Prevalence of Escherichia coli O157 and levels of aerobic bacteria and Enterobacteriaceae are reduced when hides are washed and treated with cetylpyridinium chloride at a commercial beef processing plant

The objective of this experiment was to test the potential of a combined water wash and cetylpyridinium chloride (CPC) treatment as a hide intervention applied to cattle in the holding pens of a processing plant immediately before stunning. Over 2 processing days, 149 control and 139 treated cattle were tested. Control cattle were processed in the normal manner. The treatment group was prewashed with water the day before harvest. Immediately before stunning, these cattle were sprayed twice with 1% CPC, first for 3 min, then for 1 min. Hides and preevisceration carcasses were sampled to determine aerobic plate counts, Enterobacteriaceae counts (EBC), and Escherichia coli O157 prevalence. The treatment reduced the prevalence of E. coli O157 on hides from 56% to 34% and the prevalence on preevisceration carcasses from 23% to 3%. The treatment decreased aerobic plate counts from 4.9 log CFU/100 cm2 to 3.4 log CFU/100 cm2 and EBC from 3.1 log CFU/100 cm2 to 2.0 log CFU/100 cm2 on preevisceration carcasses. The treatment of hides did not result in any detectable CPC contamination of the chilled carcasses. These data indicated that a 1% CPC treatment preceded by a water wash was capable of reducing hide prevalence of E. coli O157 from as high as 80% to less than 50%, resulting in preevisceration carcass prevalence of 5% or less. We conclude that water washing followed by an antimicrobial treatment, such as CPC, has great potential as an effective hide intervention step and should be further evaluated for implementation as a processing step after stunning and before hide removal.     

Escherichia coli O157 prevalence and enumeration of aerobic bacteria, Enterobacteriaceae, and Escherichia coli O157 at various steps in commercial beef processing plants

The effectiveness of current antimicrobial interventions used in reducing the prevalence or load of Escherichia coli O157 and indicator organisms on cattle hides and carcasses at two commercial beef processing plants was evaluated. Sponge sampling of beef cattle was performed at five locations from the initial entry of the animals to the slaughter floor to the exit of carcasses from the "hotbox" cooler. For each sample, E. coli O157 prevalence was determined and total aerobic bacteria, Enterobacteriaceae, and E. coli O157 were enumerated. E. coli O157 was found on 76% of animal hides coming into the plants, but no carcasses leaving the cooler were identified as contaminated with E. coli O157. A positive relationship was seen between the incidence of E. coli O157 in hide samples and that in preevisceration samples. Aerobic plate counts and Enterobacteriaceae counts averaged 7.8 and 6.2 log CFU/100 cm2, respectively, on hides, and 1.4 and 0.4 log CFU/100 cm2, respectively, on chilled carcasses. Aerobic plate counts and Enterobacteriaceae counts on preevisceration carcasses were significantly related to the respective levels on the corresponding hides; the carcasses of animals whose hides carried higher numbers of bacteria were more likely to carry higher numbers of bacteria. Implementation of the sampling protocol described here would allow processors to evaluate the efficacy of on-line antimicrobial interventions and allow industrywide benchmarking of hygienic practices.     

Efficacy of ozonated and electrolyzed oxidative waters to decontaminate hides of cattle before slaughter

The hides of cattle are the primary source of pathogens such as Escherichia coli O157:H7 that contaminate preevisceration carcasses during commercial beef processing. A number of interventions that reduce hide contamination and subsequent carcass contamination are currently being developed. The objective of this study was to determine the efficacy of ozonated and electrolyzed oxidizing (EO) waters to decontaminate beef hides and to compare these treatments with similar washing in water without the active antimicrobial compounds. Cattle hides draped over barrels were used as the model system. Ozonated water (2 ppm) was applied at 4,800 kPa (700 lb in2) and 15 degrees C for 10 s. Alkaline EO water and acidic EO water were sequentially applied at 60 degrees C for 10 s at 4,800 and 1,700 kPa (250 lb in2), respectively. Treatment using ozonated water reduced hide aerobic plate counts by 2.1 log CFU/100 cm2 and reduced Enterobacteriaceae counts by 3.4 log CFU/100 cm2. EO water treatment reduced aerobic plate counts by 3.5 log CFU/100 cm2 and reduced Enterobacteriaceae counts by 4.3 log CFU/100 cm2. Water controls that matched the wash conditions of the ozonated and EO treatments reduced aerobic plate counts by only 0.5 and 1.0 log CFU/100 cm2, respectively, and each reduced Enterobacteriaceae counts by 0.9 log CFU/100 cm2. The prevalence of E. coli O157 on hides was reduced from 89 to 31% following treatment with ozonated water and from 82 to 35% following EO water treatment. Control wash treatments had no significant effect on the prevalence of E. coli O157:H7. These results demonstrate that ozonated and EO waters can be used to decontaminate hides during processing and may be viable treatments for significantly reducing pathogen loads on beef hides, thereby reducing pathogens on beef carcasses.     

