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The microbiologic aspects, including diagnosis, of beta-hemolytic streptococcal and enterococcal infections.

Basic principles concerning the collection, transport, and processing of clinical specimens for the detection of Streptococcaceae are given. Identification of beta-hemolytic streptococci (S. pyogenes, S. agalactiae, Lancefield group C and G streptococci, S. anginosus) and enterococci is based on the careful observation of colony morphology and hemolytic pattern on sheep blood agar plates; subsequent genus or species confirmation is achieved by rather simple biochemical or enzymatic tests and by detection of streptococcal cell wall carbohydrate antigens (Lancefield grouping). Rapid antigen tests for the detection of group A and B streptococci directly from pharyngeal and vaginal swabs, respectively, are highly specific, thus allowing an immediate antibiotic therapy in patients with a positive test result. The reported sensitivities of these nonculture tests are too low to exclude streptococcal colonization or infection, however. The elucidation of the genetics of some major virulence factors of group A and B streptococci has contributed to knowledge of their association with disease, and molecular techniques have supplemented the traditional (mostly culture and serologic) methods for an improved understanding of the epidemiology and pathogenesis of streptococcal infections. Recently employed examples include the M protein gene typing of group A streptococci by oligonucleotide probes and the use of PCR assays for the detection of the genes encoding for the pyrogenic exotoxins. Restriction enzyme endonuclease digestions of bacterial DNA in association with DNA fragment separation by conventional or PFGE have been applied successfully to several species of Streptococcaceae (e.g., S. pyogenes, S. agalactiae). Enterococci are important pathogens in the hospital setting, exhibiting high morbidity and mortality rates in bacteremic patients with severe underlying disease. Molecular typing methods have clearly confirmed their potential to be nosocomially transmitted. E. faecalis and E. faecium still account for the majority of human infections, but some of the newer enterococcal species (at present 19 species are recognized) have been encountered as well. The definitive species identification of enterococci requires the performance of an array of biochemical tests. The increasing antimicrobial resistance of enterococci, including high-level resistance to penicillins and aminoglycosides and occasionally also to glycopeptides, has hampered standard therapeutic regimens. All enterococci isolated from serious infections should be tested for high-level gentamicin and streptomycin resistance by one of several methods evaluated; beta-lactamase production (primarily found in E. faecalis) is reliably detected by the nitrocefin test.(ABSTRACT TRUNCATED AT 400 WORDS)

Enterococcus↗

Antibiotic susceptibility testing (agar disk diffusion and agar dilution) of clinical isolates of Enterococcus faecalis and E. faecium: comparison of Mueller-Hinton, Iso-Sensitest, and Wilkins-Chalgren agar media.

Forty-two isolates of Enterococcus faecalis and 56 isolates of Enterococcus faecium, including 8 vancomycin-resistant strains, were examined for comparative susceptibility to 27 antimicrobial drugs with the agar dilution method, employing Mueller-Hinton (MHA), Iso-Sensitest (ISTA), and Wilkins-Chalgren (WCA) agar. The Bauer-Kirby agar disk diffusion method was used to comparatively test 24 of the agents in parallel. The enterococci yielded better growth on ISTA and WCA. However, WCA completely antagonized co-trimoxazole and, though less, fosfomycin. Importantly, WCA slightly reduced the activities of teicoplanin (minimal inhibitory concentrations, MICs, raised up to twofold) and vancomycin (MICs raised two- to fourfold) against enterococci and staphylococcal quality control strains. Therefore, WCA was judged unsuitable for susceptibility testing of enterococci. For E. faecalis no discrepancies between agar dilution MICs and inhibition zone diameters were encountered with augmentin, ampicillin, ampicillin-sulbactam, chloramphenicol, mupirocin, oxacillin, teicoplanin, and co-trimoxazole. Overall, MHA yielded fewer very major (category I) and major (category II) discrepancies than ISTA. However, numerous minor (category III), slight (category IV), minimal (category V), and/or negligible (category VI) discrepancies were encountered with ciprofloxacin, doxycycline, erythromycin, fosfomycin, fusidic acid, meropenem, ofloxacin and rifampin. With respect to E. faecium, only cefotaxime, mupirocin, oxacillin, and teicoplanin yielded nondiscrepant results. Several very major (I) and major (II) discrepancies were observed with augmentin, ampicillin, ampicillin-sulbactam, doxycycline, fusidic acid, imipenem, and penicillin G. Minor discrepancies (categories III-VI) were particularly numerous with augmentin, chloramphenicol, ciprofloxacin, doxycycline, and piperacillin. The largest numbers of negligible (VI) discrepancies were noted with fosfomycin, fusidic acid, and ofloxacin. It is recommended to test one cephalosporin (cefuroxime or the like) in parallel for educational purposes and to exclude fosfomycin, fusidic acid, and rifampin from test batteries because of the wide scatter of test results. The large number of minimal (V) discrepancies of ciprofloxacin against E. faecalis, the numerous minor (III) and slight (IV) discrepancies of chloramphenicol against E. faecium, and the not insignificant number of very major (I) and minor (III) discrepancies observed with meropenem against isolates of E. faecalis necessitated proposals for new disk intermediate susceptibility criteria.

