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PubMed1997

Multiplex PCR detection of vanA, vanB, vanC-1, and vanC-2/3 genes in enterococci.

Vancomycin-resistant enterococci (VRE) are increasingly isolated from clinical specimens. One hundred clinical isolates of enterococci (E. casseliflavus/E. flavescens [n = 10], E. faecalis [n = 34], E. faecium [n = 43], E. avium [n = 1], E. gallinarum [n = 11], and E. raffinosus [n = 1]) were examined for the presence of vanA, vanB, vanC-1, and vanC-2/3 genes by a single multiplex PCR performed directly with colonies from blood agar plates. Six previously characterized VRE strains which carry either vanA, vanB, vanC-1, or vanC-2 genes were used as controls. To discriminate among van genes, the PCR amplicons were digested with MspI and were electrophoresed on agarose gels. Because of significant sequence homology between vanC-2 and vanC-3 genes, this assay is unable to discriminate these genes from each other; therefore, these are referred to as vanC-2/3 genes. PCR products were detected in 63 of the 100 clinical isolates. The restriction fragment length patterns were consistent with vanA for 10 strains, vanB for 30 strains, vanC-1 for 12 strains, vanC-2 for 6 strains, and vanA and vanC-1 for 1 strain. The vancomycin MICs for the isolates with restriction fragment length patterns consistent with vanA and vanB were all > and = 64 micrograms/ml. The vancomycin MICs for the isolates with restriction fragment length patterns consistent with vanC-1 or vanC-2 were 4 to 8 micrograms/ml. The vancomycin MICs for the isolates from which no PCR amplicons were produced were 2 to 4 micrograms/ml. A PCR product was produced in four isolates (vancomycin MICs, 4 to > 256 micrograms/ml) with restriction fragment length patterns differing from those for the control vanA, vanB, vanC-1, and vanC-2 isolates. DNA sequencing of these amplicons revealed that two of the four isolates had nucleic acid sequences which were closely related to the published sequence for the vanB gene and two had nucleic acid sequences which were closely related to the published sequence for the vanC-2 and vanC-3 genes. Multiplex PCR-restriction fragment length polymorphism appears to be a useful and convenient method for rapidly detecting and discriminating genotypes for vancomycin-resistant Enterococcus spp. in the clinical laboratory. In instances in which unusual restriction fragment patterns of PCR amplicons occur, DNA sequencing can be performed to discriminate van genotypes.

Amino Acid Sequence↗

PubMed1992

In vitro activity of Ro 23-9424, a dual-action antibacterial agent, against bacterial isolates from cancer patients compared with those of other agents.

The in vitro activity of Ro 23-9424 against bacterial isolates from patients with cancer was compared with those of fleroxacin, ciprofloxacin, cefoperazone, and ceftazidime. Ro 23-9424 inhibited the majority of the members of the family Enterobacteriaceae and all Aeromonas isolates at a concentration of less than or equal to 1.0 micrograms/ml. It was also active against Acinetobacter spp. and Haemophilus influenzae, including beta-lactamase-producing strains. The MIC for 90% of isolates (MIC90) of Pseudomonas aeruginosa was 16.0 micrograms/ml. All group A and B streptococci were inhibited by less than or equal to 0.25 micrograms/ml, and 90% of group G streptococci and Streptococcus pneumoniae were inhibited by 1.0 micrograms/ml. All methicillin-susceptible strains of Staphylococcus aureus and 60% of methicillin-resistant strains were susceptible to 2.0 micrograms of Ro 23-9424 per ml, whereas the MIC90 for Staphylococcus epidermidis and Staphylococcus hominis isolates was 4.0 micrograms/ml. Staphylococcus haemolyticus and Enterococcus spp. were less susceptible; MIC90s for them were 16.0 and 32.0 micrograms/ml. Ro 23-9424 has a broad antibacterial spectrum and potential utility for therapy of infections in cancer patients.

Anti-Infective Agents↗

PubMed1997

Clinical comparison of BACTEC 9240 plus aerobic/F resin bottles and the isolator aerobic culture system for detection of bloodstream infections.

