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Vancomycin-resistant Enterococcus faecium on a pediatric oncology ward: duration of stool shedding and incidence of clinical infection.

OBJECTIVE: To determine the duration of stool shedding and incidence of clinical infection among pediatric oncology patients colonized with vancomycin-resistant Enterococcus faecium (VRE) in our institution. METHODS: Stool cultures were obtained from all patients admitted from May 15 to August 2, 1994. Patients were followed for evidence of clinical VRE infection and surveillance stool results through August 15, 1995. Genetic relatedness of stool-clinical isolate pairs and serial stool samples was evaluated using pulsed field gel electrophoresis. RESULTS: Twenty-three (32%) of 73 screened patients were colonized with VRE. Eight (35%) of the colonized patients cleared VRE from stool; 10 (43%) were persistent carriers, excreting organisms for 19 to 331 days (median, 112 days); and 5 patients had an insufficient number of stools to determine length of carriage. Persistent carriers had a median of 6 hospital readmissions; 8 of 10 were positive at first or second readmission Clinical VRE infection developed in 6 of 73 patients (annual incidence, 8.2%). Clinical cases had more days of neutropenia between colonization and infection than colonized patients during a comparable follow-up (49 vs. 16 days, P = 0.04). Five of 6 stool-clinical isolate pairs were identical by pulsed field gel electrophoresis. Serial stools from 6 of 7 patients (collected 20 to 343 days apart) were identical by pulsed field gel electrophoresis. CONCLUSION: Persistent gastrointestinal colonization with VRE is common among pediatric oncology patients. Carriage of the same VRE clone for up to 1 year was demonstrated. In the majority of cases invasive and colonizing isolates were identical by DNA fingerprinting techniques, suggesting that the colonizing VRE was the source of infection. Intermittent excretion of organisms in stool makes vigilant tracking and immediate isolation of such patients crucial to control efforts. Prolonged neutropenia may increase the risk of developing clinical infection among VRE-colonized patients.

Anti-Bacterial Agents↗

An investigation of vancomycin-resistant Enterococcus faecium within the pediatric service of a large urban medical center.

BACKGROUND: Between 1990 to 1992 and 1993 to 1995 there was a >5-fold increase (16.7% to 89.8%) in vancomycin-resistant Enterococcus faecium isolates as a percentage of all isolates of vancomycin-resistant enterococci on the pediatric units of The New York Hospital-Cornell Medical Center (NYH-CMC). A molecular epidemiologic investigation was undertaken to determine the extent to which this increase was associated with the spread of a vanA-containing clone of vancomycin-resistant E. faecium that had been previously defined in adults hospitalized at NYH-CMC or with the spread of another vanA clone that had been defined in children hospitalized on the pediatric service at Memorial Sloan-Kettering Cancer Center, which shares a common pediatric intensive care unit and pediatric house staff with NYH-CMC. METHODS: Molecular genotyping of vancomycin-resistant E. faecium isolates obtained from pediatric patients from 1993 to 1995 was performed by pulsed field gel electrophoresis of chromosomal SmaI digests. Southern hybridization was performed using vanA- and vanB-specific probes. Medical records of patients were reviewed for pertinent clinical and demographic information. RESULTS: A single vanB clone of vancomycin-resistant E. faecium was responsible for 17 (77.3%) of 22 isolates in the neonatal intensive care unit (NICU) of NYH-CMC. Two other vanB strains of vancomycin-resistant E. faecium and 2 vanA strains were identified among the 5 remaining NICU isolates. Vancomycin-resistant E. faecium isolates from the other pediatric units represented a heterogeneous population of primarily vanA strains, but vanA clonal strains previously identified from patients on adult services at NYH-CMC and from children hospitalized at Memorial Sloan-Kettering Cancer Center were not detected. CONCLUSION: A newly identified vanB clone was responsible for the increase in vancomycin-resistant E. faecium isolates in the NICU of NYH-CMC. The increase of vancomycin-resistant E. faecium among children hospitalized at NYH-CMC was unrelated to the spread of vancomycin-resistant E. faecium among adults in the same hospital or among children at an affiliated facility cared for by the same house staff and sharing a common pediatric intensive care unit.

Adolescent↗

Experience with quinupristin/dalfopristin in treating infections with vancomycin-resistant Enterococcus faecium in children.

