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Salmonella and other Enterobacteriaceae found in commercial poultry feed.

Poultry feed (mash and pelleted) and meat and bone meal samples were collected from commercial mills. All samples were analyzed for Enterobacteriaceae count (ENT) and Salmonella. The genus and species of the various Enterobacteriaceae present were also determined. The average ENT for mash, pelleted, and meal samples was log 4.1, .8, and 1.8/g, respectively. Enterobacteriaceae were present in 100, 60, and 92% and Salmonella in 58, 0, and 92% of the mash, pelleted, and meal samples, respectively. Overall, the Enterobacteriaceae most frequently isolated from all samples were Enterobacter agglomerans, Enterobacter cloacae, and Klebsiella pneumoniae. Although no Salmonella were found in the pelleted samples, the presence of other Enterobacteriaceae suggests that commercial pelleting may not totally destroy Salmonella since their heat resistance is similar to the other organisms found.

Animal Feed↗

Effects of epiphytic Enterobacteriaceae and pseudomonads on the growth of Listeria monocytogenes in model media.

Four Enterobacteriaceae (Enterobacter agglomerans and Rhanella aquatilis) and six pseudomonads (Pseudomonas fluorescens, Pseudomonas chlororaphis, Pseudomonas putida) isolated from minimally processed green endive were coinoculated at 10 degrees C with Listeria monocytogenes in a minimal medium. Pseudomonads did not modify the growth of L. monocytogenes, whereas Enterobacteriaceae reduced its maximal population by 2 to 3 log CFU/ml. The same effect was observed in a diluted yeast extract medium supplemented with amino acids and glucose, in which L. monocytogenes grown alone reached 10(9) to 10(10) CFU/ml. In the same diluted yeast extract medium, not supplemented with glucose and amino acids, the maximal population of L. monocytogenes in the presence of both Enterobacteriaceae and pseudomonads was only slightly reduced (less than 0.5 log CFU/ml). Culture filtrates of the Enterobacteriaceae had no inhibitory activity on L. monocytogenes. The effect of the Enterobacteriaceae on L. monocytogenes growth was presumably due to a competition for glucose and/or amino acids.

Amino Acids↗

Development of a PCR assay for detection of Enterobacteriaceae in foods.

A broad-range PCR assay for the detection of bacteria belonging to the Enterobacteriaceae family was developed in this study. Primers targeting the bacterial 16S rRNA gene were newly designed and used in this PCR assay. To determine the specificity of the assay, 72 different bacterial species (of 49 genera), 2 fungi, 3 animals, and 4 plants were tested. Results were positive for every tested Vibrioaceae or Enterobacteriaceae strain except Proteus mirabilis. For all other bacterial strains and eukaryotes tested, results were negative. Bacterial DNA for PCR was prepared by a simple procedure with the use of Chelex 100 resin from culture after growth in brain heart infusion medium. To test this PCR assay for the monitoring of the Enterobacteriaceae family, either Escherichia coli or Salmonella Enteritidis was inoculated into various foods as an indicator. Prior to the PCR, the inoculation of 10 to 40 CFU of bacteria per g of food was followed by a 5-h enrichment culture step, and the PCR assay allowed the detection of bacterial cells. When actual examinations of the contamination of 15 noodle foods with Enterobacteriaceae by this PCR assay were conducted, 33% (5 of 15) of the samples tested positive. These results agreed with those of the Petrifilm Enterobacteriaceae Count Plate assay. Including the enrichment culture step, the entire PCR detection process can be completed within 7 h.

Colony Count, Microbial↗

[Detection of extended-spectrum beta-lactamases and drug sensitivity test in clinical isolates of the family Enterobacteriaceae in Shanghai].