The Plus Aerobic/F resin bottle of the BACTEC 9240 automated blood culture system (Becton Dickinson Diagnostic Instrument Systems, Sparks, Md.) was compared with aerobic culture of the Isolator system (Wampole Laboratories, Cranbury, N.J.) for the detection of bloodstream microorganisms from 6,145 blood cultures collected from adult patients with suspected septicemia. The BACTEC resin bottles were incubated for 7 days, and the sediment from the Isolator tube was inoculated to sheep blood and chocolate agars which were incubated for 72 h and to inhibitory mold, brain heart infusion, and Sabouraud agars which were incubated for 21 days. A total of 622 microorganisms were recovered from 583 blood cultures. The BACTEC resin bottle recovered statistically significantly more pathogens overall than the Isolator system (P = 0.0006). When individual pathogens isolated from either system for a 7-day study period were assessed, it was determined that the BACTEC resin bottle detected statistically significantly more isolates of Staphylococcus aureus (P = 0.0113) and coagulase-negative Staphylococcus spp. (P = 0.0029) than the Isolator system. The BACTEC resin bottle also detected statistically significantly more bloodstream infections (septic episodes) caused by coagulase-negative Staphylococcus spp. (P = 0.0146). The Isolator system recovered statistically significantly more contaminants overall (P < 0.0001), and among this group of microorganisms, recovered statistically significantly more Bacillus spp. (P < 0.0001), coagulase-negative Staphylococcus spp. (P < 0.0001), and viridans group Streptococcus spp. (P = 0.0156). The Isolator system detected statistically significantly more isolates of Histoplasma capsulatum (P = 0.004), but all of these isolates were detected at > or = 7 days of incubation of fungal plates, i.e., after the system to system comparison study period (7 days). In blood culture sets which produced growth of the same pathogen in both systems, there was a statistically significant difference in median time to detection for all pathogens combined favoring the BACTEC resin bottle over the Isolator tube (P < 0.05). When assessing individual microorganisms, the median times for detection of S. aureus, Enterococcus spp., and Pseudomonas spp. were all statistically significantly less for the BACTEC system (P < 0.05). The BACTEC instrument had 79 (1.3%) false positive signals. The BACTEC system required less processing time than the Isolator system and eliminates the hands-on time for detection of positive cultures required with the Isolator system.

Adult↗

PubMed1995

Emerging multiply resistant enterococci among clinical isolates. I. Prevalence data from 97 medical center surveillance study in the United States. Enterococcus Study Group.

To assess the evolving problem of therapeutic drug resistances among enterococci, we organized a comprehensive national (United States) surveillance trial using 99 recruited microbiology laboratories in 48 of the 49 contiguous states or districts. All but two sites completed the protocol that generated information from nearly 2000 enterococci, usually isolated from blood cultures. All strains were speciated by the same method (API 20S) and were susceptibility tested by three methods (broth microdilution, disk diffusion, and Etest) against ampicillin, penicillin, vancomycin, teicoplanin, gentamicin, and streptomycin. Strains resistant to a glycopeptide or penicillin, or possessing high-level aminoglycoside resistance were referred to the monitor's laboratory for validation and additional susceptibility testing against other alternative antimicrobial agents. The most common species were Enterococcus faecalis and Enterococcus faecium. However, antimicrobial resistance occurred most often among the E. faecium isolates. Twenty-three percent of participant centers (22 sites) reported 87 vancomycin-resistant isolates, which accounts for 4.4% of the isolates evaluated. A recent audit (March 1994) of the laboratories not reporting vancomycin resistance during the study interval (October-December 1992) revealed that 61% of sites have now recognized these strains, a threefold increase in 12-15 months. Teicoplanin remained active against 28% (Van B phenotype) of vancomycin-resistant enterococci (10 E. faecalis, 13 E. faecium, and one Enterococcus spp.). Ampicillin-resistant beta-lactamase-positive strains were found only at one medical center (two strains, 0.2% of referred or validated strains); however, ampicillin-resistant strains represented 12% of all enterococcal, but nearly 60% of E. faecium strains.(ABSTRACT TRUNCATED AT 250 WORDS)

Anti-Bacterial Agents↗

PubMed2004

Specific enumeration of the probiotic strain Enterococcus faecium NCIMB 10415 in the intestinal tract and in faeces of piglets and sows.

