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Clustering of Enterococcus faecalis infections in a cardiology hospital neonatal intensive care unit.

Early identification of an outbreak is one of the main advantages of routine epidemiological surveillance. Enterococcus spp. used to be regarded as microorganisms of low pathogenicity, because they are part of the normal microbial flora of the gastrointestinal and genitourinary tract. Recently, they have emerged as important pathogenic agents, sometimes causing infections with high mortality rates. We studied a clustering of primary bloodstream infections caused by Enterococcus faecalis in a cardiology hospital neonatal intensive care unit (NICU). Four cases of primary bloodstream infection by E. faecalis were detected from April 15 to May 13, 2004, during active infection surveillance. The isolates were sensitive to glycopeptides. Some aspects of the management of these patients, including the date of insertion and placement of a central venous catheter, prescription of a specific medication, contiguity of beds, personnel attending the patients, and occurrence of diarrhea were analyzed to look for factors that might affect the spread of the microorganisms. Measures taken to hamper the spread included contact precautions throughout the unit, cleansing and disinfection of equipment and surfaces, bathing children with 2% chlorhexidine-gluconate-containing soap, professional reeducation, and reinforcement of all measures to prevent infections. We suggest that there is a need to re-evaluate preventive infection measures and to review the strategies aimed at decreasing the nosocomial infection rate in the NICU.

Bacteremia↗

Factors affecting the adsorption of bacteriocins ST194BZ and ST23LD to Lactobacillus sakei and Enterococcus sp.

Bacteriocins ST194BZ and ST23LD, produced by Lactobacillus plantarum, inhibit Gram-positive and Gram-negative bacteria. Images obtained by atomic force microscopy showed clear signs of membrane damage of Lactobacillus sakei, accompanied by the leakage of DNA and beta-galactosidase. Adsorption of the bacteriocins to cells was increased when cells were treated with buffers at pH values above neutral. An increase in bacteriocin ST194BZ adsorption to cells of Enterococcus sp. and L. sakei was observed with an increase in incubation temperatures, but at different rates for the two species. Treatment of the two species with various inorganic salts and solvents gave different results regarding the adsorption of the two bacteriocins. In general, pre-treatment of the two sensitive cells with Triton X-100, Triton X-114 and chloroform increased the adsorption of the two bacteriocins. Increased adsorption of bacteriocin ST23LD to L. sakei was recorded when the cells were pre-treated with Tris and NH4-citrate. Treatment of Enterococcus sp. and L. sakei with Na-EDTA and SDS decreased the adsorption of the two bacteriocins. Variable results were recorded with inorganic salts.

Adsorption↗

Identification and characterization of Enterococcus species isolated from forage crops and their influence on silage fermentation.

Forty-eight strains of lactic acid bacteria were isolated from forage crops, and their identification, characterization and influence on silage fermentation were studied. All isolates were Gram-positive, short-chain-forming, catalase-negative, and facultatively anaerobic cocci that did not produce gas from glucose, only formed L-lactic acid, and were able to grow at pH 9.6%, in 6.5% NaCl, or with 40% bile, but not below pH 4.5. These isolates were commonly isolated from forage crops, and they were divided into four groups (A, B, C, and D) according to sugar fermentation characteristics. Selected strains of FA 5, FA 27, FA 45, and FA 57 were identified as Enterococcus hirae, E. faecalis, E. casseliflavus, and E. mundtii, respectively, on the basis of DNA-DNA homology, Strains FA 5, FA 27, FA 45, and FA 57 and two strains from commercial inoculants, LC 10 (Lactobacillus casei) and LP 15 (L. plantarum), were used as additives to alfalfa and guinea grass silages. Alphalfa and guinea grass silages inoculated with LC 10, LP 15, FA 5 + LC 10, and FA 5 + LP 15-treatments in alfalfa and guinea grass silages had significantly lower pH values, contents of butyric acid and ammonia nitrogen, gas production, and dry matter losses compared with the control silage after 60 d of fermentation. However, the E. hirae FA 5, E. faecalis FA 27, E. casseliflavus FA 45, and E. mundtii FA 57-inoculated silages gave similar values to the control in both types of silage. The FA 27 + LC 10 and FA 27 + LP 15-inoculated silages did not differ in fermentation quality from LC 10 and LP 15-inoculated silages alone. The results confirmed that Enterococcus species were not able to improve silage quality.

