Preevisceration Carcasses Research Articles

Distribution of Extremely Heat-Resistant Escherichia coli in the Beef Production and Processing Continuum

Understanding the dynamics of stress-resistant Escherichia coli (E. coli) across the meat production and processing continuum is important for tracking sources of such microbes and devising effective modes of control. The Locus of Heat Resistance (LHR) is a ∼14–19 Kb genetic element imparting extreme heat resistance (XHR) in Enterobacteriaceae. It has been hypothesized that thermal and antimicrobial interventions applied during meat processing may select for LHR+E. coli. Thus, our goal was to study the prevalence and molecular biology of LHR+E. coli among lots of beef cattle (n = 3) from production through processing. Two hundred thirty-two generic E. coli isolated from the same animals through seven stages of the beef processing continuum (cattle in feedyards to packaged strip loins) were examined. LHR+E. coli were rare (0.6%; 1 of 180) among the early stages of the beef continuum (feces and hides at feedlot, feces and hides at harvest, and preevisceration carcasses), whereas the prevalence of LHR+E. coli on final carcasses and strip loins was remarkably higher. Half (14 of 28) of the final carcass E. coli possessed the LHR, while 79.2% (19 of 24) of the strip loin E. coli did. Eighty-five percent (29 of 34) of the LHR+E. coli presented with the XHR phenotype. The selection or enrichment of LHR+E. coli from harvest steps to the final products appeared unlikely as the LHR+E. coli isolates were effectively controlled by antimicrobial interventions typically used during beef processing. Further, whole-genome sequencing of the isolates suggested LHR+E. coli are persisting in the chilled processing environment and that horizontal LHR transfer among E. coli isolates may take place.

Presence of Presumptive Shiga Toxin–Producing Escherichia coli and Salmonella on Sheep during Harvest in Honduras

ABSTRACT Shiga toxin–producing Escherichia coli (STEC) and Salmonella are foodborne pathogens commonly harbored in the gastrointestinal tract of sheep. These pathogens can be on the hide of sheep and transferred to the carcass, causing a foodborne hazard. Salmonella can also be found in the lymph nodes of sheep, creating a biological hazard during harvest and processing. Developing countries lack baseline data on the presence of these foodborne pathogens on meat products, specifically sheep meat. Therefore, the objective of this study was to evaluate the presence of STEC and Salmonella on sheep hides, preevisceration carcasses, and final carcasses and Salmonella in subiliac lymph nodes from two small Honduran harvest facilities, plants A and B. Sponge swabs from the foreshank region of hides and carcasses and subiliac lymph node samples were collected from 96 sheep (86 at plant A; 10 at plant B). Microbial detection of STEC and Salmonella was performed by using the BAX System to screen for prevalence, and suspect samples were subjected to conventional culture isolation for confirmation. Overall baseline contamination on hides were 34.4 and 10.4% present for STEC and Salmonella; however, through implementation of sanitary procedures, preevisceration and final carcass samples were significantly decreased from the hide for both pathogens (P = 0.05). Moreover, overall plant A had significantly higher rates (P = 0.05) of STEC and Salmonella at each carcass sampling site compared with plant B. After each sampling was performed, recommendations were provided to each facility on the basis of pathogen presence, performance techniques, and contamination risks. Through recommendations and implementation of hazard analysis and critical control points, good manufacturing practices, and sanitation standard operating procedures in each facility, the meat supply in Honduras will become safer and more wholesome. Therefore, the development of a baseline with continued sampling is crucial to understand the risk of foodborne pathogens to consumers in the Honduran sheep meat supply. HIGHLIGHTS

Occurrence and characterization of Shiga toxin-producing Escherichia coli (STEC) isolated from Chinese beef processing plants

In order to investigate the prevalence, O serogroup, virulence genes and antibiotic resistance of Shiga toxin-producing Escherichia coli (STEC) in two beef plants in China, a total of 600 samples collected from 6 sites (feces, hide, pre-evisceration carcasses, post-washing carcasses, chilled carcasses and meat, 50 samples per site in each plant) were screened for the existence of Shiga toxin-encoding genes by PCR. STEC strains in positives were isolated and characterized for serogroup and antibiotic sensitivity. The PCR prevalence rate in each site was 45.0%, 31.0%, 14.0%, 13.0%, 9.0% and 18.0%, respectively. Sixteen O serogroups including O157, O146 and O76 which are associated with disease were identified. The existence of both stx1 and stx2 genes was the most common among the isolated strains (42.3%). Among the overall 26 isolates, seven and three were resistant to at least three and ten antibiotics, indicating a high antibiotic resistance in STEC strains isolated from the study.

