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PubMed2002

Monitoring transfer of recombinant and nonrecombinant plasmids between Lactococcus lactis strains and members of the human gastrointestinal microbiota in vivo--impact of donor cell number and diet.

AIMS: The generation of data of real relevance to the purported risks of DNA transfer from food-borne genetically modified microorganisms (GMMOs) using the human biota associated (HBA) rat model. Plasmid transfer between Lactococcus lactis strains and between donor strains and human gut bacteria was monitored. METHODS AND RESULTS: Transfer of the recombinant plasmid pCK1 and/or the promiscuous nonrecombinant plasmid pAMbeta1 between L. lactis strains was monitored in vivo in HBA rats. No transfer of pCK1 was observed. Transfer of pAMbeta1 was observed to Enterococcus spp. present in the HBA rats. Transconjugants persisted for 30 d and were distributed throughout the gastrointestinal tract. Both HBA rat diet and donor cell numbers impacted on transconjugant numbers. Fewer transconjugants were observed in animals fed a high-fat human type diet, while high levels of plasmid transfer were only observed at doses of donor L. lactis greater than 109 cfu. CONCLUSIONS: The utility of models of the human gut in monitoring DNA transfer events within the gut microbiota was demonstrated. SIGNIFICANCE AND IMPACT OF THE STUDY: Such findings give some confidence for the use of GMMOs with recombinant DNA borne on nonconjugative elements in fermented foods. HBA rats are a suitable model for monitoring the fate of food-borne GMMOs.

Animals↗
PubMed1999

Antimicrobial susceptibility of clinical isolates of enterococci.

To describe antimicrobial resistance patterns of Enterococcus spp., we measured minimal inhibitory concentrations (MICs) of five antimicrobial agents for 174 clinical isolates of enterococci collected in Taichung Veterans General Hospital from November, 1996 to January, 1998. Major sources included blood (48), the genitourinary tract (39), soft tissue and wounds (28), and the gynecological tract (16). The sensitivity test of vancomycin was performed using the broth microdilution method. The susceptibility of ampicillin, penicillin, gentamicin, and teicoplanin was tested by the agar dilution method. The MIC90 of penicillin, ampicillin, gentamicin, teicoplanin and vancomycin were 2, 1, > 2,048, 0.125 and 2 microg/mL, respectively. Ampicillin-resistant isolates represented only 4.6% of all enterococci tested. These results reveal that vancomycin-resistant enterococci were found in 1.7% of all enterococci isolates in this study. However, all isolates were sensitive to teicoplanin.

Adult↗
PubMed2003

The potential for disinfection of separated faecal matter by urea and by peracetic acid for hygienic nutrient recycling.

No efficient, reliable, and scale independent disinfection methods for toilet waste are available today for safe recycling of plant nutrients. Therefore, two chemical treatment methods, addition of urea or of PAA (a quaternary mixture of 15% peracetic acid, 15% hydrogen peroxide and 30% acetic acid), were evaluated for disinfection of faecal matter.Degradation of the added urea resulted in 30 g of ammonia nitrogen per kilogram of treated matter and a pH increase to approximately 9.3. This produced an efficient disinfection of E. coli, Enterococcus spp., and Salmonella spp. within 3 weeks (>6log(10) reduction) and a reduction of the chemical resistant Salmonella typhimurium 28b phage, corresponding to a decimal reduction within 7.5 days. No viable Ascaris suum eggs were found after 50 days of treatment. No reduction of spore forming Clostridia spp. was observed. Urea treatment proved to be efficient for disinfection of source separated faecal matter in a scale independent method used for safe recycling of nutrients found in the faecal matter.PAA reduced all of the above indicator organisms within 12 h after application. For this faecal material, with a dry matter content of approximately 10%, an addition of 0.5-1% of PAA (active substance, corresponding to 3.3-6.7% of the Proxitane 15 used) was required before no viable organisms were found in the material. However, this was not tested for the A. suum. No viable spore-forming bacteria or phages were detected. A high rate of bacteria regrowth occurred at 0.15% dosage and 5 days of treatment. PAA is an efficient alternative for disinfection of separated faeces if a rapid treatment is needed.

Animals↗
PubMed2007

Partial characterization of bacteriocins produced by human Lactococcus lactis and Pediococccus acidilactici isolates.

