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Phenotypes and genotypes of vancomycin-resistant enterococci isolated during long-term follow-up in a patient with recurrent bacteremia and colonization.

Twenty-seven isolates of vancomycin-resistant enterococci were obtained at monthly intervals from a bed-ridden man with hypoxic encephalopathy. During the 28-month period of the patient's hospitalization, 3 episodes of bacteremia and one episode of catheter-related infection caused by vancomycin-resistant enterococci occurred. Rectal swabs showed colonization of vancomycin-resistant enterococci for more than 2 years. Three months after termination of antimicrobial therapy, the rectal colonization for vancomycin-resistant enterococci was eradicated. Four species (Enterococcus faecium, Enterococcus gallinarum, Enterococcus faecalis, and Enterococcus casseliflavus) were identified among the 27 vancomycin-resistant enterococcus isolates. Three non-clonal related patterns were found among 17 strains of E. faecium by pulsed-field gel electrophoresis. All of the 3 E. faecalis isolates were of the VanB phenotype, but of the vanA genotype. Linezolid had the most potent in vitro activity against these vancomycin-resistant enterococcus isolates, with minimum inhibitory concentrations >2 microg/mL. Eighty-five percent of these vancomycin-resistant enterococcus isolates were susceptible to tetracycline and 66% were susceptible to quinupristin-dalfopristin. Although a high genetic correlation of E. faecium was identified in the patient with prolonged hospitalization, the isolation of 3 genetically unrelated colonized isolates suggested a lack of correlation between infection and colonization. Precautions against resistant organisms, adapted antibiotic policies, and elimination of patient carriage are useful for controlling the spread of vancomycin-resistant enterococci.

Anti-Bacterial Agents↗

Prevalence of vancomycin and high level aminoglycoside resistant enterococci among high-risk patients.

Enterococci have been recognized as clinically important pathogens in high-risk populations of hospitalized patients. The role of enterococci in nosocomial infections is being recognized with increasing frequency. The main source of these infections is usually fecal carriage of the microorganisms. In this study, gastrointestinal colonization with vancomycin resistant enterococci (VRE) and high-level aminoglycoside resistant enterococci among 316 high-risk hospitalized patients were investigated. One hundred and ninety-eight enterococci strains were isolated from stool specimens. All strains were identified to species level and 90 of the isolates were identified as Enterococcus faecalis (45%), 85 as E. faecium (21.5%), 14 as E. avium (7%), 7 as E. raffinosus (3.5%), 1 as E. durans (0.5%) and 1 as E. hirae (0.5%). Eleven of 198 strains were found to be moderately sensitive to vancomycin (MIC: 8-16 microg/ml) by the agar dilution method according to the National Committee for Clinical Laboratory Standards (NCCLS) recommendations, and the rest of these strains were found to be sensitive (MIC < or = 4 microg/ml). Twenty-eight strains showed high-level resistance to streptomycin (2,000 microg/ml) and 26 strains were found to have high-level resistance to gentamicin (500 microg/ml). Twelve of these strains had high-level resistance to both aminoglycosides. By the disk diffusion tests, 53 of 198 strains were found to be resistant to erythromycin, 51 to penicillin, 37 to ampicillin, 18 to ciprofloxacin, 14 to norfloxacin and 3 to nitrofurantoin. No beta-lactamase production was detected in 198 studied strains.

Aminoglycosides↗

[Enterococci and coliforms in yellow sheep's milk cheese].

Ninety-two samples of sheep milk yellow cheese were investigated 30 and 180 days after its production. The content of enterococci varied within a wide range--from 100 to 1,9 million per g. In 14.28% of the samples more than 10 000 enterococci per g were found. No correlation between salt content, acidity and pH and number of enterococci in the yellow cheese samples studied as well as between quantity of enterococci, period of cheese ripening and storage and the organoleptic evaluation of the product's ripeness was established. In 100% of the samples studied coli titer was above 0.1. Of the 137 differentiated enterococci strains 67.1% belonged to the species Str. durans, while 32.8%--to Str. faecium.

Animals↗

[Enterococci and coliforms in yellow sheep cheese].

