 0/ 73%), trimethoprim-sulfamethoxazole (88/ 78/ 91%), rifampicin (72/ 41/ 0%), vancomycin (0/ 0/ 36%), teicoplanin (0/ 0/ 0%), high-level-resistance for gentamicin (24/ 29/ 45%), streptomycin (34/ 56/ 55%) and kanamycin (41/ 68/ 55%). Increased vancomycin minimum inhibitory concentrations (MICs) were only detected in E. casseliflavus and E. gallinarum isolates (MIC range 8-24 microg/ml). The erm(B), catA, tet(M), aac(6')-aph(2''), aph(3')-IIIa, and ant(6)-Ia genes were detected in 91%, 32%, 86%, 98%, 100%, and 72% of the E. faecium and E. faecalis isolates resistant to erythromycin, chloramphenicol, tetracycline and high-level-resistant to gentamicin, kanamycin and streptomycin, respectively. A total of 20 unrelated pulsed-field-gel-electrophoresis patterns were found in the series of 46 high-level gentamicin-resistant E. faecalis and E. faecium isolates of this study.

Anti-Bacterial Agents↗
PubMed2003

Virulence factors and in vitro adherence of Enterococcus strains to urinary catheters.

The ability to adhere in vitro to urinary catheters and the presence of enterococcal virulence factors was determined in 30 Enterococcus urinary isolates (12 E. faecalis, 12 E. faecium, 3 E. casseliflavus, 3 E. gallinarum). Silicone, siliconized latex and polyvinyl chloride (PVC) were examined by sonication quantitative culture technique and scanning electron microscope. As compared to E. faecalis and E. faecium, E. casseliflavus and E. gallinarum displayed lower adhesion to all synthetic materials. All the tests performed showed higher adherence of all tested strains to siliconized latex and silicone than to PVC. Biofilmforming ability was observed in 5 E. faecalis but in none of the remaining strains. The gene coding enterococcal surface protein (Esp) was detected in 7 E. faecalis and 6 E. faecium strains. Gelatinase was found in 1 E. faecalis, 2 E. faecium and hemolysins were found in 6 E. faecalis and 1 E. faecium strains. All E. casseliflavus and E. gallinarum strainswere negative for these traits. Hydrophobic type of cell surface (measured by its affinity for n-hexadecane) was shown in a few isolates. Bacterial adherence was not significantly associated with the above pathogenic factors.

Bacterial Adhesion↗
PubMed2002

[Drug sensitivity of Enterococcus sp. strains isolated from clinical material or from healthy persons].

The aim of this study was to evaluate the drug susceptibility of Enterococcus sp. strains isolated in 2000-2001, from patients of five Warsaw's hospitals (154 strains) and from fecal samples of healthy persons (33 strains). On biochemical reaction profiles species of clinical enterococci were identified as: E. faecalis--66.2%, E. faecium--29.2%, E. hirae--1.9%, E. gallinarum--1.3%, E. casseliflavus--0.6% and E. avium--0.6%. The species of enterococci from stool's samples were identified as: E. faecalis--28 strains, E. durans--2 strains and single strains: E. faecium, E. gallinarum and E. casseliflavus. Susceptibility to 20 antibiotics was tested by disc diffusion method. None of these 187 enterococcal strains was vancomycin resistant; 3 strains of E. gallinarum and 1--E. casseliflavus demonstrated intermediately susceptibility to vancomycin, but they were susceptible to teicoplanin--phenotype Van C. Among clinical strains were resistant to penicillin--33.3% of E. faecalis and 100% of E. faecium, to ampicillin--over 80% of E. faecium and 1 strain of E. faecalis. None of these strains produced beta-lactamase. High-level resistance to aminoglicoside was expressed by 48 strains (47.1%) E. faecalis and 36 (80%) E. faecium isolated from clinical specimens. Both--HLR to streptomycin and gentamycin were found in 28.3% of E. faecalis and 68.9% of E. faecium. Among 33 strains isolated from fecal samples of healthy persons--3 of E. faecalis were resistant to streptomycin and one was resistant to both gentamicin and streptomycin. In general, enterococcal strains isolated from samples of healthy persons were susceptible to the most of used antibiotics. But to rifampicin none of these strains were susceptible. There were about 40% of E. faecalis strains isolated from healthy persons, resistant to tetracyline.

