Seasonal prevalence of Escherichia coli O157:H7 in beef cattle feces

Cattle are an asymptomatic reservoir of Escherichia coli O157:H7, but the bacterial colonization and shedding patterns are poorly understood. The prevalence and shedding of this human pathogen have been reported to be seasonal with rates typically increasing during warm months. The objectives of this study were (i) to assess the prevalence of E. coli O157:H7 in feces of feedlot cattle in Kansas during summer, fall, and winter months, and (ii) to characterize E. coli O157:H7 by screening for virulence factors. Of 891 fecal samples collected, 82 (9.2%) were positive for E. coli O157:H7. No significant differences in prevalence were detected among summer, fall, and winter months. The highest monthly prevalence (18.1%) was detected in February. All tested isolates were positive for stx2 (Shiga toxin 2) and eaeA (intimin) genes; 14 isolates (12.8%) also carried stx1. Our results indicate the prevalence of E. coli O157:H7 in beef cattle feces is not necessarily season dependent.     

Characterization of O157:H7 and other Escherichia coli isolates recovered from cattle hides, feces, and carcasses

In a previous study, the seasonal prevalence was reported for stx+ Escherichia coli O157:H7 in feces and on hides and carcasses of cattle at processing. Overall, 1,697 O157:H7 isolates have now been characterized for the incidence of (i) eae(O157), hlyA, stx1, and stx2 in the recovered isolates and (ii) presumptive rough and presumptive nonmotile isolates. Seven O157:H7 isolates (0.4%) lacked stx genes, although they carried eae and hlyA. All but one of the isolates carried both eae and hlyA. Approximately two-thirds of the isolates (64% when one isolate per sample was considered) carried both stx1 and stx2. E. coli O157:H7 cells that harbored both stx1 and stx2 were more often recovered from hides in the fall (79% of the fall hide isolates) and winter (84% of the winter hide isolates) than in the spring (53%) and summer (59%). Isolates recovered from preevisceration carcasses showed a similar but not statistically significant trend. Twenty-three of the 25 O157:H7 isolates carrying stx1 but not stx2 were recovered during summer. Fifteen presumptive rough and 117 presumptive nonmotile stx+ O157:H7 isolates were recovered. Ten (67%) of the presumptive rough isolates were recovered during summer. Ninety-five of the presumptive nonmotile isolates (81%) were recovered during fall. Forty-eight percent of the false-positive isolates (175 of 363) tentatively identified as O157:H7 were O157+ H7- and lacked eae(O157), hlyA, and stx. These data suggest that in beef processing samples (i) there are minor seasonal variations in the prevalence of stx genes among E. coli O157:H7 isolates, (ii) presumptive rough and presumptive nonmotile stx+ O157:H7 isolates are present, (iii) E. coli O157:H7 isolates lacking stx genes may be rare, and (iv) O157+ H7- isolates lacking stx genes can result in many false-positive results.     

Effects of a minimal hide wash cabinet on the levels and prevalence of Escherichia coli O157:H7 and Salmonella on the hides of beef cattle at slaughter

Harborage of Escherichia coli O157:H7 and Salmonella on animal hides at slaughter is the main source of beef carcass contamination during processing. Given this finding, interventions have been designed and implemented to target the hides of cattle following entry into beef processing plants. Previous interventions targeting hides have not been suitable for all beef processing plants because of cost and space restrictions. In this study, a hide wash cabinet was evaluated to determine whether it was more amenable to widespread use in the beef processing industry, especially for small and medium-size plants. Overall, 101 (35.1%) of 288 beef cattle hides sampled before entry into the hide wash cabinet harbored E. coli O157:H7 at or above the limit of detection (40 CFU/100 cm2). After passage through the hide wash cabinet, only 38 (13.2%) of 288 hides had E. coli O157:H7 levels > or =40 CFU/100 cm2. Before the hide wash cabinet, 50 (17%) of 288 hides harbored E. coli O157:H7 at levels above 100 CFU/100 cm2, with one sample as high as 20,000 CFU/100 cm2. In contrast, only 14 (5%) of 288 hides had E. coli O157:H7 levels above 100 CFU/100 cm2 after hide washing, with the highest being 2000 CFU/100 cm2. These same trends also were found for Salmonella before and after hide washing. These results indicate that the hide wash cabinet described in this study was effective and should provide small and medium-size processing plants with an affordable hide wash intervention strategy.     