Bacteria, Anaerobic↗

A case of Enterococcus faecalis endophthalmitis with corneal ulcer.

Although there have been a few reported cases of Enterococcal endophthalmitis, this is an unusual case of endophthalmitis complicated with corneal ulcer caused by Enterococcus faecalis. A 67-year-old male patient with diabetes mellitus underwent secondary intraocular lens implantation. Post-operative recovery was uneventful until a wound rupture was noted 3 weeks after the operation. On day 12 after the repair of the wound, endophthalmitis accompanied by wound necrosis and a full-thickness corneal ulcer was detected. His vision was light perception, and Enterococcus faecalis was identified by culture in samples of conjunctival sac, anterior chamber and vitreous humor. After 3 rounds of intravitreal antibiotics injection, the vitreous opacity disappeared on ultrasonographic finding but corneal opacity and corneal neovascularization still remained.

Aged↗

Enterococcus-like infections in Macrobrachium rosenbergii are exacerbated by high pH and temperature but reduced by low salinity.

Macrobrachium rosenbergii (10 to 15 g and 8 to 12 g at intermolt) were challenged with an enterococcus-like bacterium (strain KM002) previously isolated and identified as the causal agent of mortality. Challenge doses and conditions of pH, salinity and temperature were varied to determine the influence of environmental factors on the development of disease and mortality. Survival was 100% for the unchallenged control groups in all trials. In pH tests, the onset of mortality was earlier at pH 8.8 to 9.5 than at pH 4.6 to 5.2 and 7.5 to 7.7. Also, as pH 8.8 to 9.5, all challenged prawns died within 6 d in high dose challenge tests. By contrast, 20% of the prawns challenged at pH 4.6 to 5.2 and 7.5 to 7.7 survived. At low dose challenge (5 x 10(4) cfu prawn(-1)), survival increased significantly except at pH 8.8 to 9.5. In salinity tests at 2 challenge doses (1 x 10(6) and 2 x 10(7) cfu prawn(-1)), onset of mortality was earliest at 15 ppt and cumulative mortality was 100% at 15 ppt and 0 ppt. By contrast, survival was 80% at 5 and 10 ppt at the low dose challenge and 40% and 60%, respectively, at the high dose challenge. When the challenge dose was reduced to 5 x 10(4) cfu prawn(-1), survival was not significantly different at different salinity levels. In temperature tests at pH 7.2 to 7.5 and at 2 challenge doses (2 x 10(7) and 4 x 10(7) cfu prawn(-1)), the onset of mortality was earliest at 33 to 34 degrees C and total mortality occurred at 27 to 28 degrees C and 33 to 34 degrees C. By contrast, there were 40% and 20% survivors, respectively, for low and high challenge doses at 30 to 31 degrees C. Reducing the challenge dose to 5 x 10(4) cfu prawn(-1) gave higher survival in all groups. However, survival at 33 to 34 degrees C was still lowest. In similar temperature tests but at pH 8.8 to 9.5, onset of mortality was somewhat accelerated and there was 100% death for all the high challenge groups. At low challenge doses, mortality was lower but still highest in the 33 to 34 degrees C group. Results indicated that mortality of M. rosenbergii caused by this Enterococcus-like bacterium was exacerbated by environmental parameters of temperature and pH different from those known to be optimal for prawn growth. By contrast, low salinity appeared to have a beneficial effect on survival. Further work is needed to determine the mechanisms underlying these effects.

Animals↗

Daptomycin-resistant Enterococcus faecium in a patient with acute myeloid leukemia.

Daptomycin is a lipopeptide antimicrobial used for the treatment of aerobic gram-positive skin and soft tissue infections. We describe a patient with acute myeloid leukemia whose febrile neutropenia was treated with daptomycin and who later developed daptomycin-resistant Enterococcus faecium infection. Defervescence and negative blood cultures ensued after treatment with linezolid. Guidelines for testing daptomycin susceptibillities of enterococci include breakpoints only for vancomycin-susceptible Enterococcus faecalis, making interpretation of minimum inhibitory concentrations for common clinical infections difficult. No enterococcal cross-resistance has been reported among daptomycin, linezolid, or quinupristin-dalfopristin, and these agents may be viable alternatives.

