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PubMed2004

Determining incidence of extended spectrum beta-lactamase producing Enterobacteriaceae, vancomycin-resistant Enterococcus faecium and methicillin-resistant Staphylococcus aureus in 38 centres from 17 countries: the PEARLS study 2001-2002.

The PEARLS study prospectively monitored selected nosocomial pathogens from 38 centres in 13 European, three Middle Eastern countries and South Africa during 2001-2002. Extended spectrum beta-lactamase (ESBL) production rates among Escherichia coli, Klebsiella pneumoniae, and Enterobacter spp. were 5.4% (142/2609), 18.2% (401/2,206) and 8.8% (204/2,328), respectively, for all study sites. The overall ESBL production rate for the combined Enterobacteriaceae was 10.5% (747/7,143), highest in Egypt, 38.5%, and Greece, 27.4%, and lowest in The Netherlands, 2.0%, and Germany, 2.6%. IEF, PCR and DNA sequencing determined 10.7% false positives among Enterobacter spp. when using NCCLS guidelines to screen for ESBL production. The prevalence of nosocomial methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecium was 32.4% (294/908) and 8.7% (83/949), respectively. PEARLS provides baseline data against which prospective changes in resistant determinants and outcomes can be measured in this ongoing study.

Anti-Bacterial Agents↗
PubMed2006

Linezolid treatment of glycopeptide-resistant Enterococcus faecium in very low birth weight premature neonates.

Glycopeptide-resistant Enterococcus faecium has emerged as an important nosocomial pathogen with limited therapeutic options. Here we report the successful treatment of glycopeptide-resistant E. faecium infection in two very low birth weight premature infants with the new oxazolidinone linezolid. Treatment with linezolid at a dosage of 10 mg/kg every 8 h intravenously for a duration of 16 days and 14 days, respectively, was well tolerated and led to complete clinical recovery and clearance of the organism from all body sites. The two cases support the clinical efficacy and safety of linezolid in very low birth weight premature neonates with glycopeptide-resistant E. faecium infections.

Acetamides↗
PubMed2006

Molecular epidemiology of urinary tract isolates of vancomycin-resistant Enterococcus faecium from North America.

Vancomycin-resistant Enterococcus faecium (VRE) are a common cause of nosocomial infections and are important agents of gastrointestinal colonisation. As the prevalence of VRE in hospitalised patients continues to increase, implementation of appropriate infection control measures requires routine surveillance of VRE transmission patterns. The purpose of the present study was to investigate the molecular epidemiology of VRE isolates within the USA and Canada. Two hundred and eighty-three urinary tract isolates of VRE were collected in the year 2000 from ten Canadian hospitals and 28 US tertiary care medical centres representing seven of the nine geographic regions of the United States Bureau of the Census. The in vitro activity of vancomycin, teicoplanin and nine comparators was determined by broth microdilution. The genetic relatedness among isolates was evaluated by pulsed-field gel electrophoresis (PFGE). Resistance rates (intermediate and resistant) were 100% for vancomycin and 78.7% for teicoplanin. Resistance was lowest with linezolid, chloramphenicol and nitrofurantoin at 0.3%, 0.3% and 0.5%, respectively. PFGE revealed that genetic heterogeneity amongst isolates from each of the medical centres varied considerably. Despite some intracentre and intercentre VRE dissemination, the present study found no evidence for the emergence of a dominant clonal strain. This suggests that the spread of VRE within North America is a complex process involving both the horizontal transfer of glycopeptide resistance determinants and polyclonal dissemination.

Canada↗
PubMed2006

Heterogeneity of vat(E)-carrying plasmids in Enterococcus faecium recovered from human and animal sources.

