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[Neonatal meningitis due to Enterococcus spp.: presentation of four cases].

AIM: Enterococci are unusual etiologic agents of bacterial meningitis and account for only 0.3-4% of all cases. Neonatal enterococcal meningitis, which is rarely reported in the medical literature, presents characteristics that are significantly different from enterococcal meningitis affecting other age groups, particularly adults. PATIENTS AND METHODS: We retrospectively reviewed the clinical records of four newborns diagnosed with enterococcal meningitis in our center. Two were premature and two were term infants. Three were affected with early-onset meningococcal sepsis and one with late-onset sepsis. Risk factors for infection included intrapartum maternal fever in one case and prematurity in two cases, with prolonged stay in the neonatal intensive care unit and application of invasive procedures, and prior antibiotic treatment in one of infant. There were no apparent risk factors in the fourth case. Enterococcus faecalis was the causal agent in three cases and Enterococcus faecium in one. None of the enterococci were vancomycin-resistant. Antibiotic treatment included ampicillin and combinations of ampicillin and cefotaxime, ampicillin and amikacin, and vancomycin and gentamicin. None of the patients died. CONCLUSIONS: Enterococci rarely cause bacterial meningitis, though newborns seem more susceptible to this infection. With adequate bactericidal therapy, clinical outcome appears to be generally favorable.

Adult↗

[The use of the polymerase chain reaction method for the identification of representatives of the genus Enterococcus].

The possibility of the use of repetitive extragenic palindrome (REP)-specific primers to PCR-fingerprint analysis of Enterococcus genomes has been shown for the first time. Presence of the similar PCR products, and first of all the fragment 950 bp in size, has been observed in all the strains studied. Differences in PCR-product composition displayed genetical heterogeneity between Enterococcus faecium strains. These data evidence for the wide distribution of the REP, and rather high level of homology of the REP sequences among enterobacterial genomes.

Animals↗

The value of chemoprophylaxis against Enterococcus species in elective cholecystectomy: a randomized study of cefuroxime vs ampicillin-sulbactam.

HYPOTHESIS: Cephalosporins are widely used and considered to be effective as prophylaxis in biliary surgery. Nevertheless, they lack activity against enterococci. We conducted a study to compare the efficacy of ampicillin-sulbactam vs cefuroxime in preventing surgical site infections following elective cholecystectomy. DESIGN: A prospective randomized controlled trial. SETTING: A major tertiary care hospital. PATIENTS: Four hundred eighteen randomized patients (of 549 total), who from July 2002 to August 2004 underwent elective open or laparoscopic cholecystectomy with prospective assessment for development of surgical site infections for 1 month postoperatively. INTERVENTION: A single intravenous dose of 1.5 g of cefuroxime (group A, n = 207) or 3 g of ampicillin-sulbactam (group B, n = 211) was administered during induction of anesthesia. Bile and gallbladder mucosal cultures were taken intraoperatively from all patients. MAIN OUTCOME MEASURE: Number of postoperative surgical site infections. RESULTS: A postoperative surgical site infection was noted in 19 (4.5%) of 418 patients, 18 from group A and 1 from group B (P<.001). In the group that received cefuroxime, 15 (83.3%) of 18 surgical site infections were due to Enterococcus species. Intraoperative bactibilia as well as intraoperative gallbladder rupture were associated with surgical site infections (P<.001). CONCLUSIONS: A single dose of ampicillin-sulbactam favored better compared with cefuroxime for prevention of postoperative surgical site infections due to Enterococcus species after elective cholecystectomy. Ampicillin-sulbactam may be a better agent for antimicrobial prophylaxis in high-risk patients undergoing elective cholecystectomy, especially in a setting where the incidence of enterococcal infections is higher.

Ampicillin↗

Ampicillin and ampicillin-sulbactam dilution tests with mixed cultures of Bacteroides fragilis, Escherichia coli and Enterococcus.

Bacterial interactions in mixed infections may compromise antimicrobial therapy. The in vitro bactericidal activity of ampicillin and ampicillin-sulbactam against aerobic/anaerobic mixed cultures of Bacteroides fragilis, Escherichia coli and Enterococcus spp. was studied by means of broth dilution tests. The MBC of ampicillin for Enterococcus faecalis 6 was 0.25 mg/l when tested singly; in association with B. fragilis 1, however, the MBC for E. faecalis 6 was 16 mg/l. When tested singly E. coli 9 and B. fragilis 1 were both killed by ampicillin at a concentration of 4 mg/l in combination with 1 mg/l sulbactam. When both strains were associated, the MBC for B. fragilis 1 rose to > 256 mg/l. Results obtained indicate that the behaviour of bacteria as determined in pure cultures is not necessarily identical with the antibiotic susceptibility present in mixed cultures. It was shown that, besides beta-lactamase production by involved bacteria, other factors contribute to the alteration of bacterial susceptibility in mixed cultures. Synergistic and antagonistic effects between associated organisms were observed. For example, it was found that two moderately sensitive bacterial strains were resistant in mixed culture.