BACKGROUND: The emergence and spread of vancomycin-resistant Enterococcus faecium (VREF) has presented serious therapeutic difficulties because of the lack of reliably active antibiotics. Quinupristin/dalfopristin is a new injectable streptogramin antibiotic that is active against most strains of VREF. Experience with this agent in adults with VREF infections is well-documented; however, there are few reports of its use in children. We report on eight children with VREF infections who received quinupristin/dalfopristin under a compassionate use protocol. METHODS: Quinupristin/dalfopristin was administered according to the manufacturer's recommendations. Clinical and laboratory data were recorded for each patient. RESULTS: The infections treated comprised six cases of bacteremia and two of peritonitis. All patients had serious underlying conditions. Seven patients recovered fully. One patient died, having experienced a relapse of his infection after quinupristin/dalfopristin was discontinued. None of the patients experienced side effects or other adverse events. CONCLUSION: Quinupristin/dalfopristin was well-tolerated and generally effective in children with infections caused by VREF. There is increasing evidence that it may be more effective than other currently available antibiotics in some such patients.

Adolescent↗

Linezolid treatment of vancomycin-resistant Enterococcus faecium ventriculitis.

The successful treatment of a 7-month-old infant with shunt-associated ventriculitis caused by vancomycin-resistant Enterococcus faecium is presented. Linezolid was administered intravenously every 8 h; children have a greater volume of distribution and total body clearance than adults and therefore require more frequent dosing. The patient tolerated the therapy without adverse effects.

Acetamides↗

Incidence and outcome of infection by vancomycin-resistant Enterococcus following orthotopic liver transplantation.

Vancomycin-resistant Enterococcus (VRE) has become a significant nosocomial pathogen. For this study, the records of 325 patients who underwent orthotopic liver transplantation (OLT) were reviewed. Thirty-four patients were infected by VRE (incidence of 10.5%, 14% in adults vs. 5% in children, P < 0.01). Common features of patients who developed infections with VRE included previous antibiotic use (25 patients, 15 of whom received vancomycin), co-infection by other pathogens (28 patients), and relaparotomy following OLT (20 patients). Pulmonary and/or renal failure preceded infection by VRE in 11 and 4 adult patients, respectively. Biliary complications were exceedingly common in patients infected by VRE (28 patients) and significantly increased the risk of infection by VRE (21.5% vs. 3.1% for patients without biliary complications, P < 0.0001). Mortality associated with VRE infections was high (56% vs. 19% for patients not infected by VRE, P < 0.0005). The most frequent cause of death was sepsis (16 of 19 patient deaths), often polymicrobial. The high incidence of infection by VRE following OLT, the lack of effective antibiotics for the treatment of VRE, and the association of VRE with patient mortality emphasizes the need to define the risk factors associated with VRE infection. We suggest early surgical intervention to treat complications that may predispose patients to infection by VRE.

Adult↗

Selective digestive tract decontamination and vancomycin-resistant enterococcus isolation in the surgical intensive care unit.

Vancomycin-resistant Enterococcus (VRE) has emerged as a significant nosocomial pathogen in the surgical intensive care unit (SICU). We wished to test the hypothesis that the use of selective digestive tract decontamination (SDD) in the SICU affects the frequency of VRE isolation. A retrospective review of hospital records and the SICU database was performed using patients admitted to the SICU service for three or more days from January 1, 1996 to December 31, 1999 at our large tertiary-care teaching hospital. During this time use of SDD in selected patient populations decreased due to physician preference. Information gathered included length of SICU stay, presence of VRE infection or colonization, and use and duration of SDD protocol, vancomycin, and ceftazidime. There were 110 newly diagnosed VRE cases in the SICU during this time period. During the same time period 54 patients received SDD. Eight patients who received SDD had positive VRE cultures and seven had the initial positive culture after receiving SDD. Overall, 9.1% of eligible SICU patients received SDD, 18.5% of patients in the SICU for over 3 days had VRE, 7.3% of VRE patients received SDD, and 13.0% of the SICU patients who received SDD subsequently developed VRE. SDD use was not associated with VRE in univariate analysis. Logistic regression analysis showed higher odds ratios for SDD use in combination with vancomycin than for vancomycin use alone (OR=4.3 vs. 10.9). Odds ratios were over three times higher for SDD plus vancomycin plus ceftazidime use when compared to vancomycin plus ceftazidime use alone (OR=70.5 vs. 19.8). We conclude that administration of SDD alone did not correlate with increased VRE isolation, but that SDD use in conjunction with vancomycin and ceftazidime was associated with VRE isolation.

Antibiotic Prophylaxis↗

Enterococcus faecalis exacerbates burn injury-induced host responses in rats.

