Effects of the transportation of beef cattle from the feedyard to the packing plant on prevalence levels of Escherichia coli O157 and Salmonella spp

Two hundred steers and heifers from a large feedyard (65,000-head capacity) were used to determine the prevalence levels of enterohemorrhagic Escherichia coli O157 (EHEC O157) and Salmonella spp. prior to and after shipping to a commercial packing facility. Two samples, a ventral midline hide swab and a fecal sample, were aseptically collected from each animal 2 weeks prior to the date of transportation and at the packing plant immediately after exsanguination. Samples were collected from all trailers (n = 46) before animals were loaded for transport to the packing facility. The average prevalence levels of EHEC O157 on hides (18%) and in feces (9.5%) at the feedyard decreased (P > 0.05) at the packing plant to 4.5 and 5.5%, respectively. The average prevalence levels of Salmonella spp. on hides (6%) and in feces (18%) at the feedyard increased to 89 and 46%, respectively, upon arrival at the packing plant. Average prevalence levels for EHEC O157 and Salmonella spp. on the trailers were 5.43 and 59%, respectively. The results of this study demonstrate that transportation may be a potential stressor for cattle, as evidenced by the increased shedding of Salmonella spp.     

Seasonal prevalence of Shiga toxin-producing Escherichia coli, including O157:H7 and non-O157 serotypes, and Salmonella in commercial beef processing plants

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 > or = 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.     

An investigation of Escherichia coli O157 contamination of cattle during slaughter at an abattoir

The extent of contamination with Escherichia coli O157 was determined for 100 cattle during slaughter. Samples from 25 consecutively slaughtered cattle from four unrelated groups were collected from the oral cavity, hide, rumen, feces after evisceration, and pre- and postchill carcass. Ten random fecal samples were collected from the pen where each group of animals was held at the abattoir. E. coli O157 was detected using automated immunomagnetic separation (AIMS), and cell counts were determined using a combination of most probable number (MPN) and AIMS. E. coli O157 was isolated from 87 (14%) of the 606 samples collected, including 24% of 99 oral cavity samples, 44% of 100 hides, 10% of 68 fecal samples collected postevisceration, 6% of 100 prechill carcass swabs, and 15% of 40 fecal samples collected from holding pens. E. coli O157 was not isolated from rumen or postchill carcass samples. E. coli O157 was isolated from at least one sample from each group of cattle tested, and the prevalence in different groups ranged from less than 1 to 41%. The numbers of E. coli O157 differed among the animals groups. The group which contained the highest fecal (7.5 x 10(5) MPN/g) and hide (22 MPN/cm2) counts in any individual animal was the only group in which E. coli O157 was isolated from carcasses, suggesting a link between the numbers of E. coli O157 present and the risk of carcass contamination. Processing practices at this abattoir were adequate for minimizing contamination of carcasses, even when animals were heavily contaminated with E. coli O157.     

Prevalence of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella in two geographically distant commercial beef processing plants in the United States

For two large beef processing plants, one located in the southern United States (plant A) and one located in the northern United States (plant B), prevalence of Escherichia coli O157:H7, Listeria spp., Listeria monocytogenes, and Salmonella was determined for hide, carcass, and facility environmental samples over the course of 5 months. The prevalence of E. coli O157:H7 (68.1 versus 55.9%) and Salmonella (91.8 versus 50.3%) was higher (P < 0.05), and the prevalence of Listeria spp. (37.7 versus 75.5%) and L. monocytogenes (0.8 versus 18.7%) was lower (P < 0.05) for the hides of cattle slaughtered at plant A versus plant B. Similarly, the prevalence of Salmonella (52.0 versus 25.3%) was higher (P < 0.05) and the prevalence of Listeria spp. (12.0 versus 40.0%) and L. monocytogenes (1.3 versus 14.7%) was lower (P < 0.05) for the fence panels of the holding pens of plant A versus plant B. The prevalence of E. coli O157:H7 (3.1 versus 10.9%), Listeria spp. (4.5 versus 14.6%), and L. monocytogenes (0.0 versus 1.1%) was lower (P < 0.05) for preevisceration carcasses sampled at plant A versus plant B. Salmonella (both plants), Listeria spp. (plant B), and L. monocytogenes (plant B) were detected on fabrication floor conveyor belts (product contact surfaces) late during the production day. For plant B, 21 of 148 (14.2%) late-operational fabrication floor conveyor belt samples were L. monocytogenes positive. For plant B, E. coli O157:H7 and L. monocytogenes were detected in preoperational fabrication floor conveyor belt samples. Overall results suggest that there are regional differences in the prevalence of pathogens on the hides of cattle presented for harvest at commercial beef processing plants. While hide data may reflect the regional prevalence, the carcass data is indicative of differences in harvest practices and procedures in these plants.     