Frequency of Escherichia coli O157:H7 in Turkish cattle

In this study, five abattoirs in Istanbul were visited between January 2000 and April 2001. During these visits, 330 cattle were selected by a systematic sampling method. Cattle were examined clinically and breed, age, and sex were recorded. Rectal swabs were taken immediately after slaughter. Immunomagnetic separation was performed, and sorbitol-negative colonies were selected on sorbitol MacConkey agar with cefixime and tellurite (CT-SMAC agar). These colonies were checked for 4-methylenebelliferyl-beta-D-glucuronide, indol, rhamnose, and urease activity and motility. Serotypes of bacteria were determined by using antisera specific for Escherichia coli O157 and H7. All cattle selected were clinically healthy. Of 88 sorbitol-negative colonies selected on CT-SMAC agar, isolates from only 14 (4.2%) cattle reacted with anti-O157, and 13 of these isolates also reacted with anti-H7. E. coli O157:H7 was isolated from all breeds, but the numbers of isolates were largest for Holstein and Swiss Brown cows. E. coli O157:H7 was most frequently isolated from 2-year-old cattle. Similarly, it was most frequently isolated from male cattle. E. coli O157:H7 was isolated from cattle slaughtered in four of the five abattoirs studied.     

Dietary supplementation with Lactobacillus- and Propionibacterium-based direct-fed microbials and prevalence of Escherichia coli O157 in beef feedlot cattle and on hides at harvest

The objective of this study was to describe the prevalence of Escherichia coli O157 in the feces and on the hides of finishing beef cattle fed a standard diet and those fed diets supplemented with direct-fed microbials. Two hundred forty steers received one of four treatments throughout the feeding period: (i) control: no added microbials; (ii) HNP51: high dose of Lactohacillius acidophilus strain NP 51 (10(9) CFU per steer daily) and Propionibacterium freudenreichii (10(9) CFU per steer daily); (iii) HNP51+45: high dose of NP 51 (10(9) CFU per steer daily), P. freudenreichii (10(9) CFU per steer daily), and L. acidophilus NP 45 (10(6) CFU per steer daily); or (iv) LNP51+45: low dose of NP 51 (10(6) CFU per steer daily), P. freudenreichii (10(9) CFU per steer daily), and NP 45 (10(6) CFU per steer daily). Samples were collected from each animal and analyzed for the presence of E. coli O157 using immunomagnetic separation methods on day 0 (feces), 7 days before harvest (feces), and at harvest (feces and hide). At the end of the feeding period, cattle receiving HNP51 were 57% less likely to shed detectable E. coli O157 in their feces than were the controls (P < 0.01). For animals receiving HNP51+45 and LNP51+45, fecal prevalence did not differ from that of the controls. The prevalence of positive hide samples was least among cattle receiving HNP51+45 (3.3%); these animals were 79% less likely (P < 0.06) to have a positive hide sample than were the controls (prevalence = 13.8%). There was poor agreement of the culture results between fecal and hide samples collected from the same animal (kappa = 0.08; confidence interval = -0.05 to 0.2). Cattle supplemented with a high dose of NP 51 had reduced E. coli O157 prevalence in both fecal and hide samples, indicating that this treatment may be an efficacious preharvest intervention strategy.     

