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Significance of vancomycin resistant enterococci from urinary specimens at a tertiary care centre in northern India.

BACKGROUND & OBJECTIVES: There is a paucity of information on vancomycin resistance in enterococci from India. In the present study, enterococci isolated from urine specimen were screened for vancomycin resistance. METHODS: 144 enterococci isolates obtained in pure and significant numbers (>10(5) cfu/ml) from 9664 urine specimens were subjected to agar screen method of National Committee for Clinical Laboratory Standard (NCCLS) for presence/absence of vancomycin resistance. Minimum inhibitory concentration (MIC) was performerd by E-test and agar dilution method. RESULTS: Eight vancomycin resistant enterococci (VRE) were obtained, namely, 5 Enterococcus faecium, 1 each of Enterococcus faecalis, E. casseliflavus and E. pseudoavium. The MIC ranged from 8 to 32-microgm/ml. The records of these 8 patients were retrospectively reviewed for clinical details. Five patients had nosocomial urinary tract infection (UTI), while in 2 patients UTI was community acquired. One patient had asymptomatic bacteriuria. Two patients with nosocomial UTI developed clinical sepsis and died in spite of vancomycin treatment. Urinary catheterization, surgery on genitourinary tract, prior exposure to third generation cephalosporins and metronidazole were present in 6,5,3 and 2 patients respectively. INTERPRETATION & CONCLUSION: Although the frequency of isolation of VRE is not very high in our setting as compared to West, this may just be the beginning of the problem. Screening for vancomycin resistance is recommended in all patients with suspected UTI who have significant bacterial counts.

Adult↗

Control of endemic glycopeptide-resistant enterococci.

OBJECTIVE: To evaluate the epidemiology of, and control measures for, vancomycin-resistant Enterococcus (VRE) in a renal unit. DESIGN: A 3-month, prospective, prevalence culture survey of patients on a 24-bed renal unit. SETTING: A 975-bed community teaching hospital. PATIENTS: Patients admitted to the renal unit over a 3-month period. Patients identified with VRE were each matched with four patients without VRE isolated over the study period. INTERVENTIONS/CONTROL MEASURES: Resistant-organism barrier precautions. To eradicate carriage of VRE, two patients with VRE stool colonization were treated with 5 days of oral doxycycline (100 mg twice per day) and rifampin (300 mg/day). RESULTS: Seven patients with VRE (8 isolates) were identified. Five isolates were Enterococcus faecium (vancomycin MIC = 16 to 256 micrograms/mL), two were Enterococcus faecalis (MICs = 16 and 124 micrograms/mL), and one was Enterococcus gallinarum (MIC = 8.0 micrograms/mL). Eradication of carriage with VRE was accomplished in two patients treated with doxycycline and rifampin. In the final 30 days of the culture survey and at 9 months, there were no further patients with VRE identified. CONCLUSIONS: Resistant-organism precautions and elimination of patient carriage may be useful measures for controlling the spread of low-prevalence endemic vancomycin-resistant Enterococcus.

Adult↗

The challenge of vancomycin-resistant enterococci: a clinical and epidemiologic study.

BACKGROUND: Vancomycin-resistant enterococci have been recovered with increasing frequency from hospitalized patients. Risk factors, mode of nosocomial transmission, extent of colonization in hospitalized patients, and treatment options for these organisms have not been completely delineated. METHODS: We studied 53 patients (group A) with vancomycin-resistant enterococci isolated from various clinical specimens and also surveyed for vancomycin-resistant enterococci in stool specimens submitted for Clostridium difficile toxin assays (group B). Stool specimens submitted for identification of bacterial pathogens and stool specimens from hospital employees were also analyzed for vancomycin-resistant enterococci. RESULTS: Seventy-six isolates of vancomycin-resistant enterococci were recovered in group A. Five of these patients harbored vancomycin-resistant enterococci on admission. Fifty-three of 289 group B stool specimens submitted for C. difficile toxin assays yielded vancomycin-resistant enterococci. Cephalosporins and vancomycin were the most common antimicrobial agents received by both groups of patients. Enterococcus faecium isolates were more resistant than Enterococcus faecalis isolates to antimicrobial agents. All isolates exhibited high-level aminoglycoside resistance and were not beta-lactamase producers. There were at least 15 different molecular clones of E. faecium and three of E. faecalis. Vancomycin-resistant enterococcal bacteremia was associated with a 100% in-hospital mortality rate. CONCLUSIONS: Multidrug-resistant and vancomycin-resistant enterococci have become important nosocomial pathogens that are difficult to treat. Vancomycin-resistant enterococcal bacteremia was associated with a poor prognosis. We found a high rate of colonization in patients with suspected C. difficile toxin colitis. Judicious use of vancomycin and broad-spectrum antibiotics is recommended, and strict infection control measures must be implemented to prevent nosocomial transmission of these organisms.