The intestinal bacterium Enterococcus faecium NCIMB 10415 (E. faecium SF68) has been used for more than a decade as a probiotic strain in animal nutrition as well as in the prevention and treatment of diarrhoea in humans. Beneficial effects have been shown in feeding and clinical trials. However, the strain has no selective growth markers and monitoring in the intestinal tract is impossible by cultivation. Using specific nucleotide sequences, in this study a probe for colony hybridization was constructed in order to quantify this probiotic strain in feed and intestinal and faecal samples from piglets and sows. The probiotic strain showed almost constant amounts in sow faeces (1.8 x 10(5) cfu/g wet weight), while contents in digesta and piglet faeces varied on a lower level depending on gut section and piglet age. The ratio of specific probiotic counts and total enterococci was much lower than in sow faeces however the strain could be detected reliably in faeces already on the 14th day of life. The application of the colony hybridization method enables for the first time the selective detection of the widely used probiotic E. faecium NCIMB 10415 strain among total Enterococcus spp. counts of digesta, faeces and feed. It is now possible to monitor the presence of the probiotic in the intestinal tract and faeces. Results of this study have implications for the proposed modes of action of probiotics in animal nutrition.

Age Factors↗

PubMed1993

Species identification and determination of high-level aminoglycoside resistance among enterococci. Comparison study of sterile body fluid isolates, 1985-1991.

Enterococcus spp. have become the third most common cause of nosocomial infections. High-level aminoglycoside resistance (HLR), an important clinical concern, has been associated with some species of the enterococci. We evaluated the Vitek and API 20S systems for species identification and the Vitek for the detection of HLR. Enterococci from nosocomial infections (208 strains) at the University of Iowa Hospital (1985-1991) were tested by Vitek, API 20S, and reference methods. The error rate for species identification was 6.7% for the API 20S and 5.8% for the Vitek Gram-positive identification (GPI) cards. Both systems tended to incorrectly identify other enterococcal species as Enterococcus faecium. HLR was found in Enterococcus faecalis and E. faecium isolates only. The highest rates of HLR to streptomycin alone (17.9%) and with gentamicin (13.5%) was observed among E. faecalis strains, and to gentamicin alone (7.3%) was found among E. faecium isolates. No apparent differences in HLR rates were found from year-to-year over the 7-year enterococcus sample interval. Susceptibility errors for Vitek were among the streptomycin tests only. Our results demonstrated acceptable performance by the Vitek cards for enterococcal species identification and the detection of HLR. API 20S also provided an acceptable ability to speciate the enterococci within its data base, however, both systems must be improved by adding other clinical important Enterococcus species.

Aminoglycosides↗

PubMed1999

[Monitoring antibiotic resistance in Argentina. The WHONET program, 1995-1996].

The World Health Organization has implemented a surveillance program for antimicrobial resistance that is known as WHONET. In Argentina the program was developed through a network of 23 public and private hospitals that participate in national and international quality-control programs. Between January 1995 and December 1996, the antimicrobial susceptibility of 16,073 consecutive clinical isolates was determined, using the recommended standards of the National Committee for Clinical Laboratory Standards of the United States of America. More than half of the Escherichia coli urinary isolates were resistant to ampicillin and more than 30% to trimethoprim/sulfamethoxazole (SXT). When the percentage of resistant isolates from outpatients (OPs) was compared to that observed in hospitalized patients (HPs), a marked difference in antimicrobial activity was noted in the case of gentamicin (2% from OPs resistant vs. 8% from HPs resistant), norfloxacin (2% vs. 6%), and third-generation cephalosporins (7% vs. 15%). Of the Klebsiella pneumoniae isolates recovered from blood cultures, 71% and 60% showed resistance to third-generation cephalosporins and to gentamicin, respectively. The overall rate of oxacillin resistance in Staphylococcus aureus was 39%. Around half of the Enterococcus spp. isolates showed high resistance to aminoglycosides, but resistance to glycopeptides was not found. In Argentina, ampicillin and SXT were not suitable for treating diarrhea. Shigella flexneri had a higher number of isolates resistant to both of those drugs (87% and 74%, respectively) than Sh. sonnei did (47% and 71%, respectively). About 40% of the Salmonella spp. isolated in pediatric hospitals were resistant to third-generation cephalosporins. When microorganisms causing bacterial meningitis were examined, Streptococcus pneumoniae showed a resistance rate of 18% to penicillin and Haemophilus influenzae a resistance rate of 19% to ampicillin. These rates are within the intermediate range reported for other countries of the Americas and for Europe.