Impact of transportation of feedlot cattle to the harvest facility on the prevalence of Escherichia coli 0157:H7, Salmonella, and total aerobic microorganisms on hides

Prevalences of Escherichia coli O157:H7, Salmonella, and total aerobic microorganisms were determined on the hides of beef feedlot cattle before and after transport from the feedyard to the harvest facility in clean and dirty trailers. Swab samples were taken from the midline and withers of 40 animals on each of 8 days before and after shipping. After samples were collected, animals were loaded in groups of 10 on upper and lower levels of clean and dirty trailers. Animals were unloaded at the harvest facility and kept in treatment groups for sample collection after exsanguination. Salmonella was found more often on hide swabs collected from the midline than on than samples collected from the withers from animals transported in both clean and dirty trailers. Salmonella was found on significantly more hide swabs collected at harvest from both sampling locations than on those collected at the feedyard, with no differences attributed to the type of trailer. At the feedyard, clean trucks had a lower percentage of Salmonella-positive samples than did dirty trucks before animals were loaded. However, after transport, both clean and dirty trucks had a similar prevalence of Salmonella. There were no differences in Salmonella prevalence on hides collected from animals transported on the top and bottom levels of clean and dirty trucks. E. coli O157:H7 was detected on less than 2% of the samples; therefore, no practical conclusions about prevalence could be drawn. Hides sampled at harvest had higher concentrations of aerobic microorganisms than did hides sampled at the feedyard, and concentrations were higher on the midline than on the withers. Although the prevalences of Salmonella and total aerobic microorganisms increased on hides after transport from the feedyardto the plant, this increase was not related to the cleanliness of the trailers or the location of the cattle in the trailers.     

Horizontal transmission of Escherichia coli O157:H7 during cattle housing

Ruminant livestock, particularly cattle, is considered the primary reservoir of Escherichia coli O157:H7. This study examines the transmission of E. coli O157:H7 within groups of cattle during winter housing. Holstein Friesian steers were grouped in six pens of five animals. An animal inoculated with and proven to be shedding a marked strain of E. coli O157: H7 was introduced into each pen. Fecal (rectal swabs) and hide samples (900 cm2 from the right rump) were taken from the 36 animals throughout the study. Water, feed, and gate or partition samples from each pen were also examined. Within 24 h of introducing the inoculated animals into the pens, samples collected from the drinking water, pen barriers, and animal hides were positive for the pathogen. Within 48 h, the hides of 20 (66%) of 30 cohort animals from the six pens were contaminated with E. coli O157:H7. The first positive fecal samples from the noninoculated cohort animals were detected 3 days after the introduction of the inoculated steers. During the 23 days of the study, 15 of 30 cohort animals shed the marked E. coli O157:H7 strain in their feces on at least one occasion. Animal behavior in the pens was monitored during a 12-h period using closed circuit television cameras. The camera footage showed an average of 13 instances of animal grooming in each pen per hour. The study suggests that transmission of E. coli O157:H7 between animals may occur following ingestion of the pathogen at low levels and that animal hide may be an important source of transmission.     

Transportation and lairage environment effects on prevalence, numbers, and diversity of Escherichia coli O157:H7 on hides and carcasses of beef cattle at processing

Hide has been established as the main source of carcass contamination during cattle processing; therefore, it is crucial to minimize the amount of Escherichia coli O157:H7 on cattle hides before slaughter. Several potential sources of E. coli O157: H7 are encountered during transportation and in the lairage environment at beef-processing facilities that could increase the prevalence and numbers of E. coli O157:H7 on the hides of cattle. On three separate occasions, samples were obtained from cattle at the feedlot and again after cattle were stunned and exsanguinated at the processing plant (286 total animals). The prevalence of E. coli O157:H7 on hides increased from 50.3 to 94.4% between the time cattle were loaded onto tractor-trailers at the feedlot and the time hides were removed in the processing plant. Before transport, nine animals had E. coli O157:H7 in high numbers (> 0.4 CFU/cm2) on their hides. When sampled at the slaughter facility, the number of animals with high hide numbers had increased to 70. Overall, only 29% of the E. coli O157:H7 isolates collected postharvest (221 of 764) matched pulsed-field gel electrophoresis types collected before transport. The results of this study indicate that transport to and lairage at processing plants can lead to increases in the prevalence and degree of E. coli O157:H7 contamination on hides and the number of E. coli O157:H7 pulsed-field gel electrophoresis types associated with the animals. More study is needed to confirm the mechanism by which additional E. coli O157:H7 strains contaminate cattle hides during transport and lairage and to design interventions to prevent this contamination.     