AIMS: The aim of this study was to isolate bacteriocin-producing lactic acid bacteria (LAB) from human intestine. METHODS AND RESULTS: A total of 111 LAB were isolated from human adult stool and screened for their bacteriocin production. Neutralized cell-free supernatants from Lactococcus lactis subsp. lactis MM19 and Pediococcus acidilactici MM33 showed antimicrobial activity. The antimicrobials in the supernatant from a culture of L. lactis inhibited Enterococcus faecium, various species of Lactobacillus and Staphylococcus aureus; while those in the supernatant from a culture of P. acidilactici inhibited Enterococcus spp., some lactobacilli and various serotypes of Listeria monocytogenes. The antimicrobial metabolites were heat-stable and were active over a pH range of 2-10. The antimicrobial activities of the supernatants of both bacteria were inhibited by many proteases but not by catalase. The plate overlay assay allowed an approximation of size between 3.5 and 6 kDa for both antimicrobial substances. CONCLUSIONS: As the antagonistic factor(s) produced by L. lactis MM19 and P. acidilactici MM33 were sensitive to proteolytic enzymes, it could be hypothesized that bacteriocins were involved in the inhibitory activities. Inhibition spectrum and biochemical analysis showed that these bacteria produced two distinct bacteriocins. SIGNIFICANCE AND IMPACT OF THE STUDY: We are the first to isolate bacteriocin-producing strains of Pediococcus and Lactococcus from human intestine. These strains might be useful for control of enteric pathogens.

Anti-Bacterial Agents↗
PubMed2004

Extended-interval aminoglycoside dosing for treatment of enterococcal and staphylococcal osteomyelitis.

BACKGROUND: Gram-positive osteomyelitis requires long-term antibiotic therapy, much of which is often administered in the outpatient setting. Historically, synergistic aminoglycoside use in these infections requires multiple daily doses, which can be inconvenient. Data regarding extended-interval aminoglycoside dosing (EIAD), also known as once-daily dosing, in this setting are lacking. OBJECTIVE: To evaluate the safety and efficacy of EIAD in the treatment of gram-positive osteomyelitis. METHODS: Retrospective chart review of adult patients treated for documented, gram-positive osteomyelitis with EIAD at the University of New Mexico Home IV Antibiotic Clinic was conducted. The patients' medical records were reviewed by an infectious diseases clinical pharmacist. Clinical and microbiologic outcomes and the incidence of nephrotoxicity and ototoxicity were the main outcome measures. RESULTS: Fifteen patients (16 events) were included. Enterococcus spp. was the most common organism isolated. Nine patients had infected equipment or devices; 6 of these had removal of these devices in conjunction with antibiotic therapy. The median duration of antibiotic therapy was 6 weeks (range 6-31). The median duration of aminoglycoside therapy was 28 days (range 6-43). Seven patients developed nephrotoxicity, 5 of whom received an aminoglycoside in combination with vancomycin. Male patients had a higher risk of developing nephrotoxicity compared with females (p = 0.04). The mean +/- SD duration of EIAD before the development of nephrotoxicity was 34 +/- 8 days. Clinical cure was achieved in 12 (75%) patients. Three patients achieved clinical cure without hardware removal. CONCLUSIONS: Most of the patients with gram-positive osteomyelitis were successfully managed with EIAD. However, nephrotoxicity developed in a high proportion of patients and was likely related to prolonged aminoglycoside use.

Adult↗
PubMed1999

Vancomycin resistance: status quo and quo vadis.

The prevalence of vancomycin resistance is steadily rising among clinical isolates of Enterococcus spp., thereby limiting the treatment options for infections caused by vancomycin-resistant enterococci. The precise nature of the glycopeptide resistance genes has been elucidated, and many studies on gene reservoirs and strain-versus-resistance-gene epidemiology have been performed. The prevalence of vancomycin-resistant enterococci in various clinical and environmental settings in relation to nosocomial and veterinary applications of antimicrobial glycopeptides is discussed in detail in this review. Novel molecular tools for the identification of vancomycin-resistant enterococci genomes or the various resistance genes have been applied in order to expand current insight into the overall epidemiology of the resistance trait itself. The risk of the spread of vancomycin resistance to other bacterial species was recently underscored by the emergence of staphylococci showing clinical resistance to vancomycin. The topics mentioned above are elaborated on and discussed in light of the increasing medical concern on the future detection of microbial infections beyond chemotherapeutic cure.

Animal Husbandry↗
PubMed2002

[Antimicrobial susceptibility and prevalence of beta-lactamase producing clinical isolates in southern Kyushu. The results of collaborative study from 1999 to 2000].

