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PubMed2003

Enterococcus phoeniculicola sp. nov., a novel member of the enterococci isolated from the uropygial gland of the Red-billed Woodhoopoe, Phoeniculus purpureus.

A facultatively anaerobic, Gram-positive, coccoid, non-spore-forming, non-motile, catalase-negative bacterium was isolated from the uropygial (preen) gland of wild Red-billed Woodhoopoes (Phoeniculus purpureus) and designated strain JLB-1(T). Physiological and biochemical testing suggested that this homofermentative, lactic-acid-producing bacterium could belong to the genus Enterococcus or the genus Streptococcus. Comparison of the 16S rRNA gene sequence of strain JLB-1(T) with other 16S rDNA sequences in the GenBank database by BLAST analysis showed that its closest relatives are Enterococcus faecium, Enterococcus avium and Enterococcus asini. Strain JLB-1(T) may be differentiated from these species by the fact that it does not produce acid from lactose, D-mannitol, D(+)-melezitose or D-sorbitol. Furthermore, it does not hydrolyse arginine or hippurate and cannot grow in the presence of 6.5% NaCl or 40% bile. It differs from the streptococci in that it does not lyse erythrocytes. Strain JLB-1(T) is a novel member of the enterococci, for which the name Enterococcus phoeniculicola sp. nov. is proposed. The type strain is JLB-1(T) (=ATCC BAA-412(T) = DSM 14726(T)).

Animals↗

PubMed2003

Species prevalence and antibacterial resistance of enterococci isolated in Kuwait hospitals.

This study investigated the species prevalence and antibacterial resistance among enterococci isolated in Kuwait hospitals. They consisted of 415 isolates of Enterococcus faecalis (85.3 %), Enterococcus faecium (7.7 %), Enterococcus casseliflavus (4.0 %), Enterococcus avium (1.2 %), Enterococcus durans (1.0 %), Enterococcus gallinarium (0.5 %) and Enterococcus bovis (0.2 %) isolated from urine (36.6 %), blood (10.4 %), wound swabs (11.0 %), stool samples (12.0 %), high vaginal swabs (9.0 %), endocervical swabs (3.0 %) and miscellaneous sources (18.0 %). All of them were susceptible to linezolid. Fifty-two (12.5 %) isolates were ampicillin resistant but none of them produced beta-lactamase. They were resistant to erythromycin (63.3 %), tetracycline (60.5 %), ciprofloxacin (40.0 %), chloramphenicol (28.0 %), vancomycin (2.6 %), and teicoplanin (2.6 %). Fourteen, 19 and 20 % of them expressed high-level resistance to gentamicin, kanamycin and streptomycin, respectively. All of the vancomycin-resistant strains carried the vanA phenotype and genotype. There was no evidence of clonal spread of the vancomycin-resistant isolates.

Acetamides↗

PubMed2005

Evaluation of the EVIGENE VRE detection kit for detection of vanA and vanB genes in vancomycin-resistant enterococci.

The aim of this study was to evaluate the performance of the EVIGENE VRE Detection kit and compare it with PCR, considered the gold standard for detection of vancomycin-resistant enterococci (VRE). The correlation between the MIC values of vancomycin and teicoplanin using the epsilon test was also determined. In the EVIGENE VRE Detection kit, DNA probes specific for bacterial target DNA sequences are bound to microwell plates. A hundred and ten VRE (104 Enterococcus faecium and six Enterococcus faecalis) and 45 vancomycin-susceptible E. faecium were tested. All VRE strains were found to be positive for the vanA genotype using the EVIGENE VRE Detection kit. All results obtained with the EVIGENE VRE Detection kit were confirmed by PCR. MIC results for the strains also correlated highly with the PCR and kit results. The EVIGENE VRE Detection kit should be used in preference to other methods for detecting resistance genes in all strains, since it is less time-consuming, does not require the handling of hazardous chemicals and has the same specificity as PCR.

Bacterial Proteins↗

PubMed2005

Occurrence of vancomycin-resistant enterococci in humans and animals in the Czech Republic between 2002 and 2004.

