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[Characteristics of Enterococcus strains isolated from neutropenic patients at the National Bone-Marrow Transplantation Center of Tunis].

The frequency of digestive colonization of neutropenic patients by Enterococci during the phase of pre-transplant and post-transplant of bone marrow is important indeed as 441 Enterococcus spp strains have been isolated from stool-cultures and other specimens whithin a period of 35 months in 80 patients. A quantitative stool culture was done on appropriate media. Simple bile-esculine agar (BE) and bile esculine-agar additionned with 6 mg/l of vancomycine (BEV) were used for detecting Enterococci. These organisms were taken into account, when then numeration was > 10(3) UFC/g of fecal sample on BE and in all cases on BEV. Species isolated were essentially Enterococcus faecalis (39.4%), Enterococcus faecium (34.4%) and Enterococcus casseliflavus (17%). These strains were characterized by a high frequency of high-level resistance to gentamicin (40.8%). Resistance to amoxicillin concerns 40% of E. faecium strains. Seventeen multiresistant strains of E. faecium isolated from 7 patients (colonized during a long period and receiving a gut decontamination treatment) were the subject of a phenotypic analysis (biotyping, antibiotyping and determination of MICs to beta-lactam, aminoglycosides and glycopeptides). This analysis showed that these strains are distinguishable, endogenic and specific for each patient, the common multiresistance trait resulted from the selective pressure of decontamination treatment used for our patient.

Adolescent↗

[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↗

[Evaluation of a rapid method for the identification of the Enterococcus genus].

BACKGROUND: The evaluation of the reliability of a group of three tests: pyrrolidonil-arilamidase, rapid test in NaCl esculin broth and group D antistreptococci agglutination for the rapid identification of the Enterococcus genus were performed. METHODS: One hundred fifty-eight strains of streptococci: 110 strains of Enterococcus faecalis, 8 E. faecium, 3 E. avium, 16 Streptococcus intermedius, 7 S. salivarius, 4 S. bovis, 4 S. constellatus, 2 S. sanguis and 1 strain of S. equinus, S. mitis, S. uberis and S. mutans were studied. The tests used were: rapid test in NaCl- esculin broth, pyrrolidonyl-arilamidase and antigroup D streptococcus agglutination (Patho Dx Strep D/PYR; Diagnostic Product Corporation). All catalase-negative gram-positive cocci or coccibacilli which were positive in the three tests were considered to belong to the Enterococcus genus. The usefulness of Gram staining in culture in thioglycolate broth was also studied comparing it with the results obtained with solid medium. RESULTS: The results of the proposed identification scheme were evaluated calculating different parameters: sensitivity 98%, specificity 100%, positive predictive value, 100%, and negative predictive value 94%. No significant differences were observed between the results obtained with Gram staining with thioglycolate broth and those obtained with solid medium except in the case of the Streptococcus mutans strain. CONCLUSIONS: The application of a combination of tests: pyrroidonyl-arilamidase, antistreptococci group D agglutination and rapid test in NaCl-esculin broth to catalase-negative gram-positive cocci and coccobacilli allows reliable identification in a maximum of three hours of strains belonging to the Enterococcus genus.

Bacteriological Techniques↗

Susceptibility patterns of Enterococcus spp. isolated in Poland during 1996.

Susceptibility of Enterococcus spp. isolated from various clinical specimens to different antimicrobial agents was evaluated. Of the 346 enterococcal isolates obtained from four regional Polish hospitals during 6 months of 1996, 261 (75.4%) were identified as Enterococcus faecalis, 75 (21.7%) as Enterococcus faecium and ten (2.9%) as other enterococcal species. High-level resistance to gentamicin was expressed by 33.4% of E. faecalis and 86.5% of E. faecium strains and corresponding streptomycin resistance by 43.9 and 82.4%, respectively. Over 80% of E. faecium isolates were resistant to ampicillin. None of the isolates was resistant to teicoplanin, however 7.9% of E. fecalis and 1.4% of E. faecium strains were moderately susceptible to vancomycin.

Anti-Bacterial Agents↗

[Genotypes of aminoglycoside-modifying enzyme and clinical study of high-level gentamycin resistant enterococcus].

