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PubMed2002

In vitro activity of a new cephalosporin, RWJ-54428, against streptococci, enterococci and staphylococci, including glycopeptide-intermediate Staphylococcus aureus.

The need for new antimicrobial agents with activity against Gram-positive organisms has become increasingly important because of emerging resistance. We compared the activity of a new b-lactam antimicrobial agent, RWJ-54428 (MC-02 479), with representatives of other classes of antimicrobial agents against 76 Staphylococcus aureus (including four glycopeptide- intermediate strains), 50 coagulase-negative staphylococci, 20 Enterococcus faecalis, 20 Enterococcus faecium, 10 Enterococcus gallinarum/Enterococcus casseliflavus, 54 Streptococcus pneumoniae and 22 viridans streptococcal isolates. The MIC(90) of RWJ-54,428 was < or = 2 mg/L for all groups of bacteria tested except E. faecium. The activity against four strains of glycopeptide-intermediate S. aureus was similar to that for other methicillin-resistant S. aureus isolates (range 0.5-2.0 mg/L).

Acetamides↗

PubMed1995

DNA hybridization and contour-clamped homogeneous electric field electrophoresis for identification of enterococci to the species level.

In this study, 113 Enterococcus faecium, 37 Enterococcus faecalis, 24 Enterococcus gallinarum, 15 Enterococcus raffinosus, and 13 Enterococcus casseliflavus clinical isolates and American Type Culture Collection (ATCC) strains were evaluated by contour-clamped homogeneous electric field electrophoresis. Thirty-one of the E. faecium, 22 of the E. faecalis, 24 of the E. gallinarum, 15 of the E. raffinosus, and 13 of the E. casseliflavus isolates were also evaluated by DNA-DNA hybridization. Genomic DNAs from type strains E. faecalis ATCC 19433, E. faecium ATCC 19434, E. gallinarum ATCC 49573, E. raffinosus ATCC 49427, and E. casseliflavus ATCC 25788 were labeled with biotin for use as probes. E. faecalis differed from all other species in always having a largest fragment of > 400 kb. E. gallinarum was different from all other species in having all SmaI fragments of < 200 kb. Biotin-labeled probes showed a high degree of hybridization with genomic DNA from the same species and a low degree of hybridization when hybridized to genomic DNA from different species for all isolates tested except for four isolates identified as E. faecium by conventional biochemical methods. The DNA from these four isolates hybridized strongly to DNA from E. gallinarum ATCC 49573 and weakly to E. faecium ATCC 19434 DNA and had all SmaI fragments of < 200 kb in size. These data suggest that these isolates are nonmotile E. gallinarum. DNA from each ATCC type strain hybridized strongly with itself and had only a low degree of hybridization with DNA from other ATCC type strains tested. These results suggest that contour-clamped homogeneous electric field electrophoresis patterns and DNA-DNA hybridization with biotin-labeled probes may be of use for species differentiation of some enterococci.

Blotting, Southern↗

PubMed2026

Antimicrobial Resistance in Nontyphoidal Salmonella and Clinically Relevant Enterococcus From Faecal Samples of Conservation-Priority Captive Ungulates in a United Arab Emirates Urban Zoo: A Cross-Sectional Baseline Study.

Antimicrobial resistance (AMR) is a One Health challenge driven by microbial exchange among humans, animals and the environment. Zoological institutions offer useful settings for environmental AMR surveillance. This single-zoo cross-sectional study examined the occurrence, antimicrobial susceptibility and genomic characteristics of nontyphoidal Salmonella enterica (NTS) and clinically relevant Enterococcus spp. in faecal samples from 101 clinically healthy captive ungulates representing seven conservation-priority species at a major urban zoo in the United Arab Emirates. NTS was detected in 4/101 samples (3.9%), including serovars Schwarzengrund (n = 2), Kentucky (n = 1) and Chester (n = 1). Among the four recovered NTS isolates, all met the study MDR definition within the tested panel, including a Salmonella Kentucky ST198 isolate carrying multiple resistance genes and quinolone-associated mutations. Enterococcus spp. were detected in 77/101 samples (76.2%), dominated by Enterococcus faecium and Enterococcus casseliflavus (each 41.5%). Among 33 E. faecium/Enterococcus faecalis isolates tested phenotypically, resistance was generally low, with erythromycin and ciprofloxacin resistance each observed in 9.1%. One clinically important E. faecium isolate showed glycopeptide resistance and genetic markers associated with reduced daptomycin susceptibility. These single-institution cross-sectional data provide an initial regional baseline for AMR-relevant enteric bacteria in conservation-managed ungulates and identify priorities for broader longitudinal and interface-based surveillance.