Development and evaluation of an on-line hide decontamination procedure for use in a commercial beef processing plantt

The hides of cattle are the source of Escherichia coli O157:H7 that contaminates beef carcasses during commercial beef processing. Therefore, effective interventions that reduce hide contamination should reduce subsequent carcass contamination. The first objective of this study was to identify the most effective reagents for decontamination of beef hides. Cattle hides draped over barrels were used for in vitro experiments to compare the efficacy of washes using 1.6% sodium hydroxide, 4% trisodium phosphate, 4% chlorofoam, or 4% phosphoric acid, each followed by a rinse step using either water or acidified (pH 7.0) chlorine at 200 or 500 ppm. All treatments using a water rinse reduced hide coliform counts by 1.5 to 2.5 log CFU/ 100 cm2. Compared with water rinses, 200 and 500 ppm acidified chlorine rinses increased efficacy by approximately 1.0 and 2.0 log CFU/100 cm2, respectively. Vacuuming of the treated areas to remove excess liquid improved hide cleanliness by an average of an additional 1.0 log CFU/100 cm2. The second objective was to evaluate the use of an on-line hide-wash cabinet that used a sodium hydroxide wash and a chlorinated (1 ppm) water rinse. Hides sampled before entering and after exiting the cabinet had aerobic plate counts and Enterobacteriaceae counts that were reduced by 2.1 and 3.4 log CFU/100 cm2, respectively, and the prevalence of E. coli O157 on hides was reduced from 44 to 17% when the cabinet was in use. Preevisceration carcass aerobic plate counts and Enterobacteriaceae counts were both reduced by 0.8 log CFU/100 cm2, and the prevalence of E. coli O157 on preevisceration carcasses was reduced from 17 to 2% when the cabinet was in use. These results support decontamination of hides as an effective means to reduce pathogen contamination of cattle carcasses during processing.     

Treatments using hot water instead of lactic acid reduce levels of aerobic bacteria and Enterobacteriaceae and reduce the prevalence of Escherichia coil O157:H7 on preevisceration beef carcasses

Lactic acid has become the most commonly used organic acid for treatment of postevisceration beef carcasses. Many processors have also implemented 2% lactic acid washes on preevisceration carcasses. We previously demonstrated that hot water washing and steam vacuuming are effective carcass interventions. Because of the effectiveness of hot water, we compared its use with that of lactic acid as a preevisceration wash in a commercial setting. A commercial hot water carcass wash cabinet applying 74 degrees C (165 degrees F) water for 5.5 s reduced both aerobic plate counts and Enterobacteriaceae counts by 2.7 log CFU/100 cm2 on preevisceration carcasses. A commercial lactic acid spray cabinet that applied 2% L-lactic acid at approximately 42 degrees C (105 to 110 degrees F) to preevisceration carcasses reduced aerobic plate counts by 1.6 log CFU/100 cm2 and Enterobacteriaceae counts by 1.0 log CFU/100 cm2. When the two cabinets were in use sequentially, i.e., hot water followed by lactic acid, aerobic plate counts were reduced by 2.2 log CFU/100 cm2 and Enterobacteriaceae counts were reduced by 2.5 log CFU/100 cm2. Hot water treatments reduced Escherichia coli O157:H7 prevalence by 81%, and lactic acid treatments reduced E. coli O157:H7 prevalence by 35%, but the two treatments in combination produced a 79% reduction in E. coli O157:H7, a result that was no better than that achieved with hot water alone. These results suggest that hot water would be more beneficial than lactic acid for decontamination of preevisceration beef carcasses.     

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.     

Protocol for evaluating the efficacy of cetylpyridinium chloride as a beef hide intervention

The objective of this study was to establish the necessary protocols and assess the efficacy of cetylpyridinium chloride (CPC) as an antimicrobial intervention on beef cattle hides. Experiments using CPC were conducted to determine (i) the methods of neutralization needed to obtain valid efficacy measurements, (ii) the effect of concentration and dwell time after treatment, (iii) the effect of CPC on hide and carcass microbial populations when cattle were treated at a feedlot and then transported to a processing facility for harvest, and (iv) the effectiveness of spray pressure and two-spray combinations of CPC and water to reduce hide microbial populations. Residual CPC in hide sponge samples prevented bacterial growth. Dey-Engley neutralization media at 7.8% and a centrifugation step were necessary to overcome this problem. All dwell times, ranging from 30 s to 4 h, after 1% CPC application to cattle hides resulted in aerobic plate counts and Enterobacteriaceae counts 1.5 log CFU/100 cm2 lower than controls. The most effective dose of CPC was 1%, which reduced aerobic plate counts and Enterobacteriaceae counts 2 and 1 log CFU/100 cm2, respectively. Low-pressure application of 1% CPC at the feedlot, transport to the processing facility, and harvest within 5 h of application resulted in no effect on Escherichia coli O157 prevalence on hides or preevisceration carcasses. Two high-pressure CPC washes lowered aerobic plate counts and Enterobacteriaceae counts by 4 log CFU/100 cm2, and two medium-pressure CPC washes were only slightly less effective. These results indicate that under the proper conditions, CPC may still be effective for reducing microbial populations on cattle hides. Further study is warranted to determine if this effect will result in reduction of hide-to-carcass contamination during processing.     