Ampicillin↗

Bactericidal activity of daptomycin against vancomycin-resistant Enterococcus faecium in an in vitro pharmacokinetic model.

A dynamic in vitro model was used to determine the killing kinetics of daptomycin against 15 vancomycin-resistant clinical isolates of Enterococcus faecium. Concentration profiles simulating those observed in serum following administration of both low-dose (2 mg/kg) and high-dose (6 mg/kg) daptomycin were bactericidal within 5.5 and 2.8 h, respectively. In contrast, when albumin was added to the growth medium, the corresponding bacterial killing times were slowed to greater than 24 h and 7 h; these results suggest that in the clinical setting, daptomycin dosages of approximately 6 mg/kg are required to achieve bactericidal activity against vancomycin-resistant Enterococcus faecium.

Albumins↗

Enterococcus faecalis endocarditis presenting as meningitis.

Although Enterococcus faecalis is a relatively common cause of infective endocarditis, it rarely causes meningitis. A case of Enterococcus faecalis endocarditis presenting as meningitis in a 74-year-old diabetic man on chronic hemodialysis is reported. A review of the literature showed that the association of enterococcal meningitis and endocarditis has rarely been reported. This clinical association may be more common than previously recognized and it is suggested that echocardiography be considered for all patients with enterococcal hematogenous meningitis in order to rule out endocarditis.

Endocarditis, Bacterial↗

Recovery from a single blood culture of two enterococcus gallinarum isolates carrying both vanC-1 and vanA cluster genes and differing in glycopeptide susceptibility.

Two Enterococcus gallinarum isolates distinguished by different colony sizes were recovered from the same blood culture from a woman with acute myeloid leukemia. They were designated E31 (the one with larger colonies) and E32 (the one with smaller colonies). Both isolates were glycopeptide resistant, but the MICs of vancomycin and teicoplanin for E31 (32 and 2 microg/ml, respectively, consistent with the VanC phenotype) and E32 (128 and 16 microg/ml, respectively, consistent with the VanA phenotype) were different. E31 and E32 had the same plasmid profile and showed identical pulsed-field gel electrophoresis patterns after digestion of total DNA with NotI and a two-band variation after digestion with SmaI. Polymerase chain reaction experiments showed that both isolates had both the vanC-1 and vanA genes and carried a Tn1546-related transposon lacking orf1, vanY, and vanZ. The absence of these three genes was confirmed by Southern analysis with appropriate probes. Southern hybridization experiments using a vanA probe showed that this atypical Tn1546-related element appeared to be located on the chromosome. In both E31 and E32, the vanA probe hybridized to EcoRV and HindIII fragments larger in size than the hybridizing fragments observed in the VanA prototype strain Enterococcus faecium BM4147, suggesting the lack of the relevant EcoRV and HindIII restriction sites.

Anti-Bacterial Agents↗

Phenotypic and genotypic characterization of Enterococcus spp. of different origins.

Sixty-four strains of Enterococcus spp. of different origin were identified using traditional phenotypic, biochemical and cultural tests, together with molecular tests. API 20 STREP tests identified the species at a preliminary level, only one strain remaining unidentified. Strains belonging to the genus Enterococcus were tested with genus-specific primers, while species-level identification was carried out with the 16S-23S rDNA intergenic region (ITS) and species-specific primers for E. faecium and E. faecalis. Those strains found to be negative with the species-specific primers were subjected to 16S rDNA partial sequencing.

Animals↗

Enterococcus seriolicida is a junior synonym of Lactococcus garvieae, a causative agent of septicemia and meningoencephalitis in fish.

The reference strains of Enterococcus seriolicida (ATCC 49156T) (T = type strain) and of Lactococcus garvieae (ATCC 43921T) and 30 field strains of Gram-positive cocci isolated from diseased rainbow trout in Italy were found to be phenotypically (API 20 STREPT and API 50 CH) and genetically (DNA-DNA hybridization) similar. The high DNA-DNA homologies (70-100%) and the low delta Tme (less than 1.1 degrees C) among these strains showed that Enterococcus seriolicida and Lactococcus garvieae are synonyms, describing a single bacterial species. E. seriolicida strains should be classified as L. garvieae, which must be considered as a major pathogen of freshwater and salt water fish with a world-wide distribution.

Animals↗

Growth inhibition of Pseudomonas aeruginosa and Enterococcus faecalis by crude extractives of Mansonia altisima timber sawdust.