Acute Disease↗

Retinopathy associated with enterococcus enteropathy in the neonatal rat.

PURPOSE: Preretinal neovascularization has been previously observed in neonatal rats with spontaneously occurring diarrhea. This neovascularization appears analogous to retinopathy of prematurity (ROP), which occurs in human neonates. A new enterococcus species, designated Enterococcus rattus, has been isolated from the duodenum of these rats. In the present controlled study, the effect of the enteropathy induced by this organism on the retinal vasculature in the neonatal rat was further investigated. METHODS: One hundred fifty newborn Sprague-Dawley rats were randomly assigned to 6 expanded litters (n = 25). On the second day of life, animals were gavaged with either 100 microl of E. rattus suspension (1.0 X 10(7) colony forming units, inoculated group, n = 100 rats) or 100 microl saline (control group, n = 50 rats). All rats were raised in room air and were killed on day 13 of life. Duodenal and blood samples were cultured. The retinal vasculature was assessed using fluorescent microscopy and ADPase staining in a masked manner. Two additional inoculated litters and one control litter were studied for evaluation of arterial blood gases and validation of the grading method for preretinal neovascularization. RESULTS: One hundred percent of rats in the inoculated group developed severe diarrhea and had duodenal cultures positive for E. rattus compared with 0% in the control group. Preretinal neovascularization similar to ROP occurred in 55% of rats in the inoculated group compared with 2% in the control group (P = 0.001). Retinal vascular areas were reduced in the inoculated group (mean +/- SD, 89% +/- 5% versus 96% +/- 2%; P < 0.001). Rats in the inoculated group demonstrated severe growth retardation (final weight, 9.7 +/- 2.2 versus 16.7 +/- 2.7 g, P < 0.001). Inoculated animals also experienced acidosis (pH 7.31 +/- 0.06 versus 7.39 +/- 0.06 control, P = 0.04). CONCLUSIONS: A previously undescribed enterococcal enteropathy was associated with preretinal neovascularization similar to ROP in the neonatal rat. This supports an independent role for factors other than inspired oxygen in the development of ROP.

Animals↗

[Evolution of resistance in the genus Enterococcus in strains isolated from blood].

The objective of this study was to determine the evolution of the species distribution and the prevalence of resistance to the Enterococcus genus. We studied 281 strains of enterococcus isolated from blood samples: 90 throughout 1984 and 791 from the years 1994 to 1996. identification was made using PosCombo 4Y Microscan-Baxter dehydrated panels and the Rapid ID 32 Strep system (bioMerieux). The MICs were calculated using the agar dilution method according to recommendations of the NCCLS for the following antibiotics: ampicillin, vancomycin, teicoplanin, gentamicin, kanamycin and streptomycin. The production of betalactamases were evaluated using a paper disk with nitrocefin for all the strains. The genotypes with resistance to glycopeptides were determined using PCR. The percentage of E. faecalis for 1984-1986/1994-1996 was 82.2/79.4; of E. faecium 4.4/16.4; and other species 12.214.3. The resistance to ampicillin went from 1.1% to 5.8%; high level resistance to glycopeptides went from 0% to 9.9%; for low level from 7.7% to 2.6%; resistance to a high charge of gentamicin went from 27.7% to 40.8%; and that for kanamycin from 45.5% to 62.8%. Resistance to streptomycin remained constant (45.5%). No strains produced betalactamases. For the species E. faecium, a statistically significant increase was detected for global resistance to ampicillin, gentamicin and kanamycin, with resistance to streptomycin remaining at similar percentages. No high level resistance to glycopeptides was detected in the first time period, but the low level resistance was greater.

Ampicillin Resistance↗

In vitro activity of quinupristin/dalfopristin and other antibiotics against ampicillin-resistant enterococcus faecium.