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.     

Efficacy of hypobromous acid as a hide-on carcass antimicrobial intervention

Escherichia coli O157:H7 and Salmonella on cattle hides at slaughter are the main source of beef carcass contamination by these foodborne pathogens during processing. Hypobromous acid (HOBr) has been approved for various applications in meat processing, but the efficacy of HOBr as a hide antimicrobial has not been determined. In this study, the antimicrobial properties of HOBr were determined by spraying cattle hides at either of two concentrations, 220 or 500 ppm. Treatment of hides with 220 ppm of HOBr reduced the prevalence of E. coli O157:H7 on hides from 25.3 to 10.1% (P < 0.05) and reduced the prevalence of Salmonella from 28.3 to 7.1% (P < 0.05). Treatment of hides with 500 ppm of HOBr reduced (P < 0.05) the prevalence of E. coli O157:H7 on hides from 21.2 to 10.1% and the prevalence of Salmonella from 33.3 to 8.1%. The application of 220 ppm of HOBr reduced (P < 0.05) aerobic plate counts, total coliform counts, and E. coli counts on hides by 2.2 log CFU/ 100 cm(2). The use of 500 ppm of HOBr resulted in reductions (P < 0.05) of aerobic plate counts, total coliform counts, and E. coli counts by 3.3, 3.7, and 3.8 log CFU/100 cm(2), respectively, demonstrating that the use of higher concentrations of HOBr on hides resulted in additional antimicrobial activity. These results indicate that the adoption of a HOBr hide wash will reduce hide concentrations of spoilage bacteria and pathogen prevalence, resulting in a lower risk of carcass contamination.     

Treatment of cattle hides with Shellac solution to reduce hide-to-beef microbial transfer

The hide-to-beef microbial transfer-reducing effects of a novel Shellac treatment of hides (based on "on-hair immobilization" of microorganisms) were evaluated. In the hide-to-meat direct contact laboratory-based experiments, treatment of hides (of varying visual cleanliness) with Shellac produced significant microbial reductions on beef: up to 3.6 log(10) CFU/cm(2) of total viable count of bacteria (TVC), up to 2.5 log(10) CFU/cm(2) of Enterobacteriaceae (EC) and up to 1.7 log(10) CFU/cm(2) of generic Escherichia coli (GEC). In a small commercial abattoir under "bad-case" conditions (slaughtering dirty cattle, inadequate process hygiene), treatment of hides with Shellac produced significant microbial reductions on beef carcasses: 1.7 log(10) CFU/cm(2), 1.4 log(10) CFU/cm(2) and 1.3 log(10) CFU/cm(2) of TVC, EC and GEC, respectively. In both laboratory- and abattoir-based trials, TVC reductions on beef achieved by the Shellac hide treatment were superior to those achieved by the comparative sanitizer rinse-vacuum hide treatment, but reductions of EC and GEC did not differ significantly between the two hide treatments.     

Studies to evaluate chemicals and conditions with low-pressure applications for reducing microbial counts on cattle hides

Studies were conducted to identify effective antimicrobials and application parameters that could be used as decontamination interventions to reduce microbial loads on cattle hides before removal from carcasses. In study I, hide swatches inoculated with Escherichia coli O157:H7 were sprayed with 10% acetic acid (at 23 and 55 degrees C), 10% lactic acid (at 23 and 55 degrees C), 3% sodium hydroxide (at 23 degrees C) or 4 and 5% sodium metasilicate (at 23 degrees C). All antimicrobials were evaluated independently after being applied alone, being applied after a water rinse, or being followed by a water rinse. Antimicrobial treatments followed by a water rinse lowered E. coli O157:H7 populations by 0.6 to 2.4 log CFU/cm2 and resulted in hides with a surface pH of 6.3 to 9.2. Treatments in which a water rinse was followed by antimicrobial application lowered E. coli O157:H7 populations by 1.5 to 5.1 log CFU/cm2 but resulted in hides with a surface pH of 3.9 to 10.5. In study II, whole hides were treated with one of four antimicrobials (acetic acid, lactic acid, sodium hydroxide, or sodium metasilicate) followed by a water rinse. Hides were evaluated for aerobic bacterial counts, total coliform counts, and E. coli counts. Generally, all antimicrobials resulted in greater reductions (P < 0.05) of E. coli counts when compared with the control; however, only acetic and lactic acids resulted in greater reductions (P < 0.05) of aerobic bacterial counts and total coliform counts compared with the controls. These antimicrobials could be used to reduce microbial contamination on hides, potentially reducing microbiological contamination transferred to carcasses or to the plant environment.     