Molecular characterization of Escherichia coli O157:H7 hide contamination routes: feedlot to harvest

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.     

Soil solarization reduces Escherichia coli O157:H7 and total Escherichia coli on cattle feedlot pen surfaces

Feedlot pen soil is a source for transmission of Escherichia coli O157:H7, and therefore a target for preharvest strategies to reduce this pathogen in cattle. The objective of this study was to determine the ability of soil solarization to reduce E. coli O157:H7 in feedlot surface material (FSM). A feedlot pen was identified in which naturally occurring E. coli O157:H7 was prevalent and evenly distributed in the FSM. Forty plots 3 by 3 m were randomly assigned such that five plots of each of the solarization times of 0, 1, 2, 3, 4, 6, 8, and 10 weeks were examined. Temperature loggers were placed 7.5 cm below the surface of each plot, and plots to be solarized were covered with clear 6-mil polyethylene. At each sampling time, five FSM samples were collected from each of five solarized and five unsolarized plots. E. coli concentrations and E. coli O157:H7 presence by immunomagnetic separation and plating were determined for each FSM sample. Initial percentages of E. coli O157:H7-positive samples in control and solarized FSM were 84 and 80%, respectively, and did not differ (P > 0.05). E. coli O157:H7 was no longer detectable by 8 weeks of solarization, but was still detected in unsolarized FSM at 10 weeks. The average initial concentration of E. coli in FSM was 5.56 log CFU/g and did not differ between treatments (P > 0.05). There was a 2.0-log decrease of E. coli after 1 week of solarization, and a >3.0-log reduction of E. coli by week 6 of solarization (P, 0.05). E. coli levels remained unchanged in unsolarized FSM (P > 0.05). Daily peak FSM temperatures were on average 8.7°C higher for solarized FSM compared with unsolarized FSM, and reached temperatures as high as 57°C. Because soil solarization reduces E. coli O157:H7, this technique may be useful for reduction of persistence and transmission of this pathogen in cattle production, in addition to remediation of E. coli O157:H7-contaminated soil used to grow food crops.     

Escherichia coli O157:H7 excretion by commercial feedlot cattle fed either barley- or corn-based finishing diets

Effective preharvest control measures for Escherichia coli O157:H7 in cattle may significantly reduce the incidence of human disease caused by this organism. The prevalence and magnitude of fecal E. coli O157:H7 excretion was evaluated in 15 pens (300 to 500 cattle per pen) of commercial feedlot cattle fed a barley-based finishing ration and compared with that in 15 pens of cattle fed a corn-based ration. Average E. coli O157:H7 prevalence was 2.4% in barley-fed cattle and 1.3% in the corn-fed cattle (P < 0.05), and average magnitude of fecal E. coli O157:H7 excretion was 3.3 log CFU/g in the barley-fed cattle and 3.0 log CFU/g in the corn-fed cattle (P < 0.01). Corn-fed cattle had lower average fecal pH values (5.85) than did barley-fed cattle (6.51) (P < 0.01), and the average total generic fecal E. coli concentration in this group of animals (6.24 log CFU/g) was greater than that in the barley-fed cattle (5.55 log CFU/g) (P < 0.01). Specific feed ingredients may impact the frequency and magnitude of fecal excretion of E. coli O157:H7 by cattle.     

