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Sequence and genetic organization of the 19.3-kb erythromycin- and dalfopristin-resistance plasmid pLME300 from Lactobacillus fermentum ROT1.

Lactobacillus fermentum ROT1 was isolated from a raw milk dairy product. It is resistant to novobiocin, tetracycline, erythromycin and dalfopristin. A chromosomal tetracycline-resistance determinant was identified as tetM. A 19,398-bp plasmid (pLME300), present in several erythromycin-resistant strains of Lb. fermentum, was isolated from strain ROT1 and completely sequenced. Based on putative open reading frames, pLME300 contains at least four different functional regions. In region I, ORF1 shows high homologies to replication proteins of different theta-replicating plasmids. In addition, a tandem repeat of a 22-bp sequence appears 4.5 times. In region II, ORF3 may code for a methylase, and ORF4 has homologies to Mrr restriction system proteins of Deinococcus radiodurans and Escherichia coli suggesting a restriction-modification system. Region III harbours antibiotic-resistance genes, coding for a macrolide-lincosamide-streptogramin B (MLS) methylase Erm(LF) and the streptogramin A acetyltransferase Vat(E), which is identical to Vat(E) from Enterococcus faecium. Furthermore, region III shows a 91% nucleotide sequence identity to an erm-vat linkage of E. faecium. Region IV carries ORFs that appear to be involved in plasmid mobilization as characterized by a putative origin of transfer and a mobilization protein. pLME300 is the largest completely sequenced multi-resistance plasmid isolated from any Lactobacillus strain so far.

Anti-Bacterial Agents↗

Activity of newer fluoroquinolones in vitro against gram-positive bacteria.

Several newer fluoroquinolones, which have been recently introduced or are under investigation, display substantially greater potency against gram-positive organisms than the older generation agents of this class. Nevertheless, for problem organisms including methicillin-resistant strains of Staphylococcus aureus and many Enterococcus faecium, concentrations of newer antimicrobials required to inhibit 90% of organisms in the collections studied remain above those that are projected to be achievable with clinical use. Nevertheless, enhanced potency of several newer quinolones may result in a favourable pharmacodynamic profile leading to improved outcomes against gram-positive infections and possibly to the delayed or diminished emergence of resistance to these agents.

Anti-Infective Agents↗

Bacteraemia and antibiotic resistance of its pathogens reported in England and Wales between 1990 and 1998: trend analysis.

OBJECTIVES: Determination of causes, trends, and antibiotic resistance in reports of bacterial pathogens isolated from blood in England and Wales from 1990 to 1998. DESIGN: Description of bacterial isolates from blood, judged to be clinically significant by microbiology staff, reported to the Communicable Disease Surveillance Centre. SETTING: Microbiology laboratories in England and Wales. SUBJECTS: Patients yielding clinically significant isolates from blood. MAIN OUTCOME MEASURES: Frequency and Poisson regression analyses for trend of reported causes of bacteraemia and proportions of antibiotic resistant isolates. RESULTS: There was an upward trend in total numbers of reports of bacteraemia. The five most cited organisms accounted for over 60% of reports each year. There was a substantial increase in the proportion of reports of Staphylococcus aureus resistant to methicillin, Streptococcus pneumoniae resistance to penicillin and erythromycin, and Enterococcus faecalis and Enterococcus faecium resistance to vancomycin. No increase was seen in resistance of Escherichia coli to gentamicin. CONCLUSIONS: Reports from laboratories provide valuable information on trends and antibiotic resistance in bacteraemia and show a worrying increase in resistance to important antibiotics.

Bacteremia↗

In vitro antimicrobial activities of novel anilinouracils which selectively inhibit DNA polymerase III of gram-positive bacteria.