Agar↗

Antibiotic resistant fecal isolates of Enterococci among unselected patients outside the clinical sector: an epidemiological study from Southern Germany.

PURPOSE: The aim of the study was to assess the prevalence and determinants of antibiotic-resistant Enterococci in a large group of outpatients in Southern Germany. METHODS: Stool samples were collected from 497 unselected patients aged 40-75 years attending general practitioners. Enterococcus faecium (E. faecium) and Enterococcus faecalis (E. faecalis) were cultured and minimal inhibitory concentrations of antibiotics used inside and outside the clinical sector were tested. RESULTS: E. faecium and E. faecalis could be identified and cultured in 60 (12.4%) and 205 (41.2%) of the stool samples, respectively. Under non-selective culture conditions no vancomycin-resistant Enterococcus (VRE) isolate was found. Only E. faecium isolates showed resistance to fluoroquinolones, 40% were resistant to ciprofloxacin. The prevalences of E. faecium resistance to ampicillin and doxycycline were 3.3% and 13.3%, respectively, whereas 0.5% and 29.6% of the E. faecalis isolates were resistant to ampicillin and doxycycline, respectively. Antibiotic use during the last 3 months was significantly associated with antibiotic resistance (to either ampicillin, imipenem, or doxycycline) of E. faecalis isolates (OR: 2.9; CI: 1.2-6.8). CONCLUSIONS: Prevalences of resistance were generally lower than and patterns of resistance were quite different from previous investigations in the clinical setting. Recent antibiotic use was associated with increased colonization with resistant strains.

Adult↗

Emergence of unusual species of enterococci causing infections, South India.

BACKGROUND: Enterococci tend to be one of the leading causes of nosocomial infections, with E. faecalis and E. faecium accounting up to 90% of the clinical isolates. Nevertheless, the incidence of other species of enterococci from clinical sources shows an alarming increase with the properties of intrinsic resistance to several antibiotics including beta-lactams and glycopeptides. Thus proper identification of enterococci to species level is quintessential for management and prevention of these bacteria in any healthcare facility. Hence this work was undertaken to study the prevalence of unusual species of enterococci causing human infections, in a tertiary care hospital in South India. METHODS: The study was conducted in a tertiary care hospital in South India from July 2001 to June 2003. Isolates of enterococci were collected from various clinical specimens and speciated using extensive phenotypic and physiological tests. Antimicrobial susceptibility testing were performed and interpreted as per NCCLS guidelines. Whole cell protein (WCP) fingerprinting of enterococci were done for species validation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and analyzed computationally. RESULTS: Our study showed the prevalence of unusual (non-faecalis and non-faecium enterococci) and atypical (biochemical variant) species of enterococci as 19% (46 isolates) and 5% (12 isolates) respectively. The 7 unusual species (46 isolates) isolated and confirmed by phenotypic characterization includes: 15 E. gallinarum (6.2%), 10 E. avium (4.1%), 6 E. raffinosus (2.5%), 6 E. hirae (2.5%), 4 E. mundtii (1.7%), 3 E. casseliflavus-including the two atypical isolates (1.2%) and 2 E. durans (0.8%). The 12 atypical enterococcal species (5%) that showed aberrant sugar reactions in conventional phenotyping were confirmed as E. faecalis, E. faecium and E. casseliflavus respectively by WCP fingerprinting. The antimicrobial susceptibility testing depicted the emergence of high-level aminoglycoside and beta-lactam resistance among different species apart from intrinsic vancomycin resistance by some species, while all the species tested were susceptible for linezolid and teicoplanin. CONCLUSION: Our study reveals the emergence of multi-drug resistance among unusual species of enterococci posing a serious therapeutic challenge. Precise identification of enterococci to species level enables us to access the species-specific antimicrobial resistance characteristics, apart from knowing the epidemiological pattern and their clinical significance in human infections.

