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PubMed2006

Outbreak of vancomycin-resistant Enterococcus spp. in an Italian general intensive care unit.

Following the identification of two clinical isolates of vancomycin-resistant enterococci (VRE) from intensive care unit (ICU) patients, a surveillance programme detected that six of eight ICU patients were colonised by VRE. Standard epidemic control measures were instituted in the ICU. During a 16-month period, 13 (2.5%) of 509 ICU patients had VRE-positive swabs upon admission, and 43 (8.7%) of 496 VRE-negative patients were colonised by VRE in the ICU. Patients who acquired VRE in the ICU had a longer ICU stay (p < 0.0001). No other statistically significant differences were demonstrated. Two patients had documented infection (infection/colonisation index, 3.6%; overall VRE infection frequency, 0.4%), but both recovered and were discharged. VRE colonisation did not increase the mortality rate. Automated ribotyping identified three clusters containing, respectively, the first 52 Enterococcus faecium isolates, two Enterococcus faecalis isolates, and two further isolates of E. faecium. Multilocus sequence typing demonstrated that two E. faecium isolates representative of the two ribotypes belonged to sequence types 78 and 18, and that these two isolates belonged to the epidemic lineage C1, which includes isolates with a wide circulation in northern Italy. The outbreak was controlled by continuous implementation of the infection control programme, and by the opening of a new unit with an improved structural design and hand-washing facilities.

Adult↗
PubMed1999

Proficiency of clinical laboratories in Spain in detecting vancomycin-resistant Enterococcus spp. The Spanish VRE Study Group.

Studies in a variety of U.S. clinical laboratories have demonstrated difficulty in detecting intermediate and low-level vancomycin-resistant enterococci (VRE). The misclassification of "at least intermediate resistant isolates" as vancomycin susceptible may have both clinical implications and a negative impact on measures to control the spread of VRE. No published study has assessed the ability of clinical laboratories in Europe to detect VRE. So, the apparent low prevalence of VRE in European hospitals may be, in part, secondary to the inability of these laboratories to detect all VRE. In an effort to assess European laboratories' proficiency in detecting VRE, we identified 22 laboratories in Spain and asked them to test four VRE strains and one susceptible enterococcal strain from the Centers for Disease Control and Prevention collection. Each organism was tested by the routine antimicrobial susceptibility testing method used by each laboratory. Overall, VRE were correctly identified in 61 of 88 (69.1%) instances. The accuracy of VRE detection varied with the level of resistance and the antimicrobial susceptibility method. The high-level-resistant strain (Enterococcus faecium; MIC, 512 microg/ml) was accurately detected in 20 of 22 (91. 3%) instances, whereas the intermediate-resistant isolate (Enterococcus gallinarum; MIC, 8 microg/ml) was accurately detected in only 11 of 22 (50%) instances. Classification errors occurred in 27 of 88 (30.9%) instances. Misclassification as vancomycin susceptible was the most common error (16 of 27 [59.3%] instances). Our study shows that the participating Spanish laboratories had an overall acceptable proficiency in detecting VRE but that a substantial proportion of VRE isolates with low or intermediate levels of resistance were not detected. We recommend that studies be conducted to validate laboratory proficiency testing as an important step in the prevention and control of the spread of antimicrobial resistance.

Centers for Disease Control and Prevention, U.S.↗
PubMed1997

Increasing vancomycin resistance in Enterococcus spp. in Australia: facing the challenge in the laboratory.

High level vancomycin resistance in enterococci (VRE) is increasing in Australia. If the spread of VRE is to be checked, then it is vital for laboratories to be able to detect it promptly and accurately. We have established a rapid and accurate screening method using Enterococcosel agar supplemented with 6 mg/l vancomycin. It can recover VRE from spiked feces, it may be used to screen feces for carriage of VRE and it is not subject to the confounding effects which feces introduce to broth enrichment culture.

Australia↗
PubMed1997

Study to determine the ability of clinical laboratories to detect antimicrobial-resistant Enterococcus spp. in Buenos Aires, Argentina.

Few reports of vancomycin-resistant enterococci have appeared outside the USA. Therefore, we evaluated the ability of five laboratories in Buenos Aires, Argentina, to perform susceptibility testing using the disk diffusion method. Laboratories had difficulty identifying the low- and intermediate-level vancomycin-resistant phenotypes. This suggests that the disk diffusion method used by laboratories abroad may fail to detect some vancomycin-resistant enterococci.

Anti-Bacterial Agents↗
PubMed2004

Antimicrobial resistance of Enterococcus spp. isolates from raw beef and meat products.

