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PubMed2000

Simple method to identify bacteriocin induction peptides and to auto-induce bacteriocin production at low cell density.

The production of some bacteriocins by lactic acid bacteria is regulated by induction peptides (IPs) that are secreted by a dedicated secretion system. The IP gene cbaX, for carnobacteriocin A production by Carnobacterium piscicola LV17A, and a presumptive IP gene (orf6), associated with the genetic locus for enterocin B production in Enterococcus faecium BFE 900, were fused to the signal peptide of the bacteriocin divergicin A from Carnobacterium divergens LV13 to access the general secretory pathway. The culture supernatants of C. piscicola UAL26 and Lactococcus lactis MG1363 containing either of these constructs were used to induce bacteriocin production by Bac(-) cultures of C. piscicola LV17A or E. faecium CTC492. The cbaX fusion product induced bacteriocin production by Bac(-) C. piscicola LV17A, but the orf6 fusion product did not induce bacteriocin production by E. faecium CTC492. This represents a relatively simple method of confirming the role of presumptive IPs. The transformation of C. piscicola LV17A with the CbaX gene under expression of the P32 promoter from L. lactis resulted in constitutive production of bacteriocin by either the dedicated transport apparatus or the general secretory pathway.

Amino Acid Sequence↗

PubMed2007

[Septic shock due to vancomycin-resistant enterococci infection. Tigecycline monotherapy].

Adequate antimicrobial therapy is of crucial importance for the survival of critically ill patients with severe nosocomial infections. Tigecycline is an important therapeutic option for the treatment of infections caused by multi-resistant Gram-positive and Gram-negative bacteria including vancomycin-resistant enterococci (VRE). A large randomised study (patients with APACHE-II-score >30 excluded/mean APACHE-II-score 6) demonstrated that tigecycline is not inferior to imipenem/cilastatin for treatment of complicated intra-abdominal infections. However, no case has been reported with microbiological eradication and clinical cure in a patient with septic shock due to peritonitis caused by VRE and treatment with tigecycline monotherapy. Clinical details of a patient suffering from postoperative peritonitis are presented. The patient developed severe septic shock after pancreatic surgery (multiple organ failure, APACHE-II-score 34). As the site of anastomotic leakage was very small and could not be exactly identified, irrigation-suction drains were placed followed by closed postoperative continuous lavage. The pathogen responsible was identified as a vancomycin-resistant Enterococcus faecium, therefore monotherapy with tigecycline was started which resulted in microbiological response and clinical cure. Tigecycline is a new therapeutic option for the treatment of intra-abdominal infections and from an economic point of view financially rewarding when used as monotherapy.

APACHE↗

PubMed1995

Stool carriage, clinical isolation, and mortality during an outbreak of vancomycin-resistant enterococci in hospitalized medical and/or surgical patients.

During a nosocomial outbreak of infection due to vancomycin-resistant enterococci (VRE), rectal swabs that were collected weekly were used to identify and isolate VRE carriers. Over 6 months, 1,458 stool specimens from 724 high-risk patients were cultured, and 187 VRE isolates were recovered from 61 patients; 96% of the isolates were Enterococcus faecium. VRE tended to be isolated from clinical specimens from patients identified as VRE carriers by stool surveillance (P < .01). However, isolation of VRE from surveillance cultures preceded clinical isolation for only approximately 50% of the patients from whom a clinical VRE isolate was recovered. Mortality was greater (P < .05) among patients from whom a clinical VRE isolate was recovered than among patients from whom VRE was isolated only by stool surveillance. The mortality (1[17%] of 6) among patients for whom VRE was isolated from blood was similar to that (10 [27%] of 37) among patients for whom vancomycin-susceptible enterococcus was isolated from blood (P = .97). Despite prompt initiation of contact precautions for VRE carriers, the incidence of fecal carriage of VRE remained approximately 8% among this patient population for the 6-month period of the study.

Adult↗

PubMed1992

Evidence for in vivo incorporation of D-lactate into peptidoglycan precursors of vancomycin-resistant enterococci.

The VanA ligase encoded by the vancomycin resistance plasmid pIP816 of Enterococcus faecium BM4147 condenses D-alanine with various D-2-hydroxy and D-2-amino acids in vitro. D-Lactate added to the culture medium restored the vancomycin resistance of a strain that does not produce the VanH dehydrogenase and therefore appears to be a substrate of VanA in vivo.

Drug Resistance, Microbial↗

PubMed1992

[Cell growth inhibitory compound from the culture of intestinal lactic acid bacteria].

The influence of the culture supernatants of 1000 strains of intestinal lactic acid bacteria on the growth of HeLa cells was examined. The compound from Enterococcus faecium 3463 showing the strongest cell growth inhibiting effect in these strains was isolated and identified as tyramine. The effect of tyramine was activated by the newborn bovine sera (NBS), but weakly by the fetal bovine sera (FBS). Benzylamine and beta-phenethylamine also showed the NBS-dependent cell growth inhibitory effect against HeLa cells.