OBJECTIVE: To investigate extended-spectrum beta-lactamases (ESBLs) production in clinical isolates of the family Enterobacteriaceae in Shanghai, and compare the resistant rates of ESBLs-producing strains and non-ESBLs-producing strains to 11 antibiotics. METHODS: Double-disk synergy test was used to detect ESBLs in 1,026 strains of the family Enterobacteriaceae; Kirby-bauer agar diffusion method was used to judge drug sensitivity. RESULTS: 34.31% of strains of the family Enterobacteriaceae were considered ESBLs producers by double-disk synergy test. The prevalences of ESBLs in isolates of Klebsiella pneumoniae, Escherichia coli and Enterobacter cloacae were 37.40%, 30.21% and 41.82% respectively. Except Imipinem and Cefmetazole, the resistant rates of ESBLs-producing strains to other 9 antibiotics were much higher than that of non-ESBLs-producing strains(P < 0.01). The resistant rates of Imipinem to ESBLs-producing strains were the lowest. CONCLUSIONS: The prevalence of ESBLs in clinical isolates of the family Enterobacteriaceae in Shanghai was high; it is necessary for clinical laboratory to detect ESBLs in clinical isolates of the family Enterobacteriaceae routinely. The resistant rates of ESBLs-producing strains to most antibiotics were high. Imipinem and Cefmetazole were the effective antibiotics to infections caused by ESBLs-producing strains.

Anti-Bacterial Agents↗

[Enterobacteriaceae as indicators of the presence of Salmonella and the hygienic conditions of meat and bone meals].

The purpose of this study was to establish the relationship between Enterobacteriaceae counts and the presence of Salmonella in meat-and-bone meals. Ninety meat-and-bone meal sample units (9 lots) from five producing plants from the Greater Buenos Aires, Argentina, were analyzed. The presence of Salmonella was detected through pre-enrichment of the samples followed by selective enrichment in two broth cultures and isolations after 24 and 48 hours. For Enterobacteriaceae counts the recovery of sublethally injured cells was carried out for two hours on trypticase soy agar, followed by the addition of selective agar. The overall Enterobacteriaceae count for the 90 sample units was 3.0. Seven (77.7%) of the 9 lots were positive for Salmonella. The correlation coefficient between Enterobacteriaceae counts and Salmonella was 0.81 with a confidence limit (95%) of 0.35-0.95. The results of this study indicate that Enterobacteriaceae counts are not good indicators of the presence of Salmonella in meat-and-bone meals. However, they could be used to assess their hygienic quality.

Colony Count, Microbial↗

Evaluation of an automated system for identification of Enterobacteriaceae and nonfermenting bacilli.

The performance of the Vitek 2 (bioMérieux, France), a new fully automated system allowing rapid identification of microorganisms and susceptibility testing, and the Vitek 2 ID-GNB card (bioMérieux) was evaluated using 502 clinical isolates and stock collection strains of gram-negative rods belonging to 70 taxa. The number of isolates correctly identified to species and genus levels was 430 (85.7%) and 485 (96.6%), respectively. Clinical isolates of both Enterobacteriaceae and non-Enterobacteriaceae were better identified at the species level (95.3% and 74%, respectively) than stock collection strains (86.4% and 52.2%, respectively). The Vitek 2 ID-GNB card provides after 3 h a highly acceptable level of accuracy for identification of Enterobacteriaceae and non-Enterobacteriaceae, including most atypical strains encountered in clinical situations.

Automation↗

Susceptibility of Enterobacteriaceae to beta-lactam agents and fluoroquinolones: a 3-year survey in France.

OBJECTIVE: To assess trends in the susceptibility to beta-lactam agents and to fluoroquinolones of clinically relevant Enterobacteriaceae isolated over a 3-year period in 14 French hospital laboratories. METHODS: During the second quarter of 1996, 1997 and 1998, 180 consecutive non-duplicate isolates of Enterobacteriaceae were collected in each center. Sixteen beta-lactams and four quinolones were tested by the disk diffusion method. In addition, the double-disk synergy test was used to screen for the production of extended-spectrum beta-lactamase (ESBL). RESULTS: Totals of 2507, 2312 and 2506 clinical isolates were obtained in each period, respectively. The distribution of Enterobacteriaceae species according to clinical specimens and wards was similar in each study period. No significant variation in the susceptibility rates to beta-lactams and fluoroquinolones was observed, except in Klebsiella pneumoniae and Enterobacter aerogenes. The prevalence of ESBL-producing isolates decreased from 18% to 9% in the former, while it increased from 32% to 54% in the latter. At the same time, the susceptibility to ofloxacin and pefloxacin increased for K. pneumoniae (P < 0.003) and cephalosporinase-producing species (P < 0.05), except Enterobacter spp. CONCLUSION: Over the 3-year study period beta-lactams and fluoroquinolones remained highly active against Enterobacteriaceae clinical isolates, with the exception of E. aerogenes, probably as a result of the dissemination of multiresistant clones in French hospitals.