E. faecalis (67%) and E. faecium (13.7%) were most frequently isolated among enterococci that contaminate cooled and frozen processed meat, follow-up heat-treated meat products and unheated fermented dry salami. Most isolates of both species were resistant to cephalothin (95 and 83 %) and clindamycin (77 and 67%, respectively). Furthermore, E. faecalis and E. faecium isolates were resistant to erythromycin (44 and 72%), tetracycline (34.5 and 17.4%), and streptomycin (13.3 and 4.3%, respectively). Only a few of the isolates were resistant to ampicillin, ampicillin-sulbactam, chloramphenicol, and vancomycin while all isolates were susceptible to gentamicin, penicillin, and teicoplanin. During the production of heat-treated meat products, numbers of resistant isolates increased in spite of the decreasing enterococcal contamination of the samples. An opposite situation was found in the production of fermented dry salami.

Animals↗
PubMed2005

Evaluation of (GTG)5-PCR for identification of Enterococcus spp.

A set of reference strains and a group of previously unidentified enterococci were analysed by rep-PCR with the (GTG)(5) primer to evaluate the discriminatory power and suitability of this method for typing and identification of enterococcal species. A total of 49 strains representing all validly described species were obtained from bacterial collections. For more extensive evaluation of this identification approach 112 well-defined and identified enterococci isolated from bryndza cheese were tested. The (GTG)(5)-PCR fingerprinting assigned all strains into well-differentiated clusters representing individual species. Subsequently, a group including 44 unidentified enterococci isolated from surface waters was analysed to evaluate this method for identification of unknown isolates. Obtained band patterns allowed us to identify all the strains clearly to the species level. This study proved that rep-PCR with (GTG)(5) primer is a reliable and fast method for species identification of enterococci.

Bacterial Typing Techniques↗
PubMed2003

Molecular characterization of van genes found in vancomycin-resistant Enterococcus spp. isolated from the Hospital das Clínicas, FMUSP, São Paulo, Brazil.

Vancomycin-resistant enterococci strains (VRE) is an important pathogen related with hospital infections in many countries, presenting limited or no therapeutic options for treating serious infections. VRE has presented some different genotypes been VanA and VanB considered to be the most important in hospital environments. In the present study the authors investigated the prevalence of van genes (A, B an C) among clinical isolates of VRE in a five month period at a large tertiary hospital in Sao Paulo, Brazil. The results showed the presence of vanA, but not vanB or vanC in all 43 strains of E. faecalis and five E. faecium studied. The results bring an important issue, due to the possibility of resistance spread of vanA genes, to be monitored and solved by the hospital infection control team and the microbiology and molecular biology laboratories at tertiary Hospitals.

Bacterial Proteins↗
PubMed2003

Methods used for the isolation, enumeration, characterisation and identification of Enterococcus spp. 1. Media for isolation and enumeration.

Due to their significance in food, feed, environmental and clinical samples, the detection and enumeration of enterococci have become an important issue not only in daily routine but also in current research activities. Several media and protocols have been published for diverse purposes, but there is no single method, which universally meets all requirements. Depending on the nature of the accompanying microflora and its level, certain substrates and modifications thereof have to be used, taking into account various drawbacks and advantages. In addition to the historical applications (examination of water, different kinds of foods, intestinal and other clinical specimen), the detection of vancomycin-resistant enterococci (VRE) has become an important task, since VRE have found to be frequently involved in nosocomial infections. Moreover, contradictory methodological recommendations can be found in the literature. This paper will give a systematic survey of the different media and methods proposed during the last two decades. Emphasis is placed on compositional details and on specific applications of the media described.

Animals↗
PubMed2006

Antimicrobial resistance in Enterococcus spp. isolated in inflow, effluent and sludge from municipal sewage water treatment plants.

Antimicrobial resistance of enterococci was investigated in 42 samples of crude inflow, treated effluent and sludge collected in 14 municipal sewage treatment plants of Portugal. A total of 983 enterococci were recovered and tested, using the diffusion agar method, regarding their sensitivity to 10 different antimicrobial drugs. Multidrug resistance was present in 49.4% of the isolates. Only 3.3% and 0.6% of the investigated strains were resistant to ampicillin and vancomycin, respectively. Resistances found against rifampicin (51.5%), tetracycline (34.6%), erythromycin (24.8%) and nitrofurantoin (22.5%), are causes for substantial concern. Almost 14% of isolates were resistant to ciprofloxacin. Wastewater treatment resulted in enterococci decrease between 0.5 and 4log; nevertheless, more than 4.4 x 10(5)CFU/100ml were present in the outflow of the plants. Our data indicate that the use of antimicrobials had created a large pool of resistance genes and that sewage treatment processes are unable to avoid the dissemination of resistant enterococci into the environment.

Anti-Bacterial Agents↗
PubMed1993

Reliability of the E test for detection of ampicillin, vancomycin, and high-level aminoglycoside resistance in Enterococcus spp.