Anti-Bacterial Agents↗

Quantitative determination of gelatinase activity among enterococci.

Gelatinase, hemolysin and aggregation substance have all been reported to be virulence factors of enterococci. In this study, gelatinase production was investigated in isolates of Enterococcus faecalis (n=93), E. faecium (n=49) and E. avium (n=36) recovered from hospitalized patients. Gelatinase was detected in 45% of E. faecalis isolates, but could not be detected in E. faecium and E. avium. Gelatinase activity was then measured by radial diffusion for the 42 gelatinase-positive E. faecalis isolates. To convert gelatinase activity into proteinase K activity, a standard curve was produced by placing different concentrations of proteinase K into wells in the gelatine plate. Gelatinase activity per E. faecalis colony ranged from 2.6 x 10(-7) to 2.2 x 10(-5) microg/ml, proportionate to the activity of proteinase K. An approximately 84-fold difference in gelatinase concentration was observed between the colony producing the highest amount and that producing the lowest amount. This method may be useful for determining the virulence of given isolates in relation to gelatinase production as it is quick, easy and inexpensive to perform.

Adult↗

Antibiotic resistance and virulence factors among clinical and food enterococci isolated in Slovakia.

The resistance to antibiotics and the distribution of virulence factors in enterococci isolated from traditional Slovak sheep cheese bryndza was compared with strains from human infections. The occurrence of 4 enterococcal species was observed in 117 bryndza-cheese isolates. The majority of strains were identified as E. faecium (76 %) and E. faecalis (23 %). Several strains of E. durans and 1 strain of E. hirae were also present. More than 90 % of strains isolated from 109 clinical enterococci were E. faecalis, the rest belonged to E. faecium. The resistance to 6 antimicrobial substances (ampicillin, ciprofloxacin, higher concentration of gentamicin, nitrofurantoin, tetracycline and vancomycin) was tested in clinical and food enterococci. A higher level of resistance was found in clinical than in food strains and E. faecium had a higher resistance than E. faecalis; no resistance to vancomycin was detected. The occurrence of 3 virulence-associated genes, cylA (coding for hemolysin), gelE (coding for gelatinase) and esp (coding for surface protein) was monitored. Differences were found in the distribution of cylA gene between clinical and bryndza-cheese E. faecalis strains; in contrast to clinical strains (45 %), cylA gene was detected in 22 % of food isolates. The distribution of 2 other virulence factors, gelE and esp, was not significantly different in the two groups of E. faecalis strains. cylA and gelE genes were not detected in E. faecium but more than 70 % of clinical E. faecium were positive for esp, even thought none of the 79 E. faecium cheese isolates contained this gene.

Animals↗

Gentamicin resistance gene transfer from Enterococcus faecalis and E. faecium to Staphylococcus aureus, S. intermedius and S. hyicus.

Transfer of gentamicin resistance from Enterococcus faecalis and E. faecium to Staphylococcus aureus, S. hyicus and S. intermedius was accomplished by filter mating. Transfer from E. faecalis was plasmid mediated and a probe to the gentamicin resistance gene aac(6')-aph(2") hybridized to HindIII fragments of various sizes (7.5, 6.2 and 4.7 kbp) of these plasmids. Transfer from E. faecium was apparently to the chromosome in 9 of 10 instances with HindIII fragments of 3.3 or 3.0 kbp hybridizing to the probe. These results are consistent with transposon mediated resistance. Plasmid transfers to S. intermedius were at much lower frequencies than to S. aureus or to S. hyicus but presumed transposon transfer direct to the chromosome was equal in all three species.

Burns↗

The role of various agents in chicken amyloid arthropathy.