Benzylamines↗

PubMed1993

Chromosomally mediated high-level gentamicin resistance in Streptococcus mitis.

Four blood culture isolates of Streptococcus mitis were found to be resistant to penicillin (MIC, 16 to 32 micrograms/ml) and gentamicin (MIC, 128 or 1,000 micrograms/ml), and the two antibiotics demonstrated a lack of in vitro synergy. As shown by polymerase chain reaction assays, the structural gene known to encode high-level gentamicin resistance in Enterococcus faecalis, Enterococcus faecium, and Streptococcus agalactiae was also present in all four S. mitis strains. Attempts to isolate plasmids were unsuccessful, but an oligonucleotide probe derived from the gentamicin resistance gene hybridized to distinct restriction fragments of genomic DNA, suggesting that the resistance genes in these strains are integrated into the bacterial chromosome.

Chromosomes, Bacterial↗

PubMed1995

In vitro fructooligosaccharide utilization and inhibition of Salmonella spp. by selected bacteria.

In vitro experiments were conducted to determine: 1) inhibitory capacities of potential direct-fed microbial bacteria against Salmonella serotypes; and 2) the ability of Bifidobacterium bifidum, Enterococcus faecium, Lactobacillus casei, Lactococcus lactis, Pediococcus sp., and Salmonella spp. to grow in media containing fructooligosaccharides (FOS-50 or FOS pure formulation) as the only carbohydrate source. Thirteen bacteria (two strains of Bacillus coagulans, Bacillus licheniformis, Bacillus subtilis, B. bifidum, E. faecium, two strains of Lactobacillus acidophilus, L. casei, Pediococcus sp., Propionibacterium acidopropionici, P. jensenii, and Propionibacterium sp.) were tested for inhibition of six Salmonella serotypes (S. california, S. enteritidis, S. heidelberg, S. mission, S. senftenberg, and S. typhimurium) using a spot-the-lawn technique. Bifidobacterium bifidum, E. faecium, all lactobacilli, and Pediococcus sp. clearly inhibited growth of all Salmonella serotypes. In the growth experiments, E. faecium, L. lactis, and Pediococcus sp. grew in media with either FOS-50 or the pure formulation of FOS as the sole carbohydrate source. All tested Salmonella serotypes utilized FOS-50 for growth; however growth varied among the serotypes. In contrast, none of the Salmonella serotypes grew in media containing the pure formulation of FOS as the only carbohydrate source.

Animals↗

PubMed1997

[Transient hyperphosphatasemia observed in a boy with acute lymphoblastic leukemia].

A detailed time course of alkaline phosphatase (ALP; EC3.1.3.1) activity of transient hyperphosphatasemia (TH) in a 9-year-old boy with acute lymphoblastic leukemia (ALL) is described. The patient's serum ALP activity rose transiently to 49 times the upper limit of normal adult, without any evidences of hepatic and bone disease. The half-life of ALP activity was calculated about 10 days. We characterized ALP isoenzymes by usual electrophoresis using cellulose acetate membrane (Titan III iso-vis) and polyacrylamide disc gel (AlkPhor), and isoelectric focusing using polyacrylamide slab gel. The former two methods showed typical two bands (fast-alpha 2 and alpha 2 beta bands) and the latter one method revealed more basic bands of liver and bone, suggesting the extensive sialylation. The patient complained fever and diarrhea. Enterococcus faecium was detected from his stool. Etiologically, two more patients in the same ward showed TH in the same period. It suggested TH would be occurred by infectious states. Awareness of such benign forms of hyperphosphatasemia not related to malignancy will aid the physician in the differential diagnosis of elevated ALP activity.

Alkaline Phosphatase↗

PubMed1999

Characterization of a bacteriocin-like substance produced by a vaginal Lactobacillus salivarius strain.

A novel bacteriocin-like substance produced by vaginal Lactobacillus salivarius subsp. salivarius CRL 1328 with activity against Enterococcus faecalis, Enterococcus faecium, and Neisseria gonorrhoeae was characterized. The highest level of production of this heat-resistant peptide or protein occurred during the late exponential phase. Its mode of action was shown to be bactericidal. L. salivarius subsp. salivarius CRL 1328 could be used for the design of a probiotic to prevent urogenital infections.

Bacteriocins↗

PubMed2006

VanB-vanA incongruent VRE isolated from animals and humans in 1999.

16 chicken isolates and four clinical isolates of VanB-vanA incongruent vancomycinresistant Enterococcus faecium strains without vanS were isolated in 1999. Pulsed-field gel electrophoresis revealed only a peripheral relationship between the chicken isolates and clinical isolates, but suggested clonal spread in the chicken isolates.