Animals↗

PubMed2000

Evaluation of a new system, VITEK 2, for identification and antimicrobial susceptibility testing of enterococci.

We evaluated the new automated VITEK 2 system (bioMérieux) for the identification and antimicrobial susceptibility testing of enterococci. The results obtained with the VITEK 2 system were compared to those obtained by reference methods: standard identification by the scheme of Facklam and Sahm [R. R. Facklam and D. F. Sahm, p. 308-314, in P. R. Murray et al., ed., Manual of Clinical Microbiology, 6th ed., 1995] and with the API 20 STREP system and, for antimicrobial susceptibility testing, broth microdilution and agar dilution methods by the procedures of the National Committee for Clinical Laboratory Standards. The presence of vanA and vanB genes was determined by PCR. A total of 150 clinical isolates were studied, corresponding to 60 Enterococcus faecalis, 55 Enterococcus faecium, 26 Enterococcus gallinarum, 5 Enterococcus avium, 2 Enterococcus durans, and 2 Enterococcus raffinosus isolates. Among those isolates, 131 (87%) were correctly identified to the species level with the VITEK 2 system. Approximately half of the misidentifications were for E. faecium with low-level resistance to vancomycin, identified as E. gallinarum or E. casseliflavus; however, a motility test solved the discrepancies and increased the agreement to 94%. Among the strains studied, 66% were vancomycin resistant (57 VanA, 16 VanB, and 26 VanC strains), 23% were ampicillin resistant (MICs, >/=16 microgram/ml), 31% were high-level gentamicin resistant, and 45% were high-level streptomycin resistant. Percentages of agreement for susceptibility and resistance to ampicillin, vancomycin, and teicoplanin and for high-level gentamicin resistance and high-level streptomycin resistance were 93, 95, 97, 97, and 96%, respectively. The accuracy of identification and antimicrobial susceptibility testing of enterococci with the VITEK 2 system, together with the significant reduction in handling time, will have a positive impact on the work flow of the clinical microbiology laboratory.

Bacterial Typing Techniques↗

PubMed1989

Activity of glycopeptides against vancomycin-resistant gram-positive bacteria.

Gram-positive bacteria resistant to vancomycin are rare; but they include members of the genera Leuconostoc, Lactobacillus, and Pediococcus, as well as recently emerging vancomycin-resistant strains of Enterococcus faecium and Enterococcus faecalis. Vancomycin, teicoplanin, and several vancomycin derivatives were tested for their activities against vancomycin-resistant gram-positive bacteria. Vancomycin-resistant E. faecium and E. faecalis were generally cross-resistant to other glycopeptides, but some N-substituted vancomycin derivatives were active against the resistant strains, with MICs of 2 to 32 micrograms/ml. These vancomycin derivatives also had significant levels of activity against intrinsically vancomycin-resistant organisms such as Leuconostoc sp. While vancomycin resistance in E. faecium and E. faecalis was inducible, resistance in members of the genera Leuconostoc, Lactobacillus, and Pediococcus appeared to be expressed constitutively. Antibody to a vancomycin-induced membrane protein found in membranes of resistant enterococci did not detect a cross-reacting protein in other vancomycin-resistant species.

Drug Resistance, Microbial↗

PubMed1997

In-vitro bactericidal activity of quinupristin/dalfopristin alone and in combination against resistant strains of Enterococcus species and Staphylococcus aureus.