BACKGROUND: Enterococcus faecium constitutes approximately 10% of clinical isolates of enterococci and is noted for its antimicrobial resistance. In particular, E faecium is commonly resistant to ampicillin. The optimal treatment for severe infections caused by these multi-resistant organisms has yet to be determined. METHODS: Enterococci tested were isolated from blood, pleural fluid and cerebrospinal fluid. Ampicillin-resistant Enterococcus faecium (AREF) was identified using the API Rapid Strep Kit system. A total of 58 isolates of AREF were enrolled in this study. Ten different antibiotics were tested, including Synercid (quinupristin/dalfopristin), teicoplanin, vancomycin, ampicillin, trimethoprim/sulfamethoxazole (TMP/SMX), ciprofloxacin, gentamicin, chloramphenicol, rifampicin and tetracycline. The agar dilution method described by the National Committee for Clinical Laboratory Standards was used to determine the minimum inhibitory concentrations (MICs) of the antibiotics tested. RESULTS: Teicoplanin showed the best in vitro activity. Its MIC ranged from 0.25 to 2 micrograms/ml with an MIC90 of 1 microgram/ml. The MIC of vancomycin was 0.5-128 micrograms/ml with an MIC90 of 2 micrograms/ml. Three strains were vancomycin resistant, and they were the VanB phenotype. The MIC of quinupristin/dalfopristin was 0.5 to 8 micrograms/ml with an MIC90 of 2 micrograms/ml. Chloramphenicol and tetracycline showed moderate susceptibility. AREF showed high resistance to other antibiotics tested, including ciprofloxacin, gentamicin, TMP/SMX and rifampicin. High-level gentamicin resistance (MIC > 1,000 micrograms/ml) was found in 78% of AREF tested. CONCLUSIONS: Teicoplanin showed the best in vitro activity against AREF. Clinical studies are necessary to confirm the efficacy of quinupristin/dalfopristin in vivo.

Ampicillin Resistance↗

Characterization of a highly glycopeptide-resistant Enterococcus gallinarum isolate.

BACKGROUND AND PURPOSE: Adequate treatment of emergency infection involving antibiotic-resistant bacteria such as vancomycin-resistant Enterococcus requires a convergence of clinical and bacteriologic techniques. An isolate of Enterococcus gallinarum, designated as TSGH63, is known to be uncommonly vancomycin-resistant. This study investigated the genetic determinant for this unique characteristic. METHODS: After completing the conventional identification and sensitivity tests, the genomic content of E. gallinarum TSGH63 was extracted and analyzed by pulse-field electrophoresis. A set of specific primers for vanA, vanB, vanC1, and vanC2/C3 genes was then applied in a multiplex polymerase chain reaction (PCR) to differentiate its genetic content. To locate the determinant for high vancomycin resistance, the electrophoresis profile was further analyzed by Southern blot using the digoxigenin (DIG)-labeled vanA gene probe. Finally, interspecies transfer of the vancomycin-resistance determinant of E. gallinarum TSGH63 was tested by a conjugation experiment in vitro. RESULTS: A 50-kb plasmid was identified in the analysis of the genomic extract of E. gallinarum TSGH63 by pulse field electrophoresis. Using multiplex PCR, we demonstrated that E. gallinarum TSGH63 harbors a vanA gene in addition to a vanC1 gene. The DIG-labeled vanA gene-specific probe bound to the plasmid exclusively on the Southern blot. The plasmid-carried vanA gene, but not the vanC1 gene, was found to be transferable from TSGH63 to E. faecalis JH2-2 by conjugation in vitro. CONCLUSIONS: This is the first report of isolation of E. gallinarum with a high level of resistance to glycopeptides in Taiwan. The demonstrated interspecies transfer of the vancomycin-resistance gene highlights the importance of stringent control of the use of vancomycin.

Conjugation, Genetic↗

Gastrointestinal colonisation of vancomycin-resistant enterococcus in a Singapore teaching hospital.

Vancomycin-resistant enterococcus (VRE) has become an important nosocomial pathogen in many countries but is still rare in Singapore. A study was conducted from January to March 1997 at a 900-bed teaching hospital to determine the prevalence of intestinal colonisation of VRE in the patient population. A total of 299 consecutive stool specimens received by the microbiology laboratory for routine testing were plated onto two different selective media for comparison. VRE isolated were phenotypically characterised using minimum inhibitory concentrations (MICs) to vancomycin and teicoplanin and then typed using pulsed-field gel electrophoresis (PFGE) with Smal digestion of DNA. VRE were detected in the stool of 35 patients (12.3%). This group consisted of four isolates with VanB (one Enterococcus faecalis and three E. faecium) and 31 isolates with VanC (30 E. casseliflavus and one E. gallinarum). Two patients (0.7%) carried isolates (both VanB) with high level resistance to vancomycin (MIC > or = 256 microg/ml) while the rest had isolates of low level resistance (MIC = 8 microg/ml). Except for isolates from the same patients, PFGE patterns were diverse, suggesting that the VRE isolates were genotypically different and possibly introduced from many sources. This study demonstrates that VRE colonisation is not uncommon in the Singapore patient population.