The 6-anilinouracils are novel dGTP analogs that selectively inhibit the replication-specific DNA polymerase III of gram-positive eubacteria. Two specific derivatives, IMAU (6-[3'-iodo-4'-methylanilino]uracil) and EMAU (6-[3'-ethyl-4'-methylanilino]uracil), were substituted with either a hydroxybutyl (HB) or a methoxybutyl (MB) group at their N3 positions to produce four agents: HB-EMAU, MB-EMAU, HB-IMAU, and MB-IMAU. These four new agents inhibited Staphylococcus aureus, coagulase-negative staphylococci, Enterococcus faecalis, and Enterococcus faecium. Time-kill assays and broth dilution testing confirmed bactericidal activity. These anilinouracil derivatives represent a novel class of antimicrobials with promising activities against gram-positive bacteria that are resistant to currently available agents, validating replication-specific DNA polymerase III as a new target for antimicrobial development.

Anti-Bacterial Agents↗

Prophylactic and therapeutic efficacy of antibodies to a capsular polysaccharide shared among vancomycin-sensitive and -resistant enterococci.

Enterococci are important nosocomial pathogens that are increasingly difficult to treat due to intrinsic and acquired resistance to antibiotics, including vancomycin. A recently described capsular polysaccharide (CP) isolated from Enterococcus faecalis 12030 was used to evaluate the potential efficacy of active or passive immunotherapy regimens as adjunctive treatments. Evaluation of protective efficacy was carried out in immunocompetent mice challenged intravenously (i.v.) with live enterococci. In nonimmune mice, i.v. inoculations resulted in high levels of bacteria in kidneys, spleens, and livers 5 days after challenge. Mice immunized with four 10-microg doses of CP antigen/mouse were protected against challenge with the homologous E. faecalis strain. High-titer opsonic immunoglobulin G was also induced by immunizing rabbits with the purified CP, and passive transfer of this antiserum to mice produced significantly lower bacterial counts in organs than did normal rabbit serum or sterile saline. Antibodies to the polysaccharide isolated from E. faecalis 12030 were protective against Enterococcus faecalis OG1RF and against two serologically related, vancomycin-resistant Enterococcus faecium clinical isolates. Antibodies to this CP antigen were also effective as a therapeutic reagent in mice when passive therapy was initiated 48 h after live bacterial challenge. These data indicate that CP antigens from enterococci are potential targets of protective antibodies and that these antibodies may be useful for prophylaxis and treatment of enterococcal infections.

Animals↗

Sex pheromone response, clumping, and slime production in enterococcal strains isolated from occluded biliary stents.

Bile-resistant bacteria, particularly gram-positive Enterococcus faecalis and Enterococcus faecium, play an important role in biliary stent occlusion, because their sessile mode of growth protects them against host defenses and antimicrobial agents. Twelve E. faecalis and seven E. faecium strains isolated from occluded biliary stents have been investigated for slime production, presence of aggregation substance genes, and ability to adhere to Caco-2 cells. Ten isolates were strong producers of slime, and seven isolates produced clumps when exposed to pheromones of E. faecalis JH2-2 and/or OG1RF. The small E. faecium clumps differed from the large clumps of E. faecalis and were similar to those of E. faecium LS10(pBRG1) carrying a pheromone response plasmid. After induction with pheromones, the adhesion to Caco-2 cells of clumping-positive strains was found to increase from two- to fourfold. Amplicons of the expected size were detected in three clumping-positive and three clumping-negative E. faecalis isolates by using primers (agg) internal to a highly conserved region of the E. faecalis pheromone response plasmids pAD1, pPD1, and pCF10 and primers internal to prgB of the E. faecalis plasmid pCF10. The agg/prgB-positive E. faecalis strains were also positive in Southern hybridization experiments with a prgB-specific probe. No PCR products were obtained with the same primers from four clumping-positive isolates (one E. faecalis and three E. faecium strains), which were also Southern hybridization negative. Our results demonstrate that slime production and pheromone response are both present in isolated enterococci, suggesting that clinical strains with these features might have a selective advantage in colonizing biliary stents.