The developmental dynamic of enterococci and coliforms was followed up in the entire technologic process and the storage of kashkaval (yellow cheese of ewe milk) under the conditions of modern industrial production. It was found that during the whole industrial cycle up to the steam cooking of curd the amount of enterococci grew and reached its peak value in the cheddarized cheese curd (2.4--30 million per gram), increasing from 10 to 34 times as against its level in the initial milk used. The coliform bacteria also rose in number, and their amount reached maximum values of 10 to 120 mill/g in the processed and dipped curd, after which a slowly advancing reduction set in. Species of the Enterobacter (55.1%), Escherichia (14.1%), Citrobacter (19.2%), and Klebsiella (11.5%) genera were isolated. The steam cooking of cheddarized curd produced an unfavourable effect on enterococci (pasteurization effect of up to 98.3 to 99.9%) and a lethal effect on the coliforms. Enterococci that resisted steaming multiplied in kashkaval and reached their highest level--960 000 up to 39 mill/g--between the 30th and the 60th day of production after which their numbers dropped. Their amount in the ripened product varied from 95000 to 17.8 mill/g, and on the 240th following production--from below 100 to 1.6 mill/g. Coliform bacteria were not found in 0.1 g of the product mass during ripening and storage of kashkaval. Out of the 196 strains that were differentiated as enterococci 30.7 per cent of the fecalis subgroup, and 69.3 per cent--of the Sp. faecium-durans subgroup. After steaming 92.3 per cent of the strains were of the Str. faecium and Str. durans species.

Animals↗

[The effect of Lactobacillus salivarius administration on coliform bacteria and enterococci in the crop and cecum of broiler chickens].

A rifampicin-resistant Lactobacillus salivarius 51R was isolated from chicken caeca and administered orally to newly hatched broiler chickens. The resistance to rifampicin enabled us to differentiate the administered organism from indigenous strains. First day after inoculation, L. salivarius 51R dominated among lactobacilli in the crop and caeca of inoculated chickens and its counts were slightly over 7 log c.f.u. per 1 g of digesta even after 10 days. L. salivarius significantly (P < 0.01) lowered counts of enterococci and coliforms in the crop during the whole experimental period (10 days). Effects of L. salivarius administration on caecal counts other than lactobacilli were generally small. The influence of Lactobacillus strain used to species composition of enterococci and coliforms were also observed. 105 strains of enterococci and 96 strains of coliforms isolated from the crop and caeca of both control and experimental groups were characterized using identification sets and computer program. Regarding, enterococci, 63% was identified to the species level, 31% to the genus level, and 6% was not identified. Regarding coliforms, 48% was reported to the species level, 25% to the genus level, and 26% was not identified. The most prevalent species among the enterococci was E. faecalis and among the coliforms E. coli. 24 h after the administration of L. salivarius 51R there was significantly higher (P < 0.05) occurrence of E. faecalis (57% out of all enterococcal isolates) in experimental group than those in the control group (31% out of all enterococcal isolates). The perspectives in the control of pathogens in young chickens via probiotics was discussed.

Animals↗

Surveillance for antimicrobial resistance in enterococci.

AIM: To describe antimicrobial resistance patterns of Enterococcus species in Auckland. BACKGROUND: Antimicrobial resistant enterococci have emerged as major nosocomial pathogens in overseas hospitals. It is recommended that hospitals perform periodic surveys to determine local enterococcal resistance patterns. METHODS: Enterococcal isolates from four patient groups were tested: group I were recovered from routine clinical specimens; group II were stool isolates from patients at risk of having vancomycin resistant enterococci, eg, intensive care unit patients, patients receiving vancomycin, and immunocompromised patients receiving antibiotics; group III were enterococci from stool specimens sent for Clostridium difficile toxin testing; group IV were isolates from stool specimens submitted to a community laboratory for enteric pathogen testing. All enterococci isolated were tested for the presence of beta-lactamase, susceptibility to amoxycillin, teicoplanin, vancomycin, and for high level gentamicin and streptomycin resistance. RESULTS: There were 121 group I enterococcal isolates. 628 stool specimens were cultured. Enterococci were isolated from: 76/148 (51%) group II specimens; 166/279 (60%) group III specimens; and 70/201 (35%) of group IV specimens. Antimicrobial susceptibility testing was performed on 433 isolates; 74% were E faecalis, 12% E faecium, 6% E gallinarum/casseliflavus group and 8% other enterococcal species. No isolate produced beta-lactamase. All E faecalis were susceptible to amoxycillin. Two E faecium and one enterococcus species were resistant to amoxycillin (MICs all 16 mg/L). All isolates were susceptible to teicoplanin. Fourteen E gallinarum/casseliflavus group isolates had intermediate susceptibility to vancomycin (MICs of 8 mg/L). One E faecium had intermediate susceptibility to vancomycin (MIC 8 mg/L). High level gentamicin and streptomycin resistance occurred in 64 (15%) and 50 (12%) isolates respectively. CONCLUSION: Vancomycin resistance is rare and is essentially restricted to species that are rarely clinical pathogens, i.e., E casseliflavus and E gallinarum. Our results have established the local susceptibility profile for enterococcal isolates. This allows comparison with other locations and the detection of emerging trends of resistance.

