scholarly journals Molecular Epidemiology of Ceftiofur-Resistant Escherichia coli Isolates from Dairy Calves

2006 ◽  
Vol 72 (6) ◽  
pp. 3940-3948 ◽  
Author(s):  
Sarah C. Donaldson ◽  
Beth A. Straley ◽  
Narasimha V. Hegde ◽  
Ashish A. Sawant ◽  
Chitrita DebRoy ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gel Electrophoresis ◽  
Dairy Herd ◽  
Multidrug Resistant ◽  
Pulsed Field Gel Electrophoresis ◽  
Dairy Calves ◽  
Disk Diffusion ◽  
Pulsed Field ◽  
E Coli ◽  
Clonal Type

ABSTRACT Healthy calves (n = 96, 1 to 9 weeks old) from a dairy herd in central Pennsylvania were examined each month over a five-month period for fecal shedding of ceftiofur-resistant gram-negative bacteria. Ceftiofur-resistant Escherichia coli isolates (n = 122) were characterized by antimicrobial resistance (disk diffusion and MIC), serotype, pulsed-field gel electrophoresis subtypes, beta-lactamase genes, and virulence genes. Antibiotic disk diffusion assays showed that the isolates were resistant to ampicillin (100%), ceftiofur (100%), chloramphenicol (94%), florfenicol (93%), gentamicin (89%), spectinomycin (72%), tetracycline (98%), ticarcillin (99%), and ticarcillin-clavulanic acid (99%). All isolates were multidrug resistant and displayed elevated MICs. The E. coli isolates belonged to 42 serotypes, of which O8:H25 was the predominant serotype (49.2%). Pulsed-field gel electrophoresis classified the E. coli isolates into 27 profiles. Cluster analysis showed that 77 isolates (63.1%) belonged to one unique group. The prevalence of pathogenic E. coli was low (8%). A total of 117 ceftiofur-resistant E. coli isolates (96%) possessed the bla CMY2 gene. Based on phenotypic and genotypic characterization, the ceftiofur-resistant E. coli isolates belonged to 59 clonal types. There was no significant relationship between calf age and clonal type. The findings of this study revealed that healthy dairy calves were rapidly colonized by antibiotic-resistant strains of E. coli shortly after birth. The high prevalence of multidrug-resistant nonpathogenic E. coli in calves could be a significant source of resistance genes to other bacteria that share the same environment.

scholarly journals Molecular typing of Escherichia coli O157[ratio ]H7 (H−) isolates from cattle in Japan

1999 ◽  
Vol 122 (2) ◽  
pp. 337-341 ◽  
Author(s):  
M. AKIBA ◽  
T. MASUDA ◽  
T. SAMESHIMA ◽  
K. KATSUDA ◽  
M. NAKAZAWA
Keyword(s):  
Escherichia Coli ◽  
Molecular Typing ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Escherichia Coli O157 ◽  
Outbreak Strain ◽  
Pulsed Field ◽  
E Coli ◽  
Biological Methods ◽  
Pfge Profiles

A total of 77 Escherichia coli O157[ratio ]H7 (H−) isolates from cattle in Japan were investigated by molecular biological methods. Most of these isolates (43 isolates) possessed the stx2 gene, but not stx1. Fifteen bacteriophage types and 50 pulsed-field gel electrophoresis (PFGE) profiles were observed. One isolate was indistinguishable from the human outbreak strain by these methods. This indicates that cattle must be considered as a possible source of human E. coli O157[ratio ]H7 infection in Japan.