Pathophysiology of burn injury with complications of gram-positive infections is not well characterized. We have developed an in vivo rat model to study the effects of burn injury along with intra-abdominal inoculation of Enterococcus faecalis. We hypothesized that although burn injury or E. faecalis inoculation by itself may not induce significant pathophysiological responses, the combination of the two can lead to adverse pathophysiological consequences. Sprague-Dawley rats were divided into 4 groups: group 1(C), controls; group 2(B), burn injury on 30% total body surface area; group 3(EF), intra-abdominal implantation of bacterial pellet impregnated with E. faecalis; group 4(B+EF), burn injury plus bacterial pellet implantation. The mortality was 25% and 60% on day 1 and 2 in Group 4(B+EF), respectively; no significant mortality was observed in other groups. In group 4(B+EF), metabolic acidosis, respiratory alkalosis, and a hyperdynamic state developed on day 1, and metabolic and respiratory acidosis and a hypodynamic state on day 2. There were no significant alterations in metabolic or hemodynamic measurements in other groups. Intestinal microvascular permeability to albumin on day 1 and 2 was increased in group 4(B+EF). In group 2(B), microvascular permeability was not increased significantly. Although the permeability was increased on day 1 in group 3(EF), it declined on day 2. The metabolic and hemodynamic alterations were correlated with increased intestinal microvascular permeability to albumin. E. faecalis appeared to be involved in initiating a vicious cycle of burn injury-mediated disruption of intestinal integrity along with metabolic and hemodynamic derangements.

Albumins↗

Vancomycin-resistant enterococcus: case reviews after transplantation.

Increased and inappropriate use of vancomycin during the past 35 years has led to the development of vancomycin-resistant enterococcal (VRE) species that are difficult and expensive to treat. This article presents case reviews of patients who experienced vancomycin-resistant enterococcus faecium (VREF) infections during prolonged hospitalizations after transplantation. The risk factors for the development of VRE infection, prevention measures, and the interventions to be implemented once VRE infection is diagnosed are discussed.

Adult↗

Exacerbation of intestinal permeability in rats after a two-hit injury: burn and Enterococcus faecalis infection.

OBJECTIVE: To determine alterations in intestinal epithelial permeability to solutes in burn injured rats with and without Enterococcus faecalis infection and the role of neutrophils in the intestinal permeability changes. DESIGN: Prospective sham-controlled animal study. SETTING: University research laboratory. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: Rats were subjected to 30% total body surface burn (B group), E. faecalis infection (EF group) induced via intra-abdominal implantation of bacterial pellet, or combination of burn injury and E. faecalis infection (B+EF group). MEASUREMENTS AND MAIN RESULTS: In vivo measurements of intestinal permeability were carried out after intraluminal injection of H lactulose and C mannitol in the ileum of sham, B, EF, and B+EF groups of rats, 1 and 2 days after injury. Lactulose permeability was increased in the injured rat groups (B, EF, B+EF) on day 1 postinjury compared with sham. The combined injury group (B+EF) had the highest level of lactulose permeability. Although a significant change in lactulose permeability from day 1 to day 2 postinjury could not be demonstrated in the B and EF groups, lactulose permeability in the B+EF group on day 2 postinjury markedly decreased from day 1 but was still significantly higher than that in the sham group. Mannitol permeability was increased in all injured rat groups on day 1 postinjury; on day 2 it remained elevated post-B, decreased post-EF, and further increased after B+EF. Ex vivo measurements of lactulose movements across intestinal epithelial monolayers (IEC-18) were carried out in the presence of blood neutrophils from sham, B, EF, or B+EF rats. We also measured ex vivo transepithelial migration of neutrophils from sham, B, EF, or B+EF rat groups. Neither the transepithelial lactulose movement in the presence of neutrophils from, nor neutrophil migration in, the B or EF rats was significantly different from sham. However, a significant increase in transepithelial lactulose movement and neutrophil migration occurred in the B+EF group. Immunoblot analyses and in situ histochemical localizations of intestinal tight junction proteins, occludin and claudin-3, showed decreases in the distribution of occludin but not claudin-3 in the B, EF, and B+EF groups. CONCLUSIONS: Alterations in intestinal solute permeability and disruption of tight junction integrity after a two-hit injury with burn and E. faecalis infection, but not after individual injuries of burn or E. faecalis infection, are likely associated with heightened neutrophil flux across the intestinal epithelium.

Animals↗

Chairside sensor for rapid monitoring of Enterococcus faecalis activity.