Cross Infection↗

Enterococci. Biologic and epidemiologic characteristics and in vitro susceptibility.

Enterococci cause urinary tract infection (usually asymptomatic), 5% to 15% of cases of endocarditis, and rare cases of meningitis. Their role in polymicrobial infection in the abdomen and pelvis is difficult to assess. Ninety percent of enterococci are inhibited by 4 mg/L of penicillin G, by 2 mg/L of ampicillin, and by 6 mg/L of vancomycin. The penicillinase-resistant penicillins, cephalosporins, carbenicillin, and ticarcillin are at least fourfold less active against enterococci than penicillin G, whereas piperacillin has activity equivalent to penicillin G. The addition of an aminoglycoside to penicillin, ampicillin, vancomycin or piperacillin--which are not bactericidal against most strains of enterococci--results in more rapid and complete bacterial activity (ie, synergistic activity).

Anti-Bacterial Agents↗

Health and economic outcomes of vancomycin-resistant enterococci.

BACKGROUND: The health and economic impact of vancomycin-resistant enterococci has not been quantified. METHODS: A retrospective matched cohort study was conducted comparing the outcomes of patients with vancomycin-resistant enterococci (cases) with those of control subjects matched for length of hospital stay until inclusion in the cohort, hospital location, and calendar date. The propensity to be a vancomycin-resistant enterococci case was modeled based on patient characteristics, and included in multivariable models to adjust for confounding. Analyses included the following: (1) conditional logistic regression for mortality, surgery, intensive care unit admission, and discharge to long-term care; (2) linear regression for the logarithm of cost; and (3) accelerated failure time model for length of stay. RESULTS: A total of 233 cases were compared with 647 controls. Groups were similar in age (mean, 62 years), sex (female, 47%), and length of stay before inclusion in the cohort (mean, 8.1 days), but differed in primary diagnosis and comorbidities, past infection or colonization with methicillin sodium-resistant Staphylococcus aureus or Clostridium difficile, and treatment with cephalosporins or metronidazole. These variables were included in the propensity score, which had good to excellent prediction. Outcomes for cases vs controls and adjusted risks (relative risks [RRs]) were as follows: (1) case fatality rate, 17% vs 6% (RR, 2.13; P =.04); (2) length of stay after inclusion in the cohort, 15.1 vs 8.5 days (RR, 1.73; P<.001); (3) hospital costs, $52 449 vs $31 915 (RR, 1.40; P<.001); (4) surgery after inclusion in the cohort, 18% vs 10% (RR, 2.74; P =.001); (5) intensive care unit admission after inclusion in the cohort, 25% vs 14% (RR, 3.47; P<.001); and (6) transfer to an institution, 51% vs 35% (RR, 2.01; P =.001). CONCLUSION: Compared with a matched hospital population, a population with vancomycin-resistant enterococci was associated with severe adverse outcomes: increased mortality, morbidity, and costs.

Cohort Studies↗

Childhood meningitis caused by enterococci and viridans streptococci.