By comparison with agar dilution results, the E test was investigated for the ability to detect high-level aminoglycoside (gentamicin and streptomycin), ampicillin, and vancomycin resistance among strains representing six enterococcal species. For ampicillin and vancomycin, disk diffusion results also were obtained. No false high-level aminoglycoside resistance occurred, and no false gentamicin susceptibility was noted. With the high-range streptomycin E test (2,048 micrograms), 24% of the 38 resistant strains were falsely susceptible. However, these discordances could likely be reconciled by adjustments in incubation duration and by using broth microdilution rather than agar screen breakpoint criteria, or by using the lower-range (1,024-micrograms) strip. For ampicillin, category results obtained by E test and disk diffusion showed good agreement with agar dilution; E test MICs were generally comparable to agar dilution MICs. The E test was more sensitive than disk diffusion for detecting vancomycin-intermediate strains, but for these strains and those exhibiting low-level vancomycin resistance (MIC, 32 to 128 micrograms/ml), disk diffusion and E test inhibition zones must be interpreted with caution. Given the reliability of E test for detecting resistance to anti-enterococcal agents, the decision to use this method should be based on convenience, cost, testing frequency, and satisfaction with currently used methods.

Agar↗
PubMed2001

Human infections caused by glycopeptide-resistant Enterococcus spp: are they a zoonosis?

Following the detection of glycopeptide-resistant enterococci (GRE) in 1986 and their subsequent global dissemination during the 1990s, many studies have attempted to identify the reservoirs and lines of resistance transmission as a basis for intervention. The eradication of reservoirs and the prevention of GRE spread is of major importance for two reasons: (i) the emergence of high-level glycopeptide resistance in invasive enterococcal clinical isolates that are already multiresistant, has left clinicians with therapeutic options that are only at the experimental stage; and (ii) the resistance genes may spread to more virulent bacterial species such as Staphylococcus aureus, Streptococcus pneumoniae and Clostridium difficile. VanA-type strains, resistant to high levels of both vancomycin and teicoplanin, are the most commonly encountered enterococci with acquired glycopeptide resistance in humans. A widespread VanA-type GRE reservoir was detected early in farm animals that were exposed to the glycopeptide growth-promoter avoparcin. Numerous studies have provided indirect evidence for the transfer of VanA-type GRE and their resistance determinants from animal reservoirs to humans. The data collected have expanded our understanding of the promiscuous nature of antibiotic resistance, and have provided the groundwork for logical decision-making with the objective of deterring the dissemination of resistant bacteria and of their resistance genes.

Animals↗
PubMed2006

Antimicrobial resistance in Enterococcus spp. and Escherichia coli isolated from poultry feed and feed ingredients.

Poultry feed is at the start of the food safety chain in the "farm-to-fork" model, and might serve as a source of antimicrobial resistant bacteria present in poultry meat. Antimicrobial resistance was investigated in 1137 enterococci and 163 Escherichia coli strains recovered from 23 samples of commercial broiler feed and 66 samples of raw feeding materials taken over half a year timespan. Enumeration of enterococci and E. coli were also performed using traditional plating and fluorescent in situ hybridisation methods. Viable enterococci were detected in all feed samples and in 66% of samples of separate feed ingredients, while E. coli was present in 50% and 32% of feed and raw feeding materials, respectively. The median values (50th percentile) for plate and FISH counts for feeds were, respectively, 2.70 log CFU/g and 5.52 log cells/g for enterococci, and 0.15 log CFU/g and 6.00 log cells/g for E. coli. Among enterococci recovered from feed ingredients, resistance to rifampicin, erythromycin, nitrofurantoin, tetracycline, and ciprofloxacin was found in 59.8%, 21.6%, 21.2%, 18.0% and 6.9% of the isolates, respectively. A considerable proportion of the enterococci isolates obtained from broiler feed displayed resistance to tetracycline (69.1%), rifampicin (58.5%), erythromycin (52.9%) and nitrofurantoin (36.2%). Lower percentage of resistance was observed to chloramphenicol (4.6%), ciprofloxacin (3.9%), vancomycin (1.9%) and ampicillin (1.2%). Among E. coli recovered from feed ingredients and poultry feeds, resistance to ampicillin, tetracycline and streptomycin was found in 22.9%, 27.6% and 19.0% and in 22.4%, 41.4% and 17.0% of the isolates, respectively. These data show that feedstuffs and poultry feeds are extensively contaminated by resistant enterococci and, in a lesser extent, by E. coli, thus leading to their introduction in the farm environment.

Animal Feed↗
PubMed2000

Vancomycin resistance in Gram-positive bacteria other than Enterococcus spp.

This is a review article on vancomycin resistance on gram positive bacteria other than enterococci. Epidemiology of varying resistance, its clinical relevance and therapeutic options in infections caused by vancomycin resistant Listeria spp., Corynebacteria, streptococci and staphylocci are discussed.