The results of an inventory of field cases of amyloid arthropathy in chickens and of routine post-mortem recordings over a two years period are described. Studies were also performed to evaluate the amyloidogenic potential of arthrotropic bacterial species (Staphylococcus aureus, Escherichia coli and Salmonella enteritidis) isolated from chickens as well as several Enterococcus faecalis isolates compared to the amyloidogenic E. faecalis isolate (previously isolated from amyloidotic joints). As chicken anemia virus was also isolated from amyloidotic joints of field cases, it was also screened for its amyloidogenic potential. In another experiment, Mycoplasma synoviae, inactivated E. faecalis isolate 6085.94, Freund's adjuvant and an arthrotropic reovirus field isolate were also screened for amyloidogenicity by intra-articular injection. These studies showed that the ability to elicit extensive amyloid arthropathy is reserved primarily to E. faecalis, but that this property is not common to every E. faecalis isolate. Intra-articular application of complete Freund's adjuvant led to the formation of extensive joint amyloid deposits. Of the other micro-organisms studied, S. aureus, S. enteritidis and E. coli were also able to cause joint amyloidosis, but in very small amounts. Inactivated E. faecalis, chicken anemia virus and reovirus did not cause amyloid arthropathy after intra-articular inoculation. This study is consistent with results of the analyses of previous field cases and of the induction of amyloid arthropathy in chickens, suggesting a considerable role for E. faecalis in this clinical-pathological entity. Finally, strain typing by analysis of chromosomal DNA restriction endonuclease digests by pulsed-field gel electrophoresis (PFGE) of amyloidogenic, non-amyloidogenic, amyloid-associated and other E. faecalis isolates from various origins showed that all amyloidogenic and amyloid-associated E. faecalis isolates had similar restriction digests, suggesting clonal spread.

Amyloidosis↗

[Study of distribution of serovars of Enterococcus faecalis clinical isolates].

Serotyping was conducted of E. faecalis strains isolated from various clinical specimens, and the distribution of the serovars was investigated. To date, 21 serovars of E. faecalis have been identified. The most common serovars in Japanese 770 strains were types 2 (30.6%), 7 (13.9%), 1 (12.2%) and 4 (6.9%), and these four serovars accounted for 63.6% of the total strains. The most common serovars in USA 200 strains were types 1 (22.5%), 4 (14.5%) and 2 (14.0%). The most common serovars in UK 65 strains were types 2 (24.6%), 4 (13.8%) and 9 (7.7%). The distribution of E. faecalis serovars differ as a function of the country of origin. Next, serological classification was performed for E. faecalis strains isolated from various Japanese clinical specimens. For the strains obtained from the urine, the most common serovars were types 2 (32.7%) and 7 (14.6%). Similarly, the most common serovars of the strains isolated from the various other clinical specimens were as follows: types 7 (35.0%) and 2 (15.0%) from pus; types 7 (44.4%) and 2 (33.3%) from sputum; types 2 (22.3%) and 1 (13.1%) from blood; and types 2 (34.9%) and 1 (27.9%) from the vaginal smear. It had no definite pattern for the distribution of serovars. The distribution of serovars varied in accordance with isolated years. Accordingly, the data obtained in these studies revealed that the distributions of E. faecalis serovars differ as a function of the geographical area of origin, the clinical specimen of origin and the year of origin. This approach is useful for serological classification of E. faecalis strains, and it is thought that it will be useful for epidemiological studies of this bacterium.

Enterococcus faecalis↗

Antimicrobial resistance of Enterococcus spp. isolates from raw beef and meat products.

E. faecalis (67%) and E. faecium (13.7%) were most frequently isolated among enterococci that contaminate cooled and frozen processed meat, follow-up heat-treated meat products and unheated fermented dry salami. Most isolates of both species were resistant to cephalothin (95 and 83 %) and clindamycin (77 and 67%, respectively). Furthermore, E. faecalis and E. faecium isolates were resistant to erythromycin (44 and 72%), tetracycline (34.5 and 17.4%), and streptomycin (13.3 and 4.3%, respectively). Only a few of the isolates were resistant to ampicillin, ampicillin-sulbactam, chloramphenicol, and vancomycin while all isolates were susceptible to gentamicin, penicillin, and teicoplanin. During the production of heat-treated meat products, numbers of resistant isolates increased in spite of the decreasing enterococcal contamination of the samples. An opposite situation was found in the production of fermented dry salami.

Animals↗

Determination of antibiotic resistance and resistance plasmids of clinical Enterococcus species.