In this study, quinupristin/dalfopristin (Q/D)-resistant Enterococcus faecium isolates (33 from poultry farms and 1 from a human outpatient) with Q/D minimal inhibitory concentrations ranging from 4 microg/mL to 32 microg/mL were analysed. Polymerase chain reaction detected the presence of vat(E) in all isolates. Using pulsed-field gel electrophoresis (PFGE), 14 distinct PFGE patterns were identified. The human E. faecium isolate was distinguishable from the 33 farm isolates by PFGE. Southern hybridisation localised the vat(E) gene to an 11 kb plasmid and resulted in five plasmid hybridisation types. The vat(E)-carrying plasmid from the human isolate showed a nearly identical hybridisation pattern to a plasmid from a farm isolate. This study showed that the vat(E) gene, conferring resistance to Q/D, was carried on different plasmids in a heterogeneous group of E. faecium, some of which may be acquired by E. faecium capable of infecting humans.

Acetyltransferases↗
PubMed2006

Importance of amino acid alterations and expression of penicillin-binding protein 5 to ampicillin resistance of Enterococcus faecium in Taiwan.

The importance of the amino acid sequence in the C-terminal domain of penicillin-binding protein 5 (PBP5) and the levels of PBP5 expression to ampicillin resistance of Taiwan clinical isolates of Enterococcus faecium were studied. Sequence data revealed the existence of 12 amino acid sequence variants within the C-terminal domain of PBP5 in the 33 tested isolates (ampicillin minimum inhibitory concentrations (MICs) 1 mg/L to >256 mg/L). Western blot analyses of the levels of PBP5 showed that, with few exceptions, lower amounts of PBP5 were present in the susceptible strains than in the resistant strains. More importantly, a significant correlation (P=0.004, Fisher's exact test) between the expression of PBP5 and ampicillin resistance was detected. Point mutations in PBP5, including addition of aspartic acid or serine after position 466 and change of methionine to alanine or threonine at position 485, alanine or isoleucine to threonine at position 499 and glutamate to valine at position 629, were found to be significantly associated with ampicillin resistance. A significant correlation was obtained for the combined mutation (alleles 10 and 11), suggesting that combined mutation of PBP5 can be a marker for ampicillin resistance of E. faecium.

Amino Acid Sequence↗
PubMed2004

Antagonistic activity against Helicobacter pylori infection in vitro by a strain of Enterococcus faecium TM39.

Lactic acid bacteria (LAB) strains from infant feces were screened for anti-Helicobacter pylori use. In the beginning, we selected the strains based on their capability to adhere to the human intestinal epithelial cell (Int-407), colonial enterocyte-like Caco-2 cell, human cervical epithelioid carcinoma cell (HeLa), and human gastric carcinoma cell (TSGH 9201). Then, acid and bile salt tolerance of these LAB strains was evaluated. In addition, the ability of these LAB strains to inhibit the growth of H. pylori and to expel H. pylori cells from TSGH 9201 were studied. The spent culture supernatant (SCS) of a selected strain TM39, i.e., TM39-SCS, significantly inhibited the viability of H. pylori in vitro. It also inhibited the urease activity of H. pylori in vitro. For these antagonistic effects, in addition to pH and lactic acid, some factors in TM39-SCS might play the major role. Treatment of H. pylori with the SCS or cells of strain TM39 significant reduced its binding to TSGH 9201 cells. Although strain TM39 is identified as Enterococcus faecium, it is not vancomycin resistant and is proved to be safe through the invasion study and a 28-day feeding study with Wistar rats.

Animals↗
PubMed2005

Enterococcus species dominating in fresh modified-atmosphere-packaged, marinated broiler legs are overgrown by Carnobacterium and Lactobacillus species during storage at 6 degrees C.