OBJECTIVE: To determine the antibiotics resistance, aminoglycoside-modifying enzymes and homology of high-level gentamycin resistant enterococcus in clinical specimens. METHODS: The high-level gentamicin resistant (HLGR) isolates were screened by the agar method and the resistance of 14 antimicrobial agents was determined by K-B 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. RESULTS: The ratio of HLGR was 64.2% (68/106). Among the HLGR,there were no isolates resistant to linezolid, vancomycin and tecoplanin, and Enterococcus faecium was more resistant to beta-lactam antibiotics and quinolone than Enterococcus faecalis. The positive rate of aac(6')-Ie-aph(2')-Ia was 92.6% and 3 isolates had the resistance gene mostly similar to aph(2')-Id. And among 51 HLGR isolates from the hospitalized patients, PFGE grouped 17 E. faecalis isolates into 4 clusters (A-D), and 33 E. faecium isolates into 8 clusters (A-H) with A cluster as predominant. CONCLUSION: HLGR has become the important antibiotic resistance bacteria which results in nosocomial infection; and aac(6')-Ie-aph(2')-Ia is the main aminoglycoside-modifying enzyme gene which causes HLGR.

Drug Resistance, Bacterial↗

[Antibiotic susceptibility analysis of Enterococcus spp. isolated from urine].

Recently increase of enterococcal infections has been observed. These bacteria, mainly Enterococcus faecalis and Enterococcus faecium are members of the normal flora of gastrointestinal tract but also are typical opportunistic pathogens. Enterococci are characterized by natural resistance to numerous antibiotics (among them cephalosporins), and also by easy acquired resistance to antibiotics. Infections caused by multiresistant strains are difficult in treatment, chronic, recurrent and sometimes fatal are described. Enterococcal infections are caused often by E. faecalis, rarely by E. faecium. In the last years other species of enterococci have been isolated from different clinical materials (E. casseliflavus, E. avium, E. durans, E. gallinarum). The aim of this study was to analyze the antibiotics susceptibility of Enterococcus spp. isolated from urine of children with urinary tract infection (UTI), hospitalized in the Upper Silesian Health Center of Child and Mother or treated ambulatory. Susceptibility of the 130 strains of E. faecalis and 41 E. faecium to: penicillin, tetracycline, high level of aminoglycoside (gentamycin 120 microg and streptomycin 300 microg), glycopeptides (vancomycin and teicoplanin), ciprofloxacin and nitrofurantoin was estimated. All isolated strains of E. faecalis (100%) were sensitive to glycopeptides (vancomycin and teicoplanin) and to nitrofurantoin, 96% strains were sensitive to penicillin, 43% to ciprofloxacin and 28% to tetracycline. All strains of E. faecium were sensitive to glycopeptides (vancomycin and teicoplanin), 32% strains were sensitive to penicillin, 19% to tetracycline 14% to ciprofloxacin and 50% to nitrofurantoin. Twenty two strains of E. faecalis (17%) and twelve strains of E. faecium (29%) were demonstrated high level resistance to aminoglycosides (HLAR). Among HLAR strains of enterococci, were observed strains of high level resistance only to streptomycin, high level resistance only to gentamycin and high level resistance to both aminoglycosides. The HLAR strains of E. faecalis and E. faecium were also resistant to other antibiotics: penicillin, tetracycline and ciprofloxacin. The most frequent cause of child urinary tract infection was E. faecalis, with high sensitivity to penicillins, glycopeptides and nitrofurantoin. The increased role of E. faecium and other strains of enterococci also has been shown. These species demonstrated sensitivity to glycopeptides, but resistance to other antibiotics, routinely used for treatment of UTI as well as to nitrofurantoin.

Anti-Bacterial Agents↗

Enterococcus lactic acid bacteria strain SF68 and lactulose in hepatic encephalopathy: a controlled study.

Forty cirrhotic patients with non-advanced hepatic encephalopathy were randomly allocated into groups which were given orally either two capsules three times daily of a preparation of Enterococcus lactic acid bacteria strain SF68 or 30 ml lactulose four times daily. The patients were evaluated over a 10-day course of treatment and for 10 days post-treatment. The Enterococcus SF68 preparation proved to be as effective as lactulose in lowering blood ammonia, and in improving mental state and psychometric performance. Moreover, the effects of Enterococcus SF68, contrary to that of lactulose, persisted longer after treatment withdrawal. Some patients reported diarrhoea and abdominal pain with lactulose. Lactulose is a standard therapy in the treatment of patients with hepatic encephalopathy. In this study, however, the use of the Enterococcus SF68 preparation was shown to offer advantages over lactulose in these patients.

Adult↗

Double-blind report on the efficacy of lactic acid-producing Enterococcus SF68 in the prevention of antibiotic-associated diarrhoea and in the treatment of acute diarrhoea.