Anti-Bacterial Agents↗
PubMed2000

Comparison of antimicrobial resistance phenotypes and resistance genes in Enterococcus faecalis and Enterococcus faecium from humans in the community, broilers, and pigs in Denmark.

Enterococcus faecalis and E. faecium isolated from humans in the community (98 and 65 isolates), broilers (126 and 122), and pigs (102 and 88) during 1998 were tested for susceptibility to 12 different antimicrobial agents and for the presence of selected genes encoding resistance using PCR. Furthermore, the presence of vancomycin resistant enterococci was examined in 38 human stool samples using selective enrichment. Widespread resistance to chloramphenicol, macrolides, kanamycin, streptomycin, and tetracycline was found among isolates from all three sources. All E. faecium isolates from humans and pigs were susceptible to avilamycin, whereas 35% of isolates from broilers were resistant. All E. faecium isolates from humans were susceptible to vancomycin, whereas 10% and 17% of isolates from broilers and pigs, respectively, were resistant. A vancomycin resistant E. faecium isolate was found in one of the 38 human fecal samples examined using selective enrichment. All vancomycin resistant isolates contained the vanA gene, all chloramphenicol resistant isolates the cat(pIP501) gene, and all five gentamicin resistant isolates the aac6-aph2 gene. Sixty-one (85%) of 72 erythromycin resistant E. faecalis examined and 57 (90%) of 63 erythromycin resistant E. faecium isolates examined contained ermB. Forty (91%) of the kanamycin resistant E. faecalis and 18 (72%) of the kanamycin resistant E. faecium isolates contained aphA3. The tet(M) gene was found in 95% of the tetracycline resistant E. faecalis and E. faecium isolates of human and animal origin, examined. tet(K) was not observed, whereas tet(L) was detected in 17% of tetracycline resistant E. faecalis isolates and in 16% of the E. faecium isolates. tet(O) was not detected in any of the isolates from pigs, but was observed in 38% of E. faecalis isolates from broilers, in two E. faecalis isolates from humans and in three E. faecium isolates from broilers. tet(S) was not detected among isolates from animals, but was observed in 31% of E. faecalis and one E. faecium isolate from humans. This study showed a frequent occurrence of antimicrobial resistance and the presence of selected resistance genes in E. faecalis and E. faecium isolated from humans, broilers and pigs. Differences in the occurrence of resistance and tetracycline resistance genes were observed among isolates from the different sources. However, similar resistance patterns and resistance genes were detected frequently indicating that transmission of resistant enterococci or resistance genes takes place between humans, broilers, and pigs.

Animals↗
PubMed1999

[Drug resistance of Enterococcus species isolated from the urogenital system].

Despite low virulence of enterococci, they have become important nosocomial pathogens. This has been correlated with the increased use of broad-spectrum antibiotics, particularly cephalosporins. Many strains of enterococci exhibit multiple drug resistance; the most important being high-level resistance (HLR) to penicillin (MIC > 100 mg/l) and gentamicin (MIC > 500 mg/l and 2000 mg/l) and/or streptomycin (MIC > 2000 mg/l). The investigation was performed on 92 strains, isolated from genito-urinary tract and recognised as Enterococcus sp. All strains were obtained from several microbiological laboratories of Gdańsk, Gdynia and Tczew. On biochemical reaction profiles species of enterococci were identified as: E. faecalis (72.8%), E. faecalis varians (9.8%), E. durans (7.6%) and E. faecium (9.8%). The minimal inhibitory concentration (MICs) of penicillin, ampicillin, azlocillin, imipenem, gentamicin, amicacin, ciprofioxacin and vancomycin were determined by the agar dilution method. None of these 92 enterococcal strains was vancomycin resistant. 22.2% of E. faecium and 7.5% of E. faecalis showed high-level resistance to penicillin. None of these strains were produced beta-lactamase. High-level resistance to streptomycin and gentamicin was detected. Both--high-level resistance to streptomycin and gentamicin--were found in 6% E. faecalis; 11.1% E. faecalis varians and 22.2% E. faecium.