To determine the antibiotic resistance pattern and resistance plasmids, we studied 23 antibiotic-resistant clinical isolates of Enterococcus spp. which caused infection in Bayindir-Ankara Hospital, Turkey. Biochemical and physiological identification tests were applied by the Vitek system and compared with the results of protein profiles by SDS-PAGE. From 23 isolates, 20 were identified as E. faecalis, 2 as E. faecium and 1 as E. gallinarum. Twenty four antibiotics belong to 10 different groups were used in susceptibility tests. Multiple antibiotic resistance was determined in 10 of 23 Enterococcus spp. Overall resistance to the used antibiotics was 47.3% and low level resistance was 16.6%. Among the isolates tested, 8.7% demonstrated high level gentamicin resistance, 17.4% demonstrated high level streptomycin resistance, and 43.5% demonstrated penicillin resistance. High level vancomycin resistant Enterococcus spp. rate was 34.8%, and 60.9% exhibited low level resistance to vancomycin and teicoplanin. They contain plasmids which varied in numbers between 1 and 11 and the plasmid sizes ranged from 2.08 to 56.15 kb. In curing experiments with acriflavine, two different plasmids were shown in different molecular sizes of 33.49 and 13.6 kb while the first determined glycopeptide and penicillin resistance, the second one determined either glycopeptide or penicillin resistance in two different E. faecalis strains. On the other hand, a 22.58 kb plasmid, determining kanamycin resistance, was detected in an E. faecium strain. After the curing experiments, an elimination of 37.17 and 44.47 kDa protein bands was shown in E. faecium EFA1 and E. faecalis EFA13 in SDS-PAGE, respectively. This survey indicates the increase of antibiotic-resistant enterococci, especially to vancomycin in our hospital isolates.

Anti-Bacterial Agents↗

[Surveillance of bacterial resistance in China (1998-1999)].

OBJECTIVE: To determine the rates of bacterial resistance of clinical isolates obtained from patients with hospital infections (HI) or community acquired infections (CAI) in the study wards of 13 hospitals located at different areas in China. METHODS: Susceptibility test was performed on 2 081 strains isolated from 13 hospitals, using standard plate dilution method according to the guidelines of NCCLS (1998). MIC(50) and MIC(90) were used to show the antibacterial activity of antimicrobial agents, and data of R%, I% and S% represent resistant, intermediate and susceptible rates of bacteria to antimicrobial agents tested. RESULTS: Among total 98 strains of S. aureus, 87 were isolated from patients with CAI and 11 were from HI patients. The incidence of methicillin resistant S.aureus (MRSA) from total infections caused by S. aureus was 27.55%, while the incidence of MRSA from HI patients was 81.82%, which was significantly higher than that (21.84%) from CAI patients. The rate of penicillin resistant S. pneumoniae (PRSP) (R% + I%) was 22.50%; the rate of R (MIC >/= 2 mg/L) was 2.5%, and most of PRSP strains was intermediate, with I% of 20%. No strains of S. aureus, neither strains of E. faecalis, nor of E. faecium were found resistant to vancomycin. The intermediate rate (I%) of E. faecalis and E. faecium to vancomycin was 3.23% and 3.77%, respectively. Cross resistance of E. coli to quinolones was present obviously, and more than 60% of E. coli strains were resistant to fluoroquinolones tested. CONCLUSION: Multi-drug resistance (MDR) of bacteria is a serious problem in China, including MDR of MRSA, PRSP, and enterobacteriaceae; and the resistant rate of enterobacterinceae is the highest up-to-date.

Anti-Bacterial Agents↗

Peptidoglycan O acetylation and autolysin profile of Enterococcus faecalis in the viable but nonculturable state.

The O acetylation of peptidoglycan occurs specifically at the C-6 hydroxyl group of muramoyl residues. Using a combination of high-performance liquid chromatography-based organic acid analysis and carbohydrate analysis by high-pH anion-exchange chromatography, we determined that strains of Entercoccus durans, E. faecalis, E. faecium, and E. hirae produce O-acetylated peptidoglycan. The levels of O acetylation ranged from 19% to 72% relative to the muramic acid content, and they were found to vary with the growth phase of the culture. Increases of 10 to 40% in O acetylation were observed with cultures entering the stationary phase. Cells of E. faecalis in the viable but nonculturable (VBNC) state had the highest levels of peptidoglycan O acetylation. The presence of this modification to peptidoglycan was shown to inhibit the action of hen egg white lysozyme in a concentration-dependent manner. Zymography using sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels containing either O-acetylated or chemically de-O-acetylated peptidoglycan was used to monitor the production of specific autolysins in E. faecalis. Differences in the expression of specific autolysins were observed with the age of the culture, and VBNC E. faecalis produced the highest levels of these enzymes. This technique also permitted classification of the enterococcal autolysins into enzymes that preferentially hydrolyze either O-acetylated or non-O-acetylated peptidoglycan and enzymes that show no apparent preference for either substrate type.