A multicentre double-blind, placebo-controlled clinical trial, involving 123 patients at 10 centres, was carried out to assess the efficacy of a preparation of lactic acid-producing Enterococcus SF 68 in the prevention of antibiotic-associated diarrhoea and in the treatment of acute diarrhoea. In the prevention study, 45 patients being treated with antibiotics were given, concurrently, one capsule twice daily of either Enterococcus SF68 or placebo. Acute diarrhoea was present in 78 patients who were given the same treatment but three times daily. All treatments were continued for 7 days. Enterococcus SF 68 was shown to be effective in reducing the incidence of antibiotic-associated diarrhoea in comparison with placebo (8.7% compared with 27.2%, respectively). Patients with acute enteritis showed a significantly faster resolution of bowel abnormalities during treatment with Enterococcus SF68 compared with placebo.

Adult↗

Molecular detection of Enterococcus species in root canals of therapy-resistant endodontic infections.

OBJECTIVES: The objectives were to identify Enterococcus spp in nonhealing endodontic cases using PCR amplification and molecular sequencing, and to determine if the prevalence of enterococci is increased in diabetic patients. STUDY DESIGN: Specimens from 40 cases undergoing retreatment were incubated in prereduced thioglycollate broth at 37 degrees C. Extracted DNA had PCR amplification using primers that target the tuf gene of 14 Enterococcus spp. PCR products were directly sequenced and identified phylogenetically. RESULTS: Three cases were eliminated because the patients were on antibiotics or the tooth did not have a periradicular radiolucency. The remaining 37 specimens included 6 from diabetic patients. Eight specimens were positive for Enterococcus spp. Of these, 6 (19%) were from nondiabetic and 2 (33%) from diabetic patients (odds ratio = 2.1; chi squared, P = .45). Phylogenetically, all sequences from positive specimens matched E faecalis V583 (AE016947). Conclusion E faecalis was the only enterococcal species detected, with an overall prevalence of 22%.

Adult↗

Genotypic characterisation of vancomycin-resistant Enterococcus faecium isolates from haemato-oncological patients at Olomouc University Hospital, Czech Republic.

This study describes the first molecular characterisation of clinical isolates of vancomycin-resistant enterococci (VRE) in the Czech Republic. Of 2647 patient isolates of Enterococcus spp. from 1997-2002, 121 (4.6%) were identified as VRE. The most common isolates were VanA+ Enterococcus faecium (78%) and VanB+ Enterococcus faecalis (10%). In addition, five VanA+ E. faecium isolates were obtained from environmental and staff sampling. Macrorestriction analysis of SmaI restriction fragment length polymorphism was performed for 54 VanA+ E. faecium clinical isolates and the five VanA+ E. faecium environmental isolates. Thirty-two unique restriction endonuclease patterns were identified, including two predominant clonal types represented by five or more isolates. Two environmental VanA+ E. faecium isolates were closely related to two patient isolates, which had an identical SmaI macrorestriction pattern. The results indicated potential survival of strains in the hospital environment and possible subsequent transmission to hospitalised patients.

Cancer Care Facilities↗

Bacteriocin T8, a novel class IIa sec-dependent bacteriocin produced by Enterococcus faecium T8, isolated from vaginal secretions of children infected with human immunodeficiency virus.

Enterococcus faecium T8, isolated from vaginal secretions of children with human immunodeficiency virus, produces a class IIa sec-dependent bacteriocin that is structurally different from three other class IIa sec-dependent bacteriocins, i.e., enterocin P and an enterocin P-like bacteriocin, produced by Enterococcus faecium, and bacteriocin 31, produced by Enterococcus faecalis, and from a class III bacteriocin produced by E. faecalis. The genes encoding the bacteriocin, immunity protein, mobilization protein, and relaxase nuclease are located on a 7-kb plasmid. Bacteriocin T8 has a molecular mass of 5.1 kDa based on its DNA sequence, similar to the 5.0 kDa recorded for bacteriocin 31 but larger than the 4.6 kDa reported for enterocin P. At the amino acid level, bacteriocin T8 is 69% homologous to bacteriocin 31 and 47% homologous to enterocin P. Bacteriocin T8 is active against E. faecalis isolated from patients diagnosed with vaginosis, against Lactobacillus sakei, and against a Propionibacterium sp. The peptide is heat stable (60 min at 100 degrees C) and remains active in phosphate buffer from pH 4.0 to 10.0. The mode of activity is bactericidal, as determined with E. faecalis.

Amino Acid Sequence↗

Structure of the low-affinity penicillin-binding protein 5 PBP5fm in wild-type and highly penicillin-resistant strains of Enterococcus faecium.