Drug Resistance, Microbial↗
PubMed2000

Comparison of phenotypic methods to identify enterococci intrinsically resistant to vancomycin (VanC VRE).

Clinical laboratories must be able to differentiate between enterococci possessing acquired resistance to vancomycin (vanA and vanB genotypes) from those that are inherently resistant (vanC1 and vanC2/C3 genotypes). We compared several routine phenotypic tests to determine the species identity of clinical isolates of enterococci and a PCR assay for the van ligase genes was used to confirm identification of VanC VRE. The Vitek Gram Positive Identification card identified 53/60 (88%) Enterococcus faecalis and E. faecium isolates and 81/141 (57%) VanC VRE without additional testing. Another 32 of the VanC VRE required additional testing (e.g., motility and pigmentation) for correct identification. However, 7 of these 32 VanC VRE were nonmotile. The rapid ID 32 STREP strips identified 50/60 (83%) E. faecalis and E. faecium isolates and 102/141 (72%) VanC VRE. All E. faecalis and E. faecium isolates were nonmotile and did not acidify 1% methyl-alpha-D-glucopyranoside (MGP). Only 93/115 (81%) E. gallinarum and 21/26 (81%) E. casseliflavus/E. flavescens were motile but all 141 VanC VRE acidified MGP. MGP acidification can accurately differentiate VanC VRE from E. faecalis and E. faecium. Because some VanC VRE isolates are nonmotile, MGP acidification is preferred as a simple and less costly test for identification of these isolates.

Bacterial Proteins↗
PubMed1995

Species identification and antibiotic susceptibility of enterococci isolated from clinical specimens of hospitalized patients.

Over a 4-month period, a total of 315 enterococci were isolated from various clinical specimens of hospitalized patients. By applying an array of biochemical tests, all strains were accurately identified to the species level, and their susceptibilities to clinically relevant antibiotics were determined by a standardized agar dilution technique. E. faecalis and E. faecium accounted for 87.3% and 9.2% of isolates, respectively. E. avium (1%), E. gallinarum (1%), E. durans (0.6%), E. hirae (0.6%), and E. casseliflavus (0.3%) isolates were also identified. Eleven strains of E. faecium and 1 E. hirae isolate were resistant to ampicillin, but none of the isolates produced beta-lactamase. Twenty-three E. faecium and 3 E. faecalis strains as well as 1 E. hirae isolate revealed imipenem resistance. A total of 25.4% enterococci (60 E. faecalis and 19 E. faecium isolates, 1 E. hirae strain) were erythromycin-resistant. Twelve strains (11 E. faecium and 1 E. avium) exhibited ciprofloxacin resistance. High-level resistance to streptomycin was found in 58 (21.1%) E. faecalis, 9 (31%) E. faecium, and both E. hirae strains, whereas high-level gentamicin resistance (HLGR) was exclusively seen in the species E. faecalis (11.6% of isolates belonging to this species). A simple agar screening test containing 500 micrograms of gentamicin per ml proved to be highly reliable for detection of HLGR. The structural gene coding for HLGR was specifically amplified by the polymerase chain reaction in all isolates showing this resistance trait. Moreover, the gene was specifically detected by a nonradioactively labelled oligonucleotide probe in colony blot hybridization assays, indicating the potential application of these molecular approaches as a diagnostic tool.

Ampicillin↗
PubMed1991

Platelet aggregation by strains of enterococci.

The platelet aggregation capability of whole cells of Enterococcus faecalis, E. faecium and E. avium was tested. The optimum ratios of bacteria to platelets in E. faecalis (strain SMU-37), E. faecium (strain SMU-138) and E. avium (strain SMU-197) were 1.0, 1.2 and 2.0, respectively. During the platelet aggregation induced by the three strains of enterococci, 65-69% of total serotonin was released. The aggregation was totally inhibited by ethylenediaminetetraacetate (10 mM) and apyrase (1 mg/ml), while no effect was shown by aspirin (10 mM), indomethacin (10 mM) and quinacrine (1 mM). By pretreatment of platelet-poor plasma with heat (56 C, 30 min) or zymosan, the reactivities with platelets of each strain of species were markedly diminished. These results suggest that enterococci-induced platelet aggregation was an ion-dependent, cyclooxygenase-insensitive event, and plasma component(s) was (were) required for the reaction.

