scholarly journals Combined Use of Two Genetic Fingerprinting Methods, Pulsed-Field Gel Electrophoresis and Ribotyping, for Characterization of Escherichia coli O157 Isolates from Food Animals, Retail Meats, and Cases of Human Disease

2002 ◽  
Vol 40 (8) ◽  
pp. 2806-2812 ◽  
Author(s):  
S. M. Avery ◽  
E. Liebana ◽  
C.-A. Reid ◽  
M. J. Woodward ◽  
S. Buncic
Keyword(s):  
Escherichia Coli ◽  
Human Disease ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Escherichia Coli O157 ◽  
Genetic Fingerprinting ◽  
Combined Use ◽  
Retail Meats ◽  
Fingerprinting Methods

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 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.

scholarly journals Shedding of Escherichia coli O157:H7 in Dairy Cattle Housed in a Confined Environment following Waterborne Inoculation

2002 ◽  
Vol 68 (4) ◽  
pp. 1947-1954 ◽  
Author(s):  
J. A. Shere ◽  
C. W. Kaspar ◽  
K. J. Bartlett ◽  
S. E. Linden ◽  
B. Norell ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Water Treatment ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Bull Calves ◽  
Genomic Dnas ◽  
Confined Environment ◽  
Effective Water

ABSTRACT A study of Escherichia coli O157:H7 transmission and shedding was conducted with bull calves housed in individual pens within a confined environment. For comparative purposes, the numbers and duration of E. coli O157:H7 shedding in naturally infected calves were monitored after a single purchased calf (calf 156) tested positive prior to inoculation. During the next 8 days, the calves in adjacent pens and a pen directly across a walkway from calf 156 began to shed this serotype O157:H7 strain. Five of the eight calves in this room shed this O157:H7 strain at some time during the following 8 weeks. The numbers of E. coli O157:H7 isolates shed in these calves varied from 60 to 105 CFU/g of feces, and the duration of shedding ranged from 17 to >31 days. The genomic DNAs from isolates recovered from these calves were indistinguishable when compared by using XbaI digestion and pulsed-field gel electrophoresis. Inoculation of calves with 1 liter of water containing ca. 103 to 104 CFU of E. coli O157:H7/ml resulted in shedding in 10 of 12 calves (trial 1, 4 of 4 calves; trial 2, 6 of 8 calves). The inoculated calves shed the inoculation strain (FRIK 1275) as early as 24 h after administration. The duration of shedding varied from 18 to >43 days at levels from 102 to 106 CFU/g of feces. The numbers of doses necessary to initiate shedding varied among calves, and two calves in trial 2 never shed FRIK 1275 after four doses (ca. 106 CFU per dose). Results from this study confirm previous reports of animal-to-animal and waterborne dissemination of E. coli O157:H7 and highlight the need for an effective water treatment to reduce the spread of this pathogen in cattle.

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 Correlation between geographic distance and genetic similarity in an international collection of bovine faecal Escherichia coli O157[ratio ]H7 isolates

2003 ◽  
Vol 131 (2) ◽  
pp. 923-930 ◽  
Author(s):  
M. A. DAVIS ◽  
D. D. HANCOCK ◽  
T. E. BESSER ◽  
D. H. RICE ◽  
C. J. HOVDE ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gel Electrophoresis ◽  
Genetic Similarity ◽  
Genetic Relatedness ◽  
Geographic Distance ◽  
Similarity Coefficient ◽  
Pulsed Field Gel Electrophoresis ◽  
Dice Similarity Coefficient ◽  
Escherichia Coli O157 ◽  
Molecular Studies

Evidence from epidemiological and molecular studies of bovine Escherichia coli O157[ratio ]H7 suggests that strains are frequently transmitted across wide geographic distances. To test this hypothesis, we compared the geographic and genetic distance of a set of international bovine Escherichia coli O157[ratio ]H7 isolates using the Mantel correlation. For a measure of genetic relatedness, pulsed-field gel electrophoresis of six different restriction enzyme digests was used to generate an average Dice similarity coefficient for each isolate pair. Geographic distance was calculated using latitude and longitude data for isolate source locations. The Mantel correlation between genetic similarity and the logarithm of geographic distance in kilometers was −0·21 (P<0·001). The low magnitude of the Mantel correlation indicates that transmission over long distances is common. The occurrence of isolates from different continents on the same cluster of the dendrogram also supports the idea that Escherichia coli O157[ratio ]H7 strains can be transferred with considerable frequency over global distances.

Phenotypic and Genotypic Characterization of Escherichia coli Verotoxin-Producing Isolates from Humans and Pigs

2001 ◽  
Vol 64 (12) ◽  
pp. 1904-1911 ◽  
Author(s):  
ANNIE DesROSIERS ◽  
JOHN M. FAIRBROTHER ◽  
ROGER P. JOHNSON ◽  
CLARISSE DESAUTELS ◽  
ANN LETELLIER ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virulence Factors ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Hemorrhagic Colitis ◽  
Testing Technique ◽  
Fecal Samples ◽  
The Individual ◽  
Phenotypic And Genotypic Characterization

The aim of this study was to characterize verotoxin-producing Escherichia coli (VTEC) isolates obtained from humans and pigs in the same geographic areas and during the same period of time in order to determine whether porcine VTEC isolates could be related to human cases of diarrhea and also to detect the presence of virulence factors in these isolates. From 1,352 human and 620 porcine fecal samples, 11 human and 18 porcine verotoxin-positive isolates were obtained by the VT immunoblot or the individual colony testing technique. In addition, 52 porcine VTEC strains isolated from diseased pigs at the Faculté de médecine vétérinaire during the same period or from fecal samples collected previously isolated at slaughterhouses were characterized in this study. Antimicrobial resistance profiles were different between human and porcine isolates. In general, the serotypes observed in the two groups were different. No porcine isolate was of serotype O157:H7; however, one isolate was O91:NM, a serotype that has been associated with hemorrhagic colitis in humans. Also, one serotype (O8:H19) was found in isolates from both species; however, the O8:H19 isolates of the two groups were of different pathotypes. The pathotypes observed in the human and porcine isolates were different, with the exception of VT2vx-positive isolates; the serotypes of these isolates from the two groups were nevertheless different. Pulsed-field gel electrophoresis analysis indicated no relatedness between the human and porcine isolates. In conclusion, these results suggest that the porcine and human isolates of the present study were not genetically related. Most porcine VTEC isolates did not possess known virulence factors required to infect humans. However, certain non-O157:H7 porcine VTECs may potentially infect humans.

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