Gram-Positive Bacteria↗
PubMed1981

In vitro activity of N-formimidoyl thienamycin (MK0787) against resistant strains of Pseudomonas aeruginosa, Staphylococcus epidermidis, Serratia marcescens, and Enterococcus spp.

The in vitro activities of N-formimidoyl thienamycin (MK0787) and nine other antibiotics were tested against 129 clinical isolates, including 71 of enterococci, 34 of Staphylococcus epidermidis, 17 of Pseudomonas aeruginosa, and 7 of Serratia marcescens. These isolates exhibited a variety of resistant patterns: 97% of the enterococci were resistant to moxalactam; 71 and 81% of the S. epidermidis isolates were resistant to methicillin and penicillin, respectively; 47, 53, and 53% of the P. aeruginosa isolates were resistant to carbenicillin, cefotaxime, and moxalactam, respectively; and all S. marcescens isolates were resistant to amikacin, gentamicin, and tobramycin. With respect to concentrations required to inhibit growth of 90% of the isolates, N-formimidoyl thienamycin was more active than any compound tested. Determination of the concentration required to inhibit growth of 50% of the isolates showed N-formimidoyl thienamycin to be more active than any other agent against S. epidermidis, S. marcescens, and enterococci, but against Pseudomonas isolates it was less active than amikacin, gentamicin, and tobramycin. This preparation is potentially useful for patients with serious infection caused by resistant bacteria; enterococcal and S. epidermidis endocarditis infections may be special situations which merit clinical trials.

Anti-Bacterial Agents↗
PubMed1990

Evaluation of a commercial microtiter system (MicroScan) using both frozen and freeze-dried panels for detection of high-level aminoglycoside resistance in Enterococcus spp.

The MicroScan system was compared with agar dilution screen plates for the detection of high-level aminoglycoside resistance in 182 enterococcal isolates. Both the frozen Gram-Positive Combo Type 2 and the freeze-dried Type 5 panels were evaluated. The specificity of both panels for the detection of streptomycin and gentamicin resistance was 100%. However, the sensitivities for the detection of gentamicin and streptomycin resistance were 84 and 31%, respectively, for the Type 2 panels and 90 and 41%, respectively, for the Type 5 panels. The sensitivities of these panels for the detection of enterococcal high-level aminoglycoside resistance are inadequate for routine use.

Anti-Bacterial Agents↗
PubMedDate not supplied

Emerging role of Enterococcus spp in catheter-related infections: biofilm formation and novel mechanisms of antibiotic resistance.

Enterococci are gram-positive bacteria that are part of the normal human intestinal flora and can colonize the upper respiratory tract, biliary tract and vagina of otherwise healthy people. Although their virulence is relatively low, recently enterococci have emerged as significant nosocomial pathogens and are currently the 4th leading cause of hospital-acquired infections, including those associated with intravascular catheter and biliary stent implants. The frequent use of these medical devices is often associated with severe complications, including catheter-related bloodstream infections (CRBSIs) and biliary stent occlusions, because of microbial biofilm formation on the device surface. Furthermore, other than a high level of resistance to penicillin, ampicillin and aminoglycosides, a dramatic increase in vancomycin resistance of enterococci has been recently observed in most clinical settings. Clinical strains exhibiting novel mechanisms of acquired resistance to antimicrobials are frequently isolated. In addition, enterococci have a great ability to transmit these resistance traits to other species and even to other genera. Due to their associated morbidity and mortality, enterococcal infections related to medical devices currently represent a major challenge for clinicians, especially for the management of critically ill patients, resulting in prolonged hospitalization and additional health costs.

Journal Article↗
PubMed1999

[Peritonitis determined by the site of intra-abdominal surgery].

The aim of this study was to determine bacterial flora infecting the peritoneal cavity during intraabdominal surgery by site of operation. Three groups of patients were examined. 29 patients who underwent surgery on the stomach, duodenum, biliary tract or pancreas, 15 patients operated on because of acute appendicitis and 63 patients operated on because of colon or rectum tumours. At the end of the operation but before closure cultures were obtained by swab from the completed anastomosis site. Samples were placed into transport medium and transported promptly to the laboratory. The results of the bacteriological examinations showed that the peritoneal cavity of all patients operated on were infected with bacteria characteristic for the digestive tract, especially by Enterobacteriaceae spp., Enterococcus spp. and Bacteroides spp. From patients operated on because of rectum or colon tumours 3 or 4 bacterial species were isolated most often and they were often infected with P. aeruginosa and C. albicans. This was in contrast to patients from the other groups. In patients infected with polymicrobial flora, B. fragilis and E. coli or enterococci and E. coli and enterococci were most often seen.

Anastomosis, Surgical↗