Animals↗
PubMed2005

Acquired antimicrobial resistance in the intestinal microbiota of diverse cat populations.

The aim of this study was to investigate the prevalence of acquired antimicrobial resistance in the resident intestinal microbiota of cats and to identify significant differences between various cat populations. Escherichia coli, Enterococcus faecalis, E. faecium and Streptococcus canis were isolated as faecal indicator bacteria from rectal swabs of 47 individually owned cats, 47 cattery cats and 18 hospitalised cats, and submitted through antimicrobial sensitivity tests. The results revealed that bacteria isolated from hospitalised and/or cattery cats were more frequently resistant than those from individually owned cats. E. coli isolates from hospitalised cats were particularly resistant to ampicillin, tetracycline and sulfonamide. Both enterococci and streptococci showed high resistance to tetracycline and in somewhat lesser extent to erythromycin and tylosin. Most E. faecium isolates were resistant to lincomycin and penicillin. One E. faecalis as well as one E. faecium isolate from hospitalised cats showed 'high-level resistance' (MIC > 500 microg/ml) against gentamicin, a commonly used antimicrobial agent in case of human enterococcal infections. The results of this research demonstrate that the extent of acquired antimicrobial resistance in the intestinal microbiota of cats depends on the social environment of the investigated population. It is obvious that the flora of healthy cats may act as a reservoir of resistance genes.

Animals↗
PubMed1996

[Identification of Enterococcus species from clinical samples and their antimicrobial susceptibility].

The genus enterococcus has 12 species of which, E faecalis and E faecium are most important in human infections. A progressive resistance to penicillin and ampicillin has been detected in these species. The aim of this work was to identify Enterococcus species isolated in a hospital and to study their antimicrobial susceptibility. We studied 209 Enterococcus species coming from patients admitted to a public hospital. Their susceptibility to penicillin, ampicillin, imipenem, vancomycin, tetracycline, chloramphenicol, ciprofloxacin, gentamicin and streptomycin was determined with the agar dilution technique. Eighty seven percent of species were E faecalis and 7.1% were E fecium, other isolated species were E hirae, E casseliflaws, E avium, E solitarius and E faecalis variant. Thirty eight percent of these species were isolated from the urinary tract, 22% from the skin and 14% from surgical wounds. All E faecalis species were susceptible to penicillin, ampicillin, imipenem and vancomycin; 27.3% were susceptible to tetracycline, 54.7% to chloramphenicol and 80% to ciprofloxacin. Seventy three percent of E faecium species were susceptible to penicillin, 80% to ampicillin and 60% to imipenem. Sixty two percent of E faecalis and 42.4% of E faecium were resistant to streptomycin. It is concluded that the correct identification of Enterococcus species has therapeutic implications.

Anti-Bacterial Agents↗
PubMed2005

Characterization of antibiotic resistance genes and virulence factors in faecal enterococci of wild animals in Portugal.

Antibiotic susceptibility was tested in 140 non-selected enterococci (73 Enterococcus faecalis, 45 E. faecium and 22 of other species) recovered from faecal samples of 77 wild animals in Portugal. Susceptibility testing for 11 antibiotics (vancomycin, teicoplanin, ampicillin, streptomycin, gentamicin, kanamycin, chloramphenicol, tetracycline, erythromycin, quinupristin-dalfopristin and ciprofloxacin) was determined by disk diffusion and agar dilution methods. Forty-four isolates (31.4%) showed susceptibility to all the antibiotics tested (5.5% of E. faecalis; 62.2% of E. faecium; and 78.6% of E. hirae). Neither ampicillin-resistance nor acquired-vancomycin-resistance was detected and 1.4% of the isolates showed high-level-resistance for gentamicin or streptomycin. Tetracycline and erythromycin resistances were shown in 28.6% and 20.1% of the isolates, respectively. Antibiotic resistance genes were studied by polymerase chain reaction (PCR) and sequencing and tet(M) + tet(L), erm(B) or aac(6')-aph(2'') genes were detected in most of tetracycline-, erythromycin- or gentamicin-resistant enterococci respectively. Genes encoding virulence factors were studied by PCR and a wide variety of virulence genes were detected in most of E. faecalis isolates but were rarely found in E. faecium and not detected in the other species. The prevalence of genes encoding virulence factors in E. faecalis was as follows: cpd (98.6%), gelE (75.3%), agg (30.1%), fsr (17.8%), ace (9.6%) and esp (4.1%). Low percentages of antibiotic resistance was found in the faecal enterococci of wild animals but a wide variety of virulence genes were detected among E. faecalis isolates although were rare in the other species.