Bacterial Adhesion↗

Rapid detection of vanA and vanB genes directly from clinical specimens and enrichment broths by real-time multiplex PCR assay.

A real-time PCR assay previously developed for use on the Roche LightCycler platform was investigated as an alternative to culture for the direct detection of vancomycin-resistant enterococci (VRE) in clinical specimens. PCR primers and fluorescence resonance energy transfer hybridization probes specific for the vanA and vanB genes were combined in a multiplex real-time PCR assay performed directly with fecal material obtained by rectal swabbing and with enrichment broth samples. DNA was prepared from the rectal swabs and enrichment broths with a commercially available DNA preparation column designed specifically for use with fecal specimens. One hundred eighty duplicate rectal swabs were obtained from 42 patients who were previously found to be positive for VRE and who were being monitored for carriage of VRE. Direct and enrichment broth cultures were performed with one swab, while PCR was performed with the other swab as well as any corresponding presumptive positive enrichment broth. In total, 100 specimens from 30 patients remained positive for VRE by at least one method. The multiplex real-time PCR was positive for 88 enrichment broths of rectal swabs from 27 patients but for only 45 rectal swabs from 15 patients. Direct culture was positive for VRE for only 43 specimens from 11 patients, while enrichment broth culture was positive for VRE for 75 specimens from 22 patients. Inhibition studies for the multiplex real-time PCR assay, performed by spiking the DNA extracts from 50 negative rectal swabs and the corresponding enrichment broths with between 1 and 10 CFU of a VanB Enterococcus faecium strain, detected inhibition rates of 55.1 and 10%, respectively. PCR performed directly with enrichment broths was found to be significantly more sensitive than enrichment broth culture (P < 0.025). Negative samples were identified significantly earlier by PCR than by culture alone.

Anti-Bacterial Agents↗

Removal of peptidoglycan and inhibition of active cellular processes leads to daptomycin tolerance in Enterococcus faecalis.

Daptomycin is a cyclic lipopeptide antibiotic used in the clinic for treatment of severe enterococcal infections. Recent reports indicate that daptomycin targets active cellular processes, specifically, peptidoglycan biosynthesis. Within, we examined the efficacy of daptomycin against Enterococcus faecalis under a range of environmental growth conditions including inhibitors that target active cellular processes. Daptomycin was far less effective against cells in late stationary phase compared to cells in exponential phase, and this was independent of cellular ATP levels. Further, the addition of either the de novo protein synthesis inhibitor chloramphenicol or the fatty acid biosynthesis inhibitor cerulenin induced survival against daptomycin far better than controls. Alterations in metabolites associated with peptidoglycan synthesis correlated with protection against daptomycin. This was further supported as removal of peptidoglycan induced physiological daptomycin tolerance, a synergistic relation between daptomycin and fosfomycin, an inhibitor of the fist committed step peptidoglycan synthesis, was observed, as well as an additive effect when daptomycin was combined with ampicillin, which targets crosslinking of peptidoglycan strands. Removal of the peptidoglycan of Enterococcus faecium, Staphylococcus aureus, and Bacillus subtilis also resulted in significant protection against daptomycin in comparison to whole cells with intact cell walls. Based on these observations, we conclude that bacterial growth phase and metabolic activity, as well as the presence/absence of peptidoglycan are major contributors to the efficacy of daptomycin.

Anti-Bacterial Agents↗

Glycopeptide resistance in enterococci.

The selective pressure resulting from the extensive use of antibiotics over the last 50 years has led to the emergence of bacterial resistance and to the dissemination of resistance genes among pathogenic microorganisms. Consequently, we are now at serious risk of suffering intractable, life-threatening infections. The progressive emergence and rapid dissemination of resistance to glycopeptides, the last resort for treating nosocomial infections with enterococci resistant to usual antibiotics, constitute one of the most dramatic examples of such resistance. Enterococci are normal human commensals, but are also a frequent cause of nosocomial urinary tract infections and nosocomial bacteremia. Enterococcus faecalis causes 80 to 90% of human enterococcal infections, while Enterococcus faecium accounts for most of the remainder. During the last decade, our understanding of the genetics and biochemical basis of resistance to glycopeptides has increased greatly. Furthermore, the application of molecular methods for the diagnosis of glycopeptide-resistant enterococci has provided new insights into the epidemiology of enterococcal infections.