In-vitro activities of quinupristin/dalfopristin, a semisynthetic injectable streptogramin, and vancomycin were compared against multidrug-resistant Enterococcus faecium and Enterococcus faecalis, methicillin-susceptible Staphylococcus aureus ATCC 25923 and a methicillin-resistant S. aureus (MRSA). Combinations of quinupristin/dalfopristin and/or vancomycin with ofloxacin or gentamicin were evaluated using the chequerboard technique. The only synergy observed was that between quinupristin/dalfopristin plus vancomycin and quinupristin/dalfopristin plus gentamicin against E. faecium (FIC index < 0.5). Time-kill curves were performed over 24 h with an inoculum of 1 x 10(7) cfu/mL and clinically achievable concentrations of quinupristin/dalfopristin, vancomycin, ofloxacin and gentamicin (6, 30, 5 and 5 mg/L, respectively). In time-kill studies, combinations of quinupristin/dalfopristin plus vancomycin and quinupristin/dalfopristin plus gentamicin were additive, not synergic, against E. faecium and achieved 99.9% killing in 21.2 h and 19.6 h, respectively. None of the combination regimens suppressed the regrowth of E. faecalis. Quinupristin/dalfopristin combined with vancomycin demonstrated consistent synergy against ATCC 25923 and the MRSA, achieving 99.9% killing in 12.1 h and 11.9 h, respectively. Overall, quinupristin/dalfopristin alone demonstrated inhibitory activity against E. faecium, but not against E. faecalis, and bactericidal activity was achieved only with quinupristin/dalfopristin in combination with vancomycin or gentamicin against E. faecium. Quinupristin/dalfopristin plus vancomycin was the most potent and reliable combination against both strains of S. aureus in time-kill studies.

Anti-Bacterial Agents↗

PubMed2001

Bile salt hydrolase activity of Enterococci isolated from food: screening and quantitative determination.

One hundred seventeen enterococcal strains isolated from food (47 Enterococcus faecium, 48 Enterococcus faecalis, 16 Enterococcus durans, 2 Enterococcus gallinarum, 3 Enterococcus casseliflavus, and 1 Enterococcus malodoratus) were screened for bile salt hydrolase (BSH) activity on de Man, Rogosa, and Sharpe agar medium containing taurocholic acid and calcium chloride. The highest incidence of BSH-active strains was observed for E. faecalis (81%) followed by E. faecium (50%) and E. durans (44%). Isolates were grouped into four putative activity groups (no, low, medium, and high activity) based on the size of precipitation zones observed in the screening experiment. Our results showed that assumptions on BSH activity based on the size of bile precipitation zones in screening experiments did not correlate with actual activity as quantified by high-pressure liquid chromatography, but the screening assay is useful for assessing the presence or absence of BSH activity.

Amidohydrolases↗

PubMed2003

Screening for enterocins and detection of hemolysin and vancomycin resistance in enterococci of different origins.

The inhibitory activity of 122 out of 426 Enterococcus strains of geographically widespread origin and from different sources (food and feed, animal isolates, clinical and nonclinical human isolates) was tested against a wide range of indicator bacteria. Seventy-two strains, mainly belonging to the species Enterococcus faecium and Enterococcus faecalis were bacteriocinogenic. A remarkable variation of inhibitory spectra occurred among the strains tested, including inhibition of, for instance, only closely related enterococci, other lactic acid bacteria (LAB), food spoilage and pathogenic bacteria. No correlation could be found between the origin of the strains and the type of inhibitory spectrum, although a clustering of human isolates from both fecal and clinical origin was observed in the group of strains inhibiting lactic acid bacteria, Listeria, and either Staphylococcus or Clostridium. No relationship could be established between the presence of enterocin structural genes and the origin of the strain either, and hence no correlation seemed to exist between the presence of known enterocin genes and the activity spectra of these enterococci. The structural gene of enterocin A was widely distributed among E. faecium strains, whereas that of enterocin B only occurred in the presence of enterocin A. The vancomycin resistance phenotype as well as the presence of vancomycin resistance genes was also investigated. The vanA gene only occurred among E. faecium strains. The incidence of beta-hemolysis was not restricted to E. faecalis strains, but among the E. faecium strains the structural genes of cytolysin were not detected. beta-Hemolysis occurred in strains both from food and nonfood origin. It has been concluded that bacteriocin-producing E. faecium strains lacking hemolytic activity and not carrying cytolysin nor vancomycin resistance genes may be useful as starter cultures, cocultures, or probiotics.