Animals↗

PubMed2003

Risk factors for new detection of vancomycin-resistant enterococci in acute-care hospitals that employ strict infection control procedures.

Accurate assessment of the risk factors for colonization with vancomycin-resistant enterococci (VRE) among high-risk patients is often confounded by nosocomial VRE transmission. We undertook a 15-month prospective cohort study of adults admitted to high-risk units (hematology, renal, transplant, and intensive care) in three teaching hospitals that used identical strict infection control and isolation procedures for VRE to minimize nosocomial spread. Rectal swab specimens for culture were regularly obtained, and the results were compared with patient demographic factors and antibiotic exposure data. Compliance with screening was defined as "optimal" (100% compliance) or "acceptable" (minor protocol violations were allowed, but a negative rectal swab specimen culture was required within 1 week of becoming colonized with VRE). Colonization with VRE was detected in 1.56% (66 of 4,215) of admissions (0.45% at admission and 0.83% after admission; the acquisition time was uncertain for 0.28%), representing 1.91% of patients. No patients developed infection with VRE. The subsequent rate of new acquisition of VRE was 1.4/1,000 patient days. Renal units had the highest rate (3.23/1,000 patient days; 95% confidence interval [CI], 1.54 to 6.77/1,000 patient days). vanB Enterococcus faecium was the most common species (71%), but other species included vanB Enterococcus faecalis (21%), vanA E. faecium (6%), and vanA E. faecalis (2%). The majority of isolates were nonclonal by pulsed-field gel electrophoresis analysis. Multivariate analysis of risk factors in patients with an acceptable screening suggested that being managed by a renal unit (hazard ratio [HR] compared to the results for patients managed in an intensive care unit, 4.6; 95% CI, 1.2 to 17.0 [P = 0.02]) and recent administration of either ticarcillin-clavulanic acid (HR, 3.6; 95% CI, 1.1 to 11.6 [P = 0.03]) or carbapenems (HR, 2.8; 95% CI, 1.0, 8.0 [P = 0.05]), but not vancomycin or broad-spectrum cephalosporins, were associated with acquisition of VRE. The relatively low rates of colonization with VRE, the polyclonal nature of most isolates, and the possible association with the use of broad-spectrum antibiotics are consistent with either the endogenous emergence of VRE or the amplification of previously undetectable colonization with VRE among high-risk patients managed under conditions in which the risk of nosocomial acquisition was minimized.

Adult↗

PubMed2004

In vitro antienterococcal activity explains associations between exposures to antimicrobial agents and risk of colonization by multiresistant enterococci.

We compared ceftriaxone and piperacillin-tazobactam at doses ranging from 0.1 to 2 times the human equivalent daily dose (HEDD), to determine their impact on gastrointestinal colonization by ampicillin- and vancomycin-resistant Enterococcus faecium C68 in a mouse model. Ceftriaxone failed to promote colonization at doses up to 0.25 times the HEDD, whereas piperacillin-tazobactam promoted colonization at doses up to 0.5 times the HEDD. Ceftriaxone promoted colonization at doses at least 0.5 times the HEDD, whereas piperacillin-tazobactam inhibited colonization at doses at least 0.75 times the HEDD. Both piperacillin-tazobactam and ceftriaxone inhibited colonization by an enterococcal strain devoid of low-affinity penicillin-binding protein-5 (significantly increasing its susceptibility to these agents), at doses that promoted colonization by E. faecium C68. These results support a model in which the impact that different beta -lactam agents have on colonization by VRE is related to the level of the beta -lactam agent's intrinsic antienterococcal activity against the colonizing strain.

Animals↗

PubMed1995

Controlling vancomycin-resistant enterococci.

After controlling an epidemic of vanB-type vancomycin-resistant Enterococcus faecium (VRE), we contained a subsequent vanA E faecium outbreak by using prospective laboratory-based surveillance, placing patients with VRE in private rooms, requiring the use of both gowns and gloves by all personnel entering the patients' rooms, and conducting prevalence surveys of patients on affected wards.

Anti-Bacterial Agents↗

PubMed1999

Detection of clinical vancomycin-resistant enterococci in Denmark by multiplex PCR and sandwich hybridization.

Since the first cases of human infection with vancomycin-resistant enterococci (VRE) were reported in the late eighties, there has been a dramatic increase in VRE all over the world. So far, there have not been any reports of clinical VRE in Denmark. In this study we have investigated 131 clinically important enterococci sent to Statens Serum Institut from all over Denmark during the period July 1995 to May 1997. The susceptibility to vancomycin, teicoplanin, ampicillin and gentamicin was tested by the agar dilution method. In addition, two methods were developed to detect the different genotypes of glycopeptide resistance described in enterococci: a multiplex PCR assay for detection of vanA, vanB, vanC-1, vanC-2/3 ligase genes including 16S rRNA gene control primers and a sandwich hybridization assay to confirm vanA and vanB PCR-positive strains. The highest frequency of resistance to the tested antibiotics was found in the Enterococcus faecium group. Four strains were found with acquired resistance to glycopeptides: one E. faecium and one E. gallinarum were vanA positive, and two E. faecium isolates were vanB positive. These strains were isolated from different hospitals in different periods of time, and all patients recovered from their infections with VRE. Today, the PCR and sandwich hybridization methods are used for screening of vancomycin-resistant enterococci in humans as part of the Danish surveillance programme.