Crude, aqueous methanol and ethanol extracts of sawdust of Mansonia (Mansonia altisima) timber inhibited Pseudomonas aeruginosa and Enterococcus faecalis. The minimum inhibitory concentration ranged from 4.4 to 9.3 mg/ml, while the minimum bactericidal concentration was 10.0-25.0 mg/ml. Enterococcus faecalis was more susceptible, but no marked difference occurred in the antibacterial effect of methanol and ethanol extracts. The aqueous extract was less active.

Enterococcus faecalis↗

Sodium hypochlorite stress in Enterococcus faecalis: influence of antecedent growth conditions and induced proteins.

Compared with exponential growing bacteria, carbohydrate-starved cells of Enterococcus faecalis exhibit a high level of resistance to sodium hypochlorite with maximal resistance observed in cultures entering stationary phase. Chloramphenicol treatment, at various stages of growing phase, does not abolish the hypochlorite resistance of starved cells. However, Enterococcus faecalis conditioned by low sodium hypochlorite concentrations does not develop tolerance towards a lethal dose of the disinfectant. Two-dimensional gel analysis shows that protein synthesis is drastically turned off by hypochlorite treatment, whereas synthesis of a few proteins is enhanced by a low concentration of this chemical agent.

Bacterial Proteins↗

Control of a nosocomial outbreak of vancomycin resistant Enterococcus faecium in a paediatric oncology unit: risk factors for colonisation.

UNLABELLED: In order to determine the extent of vancomycin resistant enterococcus (VRE) colonisation within a paediatric oncology unit, the risk factors for the acquisition of the organism, the molecular epidemiology of the isolates and the impact of infection control measures, extensive patient and environmental surveillance was undertaken with identification, antibiotic susceptibility testing and pulsed-field gel electrophoresis (PFGE) of all VRE isolates. A matched case control study was carried out. Fourteen patients (19% of screened patients) with VRE colonisation were identified (12 with Enterococcus faecium). All isolates manifested the Van A phenotype. Extensive environmental contamination with VRE was present. PFGE of E. faecium isolates from 10 patients and from five of six environmental cultures revealed patterns suggesting genetic relatedness. Following comparison of the 14 cases with 41 controls matched for age (+/- 4 years) and cohabitation on the oncology unit, risk factors for colonisation with VRE included duration of neutropenia, (OR, 3.72; 95% CI, 1.0-13.1), and antibiotic therapy, (OR, 4.07; 95% CI, 1.08-15.3), the number of antibiotic agents received, (OR, 8.4; 95% CI, 1.34-34.3) and the duration of therapy with amikacin, (OR, 10.7; 95% CI, 1.4-81.5), ceftazidime, (OR, 11.5; 95% CI, 2.2 59.9) or teicoplanin, (OR, 12.3; 95% CI, 2.25-67.4). Implementation of stringent infection control measures reduced environmental contamination from 25% of samples in week 1 to none in week 11. Two additional colonised patients were identified during the subsequent 6 months. CONCLUSION: Risk factors for VRE colonization in paediatric oncology patients included duration of neutropenia, duration of any antibiotic therapy, exposure to ceftazidime, amikacin or teicoplanin and the number of antibiotics used. The study suggests that environmental contamination played an important role in patient-to-patient transmission of VRE and interventions including implementation of infection control measures were associated with a decreased incidence of gastro-intestinal colonisation.

Adolescent↗

Clinical significance of bacteriuria with low colony counts of Enterococcus species.

The clinical significance of low counts of enterococci in urine cultures remains unclear. The goal of this study was to investigate the clinical significance of enterococci growing in numbers lower than 100,000 colony-forming units per milliliter (cfu/ml) in urine samples. Clinical parameters were collected from patients whose midstream clean-catch urine samples grew Enterococcus spp. in amounts between >or=10,000 and 100,000 cfu/ml and who were not previously treated with antibiotics. Only those patients who had leukocyturia in addition to positive culture were considered to have true urinary tract infection (UTI). Of the 208 patients included in the study, 54% were diagnosed with true UTI. Patients with true UTI were older by 6 years (p=0.03), were more likely to be hospitalized (p=0.016), had higher rates of dysuria (p=0.0001), urgency (p=0.0001), and frequency (p=0.0001), and had more solid tumors (p=0.03). By multivariate analysis, urgency (OR=7.1) and hospitalization (OR=4.4) were identified as independent risk factors for true UTI with enterococci in low counts. Enterococcal counts in patients with true UTI were randomly distributed all along the scale between 10,000 and 100,000 cfu/ml, and no differential cutoff could be determined. In conclusion, more than half of the patients whose urine cultures grow Enterococcus spp. in counts lower than 100,000 cfu/ml may have true UTI, especially if they are hospitalized and have symptoms of dysuria, urgency, or frequency. Microbiology laboratories should perform a complete work-up on samples containing low counts of enterococci, and the final interpretation should be done by physicians, using additional clinical information.