In the period between September 2002 and May 2004, a total of 6023 rectal swabs from humans in the Czech Republic were evaluated and 821 Enterococcus spp. strains were isolated. Nine strains (1.1 %) were identified as vancomycin-resistant enterococci (VRE). Two strains were VanA Enterococcus faecium, one strain was VanB Enterococcus faecalis and six strains were VanC Enterococcus casseliflavus. In total, 527 Enterococcus spp. strains were isolated from poultry breeds of which 11 (2.1 %) were VRE. Most (54.5 %) were identified as VanA E. faecium. Cluster analysis of SmaI-generated macrorestriction patterns showed high variability in both human and animal VRE strains and no relatedness between strains from the two sources.

Animals↗

PubMed1996

Production of bacteriolytic enzymes as a tool for characterizing enterococci.

Bacteriolytic enzymes secreted by log-phase cultures of enterococci (Enterococcus faecalis, Ent. faecium, Ent. durans, Ent. hirae, Ent. casseliflavus, Ent. avium, Ent. mundtii) were analysed by means of a zymogram technique to resolve activities according to the size of their polypeptide component and their specificities towards different substrates. Heterogeneous patterns of lytic activity were observed with different species. For each test substrate, homogeneous patterns of lytic activities were observed with strains of the same species, except for Ent. faecalis strains, which showed heterogeneous lytic patterns even towards the same substrate, and could be divided into at least four different groups according to their lytic pattern. No lytic activity was common to all strains tested. Results of zymogram analysis of Enterococcus bacteriolytic enzymes were consistent with current knowledge on enterococcal taxonomy, indicating that this analytical approach may be a useful tool for fine-tuned characterization of different enterococcal strains.

Bacterial Typing Techniques↗

PubMed2004

Characterization of faecal enterococci from rabbits for the selection of probiotic strains.

AIMS: To characterize the facultative anaerobic intestinal microbiota of healthy rabbits, especially enterococci, for the selection of potential probiotic strains. METHODS AND RESULTS: Phenotypic and molecular methods were used to identify enterococcal isolates. Results obtained indicated that enterococcal microbiota widely varied among individuals both in size and in composition. Antibacterial and haemolytic activities, and resistance to acid and bile salts were determined. A small group of strains produced bacteriocins active against listeriae and indigenous clostridia and therefore they were selected as potential probiotics. One such strain, 8G, was assayed for colonization capacity. Results obtained suggested that the fate of the introduced strain depended on the composition of the enterococcal indigenous microbiota. CONCLUSIONS: Enterococcus faecalis and Ent. faecium are the predominant enterococcal species in the gut of rabbits. Other species of lactic acid bacteria were not recovered. SIGNIFICANCE AND IMPACT OF THE STUDY: The enterococcal intestinal microbiota of healthy rabbits has been characterized in detail. Monitoring the fate of an introduced probiotic in vivo is required in order to evaluate potential probiotic strains.

Animals↗

PubMed2005

Genetic diversity and safety aspects of enterococci from slightly fermented sausages.

AIMS: To determine the biodiversity of enterococci from slightly fermented sausages (chorizo and fuet) at species and strain level by molecular typing, while considering their safety aspects. METHODS AND RESULTS: Species-specific PCR and partial sequencing of 16S rRNA and sodA genes were used to identify enterococcal population. Enterococcus faecium was the most frequently isolated species followed by E. faecalis, E. hirae and E. durans. Randomly amplified polymorphic DNA (RAPD)-PCR revealed species-specific clusters and allowed strain typing. Sixty strains of 106 isolates exhibited different RAPD profiles indicating a high genetic variability. All the E. faecalis strains carried virulence genes (efaAfs, esp, agg and gelE) and all E. faecium isolates carried efaAfm gene. Enterococcus faecalis showed higher antibiotic resistance than the other species. Only one E. faecium strain showed vanA genotype (high-level resistance to glycopeptides) and E. gallinarum and E. casseliflavus/flavescens isolates showed vanC1 and vanC2/C3 genotypes (low-level resistance only to vancomycin) respectively. CONCLUSIONS: E. faecalis has been mainly associated with virulence factors and antimicrobial multi-resistance and, although potential risk for human health is low, the presence of this species in slightly fermented sausages should be avoided to obtain high quality products. SIGNIFICANCE AND IMPACT OF THE STUDY: The enterococcal population of slightly fermented sausages has been thoroughly characterized. Several relevant safety aspects have been revealed.

Base Sequence↗

PubMed2006

The influence of antibiotic use on the occurrence of vancomycin-resistant enterococci.