The positivity of beta-lactamase and antimicrobial susceptibility were determined in a total of 1,358 clinical isolates at 15 hospitals and clinics in four prefectures in southern Kyushu (Okinawa, Miyazaki, Kagoshima and Kumamoto) during the period from December 1999 to February 2000. The isolates collected comprised of 176 strains of S. aureus, 203 of H. influenzae, 102 of M. catarrhalis, 206 of E. coli, 153 of K. pneumoniae, 99 of E. cloacae, 95 of S. marcescens, 201 of P. aeruginosa, 79 of E. faecalis, and 44 of E. faecium. The frequency of CPDX resistance among E. coli in particular varied geographically, and was found to be higher in Kumamoto and Kagoshima. The strains of K. pneumoniae and E. cloacae resistant to common antimicrobial agents were particularly found in Kagoshima, and one strain of IPM-resistant E. cloacae was isolated in Miyazaki. Also, the geographical difference in the frequency of LVFX resistance among the isolates of E. cloacae was noted, the results indicating the higher prevalence in Okinawa and Kagoshima. Resistant isolates of P. aeruginosa were less common in Kagoshima, and four isolates of P. aeruginosa from Miyazaki were found to be resistant to CAZ and IPM. None of the isolates of S. aureus and Enterococcus spp. was resistant to VCM or TEIC at all. The isolates of E. faecalis resistant at high-level GM (500 micrograms/ml) and SM (1,000 micrograms/ml) were found in 27.8% and 22.8%, and those of E. faecium were 6.8% and 38.6%, respectively. Overall, the ratio of MRSA among S. aureus was 67.6%, and three isolates were resistant to ABK with no less than 8 micrograms/ml of MIC. The frequency of BLNAR (beta-lactamase-negative, ampicillin resistant) among H. influenzae isolated in Okinawa was markedly higher (isolation ratio, 37.9%) when compared with other prefectures, and the isolates of BLPACR (beta-lactamase-positive, AMPC/CVA resistant) were found only in Okinawa with a ratio of 41.6%. A total of 18 strains of ESBL defined by the NCCLS criteria (M100-S11) were isolated, eight strains of K. pneumoniae and 10 strains of E. coli. Of 18 isolates of ESBL, 13 were from Kagoshima and the remaining five were from Kumamoto.

Bacteria↗
PubMed1992

Analysis of peptidoglycan precursors in vancomycin-resistant enterococci.

Analysis by high-pressure liquid chromatography of the cytoplasmic peptidoglycan precursors of a high- and a low-level vancomycin-resistant Enterococcus spp. was performed before and after induction of resistance. This analysis showed a decrease of the D-Ala-D-Ala and UDP-MurNac-pentapeptide pools, an increase of the UDP-MurNac-tripeptide pool, and the appearance of new UDP-MurNac-containing material. These results lead us to suggest that the vancomycin-induced carboxypeptidase activity cleaves the D-Ala-D-Ala (L. Gutmann, D. Billot-Klein, S. Al-Obeid, I. Klare, S. Francoul, E. Collatz, and J. van Heijenoort, Antimicrob. Agents Chemother. 36:77-80, 1992), which in turn would prevent formation of the normal UDP-MurNac-pentapeptide and thereby of the vancomycin target. The novel UDP-MurNac-containing material is thought to correspond to peptidoglycan precursors which might be synthesized by an alternate pathway (T. D. H. Bugg, G. D. Wright, S. Dutka-Malen, M. Arthur, P. Courvalin, and C. T. Walsh, Biochemistry 30:10408-10415, 1991) and which would be unable to bind vancomycin in glycopeptide-resistant enterococci.

Cells, Cultured↗
PubMed2004

Comparison of phenotypic and genotypic taxonomic methods for the identification of dairy enterococci.

The purpose of the study was to assess the phenotypic and genotypic taxonomic congruence in order to allow species allocation of dairy enterococci. A total of 364 enterococci isolated from ewes'milk and cheese from four Portuguese Registered Designation of Origin areas and 25 type and reference strains of Enterococcus spp. were characterized by a polyphasic taxonomical approach involving 40 physiological and biochemical tests, whole-cell protein profiles, amplification of 16S-23S intergenic spacer regions (ITS-PCR) and subsequent restriction analysis (ARDRA). Ribotyping was also performed with reference strains and a subset of 146 isolates. Numerical hierarchic data analysis showed that single-technique identification levels increase from the physiological and biochemical tests to the protein approach, being lower with ITS/ARDRA and ribotyping. Cross-analysis confirmed a higher unmatching level in all pairwise combinations involving physiological and biochemical data. Whole-cell protein profiles followed by ITS/ARDRA identified 89% of the enterococci. Reliable identification of enterococci from milk and cheese could be obtained by analysis of whole-cell protein profiles. ITS-PCR can be used to confirm E. durans and E. faecium and ARDRA further confirms E. faecalis. Results revealed E. faecalis, E. durans, E. hirae and E. faecium as the prevalent species, although species prevalence showed some degree of variation among the areas.