Pulsed-Field Gel Electrophoresis Characterization of Shiga Toxin–Producing Escherichia coli O157 from Hides of Cattle at Slaughter

2002 ◽  
Vol 65 (7) ◽  
pp. 1172-1176 ◽  
Author(s):  
S. M. AVERY ◽  
A. SMALL ◽  
C.-A. REID ◽  
S. BUNCIC
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Gel Electrophoresis ◽  
Immunomagnetic Separation ◽  
Pulsed Field Gel Electrophoresis ◽  
Escherichia Coli O157 ◽  
Adult Cattle ◽  
Pulsed Field ◽  
E Coli ◽  
High Prevalence

Contamination of the brisket areas of the hides of healthy adult cattle with Shiga toxin–producing Escherichia coli O157 at slaughter in England was studied. In total, 73 cattle consignments comprising 584 animals delivered to one abattoir over 3 days during 1 week in July 2001 were studied: 26 cattle consignments arriving on Monday, 32 consignments arriving on Wednesday, and 15 consignments arriving on Friday. Consignment sizes ranged from 1 to 23 animals, with a mean consignment size of 8. The hide of the first animal to be slaughtered in each consignment was sampled by using a sponge swab moistened with 0.85% saline to rub an unmeasured brisket (ventral) area (ca. 30 by 30 cm). The process of isolating E. coli O157 from the swabs consisted of enrichment, screening with immunoprecipitation assay kits, and immunomagnetic separation. E. coli O157 was found on 24 of 73 (32.9%) cattle hides examined, and 21 of these 24 isolates produced Shiga toxins. The 24 E. coli O157 isolates produced six different pulsed-field gel electrophoresis profiles, and 18 (75%) of the isolates were of one prevalent clone. The high prevalence of one E. coli O157 clone on the hides of cattle at slaughter could be due to a high prevalence of that clone on the 18 farms involved (not investigated in the current study), in the postfarm transport or lairage environments, or both. Since the lairage environment, but not the farm of origin or the postfarm transport vehicle, was a factor common to all 18 cattle consignments, it could have played an important role in spreading the prevalent E. coli O157 clone to the cattle hides. Lairage pen floors and the stunning box floor were identified as the most probable sites along the unloading-to-slaughter route at which the brisket areas of cattle hides could become contaminated.

Molecular Characterization of Escherichia coli O157:H7 Hide Contamination Routes: Feedlot to Harvest

2006 ◽  
Vol 69 (6) ◽  
pp. 1240-1247 ◽  
Author(s):  
K. D. CHILDS ◽  
C. A. SIMPSON ◽  
W. WARREN-SERNA ◽  
G. BELLENGER ◽  
B. CENTRELLA ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Drinking Water ◽  
Gel Electrophoresis ◽  
Side Wall ◽  
Pulsed Field Gel Electrophoresis ◽  
Marker Genes ◽  
Escherichia Coli O157 ◽  
Pulsed Field ◽  
E Coli ◽  
Side Rail

This study was conducted to identify the origin of Escherichia coli O157:H7 contamination on steer hides at the time of harvest. Samples were collected from the feedlot, transport trailers, and packing plant holding pens and from the colons and hides of feedlot steers. A total of 50 hide samples were positive for E. coli O157:H7 in two geographical locations: the Midwest (25 positive hides) and Southwest (25 positive hides). Hide samples were screened, and the presence of E. coli O157: H7 was confirmed. E. coli O157:H7 isolates were fingerprinted by pulsed-field gel electrophoresis and subjected to multiplex PCR procedures for amplification of E. coli O157:H7 genes stx1, stx2, eaeA, fliC, rfbEO157, and hlyA. Feedlot water trough, pen floor, feed bunk, loading chute, truck trailer side wall and floor, packing plant holding pen floor and side rail, and packing plant cattle drinking water samples were positive for E. coli O157:H7. Pulsed-field gel electrophoresis banding patterns were analyzed after classifying isolates according to the marker genes present and according to packing plant. In this study, hide samples positive for E. coli O157:H7 were traced to other E. coli O157:H7–positive hide, colon, feedlot pen floor fecal, packing plant holding pen drinking water, and transport trailer side wall samples. Links were found between packing plant side rails, feedlot loading chutes, and feedlot pens and between truck trailer, different feedlots, and colons of multiple cattle. This study is the first in which genotypic matches have been made between E. coli O157:H7 isolates obtained from transport trailer side walls and those from cattle hide samples within the packing plant.

scholarly journals Emergence of Enterobacteriaceae Isolates Producing CTX-M Extended-Spectrum β-Lactamase in Austria