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.     

Comparison of sampling methods for microbiological testing of beef animal rectal/colonal feces, hides, and carcasses

This study compared sampling methods for detecting Escherichia coli O157:H7 and Salmonella in beef cattle feces and on hides and carcasses and for enumerating E. coli biotype I counts (ECC) on carcasses. Fecal samples were collected by rectal/colonal palpation and colonal sponge swabbing. Hides were sampled by sponge swabbing three sites, hair clipping, excision, rinsing, and gauze swabbing, whereas carcasses were sampled by three-site thoracic and pattern-mark sponge swabbing and tissue excision. Overall, irrespective of sampling method, 36.7, 13.3, and 0.0% of lots contained at least one E. coli O157:H7-positive hide, fecal, and carcass sample, respectively, while the corresponding prevalence of Salmonella was 70.0, 16.7, and 6.7%, respectively. For hide sampling, excision and gauze swabbing yielded the fewest (13.3%) E. coli O157:H7-positive samples, while hair clipping and sponge swabbing yielded the most (23.3%). None of the carcass-sampling methods detected E. coli O157:H7 or differed (P > 0.05) in their ability to enumerate ECC. Colonal swabbing was the most effective (10.0%) method for detecting E. coli O157:H7 in feces. No differences (P > 0.05) in Salmonella prevalence were observed between carcass-sampling methods, although three-site sponge swabbing and tissue excision detected the most (3.3%). Hide rinsing was the most effective (P < 0.05) Salmonella detection method (63.3%), but dangers associated with its application may preclude its use by industry; there were no differences (P > 0.05) among other hide-sampling methods. No differences (P > 0.05) in Salmonella detection were observed between fecal-sampling methods. Overall, three-site sponge swabbing was the most feasible and effective sampling method for the detection of E. coli O157:H7 and Salmonella on hides and carcasses.     

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.     

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.     

Incidence of enterohemorrhagic Escherichia coli, Escherichia coli O157, Salmonella, and Listeria monocytogenes in retail fresh ground beef, sprouts, and mushrooms

The objective of this study was to determine the prevalence of enterohemorrhagic Escherichia coli (EHEC), E. coli O157, Salmonella, and Listeria monocytogenes in retail food samples from Seattle, Wash. A total of 2,050 samples of ground beef (1,750 samples), mushrooms (100 samples), and sprouts (200 samples) were collected over a 12-month period and analyzed for the presence of these pathogens. PCR assays, followed by culture confirmation were used to determine the presence or absence of each organism. Of the 1,750 ground beef samples analyzed, 61 (3.5%) were positive for EHEC, and 20 (1.1%) of these were positive for E. coli O157. Salmonella was present in 67 (3.8%) of the 1,750 ground beef samples. Of 512 ground beef samples analyzed, 18 (3.5%) were positive for L. monocytogenes. EHEC was found in 12 (6.0%) of the 200 sprout samples, and 3 (1.5%) of these yielded E. coli O157. Of the 200 total sprout samples, 14 (7.0%) were positive for Salmonella and none were positive for L. monocytogenes. Among the 100 mushroom samples, 4 (4.0%) were positive for EHEC but none of these 4 samples were positive for E. coli O157. Salmonella was detected in 5 (5.0%) of the mushroom samples, and L. monocytogenes was found in 1 (1.0%) of the samples.     