Adult↗

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↗

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↗

Vancomycin-resistant enterococci in King Chulalongkorn Memorial Hospital: a 5-year study.

The emergence of hospital acquired infections with bacteria resistant to antimicrobials such as vancomycin resistant enterococci (VRE) has become a worldwide concern. The authors studied the prevalence and surveillance of 5 years study of VRE in King Chulalongkon Memorial Hospital and phenotype of these resistance strains. A total of enterococci 1854 isolates were collected from clinical specimens from 1995 to 1999. Screening vancomycin resistance was identified by the agar plated method and minimal inhibitory concentration (MIC) of vancomycin was determined for vancomycin-resistance strains by E-test. The results demonstated that 15 (0.81%) VRE were isolated from 1,854 specimens. Fourteen VRE were identified as Enterococcus faecium and one strain was Enterococcus faecalis. All of these strains, carrying the VanB phenotype, were susceptible to teicoplanin. Similar to other studies, most VRE strains are E. faecium. To the authors' knowledge, this is the first VRE study carried out in King Chulalongkorn Memorial Hospital. The results showed a low prevalence of VRE and surveillance of 5 years study demonstated a gradual increase of VRE. Therefore, it is important to continue periodic surveys of VRE to prevent the spread of VRE in hospitals.

Enterococcus↗

Susceptibility of Gram-positive bacteria from ICU patients in UK hospitals to antimicrobial agents.

Microbiologists in 25 sentinel laboratories were each asked to refer up to 100 clinically-significant Gram-positive bacteria isolated from consecutive intensive care unit (ICU) patients. A total of 1595 isolates were collected from patients in 23 hospitals; these included Staphylococcus aureus (47.6%), coagulase-negative staphylococci (CNS) (30.6%), enterococci (14.3%), pneumococci (2.8%) and other streptococci (3.5%). A few coryneforms, other bacilli and a Nocardia sp. were also collected. Rates of oxacillin resistance among S. aureus and CNS isolates were 59.3 and 78.5%, respectively. Vancomycin-resistant S. aureus were not detected, although two isolates (0.3%) were resistant to teicoplanin [minimum inhibitory concentrations (MICs) 8 mg/L]. In contrast, 13.7% of CNS were teicoplanin resistant (MICs 8-32 mg/L) and 1.2% were resistant to vancomycin. Among the enterococci, 72.5% were Enterococcus faecalis and 24.5% were Enterococcus faecium, the remainder including isolates of Enterococcus casseliflavus or Enterococcus gallinarum. Eighteen percent of E. faecium isolates were vancomycin-resistant, compared with only 3% of E. faecalis isolates. Rates of high-level gentamicin resistance in E. faecalis and E. faecium were 40 and 25%, respectively. Nine percent of pneumococci and streptococci were resistant to penicillin, with 7 and 11%, respectively, resistant to erythromycin. None of the isolates showed resistance to linezolid, with the MICs for the entire study population falling in the range of 0.5-4 mg/L.

Coagulase↗

Colonization by vancomycin-resistant enterococci of the intestinal tract of patients in intensive care units from French general hospitals.

OBJECTIVE: To evaluate the prevalence of fecal carriage of vancomycin-resistant enterococci (VRE) by patients hospitalized in intensive care units from 24 French general hospitals. METHODS: Rectal swabs were obtained from 647 patients hospitalized in intensive care units during the month of June 1994 and plated on agar medium selective for vancomycin-resistant enterococci. The glycopeptide resistance phenotypes and genotypes of the enterococci detected were characterized. RESULTS: Thirty-two of 647 patients (4.9%) carried VRE. Thirteen strains (2%) were identified as Enterococcus faecium and 19 (2.9%) as Enterococcus gallinarum or Enterococcus casseliflavus. None of these strains was highly resistant to gentamicin. The E. gallinarum and E. casseliflavus strains contained the vanC1 and vanC2 genes, respectively. The E. faecium strains were highly resistant to vancomycin and teicoplanin and carried the vanA gene. No infection due to VRE was observed during the study period. Pulsed-field gel analysis of total DNA following digestion with SmaI or KspI from 13 VanA-type E. faecium strains revealed intra- and inter-hospital strain heterogeneity. However, the finding of isolates with indistinguishable pulsed-field types within the same ward and in two medical centers suggests patient-to-patient transmission or a common source. Four E. faecium strains were isolated within 48 h after admission of patients. CONCLUSIONS: These results indicate that VRE form part of the normal flora of patients and that, despite the actual scarcity of infections due to VRE, there is a potential risk for dissemination of these strains in French hospitals.