Quantification of Bacteria Indicative of Fecal and Environmental Contamination from Hides to Carcasses.

Fecal bacteria, which reside in the gastrointestinal tract of cattle, can contaminate beef carcasses during processing. In beef cattle slaughter plants, the presence and concentrations of generic Escherichia coli, coliforms, Enterobacteriaceae (EB), and total aerobic bacteria are monitored as indicator organisms of fecal and environmental contamination. The objectives of this study were as follows: (1) to determine the concentrations of generic E. coli, coliforms, EB, and aerobic bacteria on beef carcasses at different processing points in Midwestern commercial beef slaughter plants during the summer, spring, and fall seasons; and (2) to estimate bacterial transfer on carcasses during the hide removal and evisceration processes. Hide and carcass surface sample swabs were collected from slaughtered cattle at four large commercial processing plants. At each plant visit (3 visits to each of the 4 plants) and during 3 seasons, 20 samples were collected at 5 points: hide-on (hide of animal near exsanguination pit), hide-off carcass, pre-evisceration carcass, postevisceration carcass, and postintervention carcass, for a total of 3600 samples. Bacterial concentrations were determined using 3M™ Petrifilm™ plates. Associations between season and processing plant with concentrations of E. coli, coliforms, EB, and total aerobic bacteria, overall, between hide-on and hide-off, and between pre- and post-evisceration, were evaluated using multilevel mixed-effects linear regression models. Bacterial concentrations on beef carcasses significantly decreased throughout processing. Moreover, hide removal was an important source of carcass contamination, given bacterial concentrations detected on hide-off carcass samples were the highest, and bearing in mind that carcass muscle surfaces should be sterile. Results from this study indicate that the interventions applied by the processing plants were effective, as they probably contributed to the significant reduction of bacterial concentrations of carcasses.

Prevalence and Characterization of Salmonella Present during Veal Harvest

Beef and veal products have been vehicles implicated in the transmission of Salmonella enterica, a gastroenteritis-causing bacteria. Recent regulatory samples collected from veal have indicated bob veal, or calves harvested within days of birth, have higher rates of Salmonella than samples collected from formula-fed veal, or calves raised 20 weeks on milk replacer formula before harvest. To investigate this problem, we collected samples from veal calf hides, preevisceration carcasses, and final carcasses at five veal processors that harvested bob or formula-fed veal or both. Prevalence and concentrations of Salmonella were determined, and then the isolates were characterized for serovar and antibiotic susceptibility. Salmonella was more prevalent (P < 0.05) among bob veal than formula-fed veal hides, preevisceration carcasses, and final carcass (84.2 versus 15.6%, 62.8 versus 10.1%, and 12.0 versus 0.4%, respectively). Concentrations of Salmonella could be estimated by using regression order statistics on hides and preevisceration carcasses at two veal plants, with one harvesting bob veal and the other bob and formula-fed veal. The concentration of Salmonella on bob veal hides at the plants was 1.45 ± 0.70 and 2.04 ± 1.00 log CFU/100 cm2, greater (P < 0.05) than on formula-fed veal hides, which was 1.10 ± 1.51 log CFU/100 cm2. Concentrations on carcasses, however, were very low. Seventeen Salmonella serovars were identified among 710 isolates. Salmonella serovars London, Cerro, and Muenster were most common to bob veal and made up 50.7, 18.7, and 6.3% of the isolates, respectively, while serovar Montevideo (6.8% of isolates) was most common to formula-fed veal. Although bob veal had increased prevalence and concentrations of Salmonella, one group of formula-fed veal was found to harbor human disease-related antibiotic-resistant Salmonella serovars Heidelberg and the monophasic variant of Typhimurium (1,4,[5],12:i:-). Veal processors have made changes to improve the safety of veal, but further efforts are necessary from both bob and formula-fed veal to address Salmonella.

Prevalence and Transmission of Antimicrobial Resistance in a Vertically Integrated Veal Calf Production System.

