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Biomedical subjects

B E Murray

Publications and source records attributed to B E Murray.

At least 73 records · Page 4Linked to original sources

Chromosomal DNA restriction endonuclease digestion patterns of beta-lactamase-producing Enterococcus faecalis isolates collected from a single hospital over a 7-year period.

Twenty-three beta-lactamase (beta-lac)-producing, highly gentamicin-resistant Enterococcus faecalis isolates collected over a 7-year period from the same hospital were examined by pulsed-field gel electrophoresis of SmaI-digested genomic DNA. The beta-lac+ isolates appeared to form a single clonal group, which had been previously designated the mid-Atlantic pattern. Eleven variations of the mid-Atlantic clone, differing by one to six bands, were identified; some of the changes were likely due to plasmid bands. However, a number of isolates had indistinguishable patterns, including some recovered over a 4-year period. There was a surprising lack of movement of the beta-lac determinant to other strains, although this trait was transferable in vitro by conjugation. We conclude that a single clone (the mid-Atlantic clone) of beta-lac+ E. faecalis has remained endemic in this hospital for at least 7 years. The reason(s) for the apparent lack of spread to other strains of E. faecalis is unknown.

Anti-Bacterial Agents↗

High-level resistance to aminoglycosides: comparison of community and nosocomial fecal isolates of enterococci.

Fecal carriage of enterococci highly resistant to streptomycin, gentamicin, and kanamycin was examined in 64 healthy volunteers with no exposure to hospitals and in 53 hospitalized individuals. High-level resistance to streptomycin and gentamicin was found in fecal specimens from 3% and 0, respectively, of the healthy volunteers and in fecal specimens from 41% and 15%, respectively, of the hospitalized individuals. We found that high-level resistance to kanamycin was also more common among hospitalized individuals than among healthy volunteers (36% vs. 17%). The frequent occurrence of high-level resistance to kanamycin in fecal isolates confirms that amikacin is a poor choice when attempting to achieve synergistic therapy for enterococcal infections, in particular for those infections that are nosocomially acquired.

Base Sequence↗

Incidence of hemolysin, gelatinase, and aggregation substance among enterococci isolated from patients with endocarditis and other infections and from feces of hospitalized and community-based persons.

The presence of hemolysin, gelatinase, and aggregation substance (by use of a probe known to hybridize to most pheromone-responsive plasmids) was determined in 192 isolates of Enterococcus faecalis from patients with endocarditis or other infections and fecal isolates from hospitalized patients or healthy volunteers, and in 86 non-E. faecalis isolates. Hemolysin was more common in nonendocarditis clinical isolates and in hospital fecal isolates (37% and 31%, respectively) than among endocarditis and community fecal isolates (16% and 20%, respectively). Gelatinase and aggregation substance, respectively, were found in 54% and 52% of isolates from endocarditis, in 58% and 72% of isolates from other infections, in 62% and 56% of hospital fecal isolates, and in 27% and 30% of fecal isolates from healthy volunteers. All 86 non-E. faecalis enterococcal isolates were negative for these traits. The absence of hemolysin, gelatinase, or the aggregation substance gene in > 45% of endocarditis E. faecalis isolates suggests that while these traits may play a role in virulence, other properties are also important.

Bacterial Proteins↗

Generation of auxotrophic mutants of Enterococcus faecalis.

A 22-kb segment of chromosomal DNA from Enterococcus faecalis OG1RF containing the pyrimidine biosynthesis genes pyrC and pyrD was previously detected as complementing Escherichia coli pyrC and pyrD mutations. In the present study, it was found that the E. faecalis pyrimidine biosynthetic genes in this clone (designated pKV48) are part of a larger cluster resembling that seen in Bacillus spp. Transposon insertions were isolated at a number of sites throughout the cluster and resulted in loss of the ability to complement E. coli auxotrophs. The DNA sequences of the entire pyrD gene of E. faecalis and selected parts of the rest of the cluster were determined, and computer analyses found these to be similar to genes from Bacillus subtilis and Bacillus caldolyticus pyrimidine biosynthesis operons. Five of the transposon insertions were introduced back into the E. faecalis chromosome, and all except insertions in pyrD resulted in pyrimidine auxotrophy. The prototrophy of pyrD knockouts was observed for two different insertions and suggests that E. faecalis is similar to Lactococcus lactis, which has been shown to possess two pyrD genes. A similar analysis was performed with the purL gene from E. faecalis, contained in another cosmid clone, and purine auxotrophs were isolated. In addition, a pool of random transposon insertions in pKV48, isolated in E. coli, was introduced into the E. faecalis chromosome en masse, and an auxotroph was obtained. These results demonstrate a new methodology for constructing defined knockout mutations in E. faecalis.