BACKGROUND: Several studies have documented the influence of antibiotic selective pressure, mainly from the use of glycopeptides, third-generation cephalosporins, quinolones and lincosamides, on the frequency of vancomycin-resistant enterococci (VRE) occurrence in hospitals. The aim of this study was to evaluate the relationship between VRE occurrence and antibiotic use in the Department of Hemato-Oncology of the Teaching Hospital in Olomouc (DHO), Czech Republic, over a 6-year period under standard and unchanged hygienic and epidemiological conditions. METHODS: During the period of 1998-2003, Enterococcus sp. strains and VRE were isolated by standard methods from clinical samples taken from DHO in-patients. The frequency of VRE occurrence was expressed as the number of isolated strains per 100 bed-days/year. DHO antibiotic consumption data were processed according to the anatomical therapeutic chemical (ATC)/defined daily dose (DDD) system valid in 2003 and expressed in defined daily dose per 100 bed-days (DDD/100 bed-days) for each year of the period. RESULTS: Since 1998, the occurrence of VRE decreased significantly (from 0.28 to 0.17 VRE/100 bed-days in 2001). Between 2001 and 2003, a significant (P < 0.05) increase from 0.17 to 0.38 was observed. The antibiotic use decreased from 205.2 in 1998 to 161.0 DDD/100 bed-days in 1999 and after an increase in 2001 (to 181.8 DDD/100 bed-days) it remained relatively stable. A significant decrease was observed in third-generation cephalosporins and quinolones (from 29.5 to 9.7 and from 42.2 to 30.2 DDD/100 bed-days respectively) between 1998 and 1999. In 2002-2003, the use of third-generation cephalosporins and glycopeptides increased substantially (from 10.1 to 13.9 and from 11.3 to 15.2 DDD/100 bed-days respectively). The Pearson correlation value was significantly positive (P < 0.05) for VRE occurrence and the use of glycopeptides and third-generation cephalosporins. CONCLUSIONS: While our study confirms the effect of use of glycopeptides and third-generation cephalosporins on occurrence of VRE, no influence of quinolones and lincosamides over the 6-year period was shown.

Anti-Bacterial Agents↗

PubMed1985

Monitoring for antibiotic resistance in enterococci consequent upon feeding growth promoters active against gram-positive bacteria.

Commercial chicken and pig farms, using different growth promoters, were monitored for enterococci resistant to a range of antibacterial agents. Due to inconsistent findings attributed to uncontrollable factors, chickens kept under experimental conditions were also studied. A total of 4216 isolates from all the surveys were examined. Resistant isolates were often encountered, even in control groups, suggesting complex population changes not necessarily solely connected with the influence of the growth promoters. It was concluded that comprehensive methods for differentiating strains are necessary to unravel this problem.

Animals↗

PubMed2004

Rate of nosocomial transmission of vancomycin-resistant enterococci from isolated patients.

To evaluate an isolation policy for patients colonised with vancomycin-resistant enterococci (VRE), we instituted active surveillance for transmission to uncolonised patients. Surveillance rectal swabs were taken and pulsed-field gel electrophoresis was performed on positive isolates. VRE transmission with an identical genotype occurred in 5 patients, giving a transmission rate of 3.7 per 1000 patient days, or 1 patient per ward each week. The present study provides a baseline for -assessment of VRE transmission and will be useful in evaluation of the effectiveness of infection control interventions.

Cross Infection↗

PubMed1999

Identification of enterococci and determination of their glycopeptide resistance in German and Austrian clinical microbiology laboratories.