Animals↗
PubMed2006

Genotypic diversity and epidemiology of high-level gentamicin resistant Enterococcus in a Chinese hospital.

OBJECTIVE: To investigate the antibiotics resistance of Enterococcus, the aminoglycoside-modifying enzymes (AME) and homology of high-level gentamicin resistant (HLGR) Enterococcus in clinical specimens for the implementation of effective infection control measures. METHODS: The resistance of 13 antimicrobial agents was determined by Kirby-Bauer (K-B) or agar dilution method. And the HLGR and high-level streptomycin resistant (HLSR) isolates were screened by agar screen. Production of beta-lactamases was tested by the nitrocefin disc method. The aminoglycoside-modifying enzyme genes were detected by polymerase chain reaction (PCR). Pulsed-field gel electrophoresis (PFGE) was used to analyze the homology of HLGR isolates from in-patients. RESULTS: No isolates resistant to linezolid, vancomycin and teicoplanin were found. Ampicillin-resistant isolates did not produce beta-lactamases and 68 HLGR isolates were screened at the rate of 64.2%. The positive rate of aac(6')-Ie-aph(2'')-Ia was 86.8% and 3 isolates had the new AME gene designated aph(2'')-Ie mostly similar to aph(2'')-Id. Among 51 HLGR isolates from in-patients, PFGE grouped 17 Enterococcus faecalis (E. faecalis) isolates into 4 clusters (A-D), and 33 Enterococcus faecium (E. faecium) isolates into 8 clusters (A-H), of which the A cluster is the main. CONCLUSIONS: HLGR has become the important antibiotic resistance pathogen causing nocosomial infection. And the aac(6')-Ie-aph(2'')-Ia gene was the main aminoglycoside-modifying enzyme gene leading to HLGR.

Acetyltransferases↗
PubMed2006

Phenotypic and genotypic study of gelatinase and beta-haemolysis activities in faecal enterococci of poultry in Portugal.

The detection of gelatinase and beta-haemolysis activities was carried out in 83 faecal enterococci (43 Enterococcus faecalis, 33 E. faecium, five E. durans and two E. hirae) of poultry origin. In addition, the presence of genes of the gelE-fsrABC locus and of the cyl operon (cylL(L), cylL(S), cylA, cylB and cylM) were studied by polymerase chain reaction and correlated with gelatinase and beta-haemolysis production, respectively. Most of our E. faecalis isolates were gelatinase-positive (88%), being this activity not frequent in the other enterococcal species (2.5%). Only one of the 33 E. faecium isolates showed a positive gelatinase reaction. All enterococci that showed gelatinase activity harboured the gelE and fsrABC genes, although these genes were also detected in four E. faecalis and one E. durans gelatinase-negative isolates. Most of our non-E. faecalis gelatinase-negative isolates did not harbour gelE-fsrABC genes. A high proportion of faecal enterococci of poultry origin harboured genes of the cyl operon (71%), although only 7% contained the five cyl tested genes (all of them E. faecalis). Only one isolate of our series could express beta-haemolysis, harbouring the whole cyl operon. The cylL(S) genotype was the most prevalent in our enterococci (39%) and also the most prevalent among our E. faecalis isolates (60%). Other genotypes detected were the following ones (% of enterococci): cylA + cylB + cylM (13%), cylL(L) + cylA (4%), cylL(L) (4%), cylL(L) + cylA + cylB + cylM (2%), cylL(L) + cylA + cylM (1%) and cylA + cylM (1%). Both phenotypic and genotypic assays are important to evaluate the virulence potential of enterococci.

Animals↗