In this study, optical spectroscopy was used to monitor a chromogenic, enzyme-substrate reaction for the rapid identification of Enterococcus faecalis. The detection system, comprising a miniature spectrophotometer and an accompanying data acquisition system, was placed in an incubator. During testing, a 3-ml test sample was placed in a cuvette within the spectrophotometer. This permitted online, real-time, and remote analysis of spectral signature needed to monitor the bacteria. It was observed that the absorption peak intensity increased conspicuously 3.5 h after inoculation and through the entire period of testing. A linear-regression analysis demonstrated a significant correlation between the increase in absorption peak intensity at 610 nm (r = 0.9389) and 653 nm (r = 0.9387) with the formation of colony-forming units. Optical spectroscopy-based sensing systems can pave the way for rapid, nonlaboratory-based approaches to monitor microbial status quantitatively and qualitatively from clinical samples.

Biosensing Techniques↗

Effects of sonicated Enterococcus faecalis extracts on interleukin-2 and interleukin-4 production by human T cells.

In the present authors' previous study, sonicated Enterococcus faecalis extracts were shown to suppress the cell cycle progression of human lymphocytes. To study the effect of this microorganism on the function of lymphocytes, the authors investigated the levels of interleukin-2 (IL-2) and interleukin-4 (IL-4) production by T lymphocytes before and after the addition of sonicated E. faecalis extracts. In this study, levels of IL-2 and IL-4 produced from T cells were evaluated by using the quantitative sandwich enzyme immunoassay technique. In response to phytohemagglutinin (PHA) stimulation, T cells produced increased levels of IL-2 and IL-4. However, the expressions of both cytokines were significantly inhibited when PHA-activated T cells were preexposed to 12.5 microg/ml and 25 microg/ml of sonicated E. faecalis extracts (p < 0.05). This effect was concentration-dependent, because the levels of IL-2 and IL-4 expressions were not affected by the addition of a low concentration (5 microg/ml) of sonicated extract. These findings suggest that Th1 and Th2 immunosuppression mediated by E. faecalis could be a part of the pathogenic mechanism of the endodontic failure associated with this microorganism.

Cells, Cultured↗

Antimicrobial efficacy of chlorhexidine and two calcium hydroxide formulations against Enterococcus faecalis.

The purpose of this study was to investigate the efficacy of chlorhexidine (CHX) and calcium hydroxide (Ca(OH2) against Enterococcus faecalis in vitro. Extracted single-rooted human teeth were instrumented up to size 40. After removal of the smear layer, an inoculum of E. faecalis was inserted into the root canals. After incubation, the inoculum was removed and the root canals were filled with one of three different disinfectants: Ca(OH2 paste, CHX 2%, and a mixture of CHX and Ca(OH2 paste (n = 10 in each group). Control teeth were filled with water of standardized hardness (n = 10). The teeth were then incubated for 3 days. After incubation, each root canal was instrumented, and the removed dentin was examined microbiologically. CHX was significantly more effective against E. faecalis than was Ca(OH2 paste or a mixture of CHX with Ca(OH2 paste (p < 0.05). There was no increase in the efficiency of Ca(OH2 paste when CHX was added (p > 0.05). The results suggest that CHX is effective in the elimination of E. faecalis from dentinal tubules under the conditions of this study.

Calcium Hydroxide↗

Susceptibilties of two Enterococcus faecalis phenotypes to root canal medications.

This study aimed to investigate and compare the efficacy of selected root canal irrigants and a medicament on a clinical isolate of Enterococcus faecalis grown as biofilm or planktonic suspension phenotype. A cell-dense pellet "presentation" prepared from planktonic phenotype was also tested. Each bacterial presentation was exposed to calcium hydroxide (pH 12.3), 0.2% chlorhexidine gluconate, 17% ethylene-diamine-tetra-acetic acid, 10% povidone iodine, or 3.0% sodium hypochlorite (NaOCl) for a range of time periods (1, 2, 4, 8, 15, 30, and 60 min). Phosphate buffered saline was used as a control agent. The difference in gradients of bacterial killing among the biofilm, planktonic suspension or pellet presentation was significant (p < 0.05) and dependent upon the test agent except in the case of NaOCl and calcium hydroxide where no difference could be detected. NaOCl was the most effective agent and achieved 100% kills for all presentations of E. faecalis after a 2 min contact time.

Biofilms↗

The expression of alpha4 integrins by human polymorphonuclear neutrophils in response to sonicated extracts of Enterococcus faecalis.