Selected clinical and laboratory parameters were studied respectively in patients with meningitis caused by enterococci and viridans streptococci in an academic children's hospital. During a nine-year period (1981-1989), enterococci or viridans streptococci were isolated from the cerobrospinal fluid (CSF) of 48 patients. In nine of these 48 patients, enterococci or viridans streptococci were the causative agents of meningitis. These nine children constituted 2.0% of 450 patients with bacterial meningitis in this period. All nine children suffered from underlying diseases; neurosurgical procedures were performed in six of these patients, of whom four had ventricular drains. A head trauma preceded the development of meningitis in another patient. Drainage of the lacrimal duct was associated with the development of meningitis in another patients. One child concurrently suffered from severe gastroenteritis. CSF leukocyte count and CSF protein levels were moderately elevated, whereas CSF glucose levels were either slightly decreased or within the normal range. Meningitis due to enterococci or viridans streptococci is seen predominantly in children under the age of one year. Predisposing factors, including neurosurgical procedures, head trauma and severe gastroenteritis, are usually present in these patients. The prognosis for recovery is generally good.

Blood↗

Neonatal bacteremia due to high level aminoglycoside resistant (HLAR) enterococci.

A study was conducted to determine the drug resistance pattern of enterococci isolated from cases of neonatal bacteremia. A total of 41 strains of enterococci were isolated of which 35 (85.4%) were Enterococcus faecalis and 6 (14.6%) were Enterococcus faecium. A total of 3 (8.6%) strains of E. faecalis and 2 (33.3%) E. faecium strains showed high level aminoglycoside resistance (HLAR). None of the enterococci tested were vancomycin resistant. Drug resistance was more common among E. faecium strains. All clinically significant isolates of enterococci should be tested for their antibiotic sensitivity pattern including HLAR, and for treatment, antibiotics are selected based on in vitro antibiotic sensitivity test results.

Aminoglycosides↗

Pathogenic enterococci: new developments in the 21st century.

Enterococci, traditionally viewed as Gram-positive commensal bacteria inhabiting the alimentary canals of humans and animals, are now acknowledged to be organisms capable of causing life-threatening infections in humans, especially in the nosocomial environment. The existence of enterococci in such a dual role is facilitated, at least in part, by its intrinsic and acquired resistance to virtually all antibiotics currently in use. Beginning with the initial identification of a 'streptococci of fecal origin' in the late 19th century, enterococci have been studied for over a century now. A number of comprehensive reviews during this time have addressed various aspects of enterococci, including classification, biology, virulence, antibiotic resistance and so on. This review specifically addresses the important advances in the field of enterococcal research that have occurred since the beginning of the 21st century. Most notable among these developments have been the insights into enterococcal genomes and pathogenicity.

Adhesins, Bacterial↗

The prevalence of enterococci in the dental plaque of chronic hospital patients.

Enterococci have caused 2 of 3 reported cases of infective endocarditis following dental treatment of patients receiving haemodialysis. Since this could indicate an increased carriage rate of enterococci in the mouths of such patients, the prevalence of enterococci in dental plaque was compared in four groups of subjects, chronic dialysis patients, chronic cardiac patients, medical staff and dental hospital out-patients. The isolation of enterococci was significantly increased in both chronic patient groups and this may be related to their greater exposure to antibiotic therapy.

Adult↗

Culture media for enterococci and group D-streptococci.

Lancefield group D-streptococci are contaminants of various food commodities, especially those of animal origin. They encompass the new genus Enterococcus comprising 13 known species and some species of streptococci which have their habitat in the intestine of animals, e.g. Streptococcus bovis, suis and equinus. The serologically based grouping may no longer constitute the best definition for streptococci from the food chain. Food hygiene monitoring systems using enterococci as indicators need reliable methods for selective cultivation and identification of marker strains. Up to now more than 100 modifications of selective media have been described for isolating streptococci or enterococci from various specimens. The selection of a medium requires either experience or consultation. It depends on the kind of specimen, the method of cultivation (plate count or membrane filter) and whether or not the habitat is heavily contaminated with other organisms. The choice of media is made more difficult as commercial versions of the same culture medium may vary in recipe and/or performance from producer to producer. Therefore, reviewing the literature may help in the choice of medium and confirmation tests. The selectivity and productivity of some commonly used or cited media are reported here, partly based on our own experience: citrate azide tween carbonate agar (CATC), kanamycin aesculin azide agar (KAA) and M-enterococcus agar (ME) including earlier results with aesculin bile azide agar (ABA), and thallous acetate tetrazolium glucose agar (TITG). No medium was completely selective for all group D-streptococci or for all enterococci but some media were highly selective for a single Enterococcus species, e.g., for E. faecalis which serves as indicator of human pollution. Confirmatory tests must be carried out when experience in the evaluation procedure is limited. Selective media for enterococci should be used only after or while checking in parallel their selectivity and productivity against appropriate test organisms.