Influence of vitamin D on fecal shedding of Escherichia coli O157:H7 in naturally colonized cattle

Three experiments were conducted to evaluate the influence of vitamin D on fecal shedding of Escherichia coli O157:H7 in cattle. In the first experiment, two groups of cattle (beef and dairy) were assigned to a control treatment or to receive 0.5 × 10(6) IU vitamin D per day via oral bolus for 10 days. Fecal samples were collected before and throughout the dosing period for culture of E. coli O157:H7. No differences were observed for fecal shedding of E. coli O157:H7 among treatments for either beef or dairy animals. Serum concentrations of vitamin D were markedly higher (P < 0.0001) in treated beef cattle but only tended to be higher (P = 0.09) in the dairy cattle. In the second experiment, three successive vitamin D dosages (2,400, 4,800, and 9,600 IU/day; 14 days each) were administered to 14 dairy steers (7 steers served as controls), fecal samples were collected daily, and serum samples were collected weekly throughout the 42-day experimental period. No significant differences in fecal prevalence or serum vitamin D concentrations were observed for any of the vitamin D dosages. A third experiment sampled feedlot cattle (winter and summer) to determine whether serum vitamin D concentrations were correlated with fecal shedding of E. coli O157:H7. A fecal sample and a blood sample were obtained in each season from 60 randomly selected animals (total of 120 fecal samples and 120 corresponding blood samples). As expected, season was highly correlated (r = 0.66) with serum vitamin D concentration with higher concentrations (P < 0.01) observed in the summer. E. coli O157:H7 prevalence (percentage of positive samples) was not highly correlated (r = 0.16) with season, although the correlation tended to be significant (P = 0.08). The proportion of cattle shedding E. coli O157:H7 was 16.7 and 6.7% for the summer and winter collections, respectively. Results of this research do not support a correlation between vitamin D intake and E. coli O157:H7 shedding in cattle.     

Impact of reducing the level of wet distillers grains fed to cattle prior to harvest on prevalence and levels of Escherichia coli O157:H7 in feces and on hides

Cattle fed finishing diets with wet distillers grains with solubles (WDGS) have been shown to harbor increased Escherichia coli O157:H7 populations in the feces and on the hides. To determine if feeding a lower level of WDGS at the end of the feeding period reduces E. coli O157:H7 load at harvest, 608 heifers were sorted into one of five treatments and fed 0, 40, or 70% WDGS (dry matter basis). For three of the treatments, WDGS was reduced midway through the study. Treatment 0W0W heifers (positive control) were fed a corn grain-based diet continuously, and 40W40W heifers (negative control) were fed 40% WDGS continuously. Heifers subjected to treatments 40W0W, 40W15W, and 70W15W were fed either 40 or 70% WDGS for the first 56 days and switched to 0 or 15% WDGS, respectively, for the last 56 days. Prior to the switch in diets, animals fed diets with 40 or 70% had higher prevalence and percent enumerable fecal samples for E. coli O157:H7. After the dietary switch, animals fed 40W0W, 40W15W, and 70W15W diets had fecal prevalence and percent enumerable samples (33.4 and 6.3%, 31.0 and 9.7%, and 34.9 and 8.4%, respectively) similar to those of animals fed 0W0W diets (10.2 and 3.2%, respectively; P > 0.05), whereas animals fed 40W40W had the highest fecal prevalence and percent enumerable samples (70.1 and 29.2%, respectively; P < 0.05). Similar relationships between the treatments were observed for hide samples. Time after dietary switch was important, as animals fed lower levels had significantly lower fecal prevalence and percent enumerable samples after 56 days, but not after 28 days. The study indicates that cattle can be switched to lower levels of dietary WDGS (15% or less) 56 days prior to harvest to significantly reduce E. coli O157:H7 in feces and on hides.     

Determining the prevalence of Escherichia coli O157 in cattle and beef from the feedlot to the cooler

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, postevisceration, 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.     