Animals↗

PubMed1992

[Affinities of PBPs of enterococci to cefepime and ampicillin].

The beta-lactam resistance of genus Streptococcus has been explained by the low binding affinity of penicillin-binding proteins (PBPs) to the drug. This study was carried out to resolve the mechanisms of resistance to beta-lactam antibiotics in the species of genus Enterococcus by means of binding affinities of PBPs. Streptococcus pyogenes, Enterococcus faecalis, Enterococcus faecium and Enterococcus avium were employed as assay microbes. Cefepime (CFPM) and ampicillin (ABPC) were used as representatives of cephems and penicillins, respectively. All the PBP fractions of S. pyogenes manifested high binding affinities to CFPM and ABPC, whereas PBPs 1 and 4 of E. faecalis showed high binding affinities to ABPC but not to CFPM. In E. faecium, PBPs of an exceptionally penicillin-susceptible strain manifested a high binding affinity to ABPC, but PBPs 5 and 6 showed low affinities to CFPM. beta-lactam resistant strains of E. faecium possessed PBPs 5 and 6 with low binding affinities to CFPM and ABPC. All the fractions of PBPs but PBP 1 in E. avium showed low binding affinities to CFPM. Although all the PBP fractions but PBPs 3 and 6 manifested high binding affinities to ABPC, PBPs 3 and 6 showed low binding affinities to ABPC. A strain of E. avium, which is susceptible to ABPC but moderately resistant to CFPM, lacked PBP 6. In conclusion, the resistance of E. avium to CFPM is based upon low binding affinities of the many fractions to this drug, and ABPC resistance is based upon PBPs 3 and 6 with low binding affinities to ABPC.

Ampicillin↗

PubMed2000

Antimicrobial susceptibility and species identification for clinical isolates of enterococci.

In order to describe the susceptibility patterns and determine the identities of Enterococcus spp. isolated at a regional teaching hospital in Taichung during the period from June through November 1998, 96 clinical isolates of enterococci were collected for further analysis. Major sources of these isolates included urine, wound, and pus. Species identification was performed using the API 20 Strep system and supplemental tests. The minimum inhibitory concentrations (MICs) of six antimicrobial agents were determined by E-test for each isolate. Disk diffusion tests were also performed and the results were compared with those reported by the clinical laboratory. Because gentamicin susceptibility tests showed inconsistent results in many isolates, MIC determinations by the microbroth dilution method were also performed for these isolates. All isolates were tested for beta-lactamase production using the chromogenic method. The results showed that Enterococcus faecalis was the most frequently encountered species (86.5%), followed by Enterococcus faecium (7.3%), Enterococcus avium (5.2%) and Enterococcus casseliflavus (1.0%). The MIC90 of ampicillin, penicillin, vancomycin, teicoplanin, ciprofloxacin and gentamicin for total isolates were 1, 3, 2, 0.25, 1, and more than 1024 microg/mL, respectively. All isolates were susceptible to vancomycin and teicoplanin. The same rate of resistance (3.1%) was found to penicillin, ampicillin and ciprofloxacin in all isolates. There were 50 (52%) and 48 (50%) isolates with high level streptomycin and gentamicin resistance, respectively. The MIC90 of ampicillin and penicillin for E. faecium were significantly higher than those for other species (96 and > 256 vs. < or = 1 and < or = 3 microg/mL, respectively); in fact, ampicillin or penicillin-resistance was only found in E. faecium. No organism was found to produce beta-lactamase. There were 29 isolates showing discrepant results between the study findings and clinical laboratory report for gentamicin susceptibility. Most isolates (27/ 29) reported as susceptible to gentamicin by the clinical laboratory showed a high level gentamicin resistance by MIC determinations. The inconsistent results may be due to discrepancy in interpretations for heterogeneous resistance, bias in selecting colonies for the disk diffusion test, or variation in the quality of Mueller-Hinton agar used. The results suggest that any pure isolates growing some colonies within the inhibition zone should be considered as gentamicin resistant even if the zone diameter is equal or greater than the susceptible breakpoint. In order to obtain accurate results, the gentamicin MIC should be determined by the dilution method for enterococcal isolates that yield intermediate inhibition zones or zones just above the susceptible limit.