Anti-Bacterial Agents↗

Occurrence and characterization of carbapenemase-producing Enterobacteriaceae: report from the SENTRY Antimicrobial Surveillance Program (2000-2004).

Emergence and dissemination of Enterobacteriaceae isolates harboring carbapenemases in various geographic regions represents a significant threat to the management of nosocomial infections. Enterobacteriaceae isolates from the SENTRY Antimicrobial Surveillance Program (2000-2004) demonstrating decreased susceptibility to imipenem and meropenem (minimum inhibitory concentration [MIC], > or =2 mg/L) were evaluated for the production of metallo-beta-lactamases and serine carbapenemases using disk approximation and polymerase chain reaction (PCR) tests. Carbapenemase-producing strains were epidemiologically typed by automated riboprinting and pulsed-field gel electrophoresis (PFGE) to establish clonality. Among 37,557 Enterobacteriaceae (5 genus groups) evaluated, 119 (0.32%) had increased carbapenem MIC values, and a carbapenemase was identified in 51 (42.9%) of these strains. KPC-2 and KPC-3 were the most frequently occurring carbapenemases (24 isolates, 20.2%) in the United States and were detected in Klebsiella spp, Citrobacter spp., Enterobacter spp., and Serratia marcescens strains isolated in New York, Arkansas, and Virginia. SME-2-producing S. marcescens were isolated in the New York City area, Texas, and Ohio, while NMC-A was found in one E. cloacae strain from New York. In contrast, metallo-beta-lactamases were prevalent in Europe. IMP-1-producing E. cloacae (11 isolates) were detected in Turkey, while VIM-1-producing strains were found in Italy (Enterobacter spp.) and Greece (Klebsiella pneumoniae). Clonal dissemination of carbapenemase-producing strains was observed in several medical centers on both continents. The occurrence of carbapenemases in various Enterobacteriaceae remains rare but appears to be spreading geographically (not in Latin America), mainly with metallo-beta-lactamases being found in Mediterranean Europe and KPC enzymes in the New York City area.

Anti-Bacterial Agents↗

First case of septicemia due to a strain belonging to enteric group 58 (Enterobacteriaceae) and its designation as Averyella dalhousiensis gen. nov., sp. nov., based on analysis of strains from 20 additional cases.

When enteric group 58 was first described as a distinct new group of Enterobacteriaceae in 1985, there were only five known human isolates: four from wounds and one from feces. In 1996, we investigated the first blood isolate of enteric group 58, a case of sepsis in a 33-year-old woman receiving total parenteral nutrition. Fifteen additional clinical isolates have since been identified at CDC, including several recognized from a collection of "unidentified" strains dating back to 1973. All strains were characterized with a standard set of 49 biochemical tests used for Enterobacteriaceae, and the results were analyzed to determine phenotypic relatedness and best taxonomic fit. Antibiograms were determined as a taxonomic tool. Original identifications provided by submitting laboratories encompassed a wide variety of Enterobacteriaceae, including 14 species in eight genera, the most common being Enterobacter spp., Salmonella spp., Serratia spp., Kluyvera spp., or Escherichia spp. Enteric group 58 strains have been most frequently isolated from traumatic injuries, fractures, and wounds and rarely from feces. Defining its clinical significance and distinguishing infection from colonization requires further study, but our case report indicates that serious systemic infection can occur. The vernacular name enteric group 58 was used from 1985 to 2004. In this paper, we formally name it Averyella dalhousiensis gen. nov., sp. nov., on the basis of its unique phenotype and its unique 16S rRNA gene sequence. These data indicate that enteric group 58 is not closely related to any of the existing genera or species of Enterobacteriaceae. The type strain is designated CDC 9501--97, and a phenotypic definition is given based on all 21 strains.

Adult↗

Ceftazidime and amikacin alone and in combination against Pseudomonas aeruginosa and Enterobacteriaceae.