Adult↗

In vitro assessment of the host response against Enterococcus faecalis used in probiotic preparations.

Along with other lactic acid bacteria, enterococci are used in food products and as health promoting agents. The safety of these products must be ensured, because they contain potentially pathogenic microorganisms. Here we present an in vitro opsonophagocytic assay that closely mimics the protective human immune response to Enterococcus faecalis and Enterococcus faecium. A collection of closely related E. faecalis isolates used as probiotics showed different susceptibilities to opsonic killing, suggesting that some of these isolates possess a capsule while other do not. This information may be helpful in assessing the safety of a given bacterial isolate used and could detect likely enterococcal candidates for probiotic preparations.

Bacterial Capsules↗

Production and characterization of enterocin 900, a bacteriocin produced by Enterococcus faecium BFE 900 from black olives.

Enterococcus faecium BFE 900 isolated from black olives produced a bacteriocin termed enterocin 900, which was antagonistic towards Lactobacillus sake, Clostridium butyricum, enterococci as well as Listeria spp. including Listeria monocytogenes. Enterocin 900 was inactivated by pepsin, alpha-chymotrypsin, proteinase K and trypsin but not by catalase, alpha-amylase, or other non-proteolytic enzymes tested. The bacteriocin was heat stable, retaining activity after heating at 121 degrees C for 15 min. Enterocin 900 was active at pH values ranging from 2.0-10.0, with highest activity at pH 6.0. Bacteriocin production occurred in the late logarithmic growth phase when culture density was ca. log 8.0 CFU ml-1. Enterocin 900 was produced in media with initial pH ranging from 6.0-10.0, but not in media with a pH lower than 6.0. Medium composition, especially the concentrations of peptone and yeast extract influenced bacteriocin production, with no bacteriocin being produced in the absence of either of these compounds. No plasmids could be isolated from Enterococcus faecium BFE 900, indicating that the gene for bacteriocin activity is located on the chromosome.

Bacteriocins↗

Genetic transformation of various species of Enterococcus by electroporation.

A transformation system for Enterococcus faecalis was developed which uses untreated (i.e., non-protoplasted) cells and the electroporation technique. The optimized protocol resulted in transformation efficiencies of up to 4 x 10(6) transformants per microgram of plasmid DNA. All strains of E. faecalis tested could be transformed by this method, albeit with differing transformation efficiencies. Using the protocol optimized for E. faecalis we successfully transformed Enterococcus faecium, E. hirae, E. malodoratus and E. mundtii.

Cell Division↗

The inhibition by ionophores in vitro of an Enterococcus-like pathogen of rainbow trout, Oncorhynchus mykiss.

Streptococcosis is a major disease of several fish species in Australia, Japan and South Africa. The minimum inhibitory concentration of some ionophores (lasalocid, monensin, narasin and salinomycin) was determined in vitro for an Enterococcus-like species pathogenic for rainbow trout (Oncorhynchus mykiss) in Australia. Forty isolates of the fish pathogen were tested, together with control strains of Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212 and Streptococcus bovis ATCC 9809. The minimum inhibitory concentrations (MIC) of erythromycin, the drug of choice for controlling streptococcosis, ranged between 0.1 and 0.8 microgram/ml whereas the MIC values for the ionophores ranged between 0.2 and 1.5 micrograms/ml. Of the ionophores tested, narasin was the most inhibitory (0.2-0.4 microgram/ml), while monensin was the least inhibitory (0.4-1.5 micrograms/ml). Salinomycin was marginally more inhibitory (0.4-0.8 microgram/ml) than lasalocid (0.8 microgram/ml).

Animals↗

Human neutrophil oxidative response and phagocytic killing of clinical and laboratory strains of Enterococcus faecalis.

Many clinical isolates of Enterococcus faecalis produce a hemolysin/bacteriocin that is plasmid mediated. Recent human epidemiologic studies and animal research suggest that this hemolysin/bacteriocin may enhance the pathogenicity of hemolysin-producing enterococci compared with non-hemolysin-producing strains. These studies determined that clinical strains that produce hemolysin/bacteriocin differed from non-hemolysin-producing clinical and laboratory strains in their ability to induce the production of reactive oxygen intermediates in human peripheral blood neutrophils and in their susceptibility to phagocytic killing in vitro. The induction of superoxide anion generation by neutrophils was demonstrated to be directly proportional to the presence of the hemolysin/bacteriocin plasmid and was transferable to a non-hemolysin-producing laboratory strain by transconjugation. The presence of the plasmid, however, did not effect killing by phagocytic cells in vitro. It is proposed that hemolysin/bacteriocin-producing strains of enterococcus may be more pathogenic due to reactive oxygen product-induced tissue injury in vitro.

Conjugation, Genetic↗