This study was undertaken to investigate immunopathologic mechanism of Enterococcus faecalis in relation to persistent apical periodontitis. We monitored the expression levels of alpha4 integrin in human polymorphonuclear neutophils (PMNs) after stimulated with sonicated extracts of E. faecalis (SEF) and compared with lipopolycaccarides (LPS) of Escherichia coli for various incubation time. Venous blood was collected from healthy volunteers and then PMNs were isolated and cultured with various concentrations of SEF for different periods of time. The levels of alpha4 integrin were measured by flow cytometry analysis. E. coli LPS group was used as a positive control and untreated PMNs as a negative control. Results showed that the expressions levels of alpha4 integrin were increased in human PMNs stimulated with E. coli LPS in comparison with unstimulated control cells (p < 0.05). In case of SEF stimulated group, the expression levels were decreased in time-dependent manner in comparison to E. coli LPS group (p < 0.05). Notably, after 12 h for incubation with SEF, the expression of alpha4 integrin was decreased in dose-dependent manner (p < 0.05). These findings suggest that E. faecalis seem to suppress PMNs recruiting activity by down-regulating alpha4 integrin expression, providing the possible mechanism that E. faecalis may play a crucial role in persistent apical periodontitis.

Analysis of Variance↗

The susceptibility of starved, stationary phase, and growing cells of Enterococcus faecalis to endodontic medicaments.

The purpose of this investigation was to compare the susceptibility of cells of Enterococcus faecalis during exponential growth, stationary phase and starvation phase to three endodontic medicaments. E. faecalis strains VP3-80 and A197A in different growth phases were exposed to saturated calcium hydroxide solution, 0.05% chlorhexidine digluconate and 0.0001% sodium hypochlorite. Cells in the exponential growth phase were the most sensitive to all three medicaments and were killed between 3 s and 10 min. Cells in stationary phase were more resistant and living cells could be recovered at 10 min. However, cells in starvation phase were the most resistant and were not totally eliminated by the medicaments during the 10-min test period. Number of surviving cells of E. faecalis cells to the tested medicaments increased 1000- to 10,000-folds in aging cultures.

Calcium Hydroxide↗

The role of environmental changes on monospecies biofilm formation on root canal wall by Enterococcus faecalis.

Biofilm mode of growth is a strategy in microorganisms to survive harsh growth conditions. Although previous studies have established the ability of Enterococcus faecalis to survive postendodontic environmental conditions, the effect of such conditions on the ultrastructural and physiochemical features of E. faecalis biofilm has received less attention. This study aims to evaluate the effect of different growth conditions on the characteristics of E. faecalis biofilm on root canal, and the penetration of E. faecalis into dentinal tubules. Forty-five intact noncarious human maxillary molars were experimented under nutrient-rich, nutrient-deprived, aerobic, and anaerobic conditions for a period of 21 days. Scanning Electron Microscopy with Energy Dispersive X-ray microanalysis, Laser Confocal Scanning Microscopy and Light microscopic examinations were carried out. The microscopic analysis highlighted a distinct variation in the ultrastructure of the biofilms formed under different experimental conditions. The EDX microanalysis showed a significant increase in the levels of Calcium (Ca) in the biofilm structures formed under anaerobic nutrient-deprived condition (p < 0.001). The depth of bacterial penetration was significantly greater in nutrient-rich condition (p < 0.001). This study demonstrated distinct ultrastructural and physiochemical properties of the biofilms formed and dentinal tubular penetration of E. faecalis under different conditions.

Aerobiosis↗

Vancomycin-resistant Enterococcus faecium endocarditis in a premature infant successfully treated with linezolid.

A 4 1/2-month-old, 26-week premature infant with multiple complications of prematurity required a central venous catheter for venous access and antibiotic treatment of bacterial nosocomial infections. He developed tricuspid valve endocarditis with vegetation caused by Enterococcus faecium resistant to ampicillin, vancomycin and quinupristin-dalfopristin but susceptible to linezolid. He was successfully treated with linezolid intravenously (7 weeks) and then orally (2 weeks).

Acetamides↗

Endocarditis due to vancomycin-resistant Enterococcus faecium in an immunocompromised patient: cure by administering combination therapy with quinupristin/dalfopristin and high-dose ampicillin.

A 56-year-old man with diabetes mellitus and cadaveric renal transplantation had vancomycin-resistant Enterococcus faecium tricuspid valve endocarditis. Relapse followed 6 weeks of treatment with intravenous gentamicin and high-dose ampicillin. On the basis of previous data suggesting the potential for synergistic activity of quinupristin/dalfopristin plus high-dose ampicillin, therapy with this combination was administered for 63 days. Cure was achieved and later confirmed at 2-year follow-up.

Ampicillin↗