Determining the Prevalence of Escherichia coli O157 in Cattle and Beef from the Feedlot to the Cooler

2006 ◽  
Vol 69 (12) ◽  
pp. 2824-2827 ◽  
Author(s):  
D. R. WOERNER ◽  
J. R. RANSOM ◽  
J. N. SOFOS ◽  
G. A. DEWELL ◽  
G. C. SMITH ◽  
...  

Prevalence of Escherichia coli O157 on cattle entering the slaughter floor may range from 10 to >70%. This study was conducted to determine the effect of E. coli O157 prevalence in fecal pats collected from feedlot pen floors on subsequent E. coli O157 prevalence on carcasses at various points in the slaughter process. Fecal pats from the feedlot pen floor were collected within 3 days before slaughter. During cattle processing at the slaughter facility, additional samples were collected from the hide, from the colon, and from the carcasses before and after evisceration and after final decontamination. Of 15 lots (a group of cattle from the same pen from a feedlot) sampled, 87% had at least one positive fecal pat from the feedlot floor, 47% had a positive hide sample, 73% had a positive colon/fecal sample, and 47% had a positive carcass sample preevisceration; however, only 8% of lots had a positive carcass sample postevisceration or after final intervention. Of the total samples tested (n = 1,328), 24.7, 14.7, 27.6, 10.1, 1.4, and 0.3% of fecal pats from the feedlot floor, hide, colon, preevisceration, post-evisceration, and final intervention samples, respectively, were positive for E. coli O157. Pens with greater than 20% positive fecal pats from the feedlot floor had 25.5% hide, 51.4% colon, and 14.3, 2.9, and 0.7% carcass samples positive at preevisceration, at postevisceration, and after final intervention, respectively. However, fecal pats from feedlot floor samples that contained less than 20% positive fecal samples showed lower pathogen prevalence, with 5.0% hide, 7.5% colon, and 6.3, 0, and 0% carcass positive samples at preevisceration, postevisceration, and post–final intervention, respectively. Data from this study can be used as part of risk assessment processes in order to identify mitigation strategies to minimize prevalence of E. coli O157 on fresh beef carcasses.

2008 ◽  
Vol 71 (9) ◽  
pp. 1761-1767 ◽  
Author(s):  
J. T. FOX ◽  
D. G. RENTER ◽  
M. W. SANDERSON ◽  
A. L. NUTSCH ◽  
X. SHI ◽  
...  

To quantify associations at slaughter between Escherichia coli O157 carcass contamination, fecal-positive animals, and high-shedding animals within truckloads of finished cattle, we sampled up to 32 cattle from each of 50 truckloads arriving at a commercial abattoir in the Midwest United States during a 5-week summer period. Carcass swab samples collected preevisceration and fecal samples collected postevisceration were matched within animals and analyzed for the presence of E. coli O157, using enrichment, immunomagnetic separation, and plating on selective media (IMS). In addition, a direct plating procedure was performed on feces to identify high-shedding animals. E. coli O157 was isolated from 39 (2.6%) of 1,503 carcass samples in 15 (30%) truckloads, and 127 (8.5%) of 1,495 fecal samples in 37 (74%) truckloads. Fifty-five (3.7%) high-shedding animals were detected from 26 (52%) truckloads. Truckload high-shedder (Spearman rank-order correlation coefficient [rs] = 0.68), IMS-positive (rs = 0.48), and combined fecal (rs =0.61) prevalence were significantly correlated with carcass prevalence. The probability of isolating E. coli O157 from a carcass was not significantly associated with the high-shedder or fecal IMS status of the animal from which the carcass was derived. However, the probability of carcass contamination was significantly associated with all truckload-level measures of fecal E. coli O157, particularly whether or not a high shedder was present within the truckload (odds ratio = 16.2; 95% confidence interval, 6.3–43.6). Our results suggest that high shedders within a truckload at slaughter could be a target for mitigation strategies to reduce the probability of preevisceration carcass contamination.