In order to show which of the initial lactic acid bacteria (LAB) contaminants are also causing spoilage of a modified-atmosphere-packaged (MAP), marinated broiler leg product at 6 degrees C, LAB were enumerated and identified on the 2nd and 17th days following manufacture. A total of 8 fresh and 13 spoiled packages were studied for LAB levels. In addition, aerobic mesophilic bacteria and Enterobacteriaceae were determined. The average CFU/g values in the 8 fresh packages were 1.3 x 10(3), 9.8 x 10(3) and 2.6 x 10(2) on de Man Rogosa Sharpe agar (MRS), Plate Count Agar (PCA) and Violet Red Bile Glucose agar (VRBG), respectively. The commercial shelf life for the product had been set as 12 days, and all packages analyzed on the 17th day were deemed unfit for human consumption by sensory analysis. The corresponding CFU/g averages in the spoiled product were 1.4 x 10(9), 1.1 x 10(9) and 3.9 x 10(7) on MRS, PCA and VRBG agar, respectively. For characterization of LAB population, 104 colonies originating from the fresh packages and 144 colonies from the spoiled packages were randomly picked, cultured pure and identified to species level using a 16 and 23S rDNA HindIII RFLP (ribotyping) database. The results showed that enterococci (35.7% of the LAB population) were dominating in the fresh product, whereas carnobacteria (59.7%) dominated among the spoilage LAB. Enterococcus faecalis, Carnobacterium piscicola and Carnobacterium divergens were the main species detected. In general, when the initial LAB population is compared to the spoilage LAB, a shift from homofermentative cocci towards carnobacteria, Lactobacillus sakei/curvatus and heterofermentative rods is seen in this marinated product.

Animals↗
PubMed2004

Growth inhibition of heat-injured Enterococcus faecium by oligophosphates in a cured meat model.

Cells of two heat-resistant strains of Enterococcus faecium were heated and incubated in meat suspensions containing curing ingredients. The concentrations of the curing ingredients were those frequently used for pasteurized ham-type products, except that the concentrations of the oligophosphates (triphosphate and diphosphate) varied. Heating tests at 69 degrees C were performed with inoculated meat suspensions in heat-sealed plastic pouches. Numbers of bacteria were counted immediately after heating and in parallel series of heated pouches incubated at 37 degrees C. Plating was performed in Tryptone Dextrose Yeast Meat Peptonised Milk Agar (TDYMP); in TDYMP Agar to which the curing ingredients were added; and in TDYMP Agar to which the curing ingredients except oligophosphates were added. The inclusion of oligophosphates in the heating medium increased the heat-injury sustained by the E. faecium cells, and in combination with rather severe heat treatment even completely blocked the growth of surviving organisms in the meat suspension incubated at 37 degrees C. The presence of oligophosphates in the culture medium TDYMP Agar severely reduced the counts of freshly heated cells; however, this effect disappeared after repair and growth of the surviving organisms in the meat suspension.

Animals↗
PubMed2005

Uptake of Escherichia coli, Vibrio cholerae non-O1 and Enterococcus durans by, and depuration of mussels (Mytilus galloprovincialis).

The uptakes of Escherichia coli, Vibrio cholerae non-O1 and Enterococcus durans by mussels (Mytilus galloprovincialis) and the times for depuration were investigated in order to determine the most useful indicator of vibrio contamination. The mussels were maintained in tanks of static seawater contaminated with bacteria at 5 log10 CFU/ml for bioaccumulation. Depuration was carried out by circulating fresh seawater through the tanks. Each organism was presented alone and with others to mussels, at temperatures of 14 and 21 degrees C. In water contaminated with either single or mixed organisms, the bacteria accumulated rapidly in the mussels reaching high concentrations after 1 h. With both single and mixed organisms, the maximum numbers of E. coli in mussels were 6.6 log10 CFU/g at 14 degrees C and 5.4 log10 CFU/g at 21 degrees C. Both V. cholerae non-O1 and E. durans alone or with other organisms reached a number ranging from 6.5 to 7 log10 CFU/g at both temperatures. During depuration the numbers of all the organisms slowly decreased, with E. coli alone, numbers ranged from 2.8 to 2 log10 CFU/g after 72 h at both 14 and 21 degrees C, and the organisms were undetectable after 144 h. With mixed organisms at 14 degrees C E. coli became undetectable after 168 h but at 21 degrees C no E. coli were recovered after 72 h. At 14 degrees C V. cholerae non-O1 alone also was undetectable after 168 h, but at 21 degrees C and with mixed organisms at both temperatures. V. cholerae was recovered after 168 h at numbers about 1 log10 CFU/g. After 168 h numbers of E. durans alone ranged from 2.6 log10 CFU/g at 14 degrees C to 1.5 log10 CFU/g at 21 degrees C, and with mixed organisms the numbers ranged from 2.3 to 2.0 log10 CFU/g at both temperatures. Of the three bacteria of faecal origin, E. durans is quickly acquired by mussels and released more slowly than the others, while E. coli quickly becomes undetectable. The results suggest that, for this kind of seafood, enterococci may be a more appropriate indicator than E. coli of risks to consumers from vibrios.