Cephalosporins↗

PubMed2003

[Role of linezolid in antimicrobial therapy].

The progressive emergence of multi-resistant gram-positive strains has prompted the search for new molecules (quinolones, streptogramins, oxazolidinones, ketolides, glycopeptides, daptomycin) to add to the current therapeutic arsenal. Linezolid, the first commercially available member of the oxazolidinone family, has evidenced activity against multi-resistant gram-positive strains (methicillin-resistant Staphylococcus aureus, S. aureus with decreased glycopeptide sensitivity, vancomycin-resistant Enterococcus spp., Streptococcus pneumoniae with decreased sensitivity to penicillin and cephalosporins), thereby providing a new option for treating infections by these microorganisms. This work reviews the microbiologic and pharmacologic aspects of this agent in order to establish its position among the available options for antimicrobial chemotherapy.

Acetamides↗

PubMed2000

Enterococcal glycopeptide resistance at an Italian teaching hospital.

Two thousand one hundred and thirteen strains of enterococci isolated at Pisa General Hospital in 1998 were analysed retrospectively to determine their glycopeptide resistance. Of all the microorganisms isolated in this period, 14.7% were enterococci (1405 Enterococcus faecalis, 19 Enterococcus faecium, six Enterococcus avium and 683 Enterococcus spp.). Two hundred and thirty (10.8%) of these enterococci were resistant or demonstrated reduced susceptibility to vancomycin and/or teicoplanin. The highest rate of resistance was found in outpatient enterococcal strains isolated from the urogenital tract. The frequency of enterococcal glycopeptide resistance at Pisa Hospital is higher than that reported from other areas of Italy.

Anti-Bacterial Agents↗

PubMed2002

Presence of erm genes among macrolide-resistant Gram-positive bacteria isolated from Danish farm soil.

The presence of erm genes was investigated among macrolide-resistant Gram-positive bacteria isolated from soil samples collected from four Danish farms that had been treated with animal waste. Soil samples were collected before, a few days after spread and 1 1/2 months and 5 months later. In 33% (9/27) of these isolates, an erm gene was detected using PCR. Eight isolates were positive for erm(B) and one isolate was positive for erm(C). No isolates contained erm(A), erm(D) or erm(F). The positive isolates were identified to genus level. Two erm(B) positive isolates were identified as Enterococcus spp., and the erm(C)-positive isolate as a Streptococcus spp., probably indicating survival from animal waste. The remaining six erm(B) positive isolates all belonged to the Bacillus cereus group. The erm(B) gene has not previously been identified in B. cereus but is frequently found among enterococci. This result could indicate horizontal transfer from bacteria of animal origin to indigenous soil bacteria.

Agriculture↗

PubMed1999

[Surveillance of gram positive cocci susceptibility to betalactams, glycopeptides, and other antimicrobials].