2006 ◽  
Vol 50 (2) ◽  
pp. 785-787 ◽  
Author(s):  
Alexandra Eisner ◽  
Elizabeth J. Fagan ◽  
Gebhard Feierl ◽  
Harald H. Kessler ◽  
Egon Marth ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
United Kingdom ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
E Coli ◽  
The United Kingdom ◽  
Extended Spectrum ◽  
Epidemiological Link ◽  
Esbl Producers

ABSTRACT Among 149 extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae isolates collected from patients in southeast Austria from 1998 to 2004, 38 Escherichia coli isolates and 11 Klebsiella spp. were CTX-M producers. The proportion of CTX-M-producers among all ESBL producers rose from 0% in 1998 to 58% in 2004. In general, CTX-M-producers had heterogeneous pulsed-field gel electrophoresis patterns, but one E. coli isolate was identical to a United Kingdom epidemic CTX-M-15-producing strain, although no epidemiological link with the United Kingdom was apparent.

Prevalence and Characterization of Escherichia coli O157:H7 on Pork Carcasses and in Swine Colon Contents from Provincially Licensed Abattoirs in Alberta, Canada

2020 ◽  
Vol 83 (11) ◽  
pp. 1909-1917
Author(s):  
SAIDA ESSENDOUBI ◽  
XIANQIN YANG ◽  
ROBIN KING ◽  
JULIA KEENLISIDE ◽  
JAVIER BAHAMON ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Gel Electrophoresis ◽  
Mean Value ◽  
Pulsed Field Gel Electrophoresis ◽  
Escherichia Coli O157 ◽  
Pulsed Field ◽  
E Coli ◽  
Swine Farms

ABSTRACT The objective of this study was to determine the prevalence of Shiga toxin–producing Escherichia coli (STEC) O157:H7 in colon contents and on carcasses from pigs slaughtered at provincially licensed abattoirs (PLAs) in Alberta, Canada. In 2017, carcass sponge samples and colon content samples were collected from 504 healthy market hogs at 39 PLAs and analyzed for E. coli O157:H7. Carcass samples were also analyzed for E. coli and aerobic colony count (ACC). Nine (1.8%) of 504 carcass samples were confirmed positive for E. coli O157:H7. Seven (1.4%) of 504 colon content samples were confirmed positive for E. coli O157:H7. These positives were found in 5 (12.8%) of 39 PLAs from hogs originating from eight farms. The E. coli O157:H7 isolates recovered from the positive samples (n = 1 isolate per sample) were clonal, as determined by pulsed-field gel electrophoresis. Six E. coli O157:H7 isolates obtained over 8 months from one PLA that only processed hogs and sourced hogs from one farm had indistinguishable pulsed-field gel electrophoresis patterns. All 16 E. coli O157:H7 isolates harbored eae and ehxA and were of stx2a subtype, suggesting that swine can carry E. coli O157:H7 of importance to human health. All carcass sponge swabs (100%) were positive for ACC. E. coli was present in 72% of carcass swabs. Carcasses from PLAs slaughtering both beef and hogs had a numerically higher ACC mean value but not statistically different compared with the carcasses from PLAs slaughtering only swine (2,799 and 610 CFU/cm2, respectively). E. coli showed a similar trend with a mean value of 0.88 CFU/cm2 in PLAs slaughtering both species and 0.26 CFU/cm2 in PLAs slaughtering only swine (P ≤ 0.05). This study provides evidence that healthy market hogs from different producers and farms in Alberta can carry E. coli O157:H7, and some strains of the organism may be able to establish persistence on some swine farms. HIGHLIGHTS

scholarly journals Prevalence and Dissemination of oqxAB in Escherichia coli Isolates from Animals, Farmworkers, and the Environment

2010 ◽  
Vol 54 (10) ◽  
pp. 4219-4224 ◽  
Author(s):  
Jingjing Zhao ◽  
Zhangliu Chen ◽  
Sheng Chen ◽  
Yuting Deng ◽  
Yahong Liu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Quinolone Resistance ◽  
Pfge Analysis ◽  
Pulsed Field ◽  
E Coli ◽  
Animal Farms ◽  
Food Animals