Animals↗

Rapid identification of enterococci by pyrrolidonyl aminopeptidase activity (PYRase).

Group A streptococci and enterococci can be differentiated from other streptococci by their ability to cleave pyrrolidonyl beta-naphthylamide (PYRase). We evaluated two PYRase systems [Strep-A-Chek (SAC), E-Y Laboratories, San Mateo, CA; Strep-A-Fluor (SAF), BioSpec, Inc., Dublin, CA) for the presumptive identification of enterococci. Initially, 40 enterococcal and 21 nonenterococcal streptococci were tested, retrospectively. A prospective comparison of SAC and SAF to bile-esculin reaction (BE) was then incorporated into our routine procedure for the identification of non-beta-hemolytic streptococcal colonies from cultures. All isolates were speciated using standard biochemical tests. We encountered 85 enterococcal and 26 nonenterococcal isolates. Tests were performed on colonies from primary plates whenever possible (77 isolates). Sensitivity and specificity of both SAC and SAF were greater than 96% in identifying enterococci from routine cultures. These PYRase tests were cost-effective, easily adaptable to work-flow, and yielded results within 30 min. Thus, PYRase testing appears to be a reasonable alternative for the identification of enterococci in the clinical laboratory.

Aminopeptidases↗

Vancomycin-resistant enterococci (VRE) in meat and environmental samples.

We investigated the spread of vancomycin-resistant enterococci (VRE) in strains from meat and environmental samples and the location of glycopeptide-resistance determinants in VanA isolates. VRE and VSE (vancomycin-sensitive enterococci) resistance patterns to six antimicrobials were also evaluated. A total of 59 meat isolates (35%) and 119 environmental isolates (26.5%) were glycopeptide resistant enterococci. In particular, 10.7% meat isolates belonged to the VanA, 8.3% to VanB and 16% to VanC phenotypes. Environmental samples presented 0.7% VanA, 14.5% VanB, and 11.4% VanC strains. Evident differences were not observed among the resistance patterns of VRE and VSE isolates. Neither an important difference was observed comparing the resistance patterns in enterococci from meat and environment. In particular a low incidence of beta-lactamic resistant strains was found, whereas high rates of resistance were observed for streptomycin (85.7% and 92.8%), kanamycin (79.7% and 96%) and gentamycin (85.1% and 91.7%). An intermediate rate of resistant bacteria emerged for erythromycin (35.1% and 10.5%). All VanA isolates independent of origin had more plasmids with different molecular weights. PCR amplification of the 732 bp fragment in plasmids from the VanA strains confirmed affiliation to the vanA gene cluster and the extrachromosomal location of the glycopeptide-resistance determinants. Our study suggests that food and environment play a potential role as reservoirs of resistance determinants, prompting the need to undertake epidemiological and molecular studies to evaluate the mobility of these genes.

Anti-Bacterial Agents↗

Decay of intestinal enterococci concentrations in high-energy estuarine and coastal waters: towards real-time T90 values for modelling faecal indicators in recreational waters.

Intestinal enterococci are the principal 'health-evidence-based' parameter recommended by WHO for the assessment of marine recreational water compliance. Understanding the survival characteristics of these organisms in nearshore waters is central to public health protection using robust modelling to effect real-time prediction of water quality at recreation sites as recently suggested by WHO and the Commission of the European Communities Previous models have more often focused on the coliform parameters and assumed two static day-time and night-time T90 values to characterise the decay process. The principal driver for enterococci survival is the received dose of irradiance from sunlight. In the water column, transmission of irradiance is determined by turbidity produced by suspended material. This paper reports the results of irradiated microcosm experiments using simulated sunlight to investigate the decay of intestinal enterococci in relatively turbid estuarine and coastal waters collected from the Severn Estuary and Bristol Channel, UK. High-turbidity estuarine waters produced a T90 value of 39.5 h. Low-turbidity coastal waters produced a much shorter T90 value of 6.6 h. In experiments receiving no irradiation, high-turbidity estuarine waters also produced a longer T90 of 65.1 h compared with corresponding low-turbidity coastal waters, T90 24.8 h. Irradiated T90 values were correlated with salinity, turbidity and suspended solids (r>0.8, p<0.001). The results suggest that enterococci decay in irradiated experiments with turbidity >200 NTU is similar to decay observed under dark conditions. Most significantly, these results suggest that modelling turbidity and or suspended solids offers a potential means of predicting T90 values in 'real-time' for discrete cells of a hydrodynamic model.