Transmission of antimicrobial resistance (AMR) from animal production systems to humans through the food supply is a public health concern. Currently, little is known about the prevalence of AMR among veal calves in the United States. Therefore, the objective of this prospective cohort study was to estimate the prevalence of AMR and multidrug resistance (MDR) among Escherichia coli within a vertically integrated production system. In addition, this study aimed to identify genes associated with phenotypic resistance to third- and fourth-generation cephalosporins (3GC and 4GC). Calves from four veal cohorts were randomly sampled resulting in a total of 166 farm fecal samples, 159 harvest fecal swabs, 164 preevisceration swabs, and 122 final carcass swabs. The prevalence of MDR among random-pick E. coli isolates recovered from the respective samples was 97% (161/166), 35% (55/159), 61% (51/84), and 24% (5/21). A selective isolation protocol found cefotaxime (a 3GC)-resistant isolates in 91% (127/140) of farm fecal samples, 34% (55/164) of preevisceration swabs, and 19% (23/122) of final carcass swabs tested. Isolates resistant to cefepime, a 4GC, were found among 24% (33/140), 6.7% (11/164), and 0.8% (1/122) of the same, respective samples. Isolates resistant to ciprofloxacin, a fluoroquinolone, were recovered from 75% (73/98) of farm fecal samples, 23% (38/164) of preevisceration swabs, and 6.6% (8/122) of final carcass swabs. The blaCMY-2 and blaCTX-M resistance genes were found in 89% (93/105) and 100% (42/42) of tested subsets of 3GC- and 4GC-resistant isolates, respectively. Pulsed-field gel electrophoresis (PFGE) analysis conducted on 3GC- and fluoroquinolone-resistant isolates showed three indistinguishable PFGE patterns from cefotaxime-resistant isolates recovered at farm and from two preevisceration carcass swabs. Although the prevalence of resistance declined between initial farm fecal samples and final carcass swabs, resistant bacteria recovered from carcasses illustrate the potential transmission of AMR to the human food supply.

Characterization of Enterohemorrhagic Escherichia coli on Veal Hides and Carcasses

Enterohemorrhagic Escherichia coli (EHEC) are Shiga toxin-producing E. coli associated with the most severe forms of foodborne illnesses. The U.S. Department of Agriculture, Food Safety and Inspection Service has identified a higher percentage of non-O157 EHEC compared with E. coli O157:H7-positive samples collected from veal trimmings than from products produced from other cattle slaughter classes. Therefore samples were collected from hides and preevisceration carcasses at five veal processors to assess E. coli O157:H7 and non-O157 EHEC contamination during bob veal and formula-fed veal dressing procedures. E. coli O157:H7 prevalence was measured by culture isolation and found to be on 20.3% of hides and 6.7% of carcasses. In contrast, a non-O157 EHEC molecular screening assay identified 90.3% of hides and 68.2% of carcasses as positive. Only carcass samples were taken forward to culture confirmation and 38.7% yielded one or more non-O157 EHEC isolates. The recovery of an EHEC varied by plant and sample collection date; values ranged from 2.1 to 87.8% among plants and from 4.2 to 64.2% within the same plant. Three plants were resampled after changes were made to sanitary dressing procedures. Between the two collection times at the three plants, hide-to-carcass transfer of E. coli O157:H7 and non-O157 EHEC was significantly reduced. All adulterant EHEC serogroups (O26, O45, O103, O111, O121, and O145) were isolated from veal carcasses as well as four other potentially pathogenic serogroups (O5, O84, O118, and O177). Bob veal was found to have a greater culture prevalence of E. coli O157:H7 and greater positive molecular screens for non-O157 EHEC than formula-fed veal (P < 0.05), but the percentage of culture-confirmed non-O157 EHEC was not different (P > 0.05) between the two types of calves. EHEC-O26, -O111, and -O121 were found more often in bob veal (P < 0.05), whereas EHEC-O103 was found more often in formula-fed veal (P < 0.05).

Effectiveness of 1,3-Dibromo-5,5-Dimethylhydantoin Applied in a Pre-Evisceration Wash Cabinet for Reducing Microbial Contamination on Beef Carcasses