Escherichia coli O157:H7 prevalence in fecal samples of cattle from a southeastern beef cow-calf herd

The proportion of fecal samples culture-positive for Escherichia coli O157:H7 was determined for samples collected from 296 beef cows on pasture in a single Florida herd in October, November, and December 2001. The overall proportion of samples that cultured positive was 0.03. The proportion of cows that were culture-positive on at least one occasion was 0.091. No effect of pregnancy status or nutritional regimen on the proportion of culture-positive samples for E. coli O157:H7 was detected. We detected a breed effect on the shedding of E. coli O157, with Romosinuano cows having a lower (P < 0.01) proportion of samples culture-positive than Angus or Brahman cows. This difference might have resulted from the presence of confounding variables; however, it also might represent evidence of breed-to-breed genetic variation in E. coli O157 shedding. Further research is warranted to evaluate breed as a possible risk factor for shedding of this important foodborne pathogen. Further substantiated findings could indicate that breed is a cow-calf-level critical control point of E. coli O157:H7.     

Reduction of Escherichia coli O157 and Salmonella in feces and on hides of feedlot cattle using various doses of a direct-fed microbial

In this study, the effectiveness of direct-fed microbials at reducing Escherichia coli O157 and Salmonella in beef cattle was evaluated. Steers (n=240) received one of the following four treatment concentrations: control = lactose carrier only; low = 1 X 10(7) CFU per steer daily Lactobacillus acidophilus NP51; medium = 5 x 10(8) CFU per steer daily L. acidophilus NP51; and high = 1 x 10(9) CFU per steer daily L. acidophilus NP51. Low, medium, and high diets also included 1 x 10(9) CFU per steer Propionibacterium freudenreichii NP24. Feces were collected from each animal at allocation of treatment and found to have no variation (P = 0.54) between cohorts concerning E. coli O157 recovery. Feces and hide swabs were collected at harvest and analyzed for the presence of E. coli O157 by immunomagnetic separation and Salmonella by PCR. No significant dosing effects were detected for E. coli O157 recovery from feces at the medium dose or from hides at the medium and high doses. E. coli O157 was 74% (P < 0.01) and 69% (P < 0.01) less likely to be recovered in feces from animals receiving the high and low diets, respectively, compared with controls. Compared with controls, E. coli O157 was 74% (P = 0.05) less likely to be isolated on hides of cattle receiving the low dose. No significant dosing effects were detected for Salmonella recovery from feces at the medium and low doses or from hides at any doses. Compared with controls, Salmonella was 48% (P = 0.09) less likely to be shed in feces of cattle receiving the high dose. No obvious dose-response of L. acidophilus NP51 on recovery of E. coli O157 or Salmonella was detected in our study.     

A comparison of the survival in feces and water of Escherichia coli O157:H7 grown under laboratory conditions or obtained from cattle feces

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.     

Escherichia coli O157:H7 shedding in vaccinated beef calves born to cows vaccinated prepartum with Escherichia coli O157:H7 SRP vaccine

Extensive research, intervention equipment, money, and media coverage have been directed at controlling Escherichia coli O157:H7 in beef cattle. However, much of the focus has been on controlling this pathogen postcolonization. This study was conducted to examine the performance, health, and shedding characteristics of beef calves that were vaccinated with an E. coli O157:H7 SRP bacterial extract. These calves had been born to cows vaccinated prepartum with the same vaccine. Cows and calves were assigned randomly to one of four treatments: (i) neither cows nor calves vaccinated with E. coli O157:H7 SRP (CON), (ii) cows vaccinated with E. coli O157:H7 SRP prepartum but calves not vaccinated (COWVAC), (iii) calves vaccinated with E. coli O157:H7 SRP but born to cows not vaccinated (CALFVAC), (iv) cows vaccinated with E. coli O157:H7 SRP prepartum and calves also vaccinated (BOTH). Calves born to vaccinated cows had significantly higher titers of anti-E. coli O157:H7 SRP antibodies (SRPAb) in circulation at branding time (P < 0.001). Upon entry to the feedlot, overall fecal E. coli O157:H7 prevalence was 23 % among calves, with 25 % in the CON treatment group, 19 % in the CALFVAC group, 32 % in the COWVAC group, and 15 % in the BOTH group (P > 0.05). Fecal shedding of E. coli O157 on arrival to the feedlot was not correlated with fecal shedding at slaughter (Spearman's rho = -0.02; P = 0.91). No significant effects of cow or calf E. coli O157:H7 SRP vaccination treatment were found on feedlot calf health or performance (P > 0.05), prevalence of lung lesions or liver abscess (P > 0.05), or morbidity, retreatment, or mortality numbers (P > 0.05). The findings of this study indicate that the timing of vaccination of calves against E. coli O157:H7 may be an important consideration for maximizing the field efficacy of this vaccine.     