2003 ◽  
Vol 66 (11) ◽  
pp. 1978-1986 ◽  
Author(s):  
GENEVIEVE A. BARKOCY-GALLAGHER ◽  
TERRANCE M. ARTHUR ◽  
MILDRED RIVERA-BETANCOURT ◽  
XIANGWU NOU ◽  
STEVEN D. SHACKELFORD ◽  
...  

The seasonal prevalence of Escherichia coli O157:H7, Salmonella, non-O157 E. coli (STEC), and stx-harboring cells was monitored at three Midwestern fed-beef processing plants. Overall, E. coli O157:H7 was recovered from 5.9% of fecal samples, 60.6% of hide samples, and 26.7% of carcasses sampled before the preevisceration wash. This pathogen also was recovered from 1.2% (15 of 1,232) of carcasses sampled at chilling (postintervention) at approximate levels of <3.0 cells per 100 cm2. In one case, the E. coli O157:H7 concentration dropped from ca. 1,100 cells per 320 cm2 at the preevisceration stage to a level that was undetectable on ca. 2,500 cm2 at the postintervention stage. The prevalence of E. coli O157:H7 in feces peaked in the summer, whereas its prevalence on hide was high from the spring through the fall. Overall, Salmonella was recovered from 4.4, 71.0, and 12.7% of fecal, hide, and preevisceration carcass samples, respectively. Salmonella was recovered from one postintervention carcass (of 1,016 sampled). Salmonella prevalence peaked in feces in the summer and was highest on hide and preevisceration carcasses in the summer and the fall. Non-O157 STEC prevalence also appeared to vary by season, but the efficiency in the recovery of isolates from stx-positive samples ranged from 37.5 to 83.8% and could have influenced these results. Cells harboring stx genes were detected by PCR in 34.3, 92.0, 96.6, and 16.2% of fecal, hide, preevisceration carcass, and postintervention carcass samples, respectively. The approximate level of non-O157 STEC and stx-harboring cells on postintervention carcasses was ≥3.0 cells per 100 cm2 for only 8 of 199 carcasses (4.0%). Overall, the prevalence of E. coli O157:H7, Salmonella, and non-O157 STEC varied by season, was higher on hides than in feces, and decreased dramatically, along with pathogen levels, during processing and during the application of antimicrobial interventions. These results demonstrate the effectiveness of the current interventions used by the industry and highlight the significance of hides as a major source of pathogens on beef carcasses.


2010 ◽  
Vol 73 (6) ◽  
pp. 1030-1037 ◽  
Author(s):  
M. E. JACOB ◽  
D. G. RENTER ◽  
T. G. NAGARAJA

Cattle feces and hides contribute to carcass contamination with Escherichia coli O157:H7, ultimately impacting beef safety. Primary objectives of our cross-sectional study were to evaluate associations among fecal, hide, and preevisceration carcass prevalence of E. coli O157:H7 and to assess factors affecting carcass contamination. Fecal, hide, and preevisceration carcass samples were collected from up to 32 cattle on each of 45 truckloads presented to a midwestern U.S. abattoir. Enrichment and selective culture were used to assess fecal, hide, and carcass prevalence, and direct plating was used to identify cattle shedding high levels of E. coli O157:H7 in feces. Fecal, hide, and carcass prevalence of E. coli O157:H7 within truckload were significantly correlated (P < 0.05) with each other. Enriched fecal sample prevalence was 13.8%, and high shedder prevalence was 3.3%; 38.5% of hides and 10.5% of carcasses were positive for E. coli O157:H7. We used logistic regression to assess animal- and truckload-level variables affecting the probability of carcasses testing positive for E. coli O157:H7. All truckload-level predictors significantly affected the probability of an E. coli O157:H7–positive carcass, including presence of a high shedder within the truckload (odds ratio [OR] = 4.0; confidence interval [CI], 1.6 to 10.1), high (>25%) within-truckload fecal prevalence (OR = 19.3; CI, 4.7 to 79.0), and high (>50%) within-truckload hide prevalence (OR = 7.7; CI, 3.1 to 19.6). The only significant animal-level predictor was having a positive hide (OR = 1.6; CI, 1.0 to 2.6). Our results suggest that preharvest interventions for reducing E. coli O157:H7 contamination of carcasses should focus on truckload (cohort)–level and hide mitigation strategies.


