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PubMed1992

Restriction fragment length polymorphism analysis of 16S ribosomal DNA of Streptococcus and Enterococcus species of bovine origin.

Twelve bacterial species including Streptococcus uberis, S. parauberis, S. agalactiae, S. dysgalactiae, S. bovis, S. mitis, S. salivarius, S. saccharolyticus, Enterococcus faecium, E. faecalis, E. avium, and Aerococcus viridans were examined for their 16S ribosomal DNA fingerprint patterns. Oligonucleotide primers complementary to 16S rRNA genes were used to amplify by the polymerase chain reaction 16S ribosomal gene fragments from genomic DNAs. The molecular sizes of the amplified 16S ribosomal DNA (rDNA) fragments from the 12 species examined ranged from 1,400 to 1,500 bp. Restriction fragment length polymorphism analysis of 16S rDNA was performed with 11 different restriction endonucleases. All 12 species examined could be differentiated on the basis of characteristic 16S rDNA fingerprint patterns by using the restriction endonucleases HhaI, RsaI, and MspI. A scheme for the differentiation of the 12 species is presented. Eleven isolates representing 11 species were obtained from cows with intramammary infections and were examined by 16S rDNA fingerprinting. All 11 species isolated from cows were differentiated by using HhaI, RsaI, and MspI restriction endonucleases. The results of this study demonstrate the potential application of 16S rDNA fingerprinting for the identification and differentiation of bacterial species.

Animals↗

PubMed1995

Detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant enterococci by PCR.

A PCR assay that allows simultaneous detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant enterococci (Enterococcus faecium, E. faecalis, E. gallinarum, and E. casseliflavus) was developed. This assay was based on specific amplification of internal fragments of genes encoding D-alanine:D-alanine ligases and related glycopeptide resistance proteins. The specificity of the assay was tested on 5 well-characterized glycopeptide-resistant strains and on 15 susceptible enterococcal type strains. Clinical isolates of enterococci that could not be identified to the species level by conventional methods were identified by the PCR test. This assay offers a specific and rapid alternative to antibiotic susceptibility tests, in particular for detection of low-level vancomycin resistance.

Anti-Bacterial Agents↗

PubMed2005

Identification of clinically relevant enterococcus species by direct sequencing of groES and spacer region.

We determined the groESL sequences (groES, groEL, and the intergenic spacer) of 10 clinically relevant Enterococcus species and evaluated the feasibility of identifying Enterococcus species on the basis of these sequences. Seven common clinical Enterococcus species, E. faecalis, E. faecium, E. casseliflavus, E. gallinarum, E. avium, E. raffinosus, and E. hirae, and three less common Enterococcus species, E. cecorum, E. durans, and E. mundtii, were examined in this study. We found that the groES genes of these enterococcal species are identical in length (285 nucleotides) and contain an unusual putative start codon, GTG. The lengths and sequences of the intergenic regions (spacers between the groES and groEL genes) are quite variable (17 to 57 bp in length) among Enterococcus species but are conserved in strains within each species, with only a few exceptions. Considerable variation of groES or groEL sequences was also observed. The evolutionary trees of groES or groEL sequences revealed similarities among Enterococcus species. However, the overall intraspecies variation of groES was less than that of groEL. The high interspecies variation and low intraspecies variation indicate that the groES and spacer sequences are more useful than groEL for identification of clinically relevant Enterococcus species. The sequences of these two genetic traits, groES and spacer, can be determined by a single PCR and direct sequencing and may provide important information for the differentiation of closely related species of Enterococcus.

Bacterial Typing Techniques↗

PubMedDate not supplied

[Characterization of persistence factors of enterococci].

Persistence factors is some species of enterococci (E. faecium, E. faecalis, E. gallinarum), isolated from healthy children and from children with purulent inflammatory diseases, were studied. The study revealed some interspecific and intraspecific differences in the activity of these factors, linked with the presence of purulent inflammatory process induced by enterococci.

Child↗

PubMed2003

[Etiology of purulent infections in the Tiumen' region based on hospital data].

A study of laboratory findings for 474 examined patients, who were discharged from the Surgical Department of the Tyumen Regional Clinical Hospital, showed that S. aureus caused the infectious processes in 31.7% of the examinees. Oxacyllinresistant strain S aureus made up 9.7%.e examinees, and 22.6% of pathologies were caused in them by E. coli. E. coli was frequently found to be combined with polyresistant strains E. faecalis, E. faecium, K. pneumoniae and P. aeruginosa, which could be explained by the circulation of hospital strains within a hospital and by the selection of microorganisms with a high resistance level to antibacterial drugs.