The efficacy of ceftazidime alone and combined with amikacin was studied in a rabbit model simulating closed-space infections at locally neutropenic sites. Six strains of Pseudomonas aeruginosa, and six Enterobacteriaceae (two strains each of Klebsiella pneumoniae and Serratia marcescens and one strain each of Escherichia coli and Citrobacter freundii) in pooled rabbit serum were each inoculated into separate subcutaneous semipermeable chambers. Intramuscular antibiotic therapy was begun 4 hr later with ceftazidime (50 mg/kg) alone and combined with amikacin (15 mg/kg) for Enterobacteriaceae or ceftazidime (100 mg/kg) alone and combined with amikacin (15 mg/kg) for pseudomonads every 6 hr for 16 doses. Amikacin alone was ineffective for all 12 strains. Ceftazidime alone was successful (greater than or equal to 5.5 log10 colony forming units (CFU)/ml decrease from drug-free control) in eliminating five of six Enterobacteriaceae but was not successful against any of the pseudomonads. Ceftazidime plus amikacin was successful against the same five of six Enterobacteriaceae and five of six pseudomonads. The best in vitro tests for the prediction of in vivo outcome were high inoculum (greater than or equal to 7 log10 CFU/ml) susceptibility, checkerboard synergism testing, and conventional inoculum time-kill rates at concentrations of antimicrobials simulating extravascular levels obtained in vivo.

Amikacin↗

Lomefloxacin (SC 47111 or NY-198), a new difluorinated quinolone: comparison of the in vitro activity with other broad spectrum antimicrobials against Enterobacteriaceae, Acinetobacter spp, Aeromonas spp, and Pseudomonas aeruginosa.

Using a broth microdilution method, we compared the in vitro activity of lomefloxacin to other broad spectrum antimicrobials against clinical strains of Enterobacteriaceae, Acinetobacter spp, Aeromonas spp, and P. aeruginosa. Against the Enterobacteriaceae and A. hydrophila, lomefloxacin showed excellent activity with MIC50 values ranging from 0.12-0.5 micrograms/ml and MIC90 values ranging from 0.12-1.0 micrograms/ml. By comparison, lomefloxacin had superior activity to nalidixic acid and difloxacin, comparable activity to norfloxacin and fleroxacin, and slightly less activity than ciprofloxacin. Lomefloxacin was also more active than imipenem, ceftazidime, aztreonam, ticarcillin/clavulanic acid (T-CA), and gentamicin. Lomefloxacin was less active (MIC90, 4 micrograms/ml) against Acinetobacter strains than Enterobacteriaceae strains. Against these same Acinetobacter strains, lomefloxacin was 8- to 64-fold more active than ceftazidime, aztreonam, T-CA, and gentamicin but slightly less active than imipenem. Lomefloxacin was 2- to 8-fold more active than fleroxacin, difloxacin, and nalidixic acid against strains of P. aeruginosa. However, lomefloxacin was less active than ciprofloxacin and norfloxacin. Lomefloxacin had comparable activity to imipenem but was 2- to 16-fold more active than ceftazidime, aztreonam, T-CA, and gentamicin. Thus, lomefloxacin demonstrated comparable activity to other fluorinated quinolones in that it possesses high active against clinical strains of Enterobacteriaceae and had good-to-moderate activity against most strains of Acinetobacter and P. aeruginosa.

4-Quinolones↗

Surveillance of oral cultures for Enterobacteriaceae during bone marrow transplantation.

Bone marrow-transplanted patients can suffer from severe life-threatening infections. Oral bacterial cultures were collected from a group of 40 patients prior to and following bone marrow transplantation every 3 days, following initial preparation and eradication of oral infections. The samples were grown on the Titertek-Enterobac kit specific for Enterobacteriaceae. In 426 oral cultures 30.5% grew gram-negative bacteria, 76.6% of them were Enterobacteriaceae. young male patients had 8.3% positive cultures at the study start, a percentage which constantly increased during later periods. Older patients did not follow the same pattern. Also, the allogeneic transplantation group had a higher percentage of Enterobacteriaceae than the autologous group (49.0 versus 19.5%). In blood cultures 18 out of the 94 positive ones showed the presence of Enterobacteriaceae. The most commonly found microorganisms in oral cultures were Klebsiella oxytoca (23%), Enterobacter cloacae (18%) and Klebsiella pneumoniae (15%). The decrease in the positive oral cultures from 35.0% during the pretransplantation period to 5.4% close to the transplantation, demonstrates that the preparatory protocol used for the prevention of oral infections was highly effective.