ObjectivesThe objective of this study was to evaluate the effect of a bromine-based antimicrobial (1,3-dibromo-5,5-dimethylhydantoin; DBDMH), for use in a pre-evisceration carcass wash cabinet, against naturally occurring beef carcass associated microflora.Materials and MethodsThe study was conducted in a commercial beef harvest facility. Carcasses were randomly selected prior to the pre-evisceration wash cabinet for treatment application, which included 2 concentrations of DBDMH: low (280 to 350 ppm) and high (550 to 630 ppm). Prior to treatment application, carcasses were swabbed (10 × 10 cm2) on the ventral midline region with a sampling sponge hydrated with 10 mL Dey/Engley neutralizing broth, to serve as the initial counts (untreated control). After DBDMH was applied to the carcasses, a second sample was taken, to provide remaining bacterial populations after treatment. The 2 DBDMH treatments were replicated over 2 production days per treatment (N = 80; n = 40). All sponge samples were analyzed for aerobic plate counts (APC) and Enterobacteriaceae counts (EB) using Petrifilm Aerobic Count Plates and Petrifilm Enterobacteriaceae Count Plates, respectively. Bacterial populations for all samples were converted and expressed as log CFU/cm2. The study was designed as a paired comparison conducted on 2 different test days per treatment (4 total production days), for 2 DBDMH concentration levels. Day was treated as a fixed effect due to the unpredictable variation of the microbial conditions of carcasses on each production day. Data were analyzed using the Mixed Procedure of SAS (SAS Inst. Inc., Cary, NC) and expressed as least squares means for log CFU/cm2. Differences were reported with a significance level of a = 0.05.ResultsThere were significant main effects of treatment and day for APC populations recovered from carcasses treated with DBDMH at 280 to 350 ppm (treatment: P < 0.0001; day P = 0.0181). Due to these main effects, APC results were separated by each production test day to evaluate efficacy of the treatment. The APC populations recovered from beef carcasses before treatment with the low concentration of DBDMH (280 to 350 ppm) were 3.4 and 2.8 log CFU/cm2, from d 1 and 2, respectively (Table 1). Following DBDMH application, APC were 1.4 and 1.0 log CFU/cm2 for d 1 and 2, respectively (Table 1). The EB populations obtained before and after treatment with the low DBDMH concentration were < 0.1 and < –0.6 log CFU/cm2; respectively (Table 1). For carcasses treated with DBDMH at 550 to 630 ppm, a significant interaction between treatment and day (P = 0.0028) was observed for the APC data; therefore, APC results were separated by each production test day. Prior to DBDMH application at 550 to 630 ppm, the APC were 2.7 and 2.8 log CFU/cm2 for d 1 and 2, respectively (Table 1). The APC declined (P < 0.05) after DBDMH was applied, and APC of 1.1 and 2.2 log CFU/cm2 for d 1 and 2, respectively, were obtained (Table 1). Corresponding EB populations before and after the high concentration treatment were < –0.1 and < –0.6 log CFU/cm2, respectively (Table 1).ConclusionIn conclusion 1,3-dibromo-5,5-dimethylhydantoin was effective against naturally occurring microflora present on beef carcasses when applied in a pre-evisceration wash cabinet.

Effectiveness of 1,3-Dibromo-5,5-Dimethylhydantoin Applied in a Pre-Evisceration Wash Cabinet for Reducing Microbial Contamination on Beef Carcasses

ObjectivesThe objective of this study was to evaluate the effect of a bromine-based antimicrobial (1,3-dibromo-5,5-dimethylhydantoin; DBDMH), for use in a pre-evisceration carcass wash cabinet, against naturally occurring beef carcass associated microflora.Materials and MethodsThe study was conducted in a commercial beef harvest facility. Carcasses were randomly selected prior to the pre-evisceration wash cabinet for treatment application, which included 2 concentrations of DBDMH: low (280 to 350 ppm) and high (550 to 630 ppm). Prior to treatment application, carcasses were swabbed (10 × 10 cm2) on the ventral midline region with a sampling sponge hydrated with 10 mL Dey/Engley neutralizing broth, to serve as the initial counts (untreated control). After DBDMH was applied to the carcasses, a second sample was taken, to provide remaining bacterial populations after treatment. The 2 DBDMH treatments were replicated over 2 production days per treatment (N = 80; n = 40). All sponge samples were analyzed for aerobic plate counts (APC) and Enterobacteriaceae counts (EB) using Petrifilm Aerobic Count Plates and Petrifilm Enterobacteriaceae Count Plates, respectively. Bacterial populations for all samples were converted and expressed as log CFU/cm2. The study was designed as a paired comparison conducted on 2 different test days per treatment (4 total production days), for 2 DBDMH concentration levels. Day was treated as a fixed effect due to the unpredictable variation of the microbial conditions of carcasses on each production day. Data were analyzed using the Mixed Procedure of SAS (SAS Inst. Inc., Cary, NC) and expressed as least squares means for log CFU/cm2. Differences were reported with a significance level of a = 0.05.ResultsThere were significant main effects of treatment and day for APC populations recovered from carcasses treated with DBDMH at 280 to 350 ppm (treatment: P < 0.0001; day P = 0.0181). Due to these main effects, APC results were separated by each production test day to evaluate efficacy of the treatment. The APC populations recovered from beef carcasses before treatment with the low concentration of DBDMH (280 to 350 ppm) were 3.4 and 2.8 log CFU/cm2, from d 1 and 2, respectively (Table 1). Following DBDMH application, APC were 1.4 and 1.0 log CFU/cm2 for d 1 and 2, respectively (Table 1). The EB populations obtained before and after treatment with the low DBDMH concentration were < 0.1 and < –0.6 log CFU/cm2; respectively (Table 1). For carcasses treated with DBDMH at 550 to 630 ppm, a significant interaction between treatment and day (P = 0.0028) was observed for the APC data; therefore, APC results were separated by each production test day. Prior to DBDMH application at 550 to 630 ppm, the APC were 2.7 and 2.8 log CFU/cm2 for d 1 and 2, respectively (Table 1). The APC declined (P < 0.05) after DBDMH was applied, and APC of 1.1 and 2.2 log CFU/cm2 for d 1 and 2, respectively, were obtained (Table 1). Corresponding EB populations before and after the high concentration treatment were < –0.1 and < –0.6 log CFU/cm2, respectively (Table 1).ConclusionIn conclusion 1,3-dibromo-5,5-dimethylhydantoin was effective against naturally occurring microflora present on beef carcasses when applied in a pre-evisceration wash cabinet.