Bacterial Infections↗

PubMed1992

Anaerobic facultative bacteria isolated from the gut of rabbits fed different diets.

Anaerobic facultative bacteria colonizing the intestinal tract of conventional rabbits fed three different diets (standard pellet, hay and pellet/hay mixture) were enumerated in brain heart infusion agar. Colony counts recovered from homogenized samples of small intestine, caecum and rectum differed with reference to the diet given. Among anaerobic groups, identified from rabbit fed pellet/hay mixture, Enterococci (E. faecalis, E. avium, E. faecium and E. durans) represented the predominant flora. Enterobacters (E. cloacae and E. aerogenes) accounted for about 10 to 25% of the bacteria in the rectum and colon respectively, whereas Staphylococci (S. intermedius, S. epidermidis and S. lentus) represented 11% of the bacteria isolated from colon.

Animal Feed↗

PubMed2005

The effectiveness of three instrumentation techniques on the elimination of Enterococcus faecalis from a root canal: an in vitro study.

The in vitro reduction of a bacterial population in a root canal by mechanical instrumentation using three techniques was evaluated. Root canals inoculated with a Enterococcus faecalis (E. faecalis) suspension were instrumented using hand Hedstroem files, Giromatic files, and Hero 642 rotary instruments. Irrigation was performed using sterile saline solution. Root canals were sampled before and after instrumentation. After serial dilutions, samples were plated onto Mitis-Salivarius agar and the colony forming units grown were counted. All instruments tested were able to significantly reduce the number of bacterial cells in the root canal, however, the results of this study indicated that Hedstroem files, Giromatic, and Hero 642 techniques were not significantly different in their ability to reduce intracanal bacteria.

Bicuspid↗

PubMed2000

PD-140248 (Parke-Davis & Co).

PD-140248 is an isomerase inhibitor and topoisomerase II inhibitor under development by Parke-Davis and Co as a potential treatment for bacterial infection. Although no details of active development have been published in the scientific literature since 1996, in September 1999, the company confirmed that PD-140248 is still in active development [338530]. The compound is one of two new pyrrolidinyl naphthyridines (the other compound being PD-131628) with a broad spectrum of antibacterial activity, including activity against quinolone-resistant bacteria. PD-140248 demonstrates strong in vitro activity, particularly against Gram-positive bacteria, such as quinolone-susceptible Staphylococcus aureus (S aureus), Staphylococcus epidermidis (S epidermidis), Staphylococcus haemolyticus (S haemolyticus), methicillin- and ciprofloxacin-resistant S aureus, Streptococcus species, Streptococcus pneumoniae (S pneumoniae) and Enterococcus faecalis (E faecalis). An MIC90 value of 0.015 microg/ml was obtained against Haemophilus influenzae (H influenzae) and Moraxella catarrhalis [160129]. The (3R,1S) enantiomer, PD-140248, demonstrated a 1- to 10-fold enhanced activity against Gram-positive and Gram-negative organisms in vitro and in vivo compared to its stereoisomers, and has thus been targeted for further preclinical studies [133377].

Journal Article↗

PubMed1989

[Bactericidal effect of daptomycin (LY 146032) compared with vancomycine and teicoplanin against gram-positive bacteria].

Daptomycin (LY 146032) is a semi-synthetic antibiotic whose structure consists of a cyclic peptide with a lipophilic chain. It inhibits synthesis of the bacterial wall differently than vancomycin and teicoplanin. The bactericidal activity of daptomycin was compared with that of vancomycin and of teicoplanin with regard to 54 Gram positive strains of bacteria: (S. aureus meti S and meti R; S. epidermidis meti S and meti R; E. faecalis, E. faecium, and Corynebacterium group JK). Vancomycin and teicoplanin have the same slow and time-dependent bactericidal activity. The activity of daptomycin is different: rapid and concentration-dependent, similar to that of the aminoglycosides. In the case of the Corynebacteria, however, the activity of daptomycin is comparable to that of vancomycin and of teicoplanin slow and time-dependent.