Drug Resistance, Microbial↗

PubMed1989

The enterococci: evidence of species-specific clinical and microbiologic heterogeneity.

Investigations of the clinical role of enterococci have been limited largely to enterococcal bacteremia and endocarditis and have not distinguished between the various species of enterococci. To characterize the full spectrum of enterococcal disease and to determine whether clinically important differences exist among infections and/or instances of colonization (infections/colonizations) by the various enterococcal species, cases of enterococcal infection/colonization diagnosed at the Dallas Veterans Administration Medical Center (DVAMC) in 1986 were reviewed. During this period, 220 Enterococcus faecalis, 21 Enterococcus faecium, 12 Enterococcus avium, and no Enterococcus durans isolates were identified in clinical specimens (other than stool) submitted to the DVAMC microbiology laboratory. Clinical characteristics of cases of infection/colonization by the three species of enterococci were similar and did not vary significantly when blood stream invasion occurred. Nevertheless, mortality data and therapeutic response rates suggested differences in virulence of the three enterococcal species. Enterococcal infections/colonizations (including bacteremia) frequently were polymicrobial. Although Enterobacteriaceae were the most common copathogens identified overall, Staphylococcus aureus was the most common copathogen in bloodstream infections. E. avium was more resistant than E. faecalis or E. faecium to penicillin G and ampicillin and less resistant to most other antimicrobial agents. The results of this investigation suggest that enterococci are a heterogeneous group of bacteria that should not be treated as a single entity in clinical investigations.

Anti-Bacterial Agents↗

PubMed1996

Quinupristin-dalfopristin (RP 59500): an injectable streptogramin combination.

The pharmacology, pharmacokinetics, activity, and potential clinical role of quinupristin-dalfopristin (RP 59500) are described. Quinupristin-dalfopristin is the first injectable formulation of the streptogramin antibiotics. Streptogramin drug products are each composed of two chemically distinct compounds, which when administered together act synergistically by inhibiting bacterial protein synthesis. Quinupristin-dalfopristin has shown activity in vitro against many strains of streptococci and staphylococci, including methicillin- and erythromycin-resistant strains of staphylococci. The combination is more active against Enterococcus faecium than Enterococcus faecalis. It has also shown activity in vitro against certain gram-negative organisms and anaerobes. The mean maximum blood concentration at the end of a one-hour infusion ranged from 0.95 mg/L for a 1.4-mg/kg dose to 24.2 mg/L for a 29.4-mg/kg dose; there was a linear correlation between dose and mean area under the concentration-time curve. Mean half-life ranged from 1.27 to 1.53 hours. The drug is under investigation in the United States in Phase III trials. Of 60 evaluable patients with documented bacteremia involving E. faecium resistant to vancomycin, 40 (67%) had a favorable clinical response. Of 11 patients with bacteremia caused by methicillin-resistant Staphylococcus aureus, 78% had a favorable response. The efficacy of quinupristin-dalfopristin in treating resistant infections has also been suggested by smaller studies and case reports. The drug may be useful in the prophylaxis of endocarditis. Adverse reactions are generally mild and transient. Quinupristin-dalfopristin may be useful in treating serious gram-positive infections, but more clinical study is needed.