Ampicillin↗

PubMed1992

[Studies of bacterial indicators for water pollution--growth of Escherichia coli and enterococci in limited nutrient conditions].

Bacteria, that would be an ideal indicator for fecal contamination in environmental water, should not proliferate but at the same time should survive a little longer than enteropathogenic bacteria in the environmental water. While conventional methods have recommended Escherichia coli as an indicator, enterococci is recommended as a preferable indicator for the estimation of fecal contamination based on the following observation. The relationship between the specific growth rate of organism and the concentration of peptone was determined and the value of Ks, a parameter analogous to Michaelis-Menten constant, was estimated. Ks of Escherichia coli, Enterococcus faecalis and Enterococcus faecium were 3.8, 11.1 and 12.5 mg/ml respectively, indicating that Escherichia coli will grow at lower concentration of peptone than enterococci. Escherichia coli also grew in human fecal suspension and in filtrated (0.22 microns) river water, and while in those conditions enterococci did not grow or survive longer than Escherichia coli, survival time was longer than for Salmonella typhi and Shigella sonnei. It appears that enterococci reflect more accurately actual fecal contamination.

Enterococcus faecalis↗

PubMed1998

Epidemiology and control of vancomycin-resistant enterococci (VRE) in a renal unit.

This study reports an outbreak of infection and colonization caused by vancomycin-resistant enterococci (VRE) in the renal service of a large teaching hospital. The polymerase chain reaction and pulsed-field gel electrophoresis were used to study the epidemiology of 26/34 strains of vancomycin-resistant Enterococcus faecalis and Enterococcus faecium from the outbreak in comparison with five strains from other hospitals in Edinburgh and the Borders, and three from other wards in the Royal Infirmary. The study revealed a heterogeneous population of vancomycin-resistant E. faecalis. Over 60% of E. faecium isolates had matching pulsed-field gel electrophoresis patterns and all of these were of VanA phenotype. These results suggest that clonal spread of VanA phenotype E. faecium within and possibly between hospitals is the major vancomycin-resistant enterococcal problem in Edinburgh. Screening of patients and isolation of colonized and infected patients appear to have been successful in controlling the spread of VRE.

Anti-Bacterial Agents↗

PubMed2003

Quinupristin-dalfopristin resistance in gram-positive bacteria: mechanism of resistance and epidemiology.

Antimicrobial resistance in gram-positive bacteria is a continuing problem resulting in significant morbidity, mortality, and cost. Because of this resistance, new antimicrobial agents have been needed. Quinupristin-dalfopristin is a recently approved agent for treatment of these infections. Shortly after its introduction into clinical medicine, resistance was reported. Resistance can occur by one or more of several mechanisms, including enzymatic modification, active transport of efflux mediated by an adenosine triphosphate-binding protein, and alteration of the target site. Resistance is rare in isolates of staphylococci and Enterococcus faecium from humans. Resistance is common in isolates recovered from food animals and is related to the use of virginiamicin as a feed additive. Considering the effect antimicrobial resistance has on human health, as well as its economic impact, measures to preserve the usefulness of these agents and delay the development of resistance are urgently needed.

Anti-Bacterial Agents↗

PubMed1999

Antimicrobial susceptibility patterns of enterococci causing infections in Europe. The European VRE Study Group.

In vitro susceptibilities of 4,208 enterococci (83% Enterococcus faecalis isolates, 13.6% Enterococcus faecium isolates, and 3.4% isolates of other species) from patients in 27 European countries towards 16 antibiotics were determined. High-level resistance to gentamicin varied by country (range, 1 to 49%; mean, 22.6% +/- 12. 3%) and per species (19.7% E. faecalis isolates, 13.6% E. faecium isolates, 3.4% by other species). Vancomycin resistance was detected in 0.06% E. faecalis, 3.8% E. faecium, and 19.1% isolates of other species. All enterococci were susceptible to LY 333328 and everninomicin, and 25% of E. faecalis isolates and 85% of other enterococci were susceptible to quinupristin-dalfopristin. The MIC of moxifloxacin and trovafloxacin for ciprofloxacin-susceptible E. faecalis at which 90% of the isolates were inhibited was 0.25 to 0.5 microg/ml.

Anti-Bacterial Agents↗