Anti-Bacterial Agents↗

PubMed1986

[Bacterial and clinical studies on imipenem/cilastatin sodium in urological infection].

Antibacterial activities of imipenem (MK-0787), cefazolin (CEZ), cefmenoxime (CMX) and cefoperazone (CPZ) were measured by using an agar dilution method against 243 strains of bacteria isolated from the patients with urinary tract infections. The MIC80 of MK-0787 against S. epidermidis, E. faecalis, E. coli, K. pneumoniae, E. cloacae, S. marcescens, P. mirabilis, P. vulgaris and P. aeruginosa was less than or equal to 0.10, 3.13, less than or equal to 0.10, 0.20, 0.78, 1.56, 3.13, 3.13 and 6.25 micrograms/ml, respectively. MK-0787 was more active than CEZ, CMX and CPZ against all bacterial species except Proteus spp. Imipenem/cilastatin sodium (MK-0787/MK-0791) 500 mg/500 mg two times a day for 5 days was given to 31 cases of chronic complicated urinary tract infections and a case of acute prostatitis. Overall clinical effectiveness rate judged by the criteria proposed by the Japanese UTI committee in 22 cases of chronic complicated urinary tract infection treated with MK-0787/MK-0791 was evaluated to be 86% including excellent in 6 cases, moderate in 13 cases and poor in 3 cases. As side effects, 2 cases encountered eruption or anorexia. Abnormal laboratory findings were observed in 6 cases.

Adult↗

PubMed1986

[Fundamental and clinical studies of imipenem and imipenem/cilastatin sodium in the field of obstetrics and gynecology].

Fundamental and clinical studies of imipenem (MK-0787), a new type of carbapenem antibiotic, and MK-0787 combined with cilastatin sodium (MK-0791), a renal dipeptidase inhibitor, were carried out. The results obtained were as follows: MK-0787 500 mg alone or MK-0787 500 mg with MK-0791 500 mg was administered by intravenous drip infusion over 30 minutes. Plasma levels of the drug were similar either following the administration of 500 mg of MK-0787 alone or 500 mg of MK-0787 with 500 mg of MK-0791. When MK-0787 was administered with MK-0791, MK-0787 and MK-0791 levels at 2 hours after the end of infusion in uterine arterial plasma were 6.8 micrograms/ml and 3.2 micrograms/ml, respectively, and in venous plasma were 8.4 micrograms/ml and 4.7 micrograms/ml, respectively. MK-0787 tissue levels ranged from 0.8 microgram/g to 3.8 micrograms/g at 205 minutes after the end of infusion. Based on these results, the plasma and tissue levels of MK-0787 and MK-0791 with b.i.d. dosage exceeded the MICs of the drug against clinical isolates in the field of obstetrics and gynecology such as E. faecalis, E. coli Klebsiella sp., Peptococcus sp., Peptostreptococcus sp. and B. fragilis immediately after the administration. However, it seemed that the b.i.d. dosage was insufficient to maintain the in vivo concentration of these agents high enough to inhibit the growth of the above bacteria. Eighteen patients with obstetric and gynecologic infection (12 with intrauterine infections, 2 with pelvic dead space inflammation, 2 with pelvic peritonitis, 1 with a vaginal cuff abscess and 1 with a vulvar abscess) and 1 patient with other infection (abdominal wall abscess) were evaluated, but 1 patient with pelvic peritonitis was later excluded from the efficacy evaluation because of a serious illness. MK-0787/MK-0791 was administered twice daily in a 30-minute intravenous drip infusion. Clinical results were excellent in 1 patient, good in 16 and poor in 1, for an efficacy ratio of 94.4%. No side effects were observed. Only abnormal laboratory findings observed were elevation of S-GOT and S-GPT in 1 patient which normalized 2 weeks after the treatment was discontinued. These results suggest that MK-0787/MK-0791 will be useful for the treatment of obstetric and gynecologic infections.

Adult↗

PubMed1994

[Method of DNA extraction and mecA, femA and PBP2' in multi-drug resistant bacterial strains].