BACKGROUND: During the last decade, there has been a progressive increase in the resistance of gram (+) cocci to betalactamics and other antimicrobials. Therefore, vancomycin and teicoplanin have incorporated as alternative antimicrobial drugs. AIM: To assess the susceptibility of gram (+) cocci to different antimicrobials including vancomycin and teicoplanin. MATERIAL AND METHODS: We studied 447 strains of gram (+) cocci coming from ambulatory and hospitalized patients. These included 308 Enterococcus sp strains, 99 Staphylococcus aureus strains and 40 coagulase negative Staphylococci strains. Enterococci susceptibility was measured using minimal inhibitory concentrations in agar and that of Staphylococci, through diffusion. Susceptibility to vancomycin and teicoplanin was measured using minimal inhibitory concentrations in all strains. RESULTS: Enterococcus faecalis was 100% susceptible to ampicillin, penicillin, vancomycin and teicoplanin, 23% susceptible to tetracyclin and 47% to chloramphenicol. Susceptibility of E faecium was 61% to penicillin, 49% to chloramphenicol, 41% to tetracyclin, 100% to vancomycin and teicoplanin. Of 19 Enterococcus spp strains, 90% were susceptible to ampicillin, 80% to penicillin, 55% to chloramphenicol and 45% to tetracyclin. Only one E casseiflavus strain had a low level resistance to vancomycin and was susceptible to teicoplanin. No Staphylococcus aureus strain was resistant to vancomycin or teicoplanin. CONCLUSIONS: A permanent surveillance of gram (+) cocci antimicrobial susceptibility is required to update therapeutic schemes.

Drug Resistance, Microbial↗

PubMed2006

Detection of human-derived fecal pollution in environmental waters by use of a PCR-based human polyomavirus assay.

Regulatory agencies mandate the use of fecal coliforms, Escherichia coli or Enterococcus spp., as microbial indicators of recreational water quality. These indicators of fecal pollution do not identify the specific sources of pollution and at times underestimate health risks associated with recreational water use. This study proposes the use of human polyomaviruses (HPyVs), which are widespread among human populations, as indicators of human fecal pollution. A method was developed to concentrate and extract HPyV DNA from environmental water samples and then to amplify it by nested PCR. HPyVs were detected in as little as 1 microl of sewage and were not amplified from dairy cow or pig wastes. Environmental water samples were screened for the presence of HPyVs and two additional markers of human fecal pollution: the Enterococcus faecium esp gene and the 16S rRNA gene of human-associated Bacteroides. The presence of human-specific indicators of fecal pollution was compared to fecal coliform and Enterococcus concentrations. HPyVs were detected in 19 of 20 (95%) samples containing the E. faecium esp gene and Bacteroides human markers. Weak or no correlation was observed between the presence/absence of human-associated indicators and counts of indicator bacteria. The sensitivity, specificity, and correlation with other human-associated markers suggest that the HPyV assay could be a useful predictor of human fecal pollution in environmental waters and an important component of the microbial-source-tracking "toolbox."

Animals↗

PubMed2002

Evaluation of the VITEK 2 system for identification and antimicrobial susceptibility testing of medically relevant gram-positive cocci.

A study was conducted to evaluate the new VITEK 2 system (bioMérieux) for identification and antibiotic susceptibility testing of gram-positive cocci. Clinical isolates of Staphylococcus aureus (n = 100), coagulase-negative staphylococci (CNS) (n = 100), Enterococcus spp. (n = 89), Streptococcus agalactiae (n = 29), and Streptococcus pneumoniae (n = 66) were examined with the ID-GPC identification card and with the AST-P515 (for staphylococci), AST-P516 (for enterococci and S. agalactiae) and AST-P506 (for pneumococci) susceptibility cards. The identification comparison methods were the API Staph for staphylococci and the API 20 Strep for streptococci and enterococci; for antimicrobial susceptibility testing, the agar dilution method according to the procedure of the National Committee for Clinical Laboratory Standards (NCCLS) was used. The VITEK 2 system correctly identified to the species level (only one choice or after simple supplementary tests) 99% of S. aureus, 96.5% of S. agalactiae, 96.9% of S. pneumoniae, 92.7% of Enterococcus faecalis, 91.3% of Staphylococcus haemolyticus, and 88% of Staphylococcus epidermidis but was least able to identify Enterococcus faecium (71.4% correct). More than 90% of gram-positive cocci were identified within 3 h. According to the NCCLS breakpoints, antimicrobial susceptibility testing with the VITEK 2 system gave 96% correct category agreement, 0.82% very major errors, 0.17% major errors, and 2.7% minor errors. Antimicrobial susceptibility testing showed category agreement from 94 to 100% for S. aureus, from 90 to 100% for CNS, from 91 to 100% for enterococci, from 96 to 100% for S. agalactiae, and from 91 to 100% for S. pneumoniae. Microorganism-antibiotic combinations that gave very major errors were CNS-erythromycin, CNS-oxacillin, enterococci-teicoplanin, and enterococci-high-concentration gentamicin. Major errors were observed for CNS-oxacillin and S. agalactiae-tetracycline combinations. In conclusion the results of this study indicate that the VITEK 2 system represents an accurate and acceptable means for performing identification and antibiotic susceptibility tests with medically relevant gram-positive cocci.