Animals↗

PubMed2001

Epidemiological analysis of vancomycin-resistant enterococci in a large tertiary-care hospital in Northern Italy.

The epidemiology of vancomycin-resistant enterococci (VRE) was studied in a large tertiary-care hospital in northern Italy from February 1993 to December 1999. Sixteen cases of bacteraemic and 17 cases of nonbacteraemic active infections caused by VRE were recorded. Fifteen of the bacteraemic and four of the nonbacteraemic infections occurred in patients in the haematology department, while the remainder were registered in other departments of the same hospital. Active surveillance for the presence of VRE in stools led to identification of 51 noninfected carriers over the 1994-1999 period; of these, 32 were haematology patients and the remainder were patients admitted to other departments. All VRE isolates carried the vanA gene. Forty-one Enterococcus faecium isolates and eight Enterococcus faecalis isolates collected in the 1993-1996 period were typed by pulsed-field gel electrophoresis. Twenty-nine isolates of Enterococcus faecium shared either indistinguishable or strictly or possibly related patterns. Of these, 26 were isolated from patients in the haematology department. This is believed to be the first study on the epidemiology of VRE carried out in a large hospital in Italy over a period of several consecutive years. It reports an increase in VRE due to the epidemic spread of genetically related strains and sporadic infections or colonisation by unrelated VRE. It also documents the success of surveillance and of the measures adopted for preventing the spread of VRE in patients at risk.

Bacteremia↗

PubMed1990

Enterococcal resistance to vancomycin and related cyclic glycopeptide antibiotics.

Enterococci belonging to various species resistant to vancomycin and related cyclic glycopeptide antibiotics have been isolated from hospitalized patients in France, the UK and the USA. All such strains examined display inducible synthesis of a membrane protein associated with resistance. The mechanism by which the membrane protein acts has not been definitively established, but it may block the access of the antibiotic to its peptidoglycan target. That the protein could be a bypass enzyme has not been ruled out. Transfer of glycopeptide resistance by conjugation to either Enterococcus faecium or Enterococcus faecalis and by transformation of Streptococcus sanguis Challis has been reported. The structural and regulatory genes encoding this resistance can be localized on plasmid and, apparently, chromosomal DNA. The plasmids encoding this resistance appear to differ from each other and have variable host ranges, but share at least some DNA sequence homology.

Aminoglycosides↗

PubMed2000

Cost-effective algorithm for detection and identification of vancomycin-resistant enterococci in surveillance cultures.

A study was undertaken to develop an easy-to-use and cost-effective algorithm for the detection and identification of vancomycin-resistant enterococci (VRE) in surveillance cultures, because the incidence of VRE outbreaks in institutions across Canada has made continuous surveillance a necessity. Enterococcus faecium and Enterococcus faecalis carry transferable resistance genes and hence are a problem for infection control. In laboratory surveillance, however, Enterococcus gallinarum and Enterococcus casseliflavus, which exhibit low-level nontransferable resistance, are also encountered and often create confusion in identification. Included in this study were a total of 218 strains of enterococci and other streptococci isolated from surveillance cultures. Conventional methods were used to determine their biochemical activities, and speciation was attempted in 121 strains using a rapid multiplex polymerase chain reaction (PCR) method that utilized primers for the vanA, B, C1, C2/C3 and ddl genes. The results indicated that by using only a few tests (Gram stain, pyrrolidonyl arylamidase activity, motility, xylose and methyl-alpha-D-glucopyranoside utilization), Enterococcus faecium/faecalis strains could be accurately differentiated from Enterococcus gallinarum/casseliflavus strains. For the 121 strains on which PCR was performed, there was a 100% correlation with the biochemical identification, with the added advantage that the presence of van genes could be determined at the same time. The cost of identification using minimal biochemical testing and PCR was less than that of identification using automated systems or a battery of conventional biochemical methods. The algorithm presented here may be used in the microbiology laboratory.

Algorithms↗

PubMedDate not supplied

In vitro activity of sparfloxacin and clinafloxacin against multidrug-resistant enterococci.