Base Sequence↗

Biochemical fingerprinting compared with ribotyping and pulsed-field gel electrophoresis of DNA for epidemiological typing of enterococci.

The Phene Plate (PhP) biochemical fingerprinting system for bacteria is based on measurements of the kinetics of bacterial biochemical reactions. This system was modified for typing of enterococci and was compared with DNA typing by pulsed-field gel electrophoresis and with ribotyping by using 45 Enterococcus faecalis isolates from international collections. It was also used to study 170 fecal enterococcal isolates from healthy individuals and 28 isolates of E. faecalis from the blood of neonates. The PhP system showed a high degree of discriminatory power for unrelated enterococcal isolates. Among the 170 unrelated fecal isolates, 107 isolates from international collections, PhP typing discriminated 19 types, and ribotyping discriminated 5 types. In most cases, when isolates were of the same DNA type, they were also of the same PhP type, and the level of agreement between these two methods was high (96%). A combination of PhP typing and DNA typing identified 34 different types, but ribotyping did not yield any further discrimination. PhP typing of E. faecalis isolates from healthy individuals (n = 89) and from the blood of neonates with septicemia (n = 28) yielded a diversity of 0.93 for both populations and similar major PhP types in both populations. Thus, the isolates from blood seemed to consist of a normal E. faecalis population, without a dominance of certain strains associated with virulence. We conclude that the PhP system is useful for epidemiological studies of enterococcal isolates, yielding results similar to those obtained with DNA typing by pulsed-field gel electrophoresis. Since PhP typing is a method that is simple and rapid and that is based on automatic evaluation of the data, it is suitable for analyzing large numbers of isolates and can be used alone or in combination with DNA typing or epidemiological and ecological studies of enterococci.

Bacterial Typing Techniques↗

Analysis of Enterococcus faecalis isolates from intercontinental sources by multilocus enzyme electrophoresis and pulsed-field gel electrophoresis.

Pulsed-field gel electrophoresis (PFGE) and multilocus enzyme electrophoresis (MLEE) were compared in this study of 65 Enterococcus faecalis isolates recovered over a 20-year period from diverse geographic sources. Clonal relationships recognized by PFGE were also recognized by MLEE; however, MLEE recognized as greater number of isolates as belonging to clonal groups than did PFGE. Both techniques were reproducible and discriminatory, but MLEE more readily recognized relationships among large numbers of isolates. MLEE confirmed the previously reported clonal spread of beta-lactamase-producing E. faecalis to six hospitals in five states. MLEE provided a useful population framework of the E. faecalis isolates in this sample, while PFGE was able to differentiate among isolates within some MLEE clonal groups.

Alleles↗

DU-6859a, a new fluoroquinolone agent. Comparative in vitro activity against enteric pathogens and multiresistant outpatient Escherichia coli.

The activity of DU-6859a, a new fluoroquinolone antimicrobial agent, was compared with that of ciprofloxacin by agar dilution susceptibility testing against enteric pathogens and multiresistant Escherichia coli. The results indicate that DU-6859a inhibits most of these organisms at concentrations similar to those of ciprofloxacin. DU-6859a showed increased activity compared to ciprofloxacin against Campylobacter species isolates.

Anti-Infective Agents↗

Recurrent group B streptococcal infections in infants: clinical and microbiologic aspects.