Among its penicillin-binding proteins (PBPs), Enterococcus faecium possesses a low-affinity PBP5, PBP5fm, which is the main target involved in beta-lactam resistance. A 7.7-kb EcoRI chromosomal fragment of E. faecium D63r containing the pbp5fm gene was cloned and sequenced. Two open reading frames (ORFs) were found. A 2,037-bp ORF encoded the deduced 73.8-kDa PBP5fm, the amino acid sequences of which were, respectively, 99.8, 78.5, and 62% homologous to those of the low-affinity plasmid-encoded PBP3r of Enterococcus hirae S185r and the chromosome-encoded PBP5 of E. hirae R40 and Enterococcus faecalis 56R. A second 597-bp ORF, designated psrfm, was found 2.3 kb upstream of pbp5fm. It appeared to be 285 bp shorter than and 74% homologous with the regulatory gene psr of E. hirae ATCC 9790. Different clinical isolates of E. faecium, for which a wide range of benzylpenicillin MICs were observed, showed that the increases in MICs were related to two mechanisms. For some strains of intermediate resistance (MICs of 16 to 64 micrograms/ml), the increased level of resistance could be explained by the presence of larger quantities of PBP5fm which had an affinity for benzylpenicillin (second-order rate constant of protein acylation [k+2/K] values of 17 to 25 M(-1) s(-1)) that remained unchanged. For the two most highly resistant strains, EFM-1 (MIC, 90 micrograms/ml) and H80721 (MIC, 512 micrograms/ml), the resistance was related to different amino acid substitutions yielding very-low-affinity PBP5fm variants (k+2/K < or = 1.5 M(-1) s(-1)) which were synthesized in small quantities. More specifically, it appeared, with a three-dimensional model of the C-terminal domain of PBP5fm, that the substitutions of Met-485, located in the third position after the conserved SDN triad, by Thr in EFM-1 and by Ala in H80721 were the most likely cause of the decreasing affinity of PBP5fm observed in these strains.

Amino Acid Sequence↗

Evaluation of the revised MicroScan dried overnight gram-positive identification panel to identify Enterococcus species.

The revised MicroScan Dried Overnight Gram-Positive Identification panel was evaluated for its efficacy at identifying Enterococcus species in comparison with conventional biochemical tests. Supplemental testing of ampicillin-susceptible Enterococcus faecium for motility and the ability to acidify methyl-alpha-D-glucopyranoside helped recognize E. gallinarum and increased the accuracy of the panel for identifying Enterococcus species to 98.5%.

Ampicillin Resistance↗

In-vitro activity of nitrofurantoin in enterococcus urinary tract infection.

OBJECTIVE: To compare in-vitro antimicrobial susceptibility of nitrofurantoin against commonly used antimicrobials for the treatment of urinary tract infection caused by enterococci. METHODS: A total of 144 enterococci (Enterococcus faecalis, n=130, Enterococcus faecium, n=14) isolated and confirmed by biochemical tests at the department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi, for a period of three years were included in the study. The antimicrobial susceptibility tests of isolated organisms were performed by standard disc diffusion method as recommended by NCCLS. In addition to nitrofurantoin (NIT), doxycycline (DOX), co-trimoxazole (COT), ampicillin (AMP), imipenem (IPM) and vancomycin (VAN) were used for antimicrobial susceptibility testing. RESULTS: One hundred and twenty seven (88%) isolates of Enterococci were susceptible to NIT while 72% (n = 103) to AMP, 13% to COT, 11% to DOX, 91% to IPM and 99% to VAN. Among the strains resistant to AMP, COT, DOX, IPM and VAN, 50-88% were susceptible to NIT. CONCLUSION: Nitrofurantoin is an effective antimicrobial in vitro and can be used for treatment of Enterococcus urinary tract infections.

Anti-Bacterial Agents↗

Third-generation cephalosporins and vancomycin as risk factors for postoperative vancomycin-resistant enterococcus infection.

OBJECTIVE: To examine use of third-generation cephalosporins (3GCs) alone and in association with vancomycin hydrochloride as a risk factor for vancomycin-resistant enterococcus (VRE) infection in surgical patients. DESIGN: Case-control retrospective study analyzing antibiotic use in the 30 days preceding culture of VRE or vancomycin-sensitive enterococcus from an infected site. SETTING: A large tertiary care teaching hospital. PATIENTS: Surgical inpatients with VRE infections between September 3, 1993, and January 29, 1997, were matched with patients with vancomycin-sensitive enterococcus infections. Matches were based on surgical procedure, initial infection site, and immunosuppression. Matches were found for 32 of 50 surgical patients with VRE. Twenty matched pairs of patients were recipients of solid organ transplants. MAIN OUTCOME MEASURES: Multivariate logistic regression analysis was done to examine 3GCs and vancomycin as risk factors for VRE infection. Univariate analysis of use of other antibiotic agents and demographic data was also performed. RESULTS: Multivariate analysis showed significant differences in the use of 3GCs both alone and concurrently with vancomycin. Univariate analysis also showed higher use of metronidazole, concurrent vancomycin and metronidazole, concurrent vancomycin and ceftazidime, and all antibiotics combined in patients with VRE infections. CONCLUSIONS: This matched control study showed that use of 3GCs, alone (P=.05) or concurrently with vancomycin (P=.05), was a risk factor for VRE infection in surgical patients. Judicious administration of third-generation antibiotics is warranted in surgical patients with other risk factors for VRE.