Prevalence and characterization of Escherichia coli O157:H7 from samples along the production line in Chinese beef-processing plants

This work investigated the prevalence of Escherichia coli O157:H7 and the antibiotic susceptibility, genetic diversity and virulence genes of isolated strains from four beef slaughter plants in China. A total of 510 samples (feces, hide, carcasses and raw meat) were tested for E. coli O157:H7 using enrichment, immunomagnetic separation, and plating on selective media. The prevalence of E. coli O157:H7 in the feces, hide, pre-evisceration carcass, post-evisceration carcass, post-washing carcass, chilled carcass, and raw meat samples was 1.43%, 1.43%, 0%, 0%, 0%, 1.25%, and 0%, respectively. Multiplex PCR assays were used for serotype confirmation and virulence gene detection. stx2, eaeA and EHEC-hlyA genes were present in all six of the strains, and the stx1 gene was not present. Fluorescence amplified fragment length polymorphism (FAFLP) was used to determine the genetic diversity of E. coli O157:H7 and revealed a high similarity between the strains isolated from feces and those isolated from carcasses. None of the isolated strains were found to be resistant to sixteen commonly used antimicrobial agents. The results of this study indicate that although E. coli O157:H7 contamination in the Chinese beef industry is sporadic and not as common as reported in other counties, all of the isolates contained three major virulence genes, presenting a high risk of disease for humans. The current research provides baseline information on E. coli O157:H7 prevalence and character profiles in Chinese beef-processing plants that can be used for future studies.

Occurrence of Antimicrobial-Resistant Escherichia coli and Salmonella enterica in the Beef Cattle Production and Processing Continuum.

Specific concerns have been raised that third-generation cephalosporin-resistant (3GC(r)) Escherichia coli, trimethoprim-sulfamethoxazole-resistant (COT(r)) E. coli, 3GC(r) Salmonella enterica, and nalidixic acid-resistant (NAL(r)) S. enterica may be present in cattle production environments, persist through beef processing, and contaminate final products. The prevalences and concentrations of these organisms were determined in feces and hides (at feedlot and processing plant), pre-evisceration carcasses, and final carcasses from three lots of fed cattle (n = 184). The prevalences and concentrations were further determined for strip loins from 103 of the carcasses. 3GC(r) Salmonella was detected on 7.6% of hides during processing and was not detected on the final carcasses or strip loins. NAL(r) S. enterica was detected on only one hide. 3GC(r) E. coli and COT(r) E. coli were detected on 100.0% of hides during processing. Concentrations of 3GC(r) E. coli and COT(r) E. coli on hides were correlated with pre-evisceration carcass contamination. 3GC(r) E. coli and COT(r) E. coli were each detected on only 0.5% of final carcasses and were not detected on strip loins. Five hundred and 42 isolates were screened for extraintestinal pathogenic E. coli (ExPEC) virulence-associated markers. Only two COT(r) E. coli isolates from hides were ExPEC, indicating that fed cattle products are not a significant source of ExPEC causing human urinary tract infections. The very low prevalences of these organisms on final carcasses and their absence on strip loins demonstrate that current sanitary dressing procedures and processing interventions are effective against antimicrobial-resistant bacteria.