Animals↗
PubMed2005

Cloning, production and functional expression of enterocin P, a sec-dependent bacteriocin produced by Enterococcus faecium P13, in Escherichia coli.

The cloning and expression of enterocin P (EntP), a sec-dependent bacteriocin produced by Enterococcus faecium P13, was studied in Escherichia coli. PCR-amplified products of the preenterocin P gene (entP) or entP plus the putative EntP immunity gene (entiP), were cloned in plasmid pETBlue-1 under the control of the inducible T7lac promoter. Although target genes in derivative plasmids pJG01 (entP) and pJG02 (entP plus entiP) did not generate products with antimicrobial activity after an in vitro combined transcription/translation reaction, they were expressed as biologically active products following transformation and induction in the E. coli Tuner(DE3)pLacI host. The use of specific antibodies and an ELISA permitted the detection and quantification of EntP in the supernatant (SN), cellular soluble protein fraction (CSF), and inclusion bodies (IB) of E. coli Tuner(DE3)pLacI cells transformed with either pJG01 or pJG02. Functional EntP from the supernatants of E. coli Tuner(DE3)pLacI (pJG01) cultures grown in a complex medium was recovered, at a high efficiency, by immunoaffinity chromatography in a single step. A purification method based on hydrophobic adsorption and reverse-phase chromatographies also permitted the recovery of active EntP from the supernatants of the same cultures grown in a minimally defined medium. The E. coli Tuner(DE3)pLacI (pJG01) cells would merit consideration as an alternative experimental model for the heterologous production and functional expression of EntP, as well as for the fast and efficient recovery of this bacteriocin from the supernatant of this recombinant producer.

Bacteriocins↗
PubMed2005

Enterocin A production by Enterococcus faecium FAIR-E 406 is characterised by a temperature- and pH-dependent switch-off mechanism when growth is limited due to nutrient depletion.

The biokinetics of cell growth and bacteriocin production of Enterococcus faecium FAIR-E 406 was studied as a function of temperature (20-45 degrees C) and pH (5.5-8.5) using de Man-Rogosa-Sharpe medium. Growth of E. faecium FAIR-E 406 was characterized by three successive growth phases and was modelled with the mechanistic nutrient depletion model. Bacteriocin production showed primary metabolite kinetics but was limited to the early growth phase. The critical biomass for switching off bacteriocin production was dependent on medium pH and incubation temperature, and was inversely correlated with the specific bacteriocin production. Doubling the concentration of the nitrogen source as well as a step-wise pH increase shifted the bacteriocin production towards a higher switch-off cell density.

Bacteriocins↗
PubMed2006

Enterococcus and Lactobacillus contamination of raw milk in a farm dairy environment.