Recently, it has become more important to establish rapid and reliable methods for identifying bacterial species and their drug resistances because of the high incidence of nosocomial infection caused by multi-drug resistant bacteria. At first, several enzymes were tested to evaluate the efficacy of DNA extraction. N-acetylmuramidase, lysozyme and achromopeptidase were most effective to extract DNA from E. faecalis, E. faecium and S. aureus, respectively. After achromopeptidase extraction, the mecA gene was amplified by polymerase chain reaction (PCR) in clinically isolated MRSA (95 strains), MSSA (66), MRSE (methicillin-resistant S. epidermidis, 37), MSSE (methicillin-sensitive S. epidermidis, 1), S haemolyticus(5), S.hominis(1) and others (Enterococcus, Pseudomonas, and Streptococcus, total 45). PCR products were analyzed by agarose gel electrophoresis. The positive rates were 95% (MRSA), 1.5% (MSSA), and 97% (MRSE). The mecA gene was also positive in 2/2 of methicillin-resistant S.haemolyticus and 2/3 of methicillin-sensitive S.haemolyticus and 1/1 of methicillin-resistant S.hominis. The mecA gene was not detected in 45 non-Staphylococcal strains. MecA and femA gene by PCR and PBP 2' by IRMA were further detected in newly isolated MRSA (20 strains), MSSA(20), MRSE(14), MSSE(1) and S.simulans(2). Complete correlation between MPIPC susceptibility and mecA were found. The femA gene was positive in 39/40 of S. aureus, and 0/14 of S.epidermidis, 2/3 of S.simulans. PBP2' were positive in 20/20 of MRSA, 0/20 of MSSA, 14/14 of MRSE, 0/1 of MSSE, 1/1 of methicillin-resistant S.simulans, and 0/2 of methicillin-sensitive S.simulans. In conclusion, diagnoses of MRSA by simultaneous detection of mecA and femA gene as well as PBP2' are similarly useful because of their specificity and rapidity.

Bacterial Proteins↗

PubMed1997

Synthesis and activities of 5-substituted-2-(p-substituted phenyl)-1-dialkylaminomethyl benzimidazole derivatives.

Nine 1,2,5-trisubstituted benzimidazole derivatives were prepared and their structure have been elucidated by IR, NMR spectral data and elemental analyses. Analgesic activity of the compounds prepared was investigated in mice by modified KOSTER test. Anti-inflammatory activity of these compounds was investigated by a carregeenan-induced hind paw edema model in mice. Their antibacterial activities were examined against S. aureus, E. faecalis, E. coli, P. aeruginosa, and antifungal activity against three kinds of yeast-like fungi (C. albicans, C. parapsilosis, C. stellatoidea).

Analgesics, Non-Narcotic↗

PubMed1998

[Detection of bacterial biofilm on medical biomaterials].

This study was performed to assess the value TTC assay in the diagnosis of biomaterial-associated infections. In this assay, soluble colourless TTC is reduced to insoluble red formazan by electron transfer associated with active oxidative bacterial metabolism and is precipitated intracellularly. Microbial adhesion and biofilm formation on the surface of medical prosthetic devices (vesicular and urinary catheters) made of various polymers (PTFE, H-PE, PCW, SL), were determined. The microorganisms which are most often isolated in medical device-associated infections: S. aureus, S. epidermidis, E. faecalis, E. coli, P. vulgaris, P. aeruginosa, C. albicans, were included into the study. The obtained results indicate that the assay using TTC as a metabolic indicator of bacterial biofilm presence, is technically simple to conduct with minimal setup time. Even when classical cultures yielded no bacterial growth, TTC assessments demonstrated bacterial biofilms. TTC assay could be recommended as a quick routine method for confirmation of biomaterial device-associated infection.

Animals↗

PubMed1998

[Enterococci as uropathogens. Frequency of isolation and sensitivity to antibacterial agents].

221 clinically significant enterococcal strains (191: E. faecalis, 27: E. faecium, 3 others) were isolated from the urine of patients hospitalised with UTI over an 18-month period (1995-1996). The susceptibility of the isolates to 8 antimicrobial agents was determined by agar dilution method (NCCLS). All enterococci were sensitive to vancomycin and most of them (over 92%) to penicillin and ampicillin. Only 3.14% of E. faecalis and 3.70% of E. faecium were resistant to ciprofloxacin. A relatively high incidence of resistance of enterococci to aminoglycosides was observed: 46.07% and 51.85% to streptomycin, 42.41% and 44.44% to gentamicin, 70.68% and 77.78% to amikacin for E. faecalis and E. faecium, respectively. Among the resistant to penicillin and amino glycosides enterococci of the two species was found a great percent "high level" resistance. There were no beta-lactamase producers among our strains. The established multiresistance accompanied by "high level" resistance requires careful consideration of antimicrobial therapy of enterococcal UTI.