Evaluation of various antimicrobial interventions for the reduction of Escherichia coli O157:H7 on bovine heads during processing

The effectiveness of electrolyzed oxidizing water, FreshFx, hot water, DL-lactic acid, and ozonated water was determined using a model carcass spray-washing cabinet. A total of 140 beef heads obtained from a commercial processing line were inoculated with Escherichia coli O157:H7 on the cheek areas. Each head was exposed to a simulated preevisceration wash and then had antimicrobial wash treatments. Hot water, lactic acid, and FreshFx treatments reduced E. coli O157:H7 on inoculated beef heads by 1.72, 1.52, and 1.06 log CFU/cm2, respectively, relative to the simulated preevisceration wash. Electrolyzed oxidizing water and ozonated water reduced E. coli O157:H7 less than 0.50 log CFU/cm2. Hot water, lactic acid, and FreshFx could be used as decontamination washes for the reduction of E. coli O157:H7 on bovine head and cheek meat.     

Rapid enrichment strategy for isolation of Listeria from bovine hide, carcass, and meat samples

Since the outbreak of foodborne illness linked to Escherichia coli O157:H7 bacteria in ground beef in the early 1980s, the beef processing industry has focused on increasing the safety of beef products by implementing procedures for surveying live cattle, carcasses, and beef products for bacterial pathogens. Effective methods are in place for screening cattle and beef products for the presence of E. coli O157:H7 contamination, and recent work has established the acceptability of these methods for surveillance of Salmonella. In keeping with the need to continually improve the food safety of beef products, new work investigating pathogen prevalence now includes surveillance for Listeria monocytogenes. Tryptic soy broth (TSB) has been documented as a robust nonselective medium for the enrichment of both E. coli and Salmonella from bovine hide, carcass, and meat samples. The University of Vermont modification medium is most often used as the primary enrichment medium for surveillance of Listeria spp. In this study, samples from bovine hides (n=50), preevisceration carcasses (n=50), and beef trim (n=193) were used to evaluate TSB as a primary enrichment medium for the isolation of Listeria spp., including L. monocytogenes. No significant difference (P > 0.05) between TSB and the University of Vermont modification medium was observed when all three sample types underwent primary enrichment for the isolation of Listeria spp. Furthermore, the standard secondary enrichment ratio for Fraser broth used for Listeria recovery can be modified to accommodate a high-throughput method for processing multiple samples.     

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.     

Microbiological characterization of lamb carcasses at commercial processing plants in the United States

Although the United States produces 203 million lb (ca. 92.1 kg) of domestic lamb and mutton each year, thorough studies of the microbiological safety during lamb processing are lacking. To address this missing information, a total of 2,548 sponge samples from pelts, preevisceration carcasses, and postintervention carcasses were collected from multiple large commercial lamb processing plants to determine aerobic plate counts, the prevalences of Escherichia coli O157:H7, non-O157 Shiga toxin-producing E. coli (STEC), and Salmonella. The averages of the aerobic plate counts from pelts, the preevisceration carcasses, and the postintervention carcasses were 6.3, 4.4, and 2.4 log CFU/100 cm2, respectively. The prevalences of E. coli O157:H7 from the pelts, the preevisceration carcasses, and the postintervention carcasses were 12.8, 1.6, and 2.9%, respectively. The average Salmonella prevalences were 14.4, 4.3, and 1.8% for pelts, preevisceration carcasses, and postintervention carcasses, respectively. The most frequently identified Salmonella serotype was Heidelberg. The prevalences of non-O157 STEC from pelts, preevisceration carcasses, and postintervention carcasses averaged 86.2, 78.6, and 81.6%, respectively. A total of 488 non-O157 S0TEC strains were isolated from postintervention carcasses. Sixty-nine different serotypes of non-O157 STEC were identified. The most frequently detected serotypes were O91:H14 (40.8%), followed by O5:H19 (18.4%). A small number of STEC serotypes associated with severe human illness were isolated from postintervention carcasses. These were serotypes O76:H19, O128:H2 (0.8%), O146:H8 (2.1%), ) O146:H21, O163:H19, and O174:H8 (1.3%). The results of this study establish a baseline for microbiological quality and prevalences of Salmonella, E. coli O157:H7, and STEC in U.S. lamb processing plants.     

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.     

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.     

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.     

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.     

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.