Animals↗

Absence of synergistic activity between ampicillin and vancomycin against highly vancomycin-resistant enterococci.

The emergence of clinical enterococcal isolates resistant to both ampicillin and vancomycin is a cause of great concern, as there are few therapeutic alternatives for treatment of infections caused by such organisms. We evaluated the effects of the combination of ampicillin with vancomycin against vancomycin-resistant clinical enterococcal isolates. Using both the checkerboard technique and time-kill curves, we examined 28 strains of enterococci (17 Enterococcus faecalis and 11 Enterococcus faecium strains) with different levels of resistance to vancomycin. Of these, 15 strains were also highly gentamicin resistant, and 9 demonstrated resistance to ampicillin. Only seven strains of E. faecalis were inhibited synergistically by the combination of vancomycin with ampicillin, and even then, the concentrations of vancomycin at which synergism was demonstrated were above levels achievable in serum. None of the ampicillin-resistant isolates (all E. faecium) were inhibited synergistically at any concentration of the drugs. In no instance was bactericidal synergism observed, and in most cases the combination resulted in less killing than with ampicillin alone. Antagonism was not observed at clinically relevant concentrations. The results of this study suggest that the combination of vancomycin with ampicillin has little to offer against these emerging pathogens.

Ampicillin↗

A new high-level gentamicin resistance gene, aph(2'')-Id, in Enterococcus spp.

Enterococcus casseliflavus UC73 is a clinical blood isolate with high-level resistance to gentamicin. DNA preparations from UC73 failed to hybridize with intragenic probes for aac(6')-Ie-aph(2'')-Ia and aph(2'')-Ic. A 4-kb fragment from UC73 was cloned and found to confer resistance to gentamicin in Escherichia coli DH5alpha transformants. Nucleotide sequence analysis revealed the presence of a 906-bp open reading frame whose deduced amino acid sequence had a region with homology to the aminoglycoside-modifying enzyme APH(2'')-Ic and to the C-terminal domain of the bifunctional enzyme AAC(6')-APH(2''). The gene is designated aph(2'')-Id, and its observed phosphotransferase activity is designated APH(2'')-Id. A PCR-generated intragenic probe hybridized to the genomic DNA from 17 of 118 enterococcal clinical isolates (108 with high-level gentamicin resistance) from five hospitals. All 17 were vancomycin-resistant Enterococcus faecium isolates, and pulsed-field typing revealed three distinct clones. The combination of ampicillin plus either amikacin or neomycin exhibited synergistic killing against E. casseliflavus UC73. Screening and interpretation of high-level aminoglycoside resistance in enterococci may need to be modified to include detection of APH(2'')-Id.

Anti-Bacterial Agents↗

Evaluation of new streptococcal latex grouping kit.

AIMS: To evaluate a new streptococcal latex grouping kit (Shield Diagnostics Ltd) and compare it against an established latex agglutination method (Streptex; Wellcome Diagnostics). METHODS: Two hundred and forty seven strains of streptococci and enterococci were tested with each kit by one operator and according to the manufacturer's instructions. Strains failing to group or giving discordant results were identified to species level. RESULTS: Two discrepant grouping results were observed and 13 non-beta haemolytic streptococci failed to group with either product. The Shield kit successfully identified 232 isolates at 15 minutes of enzyme extraction incubation compared with 224 and 233 on short (15 minutes) and long (1 hour) incubations, respectively, for Streptex (p > 0.23 for both comparisons). On short incubation only, the Shield kit detected significantly more strains of Enterococcus faecium (p = 0.007). The reaction strengths were similar for both kits (p > 0.16). No cross-reactions were observed but the Streptex kit produced significantly fewer tests with visible granularity (p < 0.003). CONCLUSIONS: Although the Shield product appeared to detect group D antigen more readily, overall no important differences in performance were observed. Prospective users of the new method should first become familiar with its characteristics.