We evaluated the in vitro susceptibility of 140 clinical enterococcal isolates to the quinolones sparfloxacin and clinafloxacin. Isolates included Enterococcus faecalis (107), Enterococcus faecium (29), Enterococcus raffinosus (3), and one Enterococcus gallinarum. There were 111 isolates that showed high-level [minimum inhibitory concentrations (MICs) > or = 2000 micrograms/ml] resistance to gentamicin and were resistant to high levels of all other aminoglycosides; five isolates produced beta-lactamase; 21 isolates were resistant (MIC > or = 16 micrograms/ml) to ampicillin and were not beta-lactamase producers; and 13 strains were resistant (MIC > or = 32 micrograms/ml) to vancomycin. Most strains were susceptible to low concentrations of sparfloxacin and clinafloxacin, with MIC90S of 0.6 microgram/ml and 0.5 micrograms/ml, respectively. There were no inoculum effects. Time-kill experiments were performed with 22 strains; using 2 x MIC at 24 h, a > or = 2 log10 reduction in growth was observed with sparfloxacin and clinafloxacin for 14 and 17 strains, respectively. Time-kill synergism experiments were performed with 15 strains lacking high-level aminoglycoside resistance. In vitro bacterial synergism with the combination of sparfloxacin or clinafloxacin with streptomycin or gentamicin was observed for five and 12 isolates, respectively. The bactericidal activity of sparfloxacin and clinafloxacin suggest that these antibiotics may prove useful for therapy of multidrug resistant enterococci.

Anti-Infective Agents↗

PubMed1996

Direct application of Etest to gram-positive cocci from blood cultures: quick and reliable minimum inhibitory concentration data.

The Etest was applied directly to 100 Gram-positive bacterial strains from blood cultures to measure their minimum inhibitory concentration (MIC). Results showed 100% concordance of MIC data between a direct Etest method and the standard Etest method for Streptococcus pneumoniae, beta-hemolytic streptococci, and viridans group streptococci. In addition, direct Etest for Staphylococcus aureus, coagulase negative staphylococci, Enterococcus faecium, and Enterococcus faecalis showed 83 to 100% correlation with standard Etests. These data indicate that the Etest is useful to obtain MIC data on Gram-positive cocci (especially streptococci) directly from positive blood cultures. The advantages of a direct Etest are two fold: MIC results can be obtained 24 hours earlier than standard methods and a more representative population of the bacterial isolate is tested.

Anti-Bacterial Agents↗

PubMed2003

The PhenePlate system for studies of the diversity of enterococcal populations from the food chain and the environment.

The growing interest in ecological investigations, such as studies of the "flow" of bacteria through the food chain, has resulted in a great need for simple typing techniques for bacteria that can be used when a large number of isolates need to be analysed. The present study describes a simple method for biochemical fingerprinting of enterococci, the PhenePlate RF (PhP-RF) system. The system is based on a 96-well microplate containing eight sets of 11 dehydrated reagents, selected to have a high discriminatory power among enterococcal isolates. The PhP plates are inoculated easily by picking single colonies directly from the primary agar culture and suspending them in the first well of each row in the microplate. The kinetics of each reaction is evaluated by measuring the absorbance value of each well three times during 64 h, whereupon a biochemical fingerprint is calculated as the mean value for each reagent over the three readings. The PhP-RF method was shown to be highly reproducible, even when results were compared between different laboratories. The discriminatory power, measured as Simpson's diversity index (Di), was as high as 0.96 for all enterococci. The PhP-RF method could also be used as a preliminary species identification method, by comparing the biochemical fingerprints of unknown strains to those of a set of reference strains of known species. Most strains of Enterococcus faecalis, Enterococcus faecium and Enterococcus hirae were correctly identified using this method. We conclude that the PhenePlate RF system is useful for rapid typing of enterococcal populations, and it is especially useful as a first screening method for ecological studies, when many isolates per sample need to be analysed.

Bacterial Typing Techniques↗