OBJECTIVE: To determine exactly how German and Austrian routine laboratories perform tests for the identification of enterococci and determination of their glycopoptide resistance. METHODS: Six enterococcal test strains with different types of glycopeptide resistance (Enterococcus faecium, VanA; E. faecalis, VanA; E. faecium, VanB; E. faecalis, VanB; E. gallinarum, VanC1; E. casseliflavus, VanC2) were sent as anonymous isolates to 73 clinical microbiology laboratories (65 in Germany; eight in Austria). The participating laboratories had to identify the strains up to the species level and to determine their antibiotic susceptibilities to the glycopeptides vancomycin and teicoplanin by the test method(s) that are used daily in the corresponding laboratories. RESULTS: The analysis of the results received from 62 laboratories (56 from Germany, six from Austria) demonstrated that the most used routine method in susceptibility testing was the agar diffusion test, followed by the Etest, and the microbroth dilution procedure. The majority of participants had no difficulties in susceptibility testing with the VanA-type strains. However, the agar diffusion test was often not able to recognize clearly the VanB and VanC strains; some problems also arose with VanB isolates in the Etest. With the microbroth dilution method, the corresponding type of glycopeptide resistance was correctly determined in the majority of enterococcal test strains. Difficulties also arose in identification, especially with the VanC strains (E. gallinarum and E. casseliflavus), which were often falsely identified as E. faecium. The reasons for these errors are obviously based on the lack of important tests (such as motility and presence of a yellow pigment) in some commercially available identification test kits.

Journal Article↗

PubMed1999

MIC distribution and inoculum effect of LY333328: a study of vancomycin-susceptible and VanA-type and VanC-type enterococci obtained from intensive care unit patient surveillance cultures.

OBJECTIVE: To determine the in vitro activity and inoculum effect of LY333328, a semisynthetic glycopeptide, against vancomycin-susceptible and vancomycin-resistant enterococcal isolates. METHODS: One hundred and seventy-six enterococcal isolates (117 vancomycin-susceptible, 29 VanA-type and 30 VanC-type isolates) obtained from surveillance cultures of 139 intensive care unit patients were studied by the standard agar dilution method. Vancomycin resistance determinants were characterized by PCR. RESULTS: The activity of LY333328 was comparable (MIC range, 0.1-2 mg/L) to those of vancomycin (0.1-4 mg/L) and teicoplanin (0.06-1 mg/L) for vancomycin-susceptible isolates. LY333328 was more active (0.1-8 mg/L) than vancomycin (256 to >1024 mg/L) and teicoplanin (32-512 mg/L) against VanA-type isolates, and similar (0.2-1 mg/L) to teicoplanin (0.1-0.5 mg/L) against VanC-type isolates. The MIC distribution of LY333328 displayed a narrower range than that of vancomycin, with no clear distinction between susceptible and resistant populations. The increment in the inoculum size, from 104 to 106 CFU/spot, of susceptible isolates increased the MIC values of LY333328, vancomycin and teicoplanin by factors of 11.4, 1.6 and 3.8, respectively. The corresponding factors for LY333328 for VanA-type and VanC-type isolates were 3.5 and 6.4, respectively. CONCLUSIONS: LY333328 displays an excellent in vitro activity against vancomycin-susceptible and -resistant enterococci. Nevertheless, the inoculum size used in susceptibility tests should be carefully controlled.

Journal Article↗

PubMed2004

Emergence of vancomycin-resistant enterococci in a tertiary hospital in Crete, Greece: a cluster of cases and prevalence study on intestinal colonisation.

The aim of this study was to investigate the clinical and epidemiological characteristics of five consecutive cases of infection with vancomycin-resistant enterococci (VRE) and the prevalence of faecal carriage of VRE among patients admitted to a 700-bed university hospital where no VRE had been isolated previously. In a 2-month period, five consecutive patients infected with VRE were detected. Three VanB+ Enterococcus faecium isolates were obtained from three patients, while two VanA+ E. faecium isolates, one VanA+ Enterococcus faecalis isolate and one VanC1+ Enterococcus gallinarum isolate were obtained from the other two patients. Of 218 faecal specimens from all hospital wards, 41 (18.8%) were found to contain VRE. Forty-two isolates of VRE were obtained, comprising one (2%) E. faecalis, 11 (27%) E. faecium, 24 (57%) E. gallinarum and six (14%) Enterococcus casseliflavus/flavescens. Four isolates carried the vanA gene, eight carried vanB, 24 carried vanC1, and six carried vanC2/C3. Use of glycopeptides, the presence of central venous catheters and renal dialysis all correlated with VRE colonisation. The prevalence rates were among the highest reported in the literature.

Adult↗

PubMed2005

Vancomycin-resistant enterococci: consequences for therapy and infection control.