Cross Reactions↗

[beta-lactamase-producing activity and antimicrobial susceptibility of major pathogenic bacteria isolated from clinical samples. Japan beta-lactamase Research Group].

beta-Lactamase production and susceptibility to an assortment of antimicrobial agents were examined in 9,483 strains of organisms isolated from clinical materials obtained from inpatients and outpatients at 104 institutions throughout Japan from December 1999 to February 2000. The organisms were Staphylococcus aureus, 1,369 strains, including 847 methicillin-resistant (MRSA) strains; Enterococcus faecalis, 735 strains; Enterococcus faecium, 302 strains; Moraxella (Branhamella) catarrhalis, 730 strains; Haemophilus influenzae, 1,142 strains; Escherichia coli, 1,276 strains; Klebsiella pneumoniae, 1,058 strains; Enterobacter cloacae, 772 strains; Serratia marcescens, 847 strains; and Pseudomonas aeruginosa, 1,252 strains. The 23 antimicrobial agents used were ampicillin, sulbactam/ampicillin, clavulanic acid/amoxicillin, oxacillin, piperacillin, cefazolin, cefotiam, cefmetazole, cefoperazone, sulbactam/cefoperazone, cefotaxime, ceftazidime, cefepime, cefpodoxime, imipenem, gentamicin, arbekacin, clarithromycin, minocycline, chloramphenicol, vancomycin, teicoplanin, and levofloxacin. Antimicrobial agents appropriate for each organism were used. Among S. aureus strains, 61.9% were MRSA, and 62.3% were positive for beta-lactamase. Among the MRSA strains, none was resistant to vancomycin or teicoplanin, and only 3% were resistant to arbekacin. There was no vancomycin resistance in the Enterococcus strains. Only 0.1% of E. faecalis strains were ampicillin-resistant. Among the M. catarrhalis strains, 97.5% produced beta-lactamase, while among the H. influenzae strains, 8.5% produced beta-lactamase and 14.5% were beta-lactamase-negative and ampicillin-resistant (BLNAR). Among the Enterobacteriaceae and P. aeruginosa strains, there were 20 (E. coli; 7/1,276, K. pneumoniae; 13/1,058) that produced extended-spectrum beta-lactamases (ESBLs), and 11 that produced class B beta-lactamases. Multiple drug resistance was advanced in every species, and organisms resistant to 7 or more common antimicrobial agents were isolated. The best combination of antimicrobial agent and beta-lactamase inhibitor was sulbactam/cefoperazone. Sulbactam/cefoperazone, cefepime, and imipenem still have excellent antimicrobial activity. Rates of resistance to each antimicrobial agent differed more among institutions than among geographical regions.

Bacteria↗

United States geographic bacteria susceptibility patterns. 1997 ASCP Susceptibility Testing Group.

Antimicrobial drug resistance in bacterial pathogens continues, with 1997 seeing reports of Staphylococcus aureus no longer fully susceptible to vancomycin occurring in the United States. To better deal with this rapidly developing problem, we present the third year of United States national data that highlights the geographic nature of increasing resistance to antimicrobial agents. In 1997, we observed increasing numbers of vancomycin-resistant Enterococcus faecium, more methicillin-resistant Staphylococcus aureus (MRSA), and an apparent spreading of penicillin resistance in pneumococci. The majority of reporting sites now indicates that over 20% of Pseudomonas aeruginosa are fully resistant to ciprofloxacin, the only oral agent available for treating this pathogen. With increasing resistance coming at a time of decreasing available resources, it is more clear than ever before that the future of infectious diseases and clinical microbiology remains filled with challenge.