Journal Article↗

Enterococcal infections in a Greek intensive care unit: a 5-y study.

In this study we determined the incidence, resistance pattern, and mortality rate associated with infection caused by Enterococcus faecalis and Enterococcus faecium among patients in a multidisciplinary intensive care unit (ICU). A total of 111 patients with E. faecalis and 60 with E. faecium infections were identified during a 5-y period (1992-96). We observed an increase in the incidence of enterococcal infections (from 5.46 to 8.46 per 1000 patients-days, p = 0.0112), due mainly to the increased incidence of E. faecium (from 0.45 to 4.06 per 1000 patients-days, p = 0.002). Blood was the most common site of enterococcus isolation. E. faecium was more resistant to antibiotics than E. faecalis, but no vancomycin resistant enterococcus was isolated. Patients with E. faecium infection had a significantly higher mortality than patients with E. faecalis infection (66% vs. 41.5%, p = 0.0035 for infection from any site and 85.7 vs. 47.7%, p = 0.012 for bacteremic patients). r 4n- D I .- .- - .. . .

Adolescent↗

Vancomycin-resistant enterococci colonizing the intestinal tracts of hospitalized patients.

A point prevalence culture survey was carried out to investigate the prevalence of fecal carriage of vancomycin-resistant enterococci (VRE) among patients admitted to an 800-bed general hospital where no VRE had been isolated previously. Twenty-two of 636 patients (3.5%) were found to be VRE carriers. Eighteen strains were identified as Enterococcus faecium, three were identified as Enterococcus gallinarum, and one was identified as Enterococcus faecalis. The susceptibilities of the enterococci to ampicillin, vancomycin, and teicoplanin were determined by the disk diffusion and the agar dilution methods. High-level resistance (HLR) to gentamicin and streptomycin was determined by the agar screening method. Eighteen strains (82%) were highly resistant to vancomycin, and four strains (18%) were moderately resistant to vancomycin. Five strains were susceptible to teicoplanin (23%; MICs, < or = 8 micrograms/ml). Only one strain (4.5%, E. faecium) showed HLR to gentamicin, and six strains (27%) showed HLR to streptomycin (one E. faecalis and five E. faecium strains). All 18 E. faecium and 1 E. faecalis strain carried the vanA gene, and 3 E. gallinarum strains carried the vanC gene. An epidemiological investigation revealed several risk factors for VRE colonization: hospitalization and duration of stay in the hematology department and prior vancomycin treatment. The study demonstrates that the patient's gastrointestinal tract is a possible reservoir for VRE, even in hospitals where VRE infections have not yet been observed. Therefore, we conclude that infection control precautions and restriction of glycopeptide usage may be key issues in limiting the emergence and spread of nosocomial VRE infections.

Adult↗

Analysis of molecular epidemiological characteristics and antimicrobial susceptibility of vancomycin-resistant and linezolid-resistant Enterococcus in China.

BACKGROUND: This study investigates the distribution and characteristics of linezolid and vancomycin susceptibilities among Enterococcus faecalis (E. faecalis) and Enterococcus faecium (E. faecium) and explores the underlying resistance mechanisms. METHODS: A total of 2842 Enterococcus clinical isolates from patients were retrospectively collected, and their clinical data were further analyzed. The minimum inhibitory concentrations (MICs) of vancomycin and linezolid were validated by broth dilution method. The resistance genes optrA, cfr, vanA, vanB and vanM were investigated using polymerase chain reaction (PCR). Housekeeping genes and resistance genes were obtianed through whole-genome sequencing (WGS). RESULTS: Of the 2842 Enterococcus isolates, 88.5% (2516) originated from urine, with E. faecium accounted for 60.1% of these. The vanA gene was identified in 27/28 vancomycin resistant Enterococcus (VRE) isolates, 4 of which carried both vanA and vanM genes. The remaining strain was vanM positive. The optrA gene was identified in all E. faecalis isolates among linezolid resistant Enterococcus (LRE). E. faecium showed a higher multiple antibiotic resistance index (MAR index) compared to E. faecalis. The multi-locus sequence typing (MLST) showed the sequence type of E. faecium mainly belongs to clonal complex (CC) 17, nearly E. faecalis isolates analyzed were differentiated into 7 characteristics of sequence types (STs), among which ST16 of CC16 were the major lineage. CONCLUSION: Urine was the primary source of VRE and LRE isolates in this study. E. faecium showed higher levels of resistance compared to E. faecalis. OptrA gene was detected in 91.6% of LRE, which could explain linezolid resistance, and van genes were detected in all vancomycin resistant Enterococcus strains, while vanA was a key resistance mechanism in VRE identified in this study.