Comparison of the molecular genotypes of Escherichia coli O157:H7 from the hides of beef cattle in different regions of North America

Cattle hides become contaminated with Escherichia coli O157:H7 via pathogen transmission in the feedlot, during transport, and while in the lairage environment at the processing facility, and the bacteria can be transferred to beef carcasses during processing. Several studies have shown that E. coli O157:H7 strains possessing indistinguishable restriction digest patterns (RDPs) can be isolated from distant locations. Most of these studies, however, examined RDPs from strains isolated within a single region of the United States or Canada. The experiment described in the present study was designed to identify the molecular genotypes of E. coli O157:H7 isolates from beef cattle hides in nine major cattle-producing regions of North America. Prevalence for E. coli O157:H7 in beef cattle hide samples ranged from 9 to 85%. Pulsed-field gel electrophoresis (PFGE) analysis of XbaI-digested genomic DNA from 1,193 E. coli O157:H7 isolates resulted in 277 unique RDPs. Of the 277 unique XbaI RDPs, 54 contained isolates collected from multiple regions. After two subsequent rounds of PFGE analysis (BlnI and SpeI), there were still many isolates (n = 154) that could not be distinguished from others, even though they were collected from different regions separated by large geographical distances. On multiple occasions, strains isolated from cattle hides in Canada had RDPs that were indistinguishable after three enzyme digestions from cattle hide isolates collected in Kansas and Nebraska. This information clearly shows that strains with indistinguishable RDPs originate from multiple sources that can be separated by large distances and that this should be taken into account when the source tracking of isolates is based on PFGE.     

Shedding of escherichia coli O157:H7 by cattle fed diets containing monensin or tylosin

Monensin and tylosin have activity against gram-positive bacteria, and it has been theorized that their effects on the intestinal environment may promote proliferation of gram-negative bacteria such as Escherichia coli. Effects of these antibiotics on the shedding of E. coli O157:H7 were studied in a feedlot environment, using 32 finishing steers. A diet containing 85% barley grain, 10% barley silage, and 5% supplement was amended with 33 ppm monensin, 11 ppm tylosin, both of these additives, or no additives (control). All steers were orally inoculated with 10(10) CFU of a mixture of four strains of nalidixic acid-resistant E. coli O157:H7. Fecal (grab), oral (mouth swab) and water, water-water bowl interface, feed, and pen floor fecal pat samples were collected weekly for 12 weeks. Prevalence of E. coli O157:H7-positive fecal grab samples did not differ (P = 0.26) among treatments, nor did the rate (P = 0.81) or duration (P = 0.85) of shedding of the organism. Fecal grab samples were positive for E. coli O157:H7 more frequently (P < 0.001) than were oral swabs. More (P = 0.02) E. coli O157:H7-positive oral swabs were recovered from the tylosin group than from controls. E. coli O157:H7 was not detected in any of 47 water samples, but was present in 1 of 47 water bowl swabs, 7 of 48 feed samples, and 36 of 48 fecal pats. Pulsed-field gel electrophoresis suggested that differences existed among inoculated strains in their ability to persist in animals and in the environment. However, this study revealed no evidence that dietary inclusion of monensin or tylosin, alone or in combination, increased fecal shedding of E. coli O157:H7 or its persistence in the environment.     