Animals↗
PubMed1993

Identification and characterization of multiple species of vancomycin-resistant enterococci, including an evaluation of Vitek software version 7.1.

A total of 374 clinical isolates of Enterococcus spp. were characterized to determine the species distribution and vancomycin resistance. The ability of the Vitek system (bioMerieux Inc, Hazelwood, St. Louis, Mo.) to identify enterococci to the species level and to recognize vancomycin resistance by using computer software version 7.1 was evaluated. Conventional methods were used for identification and agar dilution was used for susceptibility testing, the results of which were as follows (presented as number of vancomycin-resistant isolates/number of members of that species identified): 219/234 E. faecium, 9/112 E. faecalis, 2/3 E. mundtii, 0/1 E. durans, 0/1 E. hirae, 0/1 E. raffinosis, and 0/1 E. avium. Ten enterococci were in the vancomycin-intermediate category (six E. gallinarum, two E. casseliflavus, and one each of E. faecium and E. faecalis). The Vitek GPI card correctly identified 98% of E. faecium isolates, 99% of E. faecalis isolates, and only two isolates of other enterococcal species. The GPS-TA card was 98% sensitive and 95% specific for the detection of vancomycin resistance, generating a total of four very major (1.6%) and five minor errors (1.3%).

Drug Resistance, Microbial↗
PubMed2005

Antimicrobial resistance of Enterococcus sp. isolated from the intestinal tract of patients from a university hospital in Brazil.

This study reports the results about antimicrobial resistance of Enterococcus spp. isolated from intestinal tract of patients from a university hospital in Brazil. The identification of strains at species level was performed by conventional biochemical tests, API 20 Strep (bioMérieux), and polymerase chain reaction assay. The species distribution was E. faecium (34%), followed by E. faecalis (33%), E. gallinarum (23.7%), E. casseliflavus (5.2%), E. avium (1%), and E. hirae (1%). Intrinsic resistance to vancomycin characterized by presence of vanC genes was found in E. gallinarum and E. casseliflavus. The high prevalence of VanC phenotype enterococci is very important because these species have been reported as causing a wide variety of infections. Vancomycin-resistant E. faecium or E. faecalis were not found and no one isolate of these species was a beta-lactamase producer. Thirteen clinical isolates of enterococci (13.4%) showed multiresistance patterns, which were defined by resistance to three classes of antibiotics plus resistance to at least one aminoglycoside (gentamicin and/or streptomycin). The resistance to several antimicrobials shown by enterococcal strains obtained in this study is of concern because of the decrease in the therapeutic options for treatment of infections caused by enterococci.

Anti-Bacterial Agents↗
PubMed2006

Antimicrobial susceptibility of enterococci strains isolated from slaughter animals on the data of Hungarian resistance monitoring system from 2001 to 2004.

Isolates of Enterococcus spp. were collected from January 2001 to December 2004 from caecal samples of slaughtered poultry, swine and cattle in Hungary. The isolates were identified by their growth and biochemical properties and with PCR. The antibiotic susceptibility of a total number of 1272 isolates was tested with disk diffusion test to ampicillin, gentamicin, streptomycin, tetracycline, erythromycin and vancomycin. It was established that although ampicillin and amoxicillin are often used in veterinary practice its resistance rate was relatively low. In the case of tetracyclines and macrolides, a high incidence of resistance was found. Susceptibility of strains to tetracyclines and/or macrolides reduced in both 2003 and 2004 in all animal species, which may be due to the more frequent usage of these drugs in the veterinary practice following the ban of growth promoters. The annual data of vancomycin resistance point to an association between the recovery of vancomycin-resistant enterococci (VRE) isolates and the use of avoparcin. This study indicates that reducing antimicrobial resistance in food animals could be possible with lower usage of antibiotics, although variations can occur with different strains.

Abattoirs↗
PubMed2005

[An alarming problem in the therapy of infective endocarditis: the development of antibiotic-resistant strains].