Adult↗

Antimicrobial susceptibility pattern of Enterococcus species from urinary tract infections.

OBJECTIVE: To determine the extent of drug-resistance among Enterococcus species in, in vitro experiments. Test strains were isolated from cases with urinary tract infection. Antimicrobial sensitivity was determined with particular reference to the beta-lactams, glycopeptide and aminoglycoside groups of antibiotics. METHODS: Studies were carried out on pure cultures of enterococci isolated from urine specimens of 147 patients. Freshly voided, aseptically collected, midstream specimens of urine were cultured by using standard techniques. The identification and speciation of the strains of enterococci were performed by employing a battery of tests. In vitro drug-susceptibility tests of enterococci were performed against the following antibiotics: penicillin, ampicillin, ampi-sulbactum, co-amoxi-clavulanate, ciprofloxacin, vancomycin, teicoplanin, gentamicin and streptomycin, by employing the disk diffusion method. MIC values for penicillin, ampicillin, vancomycin and teicoplanin against enterococci were determined by the agar dilution technique. MIC determinations were further carried out by the E-test method, for the glycopeptide antibiotics. Prevalence of high level resistance to aminoglycoside antibiotics was determined by the agar dilution method. RESULTS: The most common species isolated was E. faecalis (87.07%), followed by E. faecium (10.88%) and E. durans (2.05%). The magnitude of resistance to both penicillin and ampicillin among the Enterococcus spp. was 23.13%, and that to ciprofloxacin was 55.78%. The three antibiotics: nitrofurantoin, co-amoxi-clavulanate, and ampi-sulbactum, showed resistance of 0.78%, 8.16% and 2.72%, respectively. High-level aminoglycoside resistance among strains of enterococci for streptomycin and gentamicin was found to be 33.84% and 36.92%, respectively. All the strains were susceptible to the glycopeptide antibiotics tested. CONCLUSION: The emergence of enterococci with alarming rates of resistance concomitantly to penicillins and aminoglycosides highlights the need for a more rational and restricted use of antimicrobials, in order to minimize the selection and spread of such strains. An early detection of these problem pathogens is also important for preventing any treatment failures.

Anti-Bacterial Agents↗

Isolation of Enterococcus species from infectious skin lesions.

Enterococcus faecalis was isolated at a frequency of 70% and was chiefly isolated from secondary infections due to ulcer/decubitus. Seven sole/predominant E. faecalis were isolated. Penicillins were more effective against E. faecalis and the sensitivities of E. faecalis to antimicrobials were higher than those of E. faecium. Some nonpredominant E. faecalis strains were sensitive to erythromycin probably due to less resistant mechanisms. The characterization of Enterococcus spp. is especially important when choosing appropriate antimicrobials for therapy.

Anti-Bacterial Agents↗

Susceptibility profile of Enterococcus faecalis isolated at the Lagos University Teaching Hospital, Nigeria.

Enterococcus faecalis is the most common of the Enterococcus genus causing infection, particularly urinary tract infections, worldwide. It is also a common cause of nosocomial infections and resistance to various antibiotics is on the increase worldwide. Thirty-five strains of E. Faecalis isolated from various clinical specimens (blood, wound swabs endocervical swabs but mostly urine) were screened for high-level aminoglycoside resistance. Their susceptibility nine antibiotics (ampicillin, gentamicin, streptomycin, vancomycin, tetracycline cotrimoxazole and chloramphenicol, ciprofloxacin and erthromycin) was also determined. All isolates were susceptible to Ampicillin and Vancomycin with MIC90 of 4microg/ml but resistant to Nalidixic acid with an MIC90>256microg/ml. Four (11%) of the isolates showed high-level resistance to Gentamicin while 11(32%) exhibited high-level resistance streptomycin after 24 hours incubation. It will appear that Ampicillin in combination with gentamicin but not streptomycin, can still be used empirically for the treatment of Enterococcal infections.