Vancomycin-resistant enterococci (VRE) have emerged as important nosocomial pathogens, initially in the USA, but now also in Europe, where hospital outbreaks are being reported with increasing frequency, although the incidence of VRE infections remains extremely low in most European countries. The recently demonstrated in-human transmission of vancomycin resistance from VRE to methicillin-resistant Staphylococcus aureus (MRSA) in two American patients underscores the potential danger of a coexisting reservoir of both pathogens. As MRSA is already endemic in many European hospital settings, prevention of endemicity with VRE seems relevant, but should be balanced against the costs associated with the implementation of effective strategies. The presence of a large community reservoir of VRE in Europe could hamper the feasibility of infection control strategies. Although the prevalence of colonisation amongst healthy subjects has apparently decreased after the ban on avoparcin use in the agricultural industry, a large proportion of admitted patients are still potential sources of VRE transmission. With no risk profile available to identify these carriers, effective screening, followed by barrier precautions for carriers, seems to be impossible. Recent studies, however, have suggested that hospital outbreaks are almost exclusively caused by specific genogroups of VRE that can be characterised phenotypically and genotypically (e.g., co-resistance to ampicillin and the presence of the variant esp gene). Based on our own experience, we propose that VRE infection control programmes should be restricted to patients colonised with these VRE strains. If such a strain is cultured from a clinical sample, surveillance amongst contact patients is recommended and barrier precautions should be implemented in the case of documented spread.

Anti-Bacterial Agents↗

PubMed2006

Genetic relatedness and antimicrobial resistance determinants among clinical isolates of enterococci from Cuba.

This study describes the genetic relationships and antimicrobial resistance determinants found among 99 clinical isolates of enterococci from 15 different hospitals in Cuba. Pulsed-field gel electrophoresis SmaI analysis demonstrated a high degree of genetic diversity. A limited number of multiresistant Enterococcus faecalis clones, showing resistance to three or more families of antimicrobial agents, were detected simultaneously in different institutions, suggesting inter-hospital circulation of selected clones, and/or selection of particular clones following their introduction into the hospital environment. Antimicrobial resistance determinants, including erm(B), aac(6')-aph(2'), aph(3'), ant(6), vanB (E. faecalis) and vanA (Enterococcus faecium) were detected by PCR in various isolates.

Aminoglycosides↗

PubMed2006

Antimicrobial resistance among faecal enterococci from healthy individuals in Portugal.

Analysis of 247 faecal enterococcal isolates from 99 healthy Portuguese individuals during 2001 revealed the presence of enterococci resistant to vancomycin (5%) and highly resistant to streptomycin (52%), kanamycin (40%) or gentamicin (11%). Most isolates were also resistant to tetracycline, erythromycin, ciprofloxacin and quinupristin-dalfopristin. The vanA (two Tn1546 types), vanC1, erm(B), aac(6')-aph(2'')-Ia, aph(3')-IIIa, vat(E) and vat(D) genes were detected. Enterococcus faecalis and Enterococcus faecium isolates with high-level resistance to gentamicin were related to Portuguese poultry isolates described previously. E. faecium isolates that were highly resistant to vancomycin or gentamicin harboured different housekeeping purK alleles associated previously with different hosts.

Animals↗

PubMed1993

Streptococci and enterococci associated with tonsils of cattle.

Streptococcus suis, Streptococcus bovis and the mastitis pathogens Streptococcus dysgalactiae and Streptococcus uberis were the most frequently occurring streptococci in tonsils of cattle. Streptococcus suis dominated in samples from calves between 1 month and 1 year of age, but was much less frequent in calves less than 1 month old. The mastitis pathogen Strep. dysgalactiae was found more often in calves than in older animals. Enterococci were relatively rare, except in the youngest age group. Nearly one third of the strains examined could not be identified to known species.

Animals↗

PubMed1995

Development of RAPD protocol for typing of strains of lactic acid bacteria and enterococci.

A protocol for typing strains of lactic acid bacteria and enterococci based on randomly amplified polymorphic DNA (RAPD) fragments has been developed. Using a single 10-mer primer, fingerprints were achieved without the need to isolate genomic DNA. Different conditions of DNA release and amplification were investigated in order to obtain reproducible results and high discrimination among strains. This RAPD protocol was successfully applied for the typing of strains belonging to the species Lactobacillus acidophilus, Lact. helveticus, Lact. casei, Lact. reuteri, Lact. plantarum, Enterococcus faecalis, Ent. faecium and Streptococcus thermophilus.

Bacterial Typing Techniques↗