OBJECTIVE: To describe the potential for recurrence of group B streptococcal (GBS) infection in infants, using pulsed-field gel electrophoresis as an epidemiologic tool. DESIGN: Retrospective review of cases identified by laboratory records and review of the literature. SETTING: Neonatal nurseries of a county hospital system. METHODS: Retrospective review of infants with second episodes of GBS bacteremia or meningitis. Digestion of chromosomal DNA with the restriction enzyme Sma I and separation of fragments by use of contour-clamped homogeneous electric field. RESULTS: Nine cases of recurrent GBS infection were identified during a 14-year period. Eight of the nine infants were born at 25 to 36 weeks of gestation, and one was born at term. The first episode of invasive GBS infection occurred at a mean age of 10.4 days (median, 3 days; range, 1 to 27 days). Parenteral antibiotic therapy was administered for a mean of 13.9 days (median, 14 days; range, 10 to 21 days). Recurrence occurred at a mean age of 42.3 days (median, 48 days; range, 23 to 68 days). One patient died during the second episode; eight infants survived to discharge home. Of seven sets of isolates analyzed from first and second GBS episodes, five were confirmed to be the same genotypically. CONCLUSION: Recurrence of GBS disease in infants may be associated with the original infecting strain or a second acquired strain.

Anti-Bacterial Agents↗

Revised estimates of enterococcal chromosomal sizes.

We previously reported that the chromosomal sizes of four strains of enterococci ranged from 2,045 to 2,761 kb. Extensive analysis and mapping subsequently confirmed the size of Enterococcus faecalis strain OG1 as 2,825 kb (prior size estimate range, 2,750-2,761 kb) (Murray et al., J. Bacteriol. 175, 5216, 1993). However, using variable conditions of electrophoresis and additional digestions, revised size estimates for the other strains are 2,852-3,093 kb for E. faecalis strain JH2-2 (prior range, 2,008-2,135 kb), 2,910-3,065 kb for E. faecalis strain HH67 (prior range, 2,170-2,288 kb), and 2,334-2,558 for E. faecium strain GE-1 (prior range, 2,045-2,155 kb). The earlier underestimations of the chromosomal sizes were due to the inconsistent presence of a large fragment, likely caused by shearing of the DNA during handling, causing it to be considered a partial digestion product, and failure to resolve multiple fragments of the same approximate size.

Chromosomes, Bacterial↗

Comparative in vitro activity of DU-6859a, a new fluoroquinolone agent, against gram-positive cocci.

The in vitro activity of DU-6859a (DU), a new fluoroquinolone agent, was evaluated against 233 gram-positive cocci and was compared with those of ciprofloxacin, vancomycin, nafcillin, and ampicillin. The MICs of DU for 90% of the staphylococci tested were < or = 0.06 microgram/ml. All of the groups A and B and viridans group streptococci were inhibited by < or = 0.125 microgram of DU per ml, which was 32-fold more active than ciprofloxacin. On the basis of MICs for 90% of the strains tested, DU was 32- and 16-fold more active than ciprofloxacin against Enterococcus faecalis and Enterococcus faecium, respectively. The bactericidal activity of DU was demonstrated by time-kill techniques against all ciprofloxacin-susceptible enterococci. DU shows promise for the treatment of infections with gram-positive cocci and warrants further evaluation by in vitro and in vivo studies.

Anti-Bacterial Agents↗

Intracellular activity of azithromycin against bacterial enteric pathogens.

Azithromycin, a new azalide antibiotic, is active in vitro against a variety of enteric bacterial pathogens. Since it is concentrated inside human neutrophils and other cells, it might be particularly useful in the treatment of infections caused by enteropathogens that invade host tissues. The intracellular activity of azithromycin against several enteric pathogens that had been phagocytosed by neutrophils was determined. Azithromycin was effective in reducing the intracellular viabilities of almost all strains tested, including representative strains of Salmonella, Shigella, and enteroinvasive, enteropathogenic, enterotoxigenic, and enterohemorrhagic Escherichia coli. Erythromycin was also effective in this model system, although azithromycin was generally more effective than erythromycin against strains of invasive enteric pathogens. Cefotaxime reduced intracellular bacterial viability to a lesser extent than either azithromycin or erythromycin. The presence of neutrophils did not significantly affect the activity of azithromycin in this system. Azithromycin may be a useful agent for the treatment of bacterial diarrhea, and clinical trials should be considered.

Azithromycin↗