Linezolid↗

Infections by ampicillin-resistant enterococci: a case-control study.

We identified 17 (20%) of 83 consecutive enterococcal isolates from hospitalized patients with documented infection as high-level ampicillin-resistant enterococci (ARE). Of these, 16 isolates were identified as Enterococcus faecium and 1 isolate as Enterococcus raffinosus. A case-control study found no significant differences with respect to underlying diseases, central venous catheterization, nosocomial acquisition of the infection and sites of infection. Patients with ARE infection were older and had a higher inhospital fatality rate than those with ampicillin-susceptible Enterococcus (ASE) infection. Hospitalization in a surgery service (usually for an abdominal procedure), prolonged hospital stay, prior treatment with antibiotics (in particular imipenem and metronidazole), were also more frequent among patients with ARE infection. ARE isolates were more frequently resistant to imipenem, ciprofloxacin and streptomycin than ASE isolates.

Adult↗

Comparative activity of quinupristin/dalfopristin and RPR 106972 and the effect of medium on in-vitro test results.

Quinupristin/dalfopristin and RPR 106972 were active in vitro against a wide range of aerobic Gram-positive organisms including Enterococcus faecium. However, most isolates of Enterococcus faecalis were resistant or of intermediate sensitivity. Against Staphylococcus aureus quinupristin/dalfopristin was more active but for all other species the range of activity of the two drugs was the same or RPR 106972 was more active. RPR 106972 was also more active against the respiratory pathogens Haemophilus influenzae and Moraxella catarrhalis. Quinupristin/dalfopristin MICs for isolates of H. influenzae (1-8 mg/L) clustered around the breakpoint. There were differences in the quality of growth, but little difference in MICs or zone diameters was obtained on three different media: Mueller-Hinton (MHA), Iso-Sensitest (ISA), and Diagnostic Sensitivity Test (DST) agars. The addition of blood to the medium increased MICs 2- to 4-fold, with MHA showing the greatest increase, and reduced zone diameters around quinupristin/dalfopristin discs by 3-4 mm, with the greatest effect on ISA.

Anti-Bacterial Agents↗

Characterization of glycopeptide-resistant enterococci from U.S. hospitals.

We examined 105 clinical isolates of glycopeptide-resistant enterococci collected from 31 U.S. hospitals in 14 states during May 1988 to July 1992. The isolates included 82 Enterococcus faecium, 8 E. faecalis, 6 Enterococcus spp., 5 E. gallinarum, 3 E. casseliflavus, and 1 E. raffinosus. The isolates were categorized into the following four phenotypes of glycopeptide resistance on the basis of their MIC patterns: (i) 70 VanA (vancomycin [Vm] MIC, > or = 64 micrograms/ml; teicoplanin [Tei] MIC, 16 to > or = 128 micrograms/ml), (ii) 26 VanB (Vm MIC, 16 to 1,024 micrograms/ml; Tei MIC, < or = 2 micrograms/ml), (iii) 5 VanC (Vm MIC, 4 to 16 micrograms/ml; Tei MIC, < or = 2 micrograms/ml) in E. gallinarum, and (iv) 3 E. casseliflavus and 1 E. raffinosus isolates for which Vm MICs were 4 to 16 micrograms/ml and Tei MICs were < or = 1 micrograms/ml were called unclassified. Of the 101 isolates with the VanA, VanB, and VanC phenotypes, 99 were confirmed by production of a specific 1,030-, 433-, or 796-bp polymerase chain reaction product, respectively, and hybridization with the respective gene probe. The vanA gene was also detected in the E. raffinosus isolate for which the Vm MIC was 16 micrograms/ml and the Tei MIC was 1 microgram/ml. The vanA gene was located on either a 34- or a 60-kb plasmid in all of the U.S. isolates examined. Pulsed-field gel electrophoresis demonstrated both intrahospital and interhospital diversity among Vmr enterococci in the United States and was more useful than plasmid analysis for epidemiologic studies.