Enterococci and lactobacilli are ubiquitously found in the intestinal microflora of humans and animals. The aim of the present study was to determine the importance of bovine faeces as a source of these organisms in raw milk. One hundred and fifty six putative enterococci and 362 lactobacilli were isolated from bovine faeces (n=26), cows' teats, raw milk, the milking machine and the milking environment on one farm. The clonal relationships of each group were investigated using Pulsed-Field Gel Electrophoresis and representatives of the different clusters were identified by repetitive DNA element (rep)-PCR fingerprinting, protein profiling, phenylalanyl-tRNA synthase (pheS) sequence analysis or 16S rDNA gene sequencing. Lactobacilli were present at approximately 3 orders of magnitude greater than enterococci in the bovine faeces. The majority of the bovine faecal enterococcal isolates were identified as Aerococcus viridans. Seven teat isolates belonged to a potential novel Aerococcus sp. and one bovine faecal isolate to a potential second novel Aerococcus sp. The lactobacilli present in the bovine faeces were predominantly Lactobacillus mucosae and Lactobacillus brevis, with small numbers of Lactobacillus plantarum. Only one Enterococcus (a strain of E. casseliflavus) out of 76 and one Lactobacillus (a strain of L. parabuchneri/kefir) out of 247 of the bovine faecal isolates was found in the milk. The major source of these bacteria in the milk was the milking equipment.

Animals↗
PubMed2006

Pheromone-inducible conjugation in Enterococcus faecalis: a model for the evolution of biological complexity?

Pheromone-inducible transfer of the plasmid pCF10 in Enterococcus faecalis is regulated using a complicated network of proteins and RNAs. The plasmid itself has been assembled from parts garnered from a variety of sources, and many aspects of the system resemble a biological kluge. Recently several new functions of various pCF10 gene products that participate in regulation of plasmid transfer have been identified. The results indicate that selective pressures controlling the evolution of the plasmid have produced a highly complex regulatory network with multiple biological functions that may serve well as a model for the evolution of biological complexity.

Bacterial Proteins↗
PubMed2005

Clivus osteomyelitis secondary to Enterococcus faecium infection in a 6-year-old girl.

A 6-year-old girl was diagnosed to suffer from clivus osteomyelitis secondary to Enterococcus faecium infection. On the basis of the magnetic resonance image, the abscess was drained via the posterior wall of the pharyngeal tract immediately. Subsequent antibiotic therapy allowed rapid improvement and long-term healing of the osteomyelitic process without any side effect. Osteomyelitis or abscess of the clivus is very rare in adult patients and extremely rare in children. Some etiopathogenetic hypotheses are discussed in this case.

Child↗
PubMed2005

Recurrent Enterococcus faecalis endophthalmitis after phacoemulsification.

We present the first reported case of late recurrence of postoperative Enterococcus faecalis endophthalmitis after cataract surgery in a 45-year-old diabetic man. Culture-positive E faecalis endophthalmitis was diagnosed 2 days after uneventful phacoemulsification. Early vitrectomy with intravitreal and subconjunctival vancomycin and amikacin and topical vancomycin resulted in apparently complete clinical resolution after 4 months and a best corrected visual acuity of 20/25. Recurrent endophthalmitis with hypopyon occurred 7 months postoperatively and resolved with intravitreal vancomycin and topical prednisolone acetate 1%. However, the patient had a similar relapse at 9 months that resulted in deterioration of visual acuity to no light perception despite a repeat vitrectomy, intraocular lens explantation, capsular bag removal, and intravitreal antibiotics. The late recurrences could have been the result of persistent sequestration of the organism in the capsular bag.

Amikacin↗
PubMed2004

Enterococcus casseliflavus bacteremia. Case report and literature review.

Motile Enterococci are rare causes of enterococcal bacteremia, usually in immunocompromised patients. We describe a case of polymicrobial bacteremia, including Enterococcus casseliflavus, Escherichia coli, and Morganella morganii, in an 80-year old woman, shortly after a liver biopsy performed to exclude cholangiocarcinoma. We further discuss the unique genetic characteristics and the clinical significance of such isolates, such as their intrinsic resistance to vancomycin, and the low virulence exhibited clinically. Further workup on the mechanisms of motile Enterococci resistance to vancomycin could probably yield useful information in achieving new ways of facing resistance attributed to the more usual, and malignant Enterococci.

Aged↗