Anti-Bacterial Agents↗

PubMed1997

Effect of norfloxacin, trimethoprim-sulfamethoxazole and nitrofurantoin on fecal flora of women with recurrent urinary tract infections.

The effects of antibiotics used for prophylaxis in women with recurrent urinary tract infections (UTIs) on the aerobic intestinal flora were investigated. Twenty-one patients with recurrent UTIs were randomly divided into three groups. The patients of each group received monotherapy with oral norfloxacin, trimethoprim-sulfamethoxazole or nitrofurantoin for one month. Urine and stool quantitative aerobic cultures were performed before prophylaxis, 2 and 4 weeks after the initiation of therapy, and 2 weeks after antibiotics were discontinued. The gram-negative aerobic flora was strongly suppressed during the administration of norfloxacin and trimethoprim-sulfamethoxazole, while Enterococcus spp. were not affected. Resistant strains of Escherichia coli were detected in two patients, one in the norfloxacin and one in the trimethoprim-sulfamethoxazole group. The aerobic intestinal flora was not affected by nitrofurantoin. These findings help in the selection of the most appropriate antimicrobial agent for prophylaxis in recurrent UTIs, so as to reduce the possibility of emergence of resistant bacterial strains.

Adult↗

PubMed2005

[Enterococcal bacteremia at Nîmes university hospital].

PURPOSE OF THE STUDY: The aim of this study was the description of enterococcal bacteremia and the evaluation of their resistance evolution to antibiotics and the virulence profile of these isolates. PATIENTS AND METHODS: We have studied all the bacteremia caused by Enterococcus spp. from the 1st of July 2003 until the 30th of June 2004 at Nîmes university hospital. For each isolate, population, clinical and microbiological data were collected. Multiplex PCR allowed for the identification of glycoprotein resistant phenotypes and furthermore, to identify the main virulence genes found in the enterococci. RESULTS: Thirty-three strains were identified from 33 patients (60.6% male) with a median age of 68 years (median: 3-96). The vital prognostic was calculated in 63.6% of the cases (MacCabe > or =1); seven patients died during their hospitalization (21.2%). The infections were nosocomial in 60.6% of the cases. No epidemic was observed. The principal comorbidities were cardiovascular and digestive pathologies. In addition, 54.5% of patients had had a surgical intervention. Enterococcus faecalis (87.9%) was the cause of infection in the majority of cases followed by Enterococcus faecium. Most of the infections were monomicrobial (75.8%). E. faecalis was sensitive to amoxicillin and to glycopeptides but was resistant to norfloxacin, erythromycin and pristinamycin. The same observations were made for the E. faecium with the exception of amoxicillin. E. faecalis had the maximum number of virulence genes and E. faecium the least. CONCLUSION: The enterococcal bacteremia had mainly a nosocomial origin. The isolates came from digestive tract flora but also from catheters. The mortality rate was 21.2%. A developing sensitivity to erythromycin and pristinamycin was observed in comparison to the data of the ONERBA (2002-2003). Only one isolate was resistant to glycopeptide. This was the first European E. faecium containing the vanD allele. This isolate proved fatal to the patient. The dark evolution of enterococcal bacteremia and their frequent nosocomial origin induce to promote studies intended for better knowing the risk factors of such infections.