Acetylation↗

Antimicrobial resistance of Enterococcus species isolated from produce.

The purpose of this study was to characterize the antibiotic resistance profiles of Enterococcus species isolated from fresh produce harvested in the southwestern United States. Among the 185 Enterococcus isolates obtained, 97 (52%) were Enterococcus faecium, 38 (21%) were Enterococcus faecalis, and 50 (27%) were other Enterococcus species. Of human clinical importance, E. faecium strains had a much higher prevalence of resistance to ciprofloxacin, tetracycline, and nitrofurantoin than E. faecalis. E. faecalis strains had a low prevalence of resistance to antibiotics used to treat E. faecalis infections of both clinical and of agricultural relevance, excluding its intrinsic resistance patterns. Thirty-four percent of the isolates had multiple-drug-resistance patterns, excluding intrinsic resistance. Data on the prevalence and types of antibiotic resistance in Enterococcus species isolated from fresh produce may be used to describe baseline antibiotic susceptibility profiles associated with Enterococcus spp. isolated from the environment. The data collected may also help elucidate the role of foods in the transmission of antibiotic-resistant strains to human populations.

Anti-Bacterial Agents↗

Susceptibility patterns of enterococci causing infections.

Enterococci are among the common organisms associated with hospital-acquired infections. We examined in vitro activities of different antibiotics to 103 enterococcal isolates. Minimal inhibitory concentrations (MICs) of penicillin G, ampicillin, gentamicin, ciprofloxacin, ofloxacin, levofloxacin, grepafloxacin, trovafloxacin and gemifloxacin were determined by broth microdilution testing method. Among the isolates 71 (69%) were identified as E. faecalis and 32 (31%) as E. faecium. While over 75% of E. faecium isolates were resistant to penicillin and ampicillin, approximately 25% of E. faecalis isolates were resistant to penicillin and ampicillin. None of the E. faecalis and E. faecium isolates were resistant to vancomycin. While 17 (52%) of E. faecium isolates exhibited high-level gentamicin resistance (HLGR), high level streptomycin resistance (HLSR) was detected in 24 (74%) of the isolates. In contrast, HLGR and HLSR rates for E. faecalis were 14 (20%) and 22 (31%), respectively. Both HLGR and HLSR were detected with higher frequency in ampicillin resistant isolates. Among fluoroquinolones, gemifloxacin and trovafloxacin were the most potent antibiotics tested. There was no increase in MIC90 values of the fluoroquinolones in ampicillin resistant isolates in comparison with ampicillin susceptible isolates. Our data suggest newer fluoroquinolones would be good alternative agents to use especially for combination drug therapy where enterococci with ampicillin resistance and HLAR are prevalent.

Anti-Bacterial Agents↗

In vitro activities of quinupristin/dalfopristin and eight other antimicrobial agents against 360 clinical isolates from Korea.

The emergence of multi-drug resistant gram-positive cocci such as methicillin-resistant (MR) staphylococci, vancomycin-resistant (VR) enterococci, and vancomycin-intermediate resistant S. aureus (VISA) has given new urgency to the development of new antimicrobial agents. One of these is quinupristin/dalfopristin (Q/D). We decided to determine the susceptibility of gram-positive cocci isolated at two university hospitals in Seoul to Q/D and compare the results with eight other antimicrobial agents. We investigated 120 isolates of S. aureus including 49 MRSAs and one VISA, 120 isolates of coagulase negative staphylococci (CNS), 64 E. faecalis and 56 E. faecium, including seven strains of VR E. faecium. Minimum inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs) for several antimicrobials, including vancomycin and Q/D, were determined by broth microdilution. All S. aureus including VISA were susceptible to Q/D. Q/D MIC90 for both methicillin-susceptible S. aureus (MSSA) and MRSA was 0.25 g/mL. 49 (87.5%) of 56 E. faecium including six of seven VR E. faecium were susceptible to Q/D. E. faecalis were not susceptible to Q/D (only 1.5% susceptible), but were inhibited by ampicillin (94% susceptible) or vancomycin (95%). CNS was susceptible to Q/D (96% susceptible) and vancomycin (100% susceptible). One of 38 staphylococci and two of 17 E. faecium were tolerant to Q/D. In conclusion, Q/D showed excellent activity against all species of gram-positive cocci including MRSA, VISA, and VR E. faecium except E. faecalis, and may provide a valuable option for the treatment of infections caused by these emerging nosocomial pathogens of gram-positive cocci.