We shall focus on infective endocarditis due to Enterococcus spp and Staphylococcus aureus, both able to develop resistance to antibiotics with different mechanisms. Vancomycin-resistant strains produce some of the most challenging nososocomial infections. Enterococci develop resistance practically to all classes of antibiotics. Vancomycin-resistant strains, in the '90s, passed from 2% to more than 25%. Five types of vancomycin-resistance were reported (from van A to van E), linked to the presence of certain classes of genes regulating the production of abnormal precursors of peptidoglycan which inhibit the action of vancomycin. Staphylococcus aureus is a fearful organism whose infections can reach a mortality rate of 80%. In 1943, as soon as penicillin G was introduced into therapy, Staphylococcus strains producers of beta-lactamase were identified. After beta-lactamase-resistant penicillins were introduced into therapy, methicillin-resistant Staphylococcus strains appeared in the '60s. In 1996 the first strain of methicillin-resistant and vancomycin-resistant Staphylococcus aureus was isolated. In 2001, in Japan, the first case of infective endocarditis due to Staphylococcus aureus resistant to methicillin and non-responsive to vancomycin was described. The resistance is connected to an increased synthesis of the cell wall, which thickens reducing the activity of vancomycin.

Drug Resistance, Bacterial↗
PubMed1995

[Bactericidal activity of combinations of ticarcillin-clavulanic acid or other beta-lactams (amoxicillin, piperacillin-tazobactam, ticarcillin) with gentamicin against Enterococcus faecalis and E. faecium].

MICs of amoxicillin, piperacillin-tazobactam, ticarcillin and ticarcillin-clavulanic acid were determined against 52 clinical strains of E. faecalis and E. faecium. Modal MICs (mg/l) of these antibiotics were 1, 4, 64 and 64, respectively. Bactericidal activity of the beta-lactams was tested at two concentrations (1/2 MIC and 100 mg/l) alone or combined with gentamicin (4 mg/l) against 10 clinical strains of Enterococcus spp. Activity of ticarcillin was not modified by the addition of clavulanic acid (4 mg/l). Time-kill curves showed that all beta-lactams at 100 mg/l were bactericidal after 18 h of incubation (decrease in bacterial counts > or = 3 log10 CFU/ml) against 6 strains of E. faecalis, with MICs of amoxicillin at 1 mg/l and 2 strains of E. faecium with MICs of amoxicillin at 1 and 2 mg/l. Addition of gentamicin resulted in a bactericidal effect at 3 or 6 hours for the great majority of the combinations studied, except against the two strains highly resistant to gentamicin. Combination of every beta-lactam at 1/2 MIC with gentamicin was synergistic (except in case of high level resistance to gentamicin) and prevented bacterial growth but was not bactericidal. Against two strains of E. faecium with high MICs of amoxicillin (8 mg/l), piperacillin-tazobactam (64 mg/l) and ticarcillin-clavulanic acid (512 mg/l), only combinations of amoxicillin 100 mg/l and gentamicin were always bactericidal. In conclusion, against E. faecalis strains, ticarcillin-clavulanic acid tested at 100 micrograms/ml had a bactericidal activity similar to that of the other beta-lactams tested, alone or combined with gentamicin.(ABSTRACT TRUNCATED AT 250 WORDS)

Amoxicillin↗
PubMed2001

Antimicrobial susceptibility patterns of enterococci in intensive care units in Sweden evaluated by different MIC breakpoint systems.

Three hundred and twenty-two (322) clinical isolates were collected from patients admitted to intensive care units (ICUs) at eight Swedish hospitals between December 1996 and December 1998. Of the isolates, 244 (76%) were Enterococcus faecalis, 74 (23%) were Enterococcus faecium and four (1%) were other Enterococcus spp. MICs of ampicillin, imipenem, meropenem, piperacillin/tazobactam, ciprofloxacin, trovafloxacin, clinafloxacin, gentamicin, streptomycin, vancomycin, teicoplanin, quinupristin/dalfopristin, linezolid and evernimicin were determined by Etest. Susceptible and resistant isolates were defined according to the species-related MIC breakpoints of the British Society for Antimicrobial Chemotherapy (BSAC), the National Committee for Clinical Laboratory Standards (NCCLS) and the Swedish Reference Group for Antibiotics (SRGA). Tentative breakpoints were applied for new/experimental antibiotics. Multidrug resistance among enterococci in ICUs is not uncommon in Sweden, particularly among E. faecium, and includes ampicillin resistance and concomitant resistance to fluoroquinolones. Almost 20% of E. faecalis isolates showed high-level resistance to gentamicin and concomitant resistance to fluoroquinolones. Vancomycin-resistant enterococci were only found sporadically. Among the new antimicrobial agents, linezolid and evernimicin showed the best activity against all enterococcal isolates. There was good concordance between the BSAC, NCCLS and SRGA breakpoints in detecting resistance. When applying the SRGA breakpoints for susceptibility, isolates were more frequently interpreted as intermediate. This might indicate earlier detection of emerging resistance using the SRGA breakpoint when the native population is considered susceptible, but with the risk that isolates belonging to the native susceptible population will be incorrectly interpreted as intermediate.