Bacteria↗

Brucellar spondylitis is associated with disturbance in gut microbiota and histamine metabolism associated inflammation.

BACKGROUND: The pathogenesis of brucellar spondylitis (BLS) has traditionally been considered to be primarily limited to local osteoarticular lesions. With the proposal of the "gut-spine axis" concept, the role of intestinal microecological dysbiosis in inflammatory spinal diseases has attracted in an increase of attention. The overactivated inflammatory cytokine network not only mediates bone destruction and intervertebral disc damage, but also forms a bidirectional interaction with gut microbiota dysbiosis through the "gut-spine axis," collectively driving disease progression. However, the inflammatory mechanism by which gut microbiota participates in the pathological process of BLS remains largely unclear. METHODS: This study recruited 20 BLS patients and 20 healthy donors. Multi-omics analysis including metagenomics, untargeted metabolomics, and targeted short-chain fatty acids (SCFAs) analysis, were used to compare the structural differences in gut microbiota between the two groups and screen for signature differential bacterial species. Plasma levels of histamine and histidine decarboxylase were measured by ELISA to clarify the role of differential histidine metabolic pathway in the disease. Additionally, plasma levels of lipopolysaccharide (LPS) and inflammatory cytokines (IL-1&#x3b2;, IL-6, IL-10, IL-17A, TNF-&#x3b1;) were detected by ELISA. The correlation between gut microbiota and inflammatory indicators was further analyzed. RESULTS: Compared to the healthy control group, the &#x3b1;-diversity of the gut microbiota in BLS patients was significantly reduced, with the microbial community structure exhibiting increased homogeneity. Beta diversity analysis revealed significant differences, suggesting that disease progression is associated with an overall imbalance in the gut microbiota and the deterioration of its specific structural composition. At the phylum level, the abundances of Actinomycetota, unclassified_d_Viruses, and Fusobacteriota were significantly increased in the gut microbiota of BLS patients compared to the control group, while the abundances of Bacillota and Pseudomonadota were significantly decreased. Further analysis revealed that, compared to the control group, the generic abundance of Enterococcus was significantly increased, while the proportions of Blautia, Faecalibacterium, Ruminococcus, Agathobacter, Roseburia, Clostridium, Eubacterium, Alistipes and Anaerobutyricum were significantly decreased. At the species level, the abundances of Enterococcus sp and Enterococcus-faecium were increased, whereas Blautia sp, Ruminococcus sp, Faecalibacterium sp, Faecalibacterium prausnitzii, Agathobacter rectalis, Eubacterium sp, Agathobacter sp, and Roseburia sp were decreased. Furthermore, untargeted metabolomics revealed that metabolites were enriched in the histidine metabolic pathway, and the levels of SCFAs including butyrate, isobutyrate, valerate, and 4-methylvalerate in the intestinal contents were reduced in BLS. Functional KEGG profiling revealed that key KOs involved in butyrate synthesis (e.g., K00074, K00172, K01640) and transport were globally downregulated in the patient group, whereas histidine decarboxylase KOs (K01693, K11755, K19787) that convert histidine to pro-inflammatory histamine were significantly enriched. The loss of butyrate-producing symbionts led to SCFAs deficiency and mucosal barrier disruption, creating ecological niches for facultatively anaerobic Enterococcus, which further exacerbated local inflammation via proteolytic fermentation and histamine production. Compared with the control group, BLS patients showed decreased plasma levels of IL-10, while levels of IL-1&#x3b2;, IL-6, IL-17A, and TNF-&#x3b1; were increased, and LPS levels were elevated. In addition, significantly elevated plasma pro-inflammatory LPS levels in patients with BLS suggest disruption of intestinal integrity and permeability. Correlation analysis indicated a close relationship between gut microbiota and inflammation. CONCLUSION: BLS is associated with gut microbiota dysbiosis and alterations in microbial metabolites, which may be linked to inflammatory responses and histamine metabolism. The differential microbial taxa identified in this study could be developed into a stool-based non-invasive diagnostic panel to facilitate early differentiation of BLS from other spinal disorders. Furthermore, restoring gut microbial balance through probiotic supplementation or dietary modulation may represent a promising adjunctive strategy to enhance the efficacy of standard antibiotic therapy and reduce disease recurrence.