Adolescent↗

PubMed1995

An analysis of 110 serious enterococcal infections. Epidemiology, antibiotic susceptibility, and outcome.

A prospective, observational study of 110 patients with serious infections due to Enterococcus spp. in 6 university and community teaching hospitals in Connecticut was conducted to define the epidemiology of community and nosocomial serious enterococcal infections and to determine risk factors, including antibiotic resistances, that contribute to outcome. Serious community and nosocomial enterococcal infections involved a variety of sites, and antibiotic resistance was common. Types of infection by major organ system were cardiovascular, 54% (catheter-related bacteremia 28%, primary bacteremia 18%, endocarditis 6%, septic thrombophlebitis 1%); intra-abdominal, 13% (including cholangitis, 6%); renal, 13%; skin and soft tissue, 5%; bone and joint, 4%; pleuropulmonary, 4%; central nervous system, 3%; deep surgical wound, 3%; and endometritis, 2%. Sixty-one percent of infections were nosocomial; 48% of these occurred in the intensive care unit. Enterococcus faecium was responsible for 20% of all infections. Antibiotic resistances among the infections included high-level gentamicin resistance (26%), ampicillin resistance (10%), and vancomycin resistance (8%). Clinical cure was achieved in 64% of patients; 6.8% of patients relapsed, 6.8% had recurrence of the infection with a different pathogen, and overall mortality was 23%. Ampicillin resistance and a high acute physiology and chronic health evaluation (APACHE) II score were highly predictive of lack of cure.

Adolescent↗

PubMed2006

A field study of the microbiological quality of fresh produce of domestic and Mexican origin.

Produce is responsible for an increasingly larger proportion of foodborne disease outbreaks. In particular, the globalization of the food supply may introduce new food safety risks and allow widespread distribution of contaminated food, particularly produce. The objectives of this study were to: (i) compare the overall quality of domestic and Mexican produce throughout the packing process; (ii) examine changes in microbiological quality of both domestic and Mexican produce at each stage of production and processing; and (iii) evaluate the prevalence of select pathogens on fresh produce, including leafy green, herbs, melons, and vegetables. Furthermore, we also sought to characterize the antibiotic resistance profiles of Enterococcus faecium and Enterococcus faecalis strains isolated from fresh produce. A total of 466 produce and matching environmental swab samples was collected from various locations in packing sheds in the southern US from November 2002 through December 2003. These samples were assayed by enumerative tests for total aerobic bacteria (APC), total coliforms, total Enterococcus, and E. coli. Produce samples were also analyzed for the presence of Salmonella, Listeria monocytogenes, Shigella, and E. coli O157:H7. A total of 112 E. faecium and E. faecalis isolates were further screened for antibiotic resistance using a panel of seventeen antibiotics. Overall, the microbiological quality of fresh produce ranged from 4.0 to 7.9 log(10) CFU/g (APC); less than 1.0 log(10) to 4.5 log(10) CFU/g (coliforms); less than 1.0 log(10) to 4.0 log(10) CFU/g (E. coli); and less than 1.0 log(10) to 5.4 log(10) CFU/g (Enterococcus). No Salmonella, Shigella, or E. coli O157:H7 were detected from the 466 25-g produce samples tested. However, three domestic cabbage samples were found to be positive for L. monocytogenes. Of the Enterococcus isolates, E. faecium had a higher degree of resistance to antibiotics in general, while Enterococcus spp. isolated from Mexican produce had a higher degree of antibiotic resistance when compared to strains isolated from produce samples of domestic origin. Despite increased attention to the role of imported produce in foodborne disease, this study does not support the assumption that domestic produce is of higher microbial quality than Mexican produce.

Anti-Bacterial Agents↗

PubMed2000

Antimicrobial susceptibility and species identification for clinical isolates of enterococci.

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

Drug Resistance, Microbial↗