2011 ◽  
Vol 74 (7) ◽  
pp. 1186-1188 ◽  
Author(s):  
M. E. JACOB ◽  
K. M. ALMES ◽  
X. SHI ◽  
J. M. SARGEANT ◽  
T. G. NAGARAJA

Escherichia coli O157:H7 causes foodborne illness in humans; cattle are considered a primary reservoir for the organism, and transmission is often through contaminated food products or water. The objective of this study was to determine the genetic diversity of E. coli O157:H7 within a single individual bovine fecal sample based on pulsed-field gel electrophoresis (PFGE) typing. Fecal samples (n = 601) were collected from dairy and beef cattle at three separate facilities, and E. coli O157:H7 was isolated by enrichment, immunomagnetic separation, and plating on selective medium. The prevalence of E. coli O157:H7 was 46 (7.7%) of 601. From each positive fecal sample, up to 10 putative colonies were tested, and isolates from samples with at least seven positive colonies were subtyped using PFGE and tested for six major virulence genes by multiplex PCR. A total of 254 E. coli O157:H7 isolates from 27 samples met these criteria and were included in PFGE analysis. Fifteen PFGE subtypes (<100% Dice similarity) were detected among the 254 isolates, and there were no common subtypes between the three locations. Seven (26%) of 27 fecal samples had E. coli O157:H7 isolates with different PFGE subtypes (mean = 2.1) within the same sample. The virulence gene profiles of different isolates from the same sample were always identical, regardless of the number of PFGE types. The results of this study suggest that determining the PFGE pattern of a single isolate from a bovine sample may not be sufficient when comparing isolates from feces, hides, or carcasses, because multiple PFGE subtypes are present.


2008 ◽  
Vol 74 (10) ◽  
pp. 3138-3142 ◽  
Author(s):  
Haiping Li ◽  
Mehrdad Tajkarimi ◽  
Bennie I. Osburn

ABSTRACT Vacuum cooling is a common practice in the California leafy green industry. This study addressed the impact of vacuum cooling on the infiltration of Escherichia coli O157:H7 into lettuce as part of the risk assessment responding to the E. coli O157:H7 outbreaks associated with leafy green produce from California. Vacuum cooling significantly increased the infiltration of E. coli O157:H7 into the lettuce tissue (2.65E+06 CFU/g) compared to the nonvacuumed condition (1.98E+05 CFU/g). A stringent surface sterilization and quadruple washing could not eliminate the internalized bacteria from lettuce. It appeared that vacuuming forcibly changed the structure of lettuce tissue such as the stomata, suggesting a possible mechanism of E. coli O157:H7 internalization. Vacuuming also caused a lower reduction rate of E. coli O157:H7 in stored lettuce leaves than that for the nonvacuumed condition.


2005 ◽  
Vol 68 (1) ◽  
pp. 26-33 ◽  
Author(s):  
K. STANFORD ◽  
S. J. BACH ◽  
T. H. MARX ◽  
S. JONES ◽  
J. R. HANSEN ◽  
...  

On-farm methods of monitoring Escherichia coli O157:H7 were assessed in 30 experimentally inoculated steers housed in four pens over a 12-week period and in 202,878 naturally colonized feedlot cattle housed in 1,160 pens on four commercial Alberta feedlots over a 1-year period. In the challenge study, yearling steers were experimentally inoculated with 1010 CFU of a four-strain mixture of nalidixic acid–resistant E. coli O157:H7. After inoculation, shedding of E. coli O157:H7 was monitored weekly by collecting rectal fecal samples (FEC), oral swabs (ORL), pooled fecal pats (PAT), manila ropes (ROP) orally accessed for 4 h, feed samples, water, and water bowl interface. Collection of FEC from all animals per pen provided superior isolation (P < 0.01) of E. coli O157:H7 compared with other methods, although labor and animal restraint requirements for fecal sample collection were high. When one sample was collected per pen of animals, E. coli O157:H7 was more likely to be detected from the ROP than from the FEC, PAT, or ORL (P < 0.001). In the commercial feedlot study, samples were limited to ROP and PAT, and E. coli O157:H7 was isolated in 18.8% of PAT and 6.8% of ROP samples. However, for animals that had been resident in the feedlot pen for at least 1 month, isolation of E. coli O157:H7 from ROP was not different from that from PAT (P = 0.35). Pens of animals on feed for <30 days were six times more likely to shed E. coli O157:H7 than were animals on feed for >30 days. However, change in diet did not affect shedding of the organism (P > 0.23) provided that animals had acclimated to the feedlot for 1 month or longer. Findings from this study indicate the importance of introduction of mitigation strategies early in the feeding period to reduce transference and the degree to which E. coli O157:H7 is shed into the environment.