Anti-Bacterial Agents↗

[Evaluation of usefulness of the PYR-Wellcome test for identifying microorganisms from the genus enterococcus].

The study was aimed at evaluation of usefulness of PYR-Wellcome test in bacteriological diagnostic. Results of identification of enterococcal strains by application of the above test were performed on 157 strains of Enterococcus spp., 15 strains of Streptococcus pyogenes and 12 of Streptococcus bovis. Application of test for PYR-ase production results in shortening of diagnostic procedure. Basing on morphological features, including type of hemolysis, lack of catalase production and presence in the bacterial cell of a peptidase hydrolyzing L-pirrolidonyl-beta-naphthylamide, it is possible to classify with high probability the investigated microorganism to the Enterococcus spp. The Shortened procedure is satisfactory for routine identification of enterococcal strains in diagnostic laboratories.

Enterococcus↗

Serum cholesterol levels in axenic mice colonized with Enterococcus faecium and Lactobacillus acidophilus.

Hypocholesterolemic effect was shown in axenic, mono, bicolonized and conventional mice: the effect was different depending on probiotic properties of intestinal microorganisms. Contamination by Enterococcus faecium CX determined the highest effect: haematic cholesterol level decrease was 16.9% in females and 7.8% in males. In mice contaminated by Lactobacillus acidophilus N5 the decrease of haematic cholesterol levels was less and not relevant in mice contaminated by conventional microflora. Enterococcus faecium CX and Lactobacillus acidophilus N5 strains were able to grow in presence of bile salts, to colonize intestinal tract, to survive at gastric conditions and to assimilate cholesterol (E. faecium more than L. acidophilus). The authors consider the possibility to associate probiotic strains with these characteristics for the health of consumers.

Animal Feed↗

Treatment of multifocal vancomycin-resistant Enterococcus faecium osteomyelitis in sickle cell disease: a preliminary report.

Repeat episodes of musculoskeletal infarction coupled with immunosuppression predispose sickle cell patients to infectious complications throughout their lives. Osteomyelitis is a familiar complication of sickle cell disease, and it may result in significant morbidity, especially when occurring in multiple sites. Staphylococcus and Salmonella remain the most common causes of osteomyelitis in sickle cell patients. Vancomycin-resistant enterococcus (VRE) infections have been reported mainly in connection with bacteremias and infections outside of the musculoskeletal system. To our knowledge, only a few cases of VRE long bone osteomyelitis have been reported in the literature. A few antimicrobial agents are available to treat VRE infections. The occurrence of VRE osteomyelitis is a major clinical concern, especially in an immunocompromised host, such as a sickle cell patient. We present a case of multiple long bone vancomycin-resistant Enterococcus faecium (mixed organisms) osteomyelitis in a sickle cell patient, and we report on a new method of using quinupristin-dalfopristin as part of the management plan to treat a complicated VRE infection successfully. We discuss the mechanism of action of anti-VRE drugs and the future direction to combat VRE in orthopedic infections.

Anemia, Sickle Cell↗

A case of endogenous Enterococcus faecalis endophthalmitis.

A case of successful treatment by vitrectomy and lensectomy of early-stage endogenous endophthalmitis caused by Enterococcus faecalis was reported. The case was a 50-year-old man with poorly controlled diabetes. Following T-tube drainage for a necrotic cholecystitis operation, he developed iridocyclitis in both eyes as well as fever. At the time of his first visit to our clinic, his right eye had already lost light perception. His left eye had visual acuity recognizing hand movement, marked uveitis, complicated cataract, and dense vitreous opacity. As gram-positive cocci were isolated from the aspirated vitreous, lensectomy and vitrectomy under irrigation with antibiotics were conducted. After the administration of systemic postoperative antibiotics and human immunoglobulin, the patient showed remarkable improvement in the state of his ocular fundus. By 60 days after the surgery, the visual acuity of his left eye recovered to 0.2. Endogenous Enterococcus faecalis endophthalmitis has rarely been reported in Japan. The diagnosis and treatment of this disease with reference to the above findings were discussed.

Cataract Extraction↗