Humans↗

The gene encoding the low-affinity penicillin-binding protein 3r in Enterococcus hirae S185R is borne on a plasmid carrying other antibiotic resistance determinants.

Two plasmid-derived NcoI DNA fragments of 14 and 4.5 kb, respectively, have been isolated from the multidrug-resistant strain Enterococcus hirae S185R and analyzed. The 14-kb fragment contains two inverted (L and R) IS1216 insertion modules of the ISS1 family. These modules define a Tn5466 transposon-like structure that contains one copy of the methylase-encoding ermAM conferring erythromycin resistance and one copy of the adenylyl-transferase-encoding aadE conferring streptomycin resistance. Immediately on the left side of IS1216L there occurs a copy of pbp3r encoding the low-affinity penicillin-binding protein (PBP) PBP3r, itself preceded by a psr-like gene (psr3r) that controls the synthesis of PBP3r. ermAM, aadE, and the transposase gene (tnp) of IS1216R have the same polarities, and these are opposite those of psr3r, pbp3r, and the tnp gene of IS1216L. The 4.5-kb fragment is a copy of the 4.5-kb sequence at the 5' end of the 14-kb fragment, although it is not a restriction product of the 14-kb fragment. It contains three genes with the same polarity: psr3r, pbp3r, and tnp in an IS1216 element. Because of the very high degree of identity (99%) with the chromosomal psrfm and pbp5fm genes of Enterococcus faecium D63R, it is proposed that both the psr3r and pbp3r genes were transferred from an E.faecium strain and inserted in a plasmid of E. hirae. E. hirae is the first known bacterial species in which a low-affinity PBP-encoding gene has been found to be plasmid borne.

Bacterial Proteins↗

[Gentamicin resistance of enterococcus genus strains isolated from clinical material].

The occurrence of High Level Gentamicin Resistance (HLGR) was tested among the strains of Enterococcus genus isolated from material potent to microbiological laboratory. The investigation included 184 strains of enterococci, and among them 178 strains of Enterococcus faecalis (96.7%) and 6 strains of Enterococcus faecium (3.3%). HLGR was detected with disks-diffusion method with disks containing 120 micrograms of gentamicin. The screen method with MHA-2 containing 500 mg/l of gentamicin in doubtful results was used. The resistance to high concentration of gentamicin was detected in 29 strains of enterococci (15.76%). The highest percentage of HLGR was proved among strains of enterococci isolated from pyogenic material of hospitalised patients (27.3%).

Anti-Bacterial Agents↗

Daptomycin in vitro susceptibility in European Gram-positive clinical isolates.

The objective of this study was to determine the activity of daptomycin, a novel lipopeptide, against European Gram-positive isolates (n = 1539). The MIC(90)-values of daptomycin against Staphylococcus aureus isolates was 0.25 mg/L, against Enterococcus faecalis 4 mg/L, against Enterococcus faecium 8 mg/L, 0.25 mg/L against Staphylococcus epidermidis, and 0.25mg/L against Streptococcus pneumoniae. Daptomycin was equally potent against antibiotic-susceptible and resistant strains within a particular species. Based on a breakpoint of 1 mg/L for S. aureus and group A streptococci, all isolates tested were susceptible to daptomycin. Based on a breakpoint of 4 mg/L for vancomcyin-susceptible E. faecalis 99.7% of these isolates were susceptible to daptomycin.

Daptomycin↗