2016 ◽  
Vol 79 (1) ◽  
pp. 66-74 ◽  
Author(s):  
P. B. SHRIDHAR ◽  
L. W. NOLL ◽  
X. SHI ◽  
B. AN ◽  
N. CERNICCHIARO ◽  
...  

ABSTRACT Shiga toxin–producing Escherichia coli (STEC) serogroups O26, O45, O103, O111, O121, and O145, called non-O157 STEC, are important foodborne pathogens. Cattle, a major reservoir, harbor the organisms in the hindgut and shed them in the feces. Although limited data exist on fecal shedding, concentrations of non-O157 STEC in feces have not been reported. The objectives of our study were (i) to develop and validate two multiplex quantitative PCR (mqPCR) assays, targeting O-antigen genes of O26, O103, and O111 (mqPCR-1) and O45, O121, and O145 (mqPCR-2); (ii) to utilize the two assays, together with a previously developed four-plex qPCR assay (mqPCR-3) targeting the O157 antigen and three virulence genes (stx1, stx2, and eae), to quantify seven serogroups and three virulence genes in cattle feces; and (iii) to compare the three mqPCR assays to a 10-plex conventional PCR (cPCR) targeting seven serogroups and three virulence genes and culture methods to detect seven E. coli serogroups in cattle feces. The two mqPCR assays (1 and 2) were shown to be specific to the target genes, and the detection limits were 4 and 2 log CFU/g of pure culture–spiked fecal samples, before and after enrichment, respectively. A total of 576 fecal samples collected from a feedlot were enriched in E. coli broth and were subjected to quantification (before enrichment) and detection (after enrichment). Of the 576 fecal samples subjected, before enrichment, to three mqPCR assays for quantification, 175 (30.4%) were quantifiable (≥4 log CFU/g) for at least one of the seven serogroups, with O157 being the most common serogroup. The three mqPCR assays detected higher proportions of postenriched fecal samples (P < 0.01) as positive for one or more serogroups compared with cPCR and culture methods. This is the first study to assess the applicability of qPCR assays to detect and quantify six non-O157 serogroups in cattle feces and to generate data on fecal concentration of the six serogroups.


2004 ◽  
Vol 67 (4) ◽  
pp. 672-678 ◽  
Author(s):  
S. J. BACH ◽  
T. A. McALLISTER ◽  
G. J. MEARS ◽  
K. S. SCHWARTZKOPF-GENSWEIN

The effects of weaning and transport on fecal shedding of Escherichia coli and on E. coli O157:H7 were investigated using 80 Angus and 94 Charolais range steer calves blocked by breed and assigned to four treatments. The calves were or were not preconditioned before transport on commercial cattle liner to the feedlot via long (15 h) or short (3 h) hauling duration, yielding preconditioned long haul (P-L; n = 44), preconditioned short haul (P-S; n = 44), nonpreconditioned long haul (NP-L; n = 43), and nonpreconditioned short haul (NP-S; n = 43). Preconditioned calves were vaccinated and weaned 29 and 13 days, respectively, before transport. Nonpreconditioned calves were weaned 1 day before long or short hauling, penned for 24 h and hauled again for 2 h, and vaccinated on arrival at the feedlot. Fecal samples were collected from calves while on pasture, at weaning, at loading for transport, on arrival at the feedlot, twice in the first week, and on days 7, 14, 21, and 28 for enumeration of total E. coli (biotype 1) and detection of E. coli O157:H7. No calves were positive for E. coli O157:H7 before transport. Following transport, more (P < 0.005) NP-L calves (6 of 43) tested positive for E. coli O157:H7 than did P-L (1 of 44), NP-S (1 of 43), or P-S (0 of 44) calves, and on days 0, 1, 7, and 21, their levels of shedding of E. coli were higher (P < 0.005). The calves' susceptibility to infection from the environment (possibly the holding facilities or feedlot pens) was likely elevated by the stresses of weaning, transport, and relocation. Lack of preconditioning and long periods of transport (NP-L) increased fecal shedding of E. coli and E. coli O157:H7. Preconditioning may serve to reduce E. coli O157:H7 shedding by range calves on arrival at the feedlot.