Low prevalence of Escherichia coli O157:H7 in horses in Ohio, USA

Manure from draft animals deposited in fields during vegetable and fruit production may serve as a potential source of preharvest pathogen contamination of foods. To better quantify this risk, we determined the prevalence of Escherichia coli O157:H7 in horses. Between June and September 2009, freshly voided fecal samples were collected from horses stabled on 242 separate premises in Ohio, USA. Overall, the prevalence of E. coli O157:H7 was 1 of 242 (0.4% prevalence, 95% confidence interval [CI] = 0.01 to 2.28). E. coli O157:H7 was recovered from none of the 107 equine fecal samples (0% prevalence, 95% CI = 0.00 to 3.39) that originated from locations without ruminant presence, and only 1 of the 135 horse fecal samples (0.7% prevalence, 95% CI = 0.02 to 4.06) from sites where ruminants were also present. The lone positive sample was collected from a horse that was costabled with a goat. Subsequent sampling at that location identified indistinguishable subtypes of E. coli O157:H7 present in the cohoused goat, in the environment, insects, sheep, and other goats housed in an adjacent field. E. coli O157:H7 was not isolated from the five subsequent samples from this horse. These data indicate that E. coli O157:H7 carriage by horses is an uncommon event.     

Cross-contamination of lettuce with Escherichia coli O157:H7

Contamination of produce by bacterial pathogens is an increasingly recognized problem. In March 1999, 72 patrons of a Nebraska restaurant were infected with enterohemorrhagic Escherichia coli (EHEC) O157:H7, and shredded iceberg lettuce was implicated as the food source. We simulated the restaurant's lettuce preparation procedure to determine the extent of possible EHEC cross-contamination and growth during handling. EHEC inoculation experiments were conducted to simulate the restaurant's cutting procedure and the subsequent storage of shredded lettuce in water in the refrigerator. All lettuce pieces were contaminated after 24 h of storage in inoculated water (2 x 10(9) CFU of EHEC per 3 liters of water) at room temperature or at 4 degrees C; EHEC levels associated with lettuce increased by > 1.5 logs on the second day of storage at 4 degrees C. All lettuce pieces were contaminated after 24 h of storage in water containing one inoculated lettuce piece (approximately 10(5) CFU of EHEC per lettuce piece) at both temperatures. The mixing of one inoculated dry lettuce piece with a large volume of dry lettuce, followed by storage at 4 degrees C or 25 degrees C for 20 h resulted in 100% contamination of the leaves tested. Microcolonies were observed on lettuce stored at 25 degrees C, while only single cells were seen on leaves stored at 4 degrees C, suggesting that bacterial growth had occurred at room temperature. Three water washes did not significantly decrease the number of contaminated leaves. Washing with 2,000 mg of calcium hypochlorite per liter significantly reduced the number of contaminated pieces but did not eliminate contamination on large numbers of leaves. Temperature abuse during storage at 25 degrees C for 20 h decreased the effectiveness of the calcium hypochlorite treatment, most likely because of bacterial growth during the storage period. These data indicate that storage of cut lettuce in water is not advisable and that strict attention must be paid to temperature control during the storage of cut lettuce.     

Incidence and toxin production ability of Escherichia coli O157:H7 isolated from cattle trucks

Twelve cattle trucks were analyzed for the presence of Escherichia coli O157:H7. Three of them had been washed prior to arrival, and the others had not. Seventy-five percent of the trailers were positive for the presence of this foodborne pathogen. A total of 54 cultures were isolated and identified as E. coli O157:H7, all from the trucks that had not been cleaned. Most of the cultures (96.4%) produced Shiga-like toxin (verotoxin). No E. coli O157:H7 was detected in cattle trucks that were cleaned before arrival at the cattle pens. The incidence of E. coli O157:H7 in transport trailers increases the potential risk of contamination of cattle and transmission from farms to feedlots and to packing plants. This contamination increases the potential of contamination of meat during harvest and the risk of foodborne illnesses.