Adolescent↗

Association of broad-spectrum antibiotic use with faecal carriage of oxyiminocephalosporin-resistant enterobacteriaceae in an intensive care unit.

The link between administration of antibiotics and detection of third-generation-cephalosporin-resistant (TGCR) enterobacteriaceae in faeces was studied in patients in a burns intensive care unit (ICU). The presence of extended-spectrum beta-lactamase producers was also determined in these isolates. At least two rectal swab samples were taken from 43 of 72 patients admitted to the ICU from January 1998 to June 1999. Antibiotic resistance tests were performed for all isolated enterobacteriaceae using the methods of the National Committee for Clinical Laboratory Standards. Only 10 out of 30 antibiotic-treated patients showed TGCR enterobacteriaceae in faeces. Fisher's exact test showed a relationship between the administration of oxyiminocephalosporins (third-generation cephalosporins) (P=0.002) or carbapenems (P=0.003) and the isolation of TGCR enterobacteriaceae from faeces. The administration of oxyiminocephalosporins led to the selection of resistant strains in the faecal flora.

Anti-Bacterial Agents↗

The effect of commercial steam pasteurization on the levels of Enterobacteriaceae and Escherichia coli on naturally contaminated beef carcasses.

The aim of the study was to assess the reduction achieved by steam pasteurization of beef carcasses of Escherichia coli, Enterobacteriaceae and total aerobic mesophilic plate counts (APCs). In total, 30 carcass halves were exposed to steam pasteurization (90 degrees C, 10 s exposure time) and the 30 corresponding carcass halves remained as untreated controls. The neck, midline and rump were sampled on each carcass half. Significant reductions in E. coli incidence (P < 0.05) and counts, 0.5 log10 CFU 1000 cm(-2) (P < 0.05), were observed on rump sites only. Significant reductions (>0.8 log10 CFU 1000 cm(-2)) of Enterobacteriaceae were observed at all carcass sites sampled (P < 0.05). Enterobacteriaceae reductions (>2 log10 CFU 1000 cm(-2)) were highly significant at the more contaminated sites (P < 0.001). Reductions in total APCs were inconsistent. Steam pasteurization significantly reduced the level of E. coli and Enterobacteriaceae at more contaminated sites, but did not result in complete decontamination. Therefore, steam pasteurization should be classed as an aid to hygienic beef processing, but not as a critical control point.

Abattoirs↗

DNA based classification of food associated Enterobacteriaceae previously identified by Biolog GN Microplates.

Enterobacteriaceae are frequently isolated from food products and it is essential to have methods for correct identification for both food hygiene and epidemiology reasons. Phenotypic methods are not always sufficient and have to be supplemented by DNA based methods. In the present study, 70 strains of Enterobacteriaceae derived from milk, fish and meat that had previously been identified by Biolog GN Microplates were genomically classified together with 15 representative type strains of species of Enterobacteriaceae. The field strains were dominated by Hafnia alvei, Serratia liquefaciens and Rahnella aquatilis. All strains were subjected to temporal temperature gel electrophoresis (TTGE) analysis using amplicons encompassing the V3, V4 and V9 variable regions of the 16S rRNA gene. Selected strains were analysed by ribotyping and partial 16S rDNA sequencing. The type strains were differentiated into 10 different TTGE groups. Two of the groups contained two type strains. Enterobacter aerogenes and Klebsiella planticola were not distinguished due to their identical sequences and Yersinia ruckeri and Citrobacter freundii showed the same migration pattern. The 70 food strains could be differentiated into 14 TTGE groups where 33 strains (47.1%) could be assigned to TTGE groups including type or reference strains. Rahnella strains were dispersed into three TTGE groups of which one group corresponded to Rahnella genomospecies 1 and one to genomospecies 3. The grouping of Rahnella strains was supported by ribotyping and phylogenetic analysis. TTGE can be a useful additional tool for identification on the species level of food related Enterobacteriaceae.

Animals↗

Efficacy of ertapenem in the treatment of serious infections caused by Enterobacteriaceae: analysis of pooled clinical trial data.