Adult↗

PubMed2005

Detection of the esp gene in high-level gentamicin resistant Enterococcus faecalis strains from pet animals in Japan.

We investigated the prevalence of the esp gene and the susceptibility to gentamicin in Enterococcus faecalis and E. faecium strains obtained from pet animals. Nine of 30 E. faecalis and 2 of 38 E. faecium strains from the pet animals had the esp gene. Three esp-positive E. faecalis strains, which were isolated from two dogs and a cat, showed gentamicin MICs of > or =256 microg/ml and harbored the high-level gentamicin resistance (HLGR) gene, aac(6')-Ie-aph(2'')-Ia. Of the nine esp-positive E. faecalis strains, five, including the three strains with the HLGR gene, were closely related by numerical analysis of PFGE patterns. Longitudinal investigation needs to elucidate whether the HLGR gene was incorporated into a subpopulation of the esp-positive E. faecalis.

Animals↗

PubMed2001

Implications of multiple antimicrobial-resistant enterococci associated with the poultry environment.

Poultry are increasingly being associated with carriage of multiresistant organisms that may cause disease in humans. Among these organisms are the enterococci, not regarded as a common cause of hospital-acquired infections. The use of antimicrobials for growth promotion in poultry production envronments may facilitate the dissemination of resistance to Enterococcus spp. that have the potential to be clinically significant. To assess descriptively the degree of multiresistant enterococci in the poultry environment of the Delmarva (Delaware-Maryland-Virginia) East Coast region of the USA, litter samples from regional commercial poultry houses and transport container swabs from processing facilities were cultured for Enterococcus spp. Using a microtiter plate adaptation of a conventional biochemical screen, the predominant species identfied were E. faecalis (61.2%), E. faecium (18.6%) and E. gallinarum (2.4%). Resistance to the cephalosporin, macrolide and tetracycline classes of antimicrobials was uniform with broader resistance to penicillin and derivatives present in a majority of E. faecium isolates. High-level streptomycin resistance was evident in close to 30% of all isolates with a majority of E. faecalis variants possessing resistance. High-level gentamicin resistance was detected at a low frequency (2.6%) only within the E. faecalis group with resistance to low-level gentamicin levels present in a majority of both the E. faecalis group and subsets of E. faecium. No unexpected vancomycin resistance was detected. Of particular interest was resistance to the streptogramin quinupristin-dalfopristin (Q-D or Synercid), which was present in 70.4% of E. faecium and E. faecium variants.

Animals↗

PubMed2006

Detection of genes encoding virulence factors and bacteriocins in fecal enterococci of poultry in Portugal.

Seventy-six Enterococcus isolates (43 E. faecalis, 30 E. faecium, two E. durans, and one E. hirae) recovered from fecal samples of poultry in a slaughterhouse (one isolate per fecal sample and one fecal sample per lot of animals) were studied for bacteriocin production and for the presence of genes encoding bacteriocins and virulence factors. The presence of genes encoding virulence factors (cpd, geE, fsr, ace, agg, and esp) and bacteriocins (entA, entB, entP, entQ, entAS-48, entL50A/B, cyl, and bac31) were studied by polymerase chain reaction in all enterococci. At least two virulence genes were detected in all 43 E. faecalis isolates, cpd and gelE being the most frequently detected genes (97.7%) followed by ace (62.8%), agg (39.5%), fsr (27.9%), and esp (2.3%). No virulence genes were detected in the other enterococcal species with the exception of one E. faecium and one E. durans isolates that harbored the gelE gene. Antimicrobial activity against eight indicator bacteria (including Listeria monocytogenes) was assayed in the enterococci, and 23 (30.3%) showed inhibitory activity against L. monocytogenes, the other 22 enterococci showing activity against indicator bacteria other than L. monocytogenes. Only the entA, entB, and cyl genes were detected in our study (entA + entB in nine E. faecium isolates and the cyl gene in seven E. faecalis isolates). A wide variety of virulence genes have been detected in fecal E. faecalis isolates from poultry, but not in the other enterococcal species. However, the presence of known bacteriocin structural genes is associated more with the E. faecium species.

Animals↗