ABSTRACT OqxAB has recently been identified as one of the mechanisms of plasmid-mediated quinolone resistance (PMQR). Compared to what is observed for other PMQR determinants, there is a paucity of data with regard to the prevalence and epidemiology of OqxAB and its contribution to resistance to different antimicrobials. In this study, the prevalence and dissemination of oqxAB and other PMQR genes in Escherichia coli isolates from animals, farmworkers, and the environment in 2002 in China were investigated. Of the 172 E. coli isolates, 39.0% carried oqxA, while only 4.1%, 2.9%, and 0.6% carried qnr (1 qnrB6 isolate, 5 qnrS1 isolates, and 1 qnrD isolate), qepA, and aac(6′)-Ib- cr, respectively. Among the 33 isolates from farmworkers, 10 (30.3%) were positive for oqxA. oqxAB was associated with IS26 and was carried on the 43- to 115-kb IncF transferable plasmid. Transconjugants carrying oqxAB showed 4- to 16-fold increases in the MICs of quinolones, 16- to 64-fold increases in the MICs of quinoxalines, 8- to 32-fold increases in the MICs of chloramphenicol and trimethoprim-sulfamethoxazole, and 4- to 8-fold increases in the MICs of florfenicol compared to the levels for the recipient. The pulsed-field gel electrophoresis (PFGE) analysis showed that the high levels of prevalence and dissemination of oqxA B in E. coli in animal farms were primarily due to the transmission of plasmids carrying oqxAB, although clonal transmission between human and swine E. coli isolates was observed. It is concluded that oqxAB was widespread in animal farms in China, which may be due to the overuse of quinoxalines in animals. This study warrants the prudent use of quinoxalines in food animals.

Source Tracking of Escherichia coli O157:H7 and Salmonella Contamination in the Lairage Environment at Commercial U.S. Beef Processing Plants and Identification of an Effective Intervention†

2008 ◽  
Vol 71 (9) ◽  
pp. 1752-1760 ◽  
Author(s):  
TERRANCE M. ARTHUR ◽  
JOSEPH M. BOSILEVAC ◽  
DAYNA M. BRICHTA-HARHAY ◽  
NORASAK KALCHAYANAND ◽  
DAVID A. KING ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gel Electrophoresis ◽  
The United States ◽  
Pulsed Field Gel Electrophoresis ◽  
Processing Plant ◽  
Escherichia Coli O157 ◽  
Pulsed Field ◽  
E Coli ◽  
Beef Processing ◽  
Processing Plants

Transportation from the feedlot and lairage at the processing plant have been identified as potential sources of Escherichia coli O157:H7 and Salmonella hide contamination. The objective of this study was to perform a comprehensive tracking analysis of E. coli O157:H7 and Salmonella associated with beef cattle from the feedlot through processing. Cattle (n = 581) were sampled in a feedlot, then transported in multiple lots to three commercial, fed beef processing plants in the United States, where they were sampled again. Samples were collected from the tractor trailers prior to loading cattle and from the lairage environment spaces prior to entry of the study cattle. Pathogen prevalence on cattle hides increased on every lot of cattle between exiting the feedlot and beginning processing. Prior to loading cattle, E. coli O157:H7 was found in 9 (64%) of 14 tractor trailers. E. coli O157:H7 was detected in over 60% of the samples from each lairage environment area, while Salmonella was detected in over 70% of the samples from each lairage environment area. E. coli O157:H7 and Salmonella isolates (n 3,645) were analyzed using pulsed-field gel electrophoresis. The results of the pulsed-field gel electrophoresis tracking indicate that the transfer of bacteria onto cattle hides that occurs in the lairage environments of U.S beef processing plants accounts for a larger proportion of the hide and carcass contamination than does the initial bacterial population found on the cattle exiting the feedlot. Finally, the results of this study indicate that hide wash cabinets are effective in removing contamination derived from the lairage environment.

scholarly journals Mapping of Escherichia coli chromosomal Tn5 and F insertions by pulsed field gel electrophoresis.