Prevalence of Escherichia coli O157:H7 and performance by beef feedlot cattle given Lactobacillus direct-fed microbials

Fecal shedding of Escherichia coli O157:H7, the prevalence of Escherichia coli O157:H7 in pens and on carcasses and hides, and cattle performance as a result of daily dietary supplementation with Lactobacillus-based direct-fed microbials (DFMs) were evaluated in a feeding trial involving 180 beef steers. Steers were evaluated for shedding of E. coli O157:H7 by an immunomagnetic separation technique on arrival at the feedlot, just before treatment with the DFMs, and every 14 days thereafter until slaughter. Composite pen fecal samples were collected every 14 days (alternating weeks with animal testing), and prevalence on hides and carcasses at slaughter was also evaluated. Feedlot performance (body weight gain and feed intake) was measured for the period during which the DFMs were fed. Gain efficiency was calculated as the ratio of weight gain to feed intake. Lactobacillus acidophilus NPC 747 decreased (P < 0.01) the shedding of E. coli O157:H7 in the feces of individual cattle during the feeding period. E. coli O157:H7 was approximately twice as likely to be detected in control animal samples as in samples from animals receiving L. acidophilus NPC 747. In addition, DFM supplementation decreased (P < 0.05) the number of E. coli O157:H7-positive hide samples at harvest and the number of pens testing positive for the pathogen. Body weight gains (on a live or carcass basis) and feed intakes during the DFM supplementation period did not differ among treatments. Gain efficiencies on a live-weight basis did not differ among treatments, but carcass-based gain/feed ratios tended (P < 0.06) to be better for animals receiving the two DFM treatments than for control animals. The results of this study suggest that the feeding of a Lactobacillus-based DFM to cattle will decrease, but not eliminate, fecal shedding of E. coli O157:H7, as well as contamination on hides, without detrimental effects on performance.     

Methods for recovering Escherichia coli O157:H7 from cattle fecal, hide, and carcass samples: sensitivity and improvements

The Meats Research Unit (MRU) methods, developed by MRU scientists of the U.S. Meat Animal Research Center, have been used to study the prevalence of Escherichia coli O157:H7 in cattle carcass, hide, and fecal samples. The sensitivity of these methods for recovery of injured E. coli O157:H7 cells from inoculated and uninoculated samples was determined, and potential improvements to these methods were evaluated. When using the conventional MRU methods, 91% of the pre-evisceration carcass samples tested positive for E. coli O157:H7 when inoculated with 5 to 10 CFU, 100% of hide samples tested positive for E. coli O157:H7 when inoculated with 30 to 50 CFU, and 96% of the fecal samples produced positive results when inoculated with 300 to 400 CFU per 10 g. The addition of a phosphate buffer to the tryptic soy broth enrichment improved recovery of E. coli O157:H7 from feces. Using the modified enrichment, 92% of the samples were identified as positive when inoculated with 10 to 30 CFU per 10 g. Substituting a commercially available wash buffer for the phosphate-buffered saline (PBS) plus Tween 20 wash buffer during immunomagnetic separation of hide samples improved recovery of the target organism at lower inoculum concentrations. When comparing uninoculated samples, substituting a PBS buffer plus a zwitterionic detergent for PBS plus Tween 20 also had a positive effect on recovery of E. coli O157:H7 from hide samples. Data presented here indicate that the MRU methods are highly effective at recovering injured E. coli O157:H7 from fecal, hide, and beef carcass samples; however, modifications can be added to increase the sensitivity.     

Distribution of Escherichia coli 0157 and Salmonella on hide surfaces, the oral cavity, and in feces of feedlot cattle

To determine the distribution of pathogens on cattle hides at the feedlot, samples were collected from six hide surface locations (back, flank, hock, neck, perineum, and ventrum), the oral cavity, the rectal-anal junction, and the feces of feedlot cattle and subjected to Escherichia coli 0157 detection via culture methods and to Salmonella detection via PCR. E. coli 0157 was isolated from one or more of the sampling locations from 31 (42.5%) of the 73 animals sampled. Location-specific prevalence of E. coli 0157 was 5% for back samples, 5% for flank samples, 12% for hock samples, 7% for neck samples, 12% for perineum samples, 8% for ventrum samples, 1% for oral cavity samples, 4% for rectal-anal junction swabs, and 23% for fecal grab samples. Salmonella was isolated from one or more of these sample locations from 100% (50 of 50 samples) of all animals sampled. Location-specific prevalence of Salmonella was 76% for back samples, 74% for flank samples, 94% for hock samples, 76% for neck samples, 88% for perineum samples, 86% for ventrum samples, 94% for oral cavity samples, 64% for rectal-anal junction swabs, and 50% for fecal grab samples. The sampling locations that maximized the likelihood of finding E. coli 0157 and Salmonella (84 and 96%, respectively) if the animal was positive at one sampling location or more were the hock, perineum, and fecal grab. These data suggest that the use of multiple sample locations is useful when isolating these pathogens from feedlot cattle. Focusing on one sampling location may underestimate the prevalence.     