Anti-Bacterial Agents↗

Esterolytic and lipolytic activities of lactic acid bacteria isolated from ewe's milk and cheese.

In the present work, we report on the esterase and lipase activities of lactic acid bacteria representing the genera Lactococcus, Leuconostoc, Lactobacillus, and Enterococcus isolated from ewe's milk and cheeses. Esterase activity was studied using alpha- and beta-naphthyl derivatives of 2 to 12 carbon atoms and postelectrophoretic detection. The lactic acid bacteria evaluated had intracellular esterase activities, which preferentially degraded the alpha- and beta-naphthyl derivatives of 2 to 6 carbon atoms. By studying postelectrophoretic patterns, it was found that some strains presented more than one esterase. Lactobacillus plantarum O236 showed four enzymes that hydrolyze carboxyl ester linkages with different specificity. Lipase activity was studied in intracellular and extracellular fractions using tributyrin, tricaprylin, triolein, and milk fat as substrates. The intracellular and extracellular fractions of Leuconostoc mesenteroides O257, Lactobacillus plantarum O236, and Lactobacillus acidophilus O177 were able to hydrolyze tributyrin. L. plantarum O186, L. acidophilus O252, Enterococcus faecium O174 and O426, and Enterococcus faecalis Ov409 showed lipase activity associated with the intracellular fraction on tributyrin. Lactococcus lactis O233, L. plantarum O155, and Lactobacillus casei O190 did not hydrolyze triglycerides. Not all strains that showed esterase activity exhibited high activity on triglycerides. Esterase and lipase activities were species- and strain-specific. Wide variations in activity between strains highlight the need for selecting appropriate starters to produce enzyme-modified cheese as well as accelerated ripened cheese.

Animals↗

Antimicrobial resistance among Gram-positive agents of bacteraemia in the UK and Ireland: trends from 2001 to 2019.

OBJECTIVES: The BSAC Bacteraemia Resistance Surveillance collected isolates from UK and Irish hospitals for central testing. Concurrent UKHSA surveillance collated English hospitals' own susceptibility data. Results were collated and compared. METHODS: BSAC Surveillance collected quotas of isolates per site annually from 2001 to 2019. MIC testing was by BSAC agar dilution, with resistance mechanisms identified by synergy tests, interpretive reading and PCR. The UKHSA sought hospitals' data on all bacteraemia isolates. RESULTS: Both surveillance systems recorded dramatic falls in MRSA, from c. 40% of bloodstream Staphylococcus aureus in 2001 to <10% by 2019. Both noted rises in the proportion of MRSA (especially) and MSSA resistant to fusidic acid, along with declines of ciprofloxacin and macrolide resistance amongst MRSA. Methicillin resistance also fell among coagulase-negative staphylococci, albeit only modestly; fusidic acid resistance rose. Shifts for pneumococci were complex, reflecting vaccine-contingent serotype displacements; resistance rates remained low, with high-dose penicillin almost universally active. Enterococcus faecium became more prevalent relative to Enterococcus faecalis; vancomycin resistance averaged 29% among E. faecium versus 2% in E. faecalis, without trend. Erythromycin resistance rose among groups B, C and G (but not group A) streptococci. Oxazolidinones, tigecycline, daptomycin and anti-PBP2' cephalosporins retained near-universal activity against target species, except that tigecycline has been compromised by breakpoint reductions for streptococci. CONCLUSIONS: Gram-positive pathogens were the dominant historical pathogens of bacteraemia. The trends seen here-with many near-universally active antibiotics-indicate little hazard of this situation returning. Nevertheless, few treatments exist in some settings, notably multi-resistant E. faecium endocarditis.

Humans↗

Evaluation of the Vitek GPS-TA card for laboratory detection of high-level gentamicin and streptomycin resistance in enterococci.

The Vitek GPS-TA card (Vitek Systems, Hazelwood, Mo.) was compared with single-concentration broth microdilution and disk diffusion methods using high-content disks for the detection of high-level resistance to gentamicin and streptomycin in 99 isolates of enterococci (81 Enterococcus faecalis isolates and 18 Enterococcus faecium isolates). The GPS-TA card accurately detected high-level resistance to gentamicin, but not streptomycin, in E. faecalis. When streptomycin is being considered for therapy, either disk diffusion or time-kill studies should be used to confirm susceptible results obtained by Vitek testing. Additional studies are needed to determine the best method for testing E. faecium isolates.

Drug Resistance, Microbial↗