Genetics ◽  
1988 ◽  
Vol 119 (2) ◽  
pp. 227-236
Author(s):  
C L Smith ◽  
R D Kolodner
Keyword(s):  
Escherichia Coli ◽  
Physical Mapping ◽  
Gel Electrophoresis ◽  
Physical Map ◽  
Pulsed Field Gel Electrophoresis ◽  
Genetic Maps ◽  
Low Resolution ◽  
Transposon Tn5 ◽  
Pulsed Field ◽  
E Coli

Abstract A low resolution Not I physical map of Escherichia coli was recently constructed. In this report we demonstrated that this map can be used to map Tn5 and F insertions physically. The transposon, Tn5, contains Not I recognition sequences in its IS50 sequences. F plasmid contains an unmapped Not I site. Hence, the location of Tn5 and F in the chromosome can be mapped by identifying the location of the introduced Not I sites using pulsed field gel electrophoresis. The physical mapping of genetically mapped Tn5 insertions confirm the previously constructed Not I map and helps align the E. coli physical and genetic maps. The use of Tn5 can assist the construction of both physical and genetic maps for microorganisms lacking such maps. Variations on this approach will facilitate physical mapping with a wide variety of organisms, enzymes, and genetic elements.

Occurrence of extended-spectrum β-lactamase, AmpC and virulence genes in Escherichia coli isolates from vegetable salads in Morocco

2017 ◽  
Vol 119 (7) ◽  
pp. 1633-1647
Author(s):  
Kaotar Nayme ◽  
Abouddihaj Barguigua ◽  
Brahim Bouchrif ◽  
Idrissa Diawara ◽  
Fatima El Otmani ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gel Electrophoresis ◽  
Virulence Genes ◽  
Phylogenetic Group ◽  
Pulsed Field Gel Electrophoresis ◽  
Resistant Bacteria ◽  
Content Type ◽  
Pulsed Field ◽  
E Coli ◽  
Extended Spectrum

Purpose The purpose of this paper is to assess the occurrence of the extended-spectrum β-lactamases (ESBLs) and AmpC β-lactamase genes in 144 Escherichia coli isolates recovered from 160 vegetable salad samples. Design/methodology/approach Among the 144 E. coli isolates recovered from 160 vegetable salads, 17 (12 percent) ceftazidime-resistant isolates were screened for ESBL production with the double disk-diffusion test. The ESBL-producing isolates were characterized for antimicrobial resistance, the presence of virulence genes and plasmid-mediated quinolone resistance (PMQR) determinants. The isolates were also subjected to phylogenetic group typing. The existence of plasmid AmpC genes and mutations in the regulatory region of the chromosomal AmpC gene was assessed using polymerase chain reaction (PCR) and sequencing. All β-lactamase isolates were further characterized by pulsed-field gel electrophoresis to determine the genetic relatedness. Findings Overall, 17 (12 percent) of the 144 E. coli isolates studied were ceftazidime resistant. Among the 17 isolates, 13 (77 percent) were multidrug resistant and four (23.5 percent) were ESBL producers. The bla CTX-M14 was the only gene detected. Of the 12 AmpC-producing isolates, three (18 percent) harbored plasmid-encoded AmpC and sequencing analysis of the chromosomal AmpC genes revealed mutations in the promoter/attenuator region. PMQR determinants were detected in 9 (52 percent) isolates. A was the most prevalent phylogenetic group (56 percent), followed by groups B1 (31 percent), D (6 percent), and B2 (6 percent). PCR showed that six (50 percent) ESBL/AmpC-producing E. coli isolates carried one and/or two virulence genes. Pulsed-field gel electrophoresis showed no epidemiological relationship between these isolates. Originality/value This study places vegetable salads within the spectrum of ecological niches that may be vehicles for antibiotic-resistant bacteria/genes with clinical interest and these findings are worthy of attention as their spread to humans by ingestion cannot be dismissed.

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