Anti-Bacterial Agents↗

Survey of genomic diversity among Enterococcus faecalis strains by microarray-based comparative genomic hybridization.

We have compared nine Enterococcus faecalis strains with E. faecalis V583 by comparative genomic hybridization using microarrays (CGH). The strains used in this study (the "test" strains) originated from various environments. CGH is a powerful and promising tool for obtaining novel information on genome diversity in bacteria. By CGH, one obtains clues about which genes are present or divergent in the strains, compared to a reference strain (here, V583). The information obtained by CGH is important from both ecological and systematic points of view. CGH of E. faecalis showed considerable diversity in gene content: Compared to V583, the percentage of divergent genes in the test strains varied from 15% to 23%, and 154 genes were divergent in all strains. The main variation was found in regions corresponding to exogenously acquired or mobile DNA in V583. Antibiotic resistance genes, virulence factors, and integrated plasmid genes dominated among the divergent genes. The strains examined showed various contents of genes corresponding to the pTEF1, pTEF2, and pTEF3 genes in V583. The extensive transport and metabolic capabilities of V583 appeared similar in the test strains; CGH indicated that the ability to transport and metabolize various carbohydrates was similar in the test strains (verified by API 50 CH assays). The contents of genes related to stress tolerance appeared similar in V583 and the nine test strains, supporting the view of E. faecalis as an organism able to resist harsh conditions.

Enterococcus faecalis↗

Comparative genomic characterization and antimicrobial resistance of bacteremia-causing Enterococcus faecium and Enterococcus faecalis in a Chinese hospital.

Enterococci are common commensals of the human gut and important opportunistic pathogens, with Enterococcus faecium and Enterococcus faecalis being the most clinically prevalent species. A significant epidemiological shift has emerged with an increasing clinical burden of E. faecium. To compare genomic evolution of E. faecium and E. faecalis, we performed whole-genome sequencing on 93 E. faecium and 32 E. faecalis isolates causing bloodstream infections at a single hospital (2022-2024). Analysis of patient demographics revealed that E. faecium infections originated from fewer sources than E. faecalis, with a higher proportion deriving from intra-abdominal infections. Multilocus sequence typing identified ST78 and ST789 as the predominant sequence types for E. faecium, whereas ST16 and ST179 were most common for E. faecalis. E. faecium carried more antimicrobial resistance genes and putative virulence marker (PVM)-type virulence genes than E. faecalis, with vancomycin resistance predominantly mediated by vanHAX (33/93, 35.5%) and a single E. faecalis isolate also carrying vanHAX (1/32, 3.1%); the structurally incomplete vanHMX gene cluster was detected in 11 E. faecium isolates. Pan-genome analysis indicated a larger core genome in E. faecalis compared to E. faecium, consistent with greater plasmid replicon diversity in the latter. Intra-host comparisons showed that two E. faecalis pairs from the same patient were clonally related, with one isolate acquiring a vanHAX plasmid conferring vancomycin resistance. In contrast, E. faecium isolates exhibited marked genomic diversity even among clonally related pairs. These findings suggest that E. faecium possesses greater genomic plasticity and adaptive potential to the clinical environment.IMPORTANCEThis study provides a detailed comparison of clinical and genomic features between Enterococcus faecium and Enterococcus faecalis from the same hospital setting. We show that E. faecium isolates, mainly ST78/ST789, carry more antimicrobial resistance genes and a higher number of putative virulence marker (PVM) genes than E. faecalis, reflecting their hospital-adapted nature. E. faecium also exhibits a smaller core genome and greater diversity of plasmid replicon types, indicating higher genomic plasticity and capacity for horizontal gene transfer. By contrast, E. faecalis retains a larger core genome and a set of classical virulence factors, and its within-host isolates are clonally related. These distinct genomic profiles help to understand how the two species adapt to clinical environments and may inform more targeted infection control strategies and resistance surveillance.

Enterococcus faecium↗