Development of methods for the recovery of Escherichia coil O157:H7 and Salmonella from beef carcass sponge samples and bovine fecal and hide samples

Culture methods were developed for the concurrent recovery of Escherichia coli O157:H7 and Salmonella from bovine carcass, hide, and fecal samples. Several enrichment conditions were tested for the overall growth of pure cultures; tryptic soy broth for 2 h at 25 degrees C and then for 6 h at 42 degrees C was the protocol selected for use. Immunomagnetic separation (IMS) was incorporated for sensitivity and selectivity, along with a post-IMS enrichment for the recovery of Salmonella as recommended by the manufacturer. Selective agars for plating after IMS were chosen on the basis of ease of target colony identification. Sorbitol MacConkey agar supplemented with cefixime and potassium tellurite and Rainbow agar supplemented with novobiocin and potassium tellurite were chosen for the recovery of E. coli O157:H7. Brilliant green agar with sulfadiazine and Hektoen enteric agar supplemented with novobiocin were selected for the recovery of Salmonella. The resulting methods were evaluated along with standard or previously used methods for the recovery of E. coli O157:H7 and Salmonella from bovine hide and fecal samples and carcass sponge samples. The Meats Research Unit (MRU) methods performed at least as well as the established methods, except that a secondary enrichment in tetrathionate (TT) broth prior to IMS was required for the optimal recovery of Salmonella from feces. Thus, the MRU and MRU-TT methods are effective in the recovery of both E. coli O157:H7 and Salmonella from a single bovine carcass, hide, or fecal sample.     

Development of an experimental model to assess the ability of Escherichia coli O157:H7-inoculated fecal pats to mimic a super shedder within a feedlot environment

This study was conducted to develop an experimental model that could assess the ability of Escherichia coli O157:H7-inoculated fecal pats to mimic a super shedder (>10(4) CFU/g of feces) within a feedlot environment. The day before the study began, 48 steers that had been negative for E. coli O157:H7 in feces for three consecutive weeks were sorted into three treatment groups, with two replicate pens per treatment and 8 steers per pen. Steers within the pens (20.50 by 10.75 m) were exposed to control feces or feces inoculated with two levels of a mixture of five strains of nalidixic acid-resistant E. coli O157:H7 (low level, 10(2) CFU/g; high level, 10(5) CFU/g). Five 300-g fecal pats were introduced into the pens twice daily (10:00 a.m. and 2:30 p.m.) on days 0 through 6 and days 14 through 20. Pats were placed in the pen at random locations to mimic defecation of a steer within the pen. Fecal grab samples, hide swab samples (500-cm2 area of the rump), natural fecal pat samples (freshly voided), and rope samples (1.22-m-long manila rope) where obtained at multiple times during the 49-day trial to evaluate the spread of nalidixic acid-resistant E. coli O157:H7 throughout the feedlot environment and among penmates. Immunomagnetic separation and selective media were used to detect E. coli O157:H7. Nalidixic acid-resistant E. coli O157:H7 was detected in 13 high-level treatment fecal grab samples, 7 high-level treatment hide swab samples, 1 low-level hide swab sample, and 2 high-level rope samples. For both fecal grab and hide swab samples, the overall prevalence of E. coli O157:H7 in the high-level group was greater (P < 0.01) than that for the pooled low-level and control groups. Addition of inoculated fecal pats to pens increased transmission of E. coli O157:H7 among penmates, but cattle that acquired E. coli O157:H7 shed the bacterium for only a short time at low levels. Transmission of E. coli O157:H7 from the feces of super shedders to naive penmates may contribute to the observed transient nature of shedding of E. coli O157:H7 among feedlot cattle.     

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.     

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.     

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

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.