Anti-Bacterial Agents↗
PubMed2000

Characterization and cloning of the genes encoding enterocin 1071A and enterocin 1071B, two antimicrobial peptides produced by Enterococcus faecalis BFE 1071.

The pH-neutral cell supernatant of Enterococcus faecalis BFE 1071, isolated from the feces of minipigs in Göttingen, inhibited the growth of Enterococcus spp. and a few other gram-positive bacteria. Ammonium sulfate precipitation and cation-exchange chromatography of the cell supernatant, followed by mass spectrometry analysis, yielded two bacteriocin-like peptides of similar molecular mass: enterocin 1071A (4.285 kDa) and enterocin 1071B (3.899 kDa). Both peptides are always isolated together. The peptides are heat resistant (100 degrees C, 60 min; 50% of activity remained after 15 min at 121 degrees C), remain active after 30 min of incubation at pH 3 to 12, and are sensitive to treatment with proteolytic enzymes. Curing experiments indicated that the genes encoding enterocins 1071A and 1071B are located on a 50-kbp plasmid (pEF1071). Conjugation of plasmid pEF1071 to E. faecalis strains FA2-2 and OGX1 resulted in the expression of two active peptides with sizes identical to those of enterocins 1071A and 1071B. Sequencing of a DNA insert of 9 to 10 kbp revealed two open reading frames, ent1071A and ent1071B, which coded for 39- and 34-amino-acid peptides, respectively. The deduced amino acid sequence of the mature Ent1071A and Ent1071B peptides showed 64 and 61% homology with the alpha and beta peptides of lactococcin G, respectively. This is the first report of two new antimicrobial peptides representative of a fourth type of E. faecalis bacteriocin.

Amino Acid Sequence↗
PubMed1993

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

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

Anti-Bacterial Agents↗
PubMed2006

Systemic inflammatory response syndrome in nosocomial bloodstream infections with Pseudomonas aeruginosa and Enterococcus Species: comparison of elderly and nonelderly patients.

OBJECTIVES: To determine whether the systemic inflammatory response syndrome (SIRS), clinical course, and outcome of monomicrobial nosocomial bloodstream infection (BSI) due to Pseudomonas aeruginosa or Enterococcus spp. is different in elderly patients than in younger patients. DESIGN: Historical cohort study. SETTING: An 820-bed tertiary care facility. PARTICIPANTS: One hundred twenty-seven adults with P. aeruginosa or enterococcal BSI. MEASUREMENTS: SIRS scores were determined 2 days before the first positive blood culture through 14 days afterwards. Elderly patients (> or =65, n=37) were compared with nonelderly patients (<65, n=90). Variables significant for predicting mortality in univariate analysis were entered into a logistic regression model. RESULTS: No difference in SIRS was detected between the two groups. No significant difference was noted in the incidence of organ failure, 7-day mortality, or overall mortality between the two groups. Univariate analysis revealed that Acute Physiology And Chronic Health Evaluation (APACHE) II score of 15 or greater at BSI onset; adjusted APACHE II score (points for age excluded) of 15 or greater at BSI onset; and respiratory, cardiovascular, renal, hematological, and hepatic failure were predictors of mortality. Age, sex, use of empirical antimicrobial therapy, and infection with imipenem-resistant P. aeruginosa or vancomycin-resistant enterococci did not predict mortality. Multivariate analysis revealed that hematological failure (odds ratio (OR)=8.1, 95% confidence interval (CI)=2.78-23.47), cardiovascular failure (OR=4.7, 95% CI=1.69-13.10), and adjusted APACHE II > or = 15 at BSI onset (OR=3.1, 95% CI=1.12-8.81) independently predicted death. CONCLUSION: Elderly patients did not differ from nonelderly patients with respect to severity of illness before or at the time of BSI. Elderly patients with pseudomonal or enterococcal BSIs did not have a greater mortality than nonelderly patients.

Age Factors↗