Dose-Response Relationship, Radiation↗

Safety aspects of enterococci from the medical point of view.

Enterococci occur in a remarkable array of environments. They can be found in soil, food, and water, and make up a significant portion of the normal gut flora of humans (10(5)-10(7)/g of stool) and animals. As other bacteria of the gut flora, enterococci can also cause infectious diseases. Most clinical isolates are Enterocococus faecalis, which account for 80-90% of clinical strains. Enterocococus faecium accounts for 5-10% of such isolates. Typical enterococcal infections occur in hospitalised patients with underlying conditions representing a wide spectrum of severity of illness and immune modulation. Enterococci today rank second to third in frequency among bacteria isolated from hospitalised patients. They are isolated from urinary tract infections, intra-abdominal and pelvic infections, bacteremias, wound and tissue infections, and endocarditis--often as part of a polymicrobial flora. Surprisingly, little is known about the factors that contribute to the ability of enterococci to cause infections. Many strains of E. faecalis produce a cytolysin (haemolysin) exhibiting tissue-damaging capacity. Further extracellular products often observed in clinical isolates are a proteinase (gelatinase), hyaluronidase, and extracellular superoxide. Furthermore, many of the clinical isolates possess the aggregation substance on the surface and an extracellular surface protein, both contributing to the adherence to eucaryotic cells. Some strains of E. faecalis, and many E. faecium strains are resistant to multiple antimicrobials. The ultimate role of all these factors in enterococcal pathogenicity remains to be determined. It was previously thought that enterococcal infections were endogenously acquired from the patient's own gut flora. A rather new concept that has emerged is that enterococcal disease is a two-stage process. There is an initial colonisation of the gastrointestinal tract by enterococcal strains possessing virulence traits and/or antibiotic resistance. Subsequently, this population spreads, often facilitated by antibiotic elimination of competitors. For a selected number of patients, there is subsequent tissue invasion from the gastrointestinal tract reservoir. From this concept, it can be deduced that enterococcal strains without virulence traits and antibiotic resistances exogenously transferred into the human gut via food products or probiotics will not represent any risk for immunocompetent individuals. In very severely immunocompromised patients, however, a risk for enterococcal disease by such strains cannot completely be excluded.

Consumer Product Safety↗

Antibiotic resistance of enterococci isolated at a teaching hospital in Kuwait.

Enterococci isolated in a teaching hospital were studied for their resistance to different antibiotics. Minimum inhibitory concentrations to high-level aminoglycosides and glycopeptide antibiotics were determined by agar dilution and E-test methods respectively. Genes encoding aminoglycoside-modifying enzymes were detected by the polymerase chain reaction (PCR). 195 enterococci were isolated from urines (54.3%), wounds (16.4%), blood (10.2%), and miscellaneous sources (18.9%). They consisted of E. faecalis (88.7%), E. faecium (9.2%), E. casseliflavus (1.5%) and E. bovis (0.5%). None of the enterococci produced penicillinase but 3.5% of them were resistant to ampicillin. They were also resistant to high-level gentamicin (15.9%), kanamycin (22.0%), streptomycin (21.0%), tetracycline (65.1%), erythromycin (62.6%), ciprofloxacin (36.1%), chloramphenicol (26.1%), vancomycin (3.0%) and teicoplanin (2.0%). Most of the high-level aminoglycoside-resistant isolates contained genes coding the bifunctional aminoglycoside modifying enzymes AAC(6')-APH(2"), APH(3') and ANT(6') but not the ANT(4') enzyme. The results demonstrated a low prevalence of vancomycin resistance among Enterococci in this hospital.

Academic Medical Centers↗