2006 ◽  
Vol 69 (1) ◽  
pp. 6-11 ◽  
Author(s):  
L. SCOTT ◽  
P. McGEE ◽  
J. J. SHERIDAN ◽  
B. EARLEY ◽  
N. LEONARD

Escherichia coli O157:H7 is an important foodborne pathogen that can cause hemorrhagic colitis and hemolytic uremic syndrome. Cattle feces and fecally contaminated water are important in the transmission of this organism on the farm. In this study, the survival of E. coli O157:H7 in feces and water was compared following passage through the animal digestive tract or preparation in the laboratory. Feces were collected from steers before and after oral inoculation with a marked strain of E. coli O157:H7. Fecal samples collected before cattle inoculation were subsequently inoculated with the marked strain of E. coli O157:H7 prepared in the laboratory. Subsamples were taken from both animal and laboratory-inoculated feces to inoculate 5-liter volumes of water. E. coli O157:H7 in feces survived up to 97 days, and survival was not affected by the method used to prepare the inoculating strain. E. coli O157:H7 survived up to 109 days in water, and the bacteria collected from inoculated cattle were detected up to 10 weeks longer than the laboratory-prepared culture. This study suggests that pathogen survival in low-nutrient conditions may be enhanced by passage through the gastrointestinal tract.


2007 ◽  
Vol 70 (10) ◽  
pp. 2230-2234 ◽  
Author(s):  
T. W. THOMPSON ◽  
T. P. STEPHENS ◽  
G. H. LONERAGAN ◽  
M. F. MILLER ◽  
M. M. BRASHEARS

Rapid enzyme-linked immunosorbent assays (ELISAs) are approved for detection of Escherichia coli O157 in beef products. However, these kits have also been used in the industry to detect this pathogen on hides or in feces of cattle, although this use has not been validated. The objective of this study was to compare commercially available ELISAs (E. coli Now, Reveal, and VIP) with immunomagnetic separation along with selective media to detect E. coli O157 on hides, in feces, and in medium- and low-level-inoculated ground beef and carcasses (simulated by using briskets) samples. Naturally infected hide and fecal samples were subjected to both the immunomagnetic separation method and ELISAs for the detection of E. coli O157. Additionally, E. coli O157 inoculated and noninoculated ground beef and beef briskets were used to simulate meat and carcass samples. When comparing the detection results from the ELISAs (E. coli Now, Reveal, and VIP) to the immunomagnetic separation method, poor agreement was observed for fecal samples (kappa = 0.10, 0.02, and 0.03 for E. coli Now, Reveal, and VIP, respectively), and fair-to-moderate agreement was observed for hide samples (kappa = 0.30, 0.51, and 0.29 for E. coli Now, Reveal, and VIP, respectively). However, there was near-perfect agreement between the immunomagnetic separation method and ELISAs for ground beef (kappa = 1, 1, and 0.80 for E. coli Now, Reveal, and VIP, respectively) and brisket (kappa = 1, 1, and 1 for E. coli Now, Reveal, and VIP, respectively) samples. Assuming immunomagnetic separation is the best available method, these data suggest that the ELISAs are not useful in detecting E. coli O157 from hide or fecal samples. However, when ELISAs are used on ground beef and beef brisket samples they can be used with a high degree of confidence.


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