OBJECTIVE: The efficacy of ertapenem, 1 g once a day, for treatment of adults with serious infections caused by Enterobacteriaceae was compared with ceftriaxone 1 g once a day [complicated urinary tract infection (CUTI) and community-acquired pneumonia (CAP)] or piperacillin-tazobactam, 3.375 g every 6 h (complicated intra-abdominal, complicated skin/skin structure and acute pelvic infections). PATIENTS AND METHODS: This combined analysis included the subgroup of all 1167 treated patients infected with Enterobacteriaceae from seven randomized double-blind studies. RESULTS: Escherichia coli was the most common pathogen, accounting for 65.3% of all Enterobacteriaceae. Among evaluable patients with deep tissue (intra-abdominal, skin and pelvic) infections, the combined clinical cure rates were 84.8% (223 of 263) for ertapenem and 82.9% (194 of 234) for piperacillin-tazobactam [95% confidence interval (CI) for the difference, adjusting for infection, -4.9% to 8.9%]. Cure rates by infection for ertapenem and piperacillin-tazobactam, respectively, were: intra-abdominal, 85.1% (143 of 168) and 79.9% (119 of 149); pelvic, 86.8% (46 of 53) and 94% (47 of 50); skin/skin structure, 81% (34 of 42) and 80% (28 of 35). Among patients with CUTI, microbiological cure rates were 90.5% (220 of 243) for ertapenem and 92% (196 of 213) for ceftriaxone (95% CI for the difference, -7.1% to 4.1%). In patients with CAP, clinical cure rates were 95% (19 of 20) for ertapenem and 88.9% (16 of 18) for ceftriaxone. CONCLUSION: Ertapenem therapy was as effective as either piperacillin-tazobactam or ceftriaxone for serious infections caused by Enterobacteriaceae.

Adult↗

Distribution of a transposon-like element carrying bla(CMY-2) among Salmonella and other Enterobacteriaceae.

OBJECTIVES: The dissemination of cephamycin resistance in Enterobacteriaceae and its correlation with a transposon-like DNA element consisting of a specific tnpA-bla(CMY-2)-blc-sugE structure were investigated. METHODS: A total of 140 enterobacterial isolates belonging to 17 species (10 genera) of Enterobacteriaceae phenotypically characterized as putative AmpC-producers were evaluated. The isolates were examined by PCR analysis, DNA-DNA hybridization and nucleotide sequencing. RESULTS: The bla(CMY-2)-carrying element was detected in 34 isolates from 10 species (9 genera), including all 14 Salmonella and 4 Shigella isolates as well as 7 of the 10 Escherichia coli isolates tested. The remaining 9 isolates were from 112 isolates of the other 14 species tested. The genetic structure of the bla(CMY-2)-carrying element was identical in 29 isolates, while in 3 E. coli and 2 Citrobacter isolates an additional insertion sequence IS1 was found inserted at various nucleotide positions close to the 3' end, either within or downstream, of tnpA. In 12 of the 14 representative isolates examined, the bla(CMY-2)-carrying element was found inserted in the finQ gene of various-sized plasmids with highly conserved 8 bp direct repeats flanking the junction regions. Among the other 106 non-CMY-2-producing isolates, plasmid-mediated ampC genes were found only in one isolate of Enterobacter aerogenes which carried a bla(DHA-1)-like gene. CONCLUSIONS: bla(CMY-2) is the most prevalent plasmid-mediated ampC gene among Enterobacteriaceae. All the bla(CMY-2) genes identified in the present study were associated with a specific transposon-like element that may be responsible for the spread of bla(CMY-2) among Enterobacteriaceae.

Cephalosporin Resistance↗

Oral and oropharyngeal prevalence of Enterobacteriaceae in humans: a review.

Members of the Enterobacteriaceae family are widely distributed in nature and exhibit substantial diversity in ecology, host range and pathogenic potential for man. While wide discrepancies in methodology exist between epidemiological studies, the available data indicate an increased prevalence of oral and/or oropharyngeal Enterobacteriaceae carriage in patients with illnesses of varying severity compared with healthy subjects. This paper reviews the prevalence of Enterobacteriaceae in the oral and oropharyngeal region of healthy human subjects and those affected by different disease entities, and discusses the complexities associated with collating and interpreting such data. The effect of antimicrobials and antiseptics on oral and oropharyngeal Enterobacteriaceae has also been reviewed, while highlighting the gaps in knowledge and future research directions.

Adolescent↗