Adult↗

Effect of hypoxia on the immune response of giant freshwater prawn Macrobrachium rosenbergii and its susceptibility to pathogen Enterococcus.

Giant freshwater prawns Macrobrachium rosenbergii (14-19 g) were challenged with Enterococcus (3 x 10(5) cfu prawn(-1)) previously incubated in TSB medium for 24 h, then placed in water having concentrations of dissolved oxygen (DO) at 7.75, 4.75, 2.75 and 1.75 mg l(-1). Onset of mortality occurred after 6 h exposure to 1.75 mg l(-1) DO, and after 12 h exposure to 2.75 mg 1(-1) DO. Cumulative mortality of prawns at 1.75 mg l(-1) DO was significantly higher than that at 4.75 and 2.75 mg l(-1) DO, and cumulative mortality of prawns at 4.75 and 2.75 mg l(-1) DO was significantly higher than that at 7.75 mg l(-1) DO after 96 h. The prawns (20-30 g) which had been placed in water for 0 to 120 h at 7.75, 4.75, 2.75 and 1.75 mg l(-1) DO were examined for the THC (total haemocyte counts), DHC (differential haemocyte counts), phenoloxidase activity, respiratory burst, percentage phagocytosis and clearance efficiency. No significant difference in semi-granular cells and granular cells of prawns was observed among four treatments. The prawns following 120 h exposure to 2.75 mg l(-1) DO decreased significantly the hyaline cells and THC by 39% and 36%, respectively. Phenoloxidase activity and respiratory burst decreased significantly by 33% and 11% when the prawns were exposed to 2.75 mg l(-1) DO after 24 h, respectively. Percentage phagocytosis and clearance efficiency to Enterococcus decreased significantly by 44% and 54% for the prawns following 12 h exposure to 2.75 mg l(-1) DO, respectively. It is concluded that DO as low as 2.75 mg l(-1) and 4.75 mg l(-1) causes depression in immune system of M. rosenbergii, and increases its susceptibility to Enterococcus infection.

Animals↗

Detection of aminoglycoside-penicillin synergy against Enterococcus faecium using high-content aminoglycoside disks.

Thirty-seven Enterococcus faecium strains were screened for high-level aminoglycoside resistance with an agar diffusion test using high-content aminoglycoside disks (300 micrograms of streptomycin and 120 micrograms of gentamicin, tobramycin, kanamycin or amikacin). The inhibition zones obtained were correlated with results of time-kill penicillin-aminoglycoside synergy studies. An 11 mm breakpoint differentiated strains susceptible or resistant to the synergy of streptomycin plus penicillin. Irrespective of the inhibition zones obtained with tobramycin and kanamycin disks, Enterococcus faecium strains never showed synergy with penicillin in combination with these aminoglycosides. Penicillin-amikacin synergy cannot be predicted by the amikacin disks. Nevertheless, even though kanamycin disks do not predict penicillin-kanamycin synergy, they can be used to predict penicillin-amikacin synergy. In summary, high-content streptomycin, gentamicin and kanamycin disks can be used to predict the susceptibility of Enterococcus faecium strains to the synergistic combination of penicillin plus one of the aminoglycosides (streptomycin, gentamicin or amikacin, respectively).

Aminoglycosides↗

In vitro activity of daptomycin-metronidazole combinations against mixed bacterial cultures: reduced activity of metronidazole against Bacteroides species in the presence of Enterococcus faecalis.

The in vitro activity of daptomycin-metronidazole combinations against mixed cultures of gram-positive facultative cocci and strains of the Bacteroides fragilis group was investigated. Metronidazole did not influence the high activity of daptomycin against strains of Staphylococcus aureus, Staphylococcus epidermidis and Enterococcus faecalis in the absence or presence of the co-cultured Bacteroides strains. In contrast, the Enterococcus faecalis isolates protected the co-cultured Bacteroides strains against the killing effect of metronidazole, even at a concentration four- or eightfold the MIC of metronidazole. Killing curve experiments confirmed this protective effect of different isolates of Enterococcus faecalis.

Anti-Bacterial Agents↗