Impact of Sampling Area and Location on Measurement of Indicator Organisms during Beef Carcass Interventions

The effect of the sponge sample collection site on the recovery of multiple indicator organisms from beef carcass surfaces was evaluated to simplify and validate our previous sampling method for ease of implementation as a general protocol. Sponge samples were collected at three beef processing plants using hot water or acidic antimicrobials as interventions. Two 4,000-cm(2) samples were collected from preevisceration carcasses (n = 248), one from the inside and outside round area (top site) and one from the navel-plate-brisket-foreshank area (bottom site). One-half of the samples (n = 124) were collected before a wash cabinet intervention and the other half after the intervention. The numbers of total aerobic bacteria, Enterobacteriaceae, coliforms, and Escherichia coli were determined for one-half of each individual sponge sample. The other halves of the sponges were combined to represent a top plus bottom 8,000-cm(2) SAMPLE: For the preintervention carcasses, 4,000-cm(2) samples collected from the top or bottom sites of the carcasses were not significantly different (P > 0.05) from each other or from the 8,000-cm(2) combined sample in recovery of the indicator organisms. Significant reductions of indicator organisms were observed in all three types of sponge samples after intervention; however, samples collected from the bottom site recovered less organisms (P < 0.05) compared with samples of the other types. These results suggested that samples collected from either the top or the bottom site of the carcasses with this method are suitable for monitoring indicator organisms as long as the same sampling site is consistently used.

Prevalence and profile of Salmonella from samples along the production line in Chinese beef processing plants

This work investigated the prevalence of Salmonella, the serotypes and antibiotic resistance of the isolated strains from four beef processing plants of China. The prevalence of Salmonella in hide (n = 70), feces (n = 70), pre-evisceration carcass (n = 70), post-evisceration carcass (n = 70), post-washing carcass (n = 70), chilled carcass (n = 80), and raw meat (n = 80) samples was 20.0%, 18.6%, 1.4%, 1.4%, 2.9%, 1.3%, and 1.3%, respectively. Among the four plants, there were significant differences in the prevalence of Salmonella on hides and in feces. During the processing, Salmonella was significantly reduced after hide removal. Seven serotypes of Salmonella were identified among the eighty-three isolates. Salmonella Agona was the dominant serotype (p < 0.05, 53.0%), followed by Salmonella Senftenberg (16.9%), Salmonella Meleagridis (10.8%), and Salmonella Derby (9.6%). None of the isolated strains were found to be resistant to sixteen commonly used antimicrobial agents. The results of this study indicate that Salmonella contamination is common in samples along the production line, with S. Agona as dominant serotype. Specific measures should be taken to prevent and/or treat Salmonella contamination in corresponding products in Chinese beef processing plants. Furthermore, the current research might provide baseline information of Salmonella prevalence profile in Chinese beef processing plant, which could be used for the future study.

Isolation and Characterization of Clostridium difficile Associated with Beef Cattle and Commercially Produced Ground Beef

The incidence of Clostridium difficile infection has recently increased in North American and European countries. This pathogen has been isolated from retail pork, turkey, and beef products and reported associated with human illness. This increase in infections has been attributed to the emergence of a toxigenic strain designated North America pulsed-field gel electrophoresis type 1 (NAP1). The NAP1 strain has been isolated from calves as well as ground meat products, leading to speculation of illness from consumption of contaminated meat products. However, information on C. difficile associated with beef cattle during processing and commercially produced ground beef is limited. To address this data gap, samples from various steps during beef production were collected. Samples from hides (n = 525), preevisceration carcasses (n = 475), postintervention carcasses (n = 471), and 956 commercial ground beef samples were collected from across the United States. The prevalence of C. difficile spores on hides was 3.2%. C. difficile spores were not detected on preevisceration and postintervention carcasses or in commercially produced ground beef. Phenotypic and genetic characterizations were carried out for all 18 isolates collected from hide samples. Twenty-two percent of the isolates were nontoxigenic strains, while 78% of the isolates were toxigenic. Toxinotyping and PCR ribotyping patterns revealed that 6 and 33% of the isolates were identified as NAP1 and NAP7 strains, respectively. This article evidences that the prevalence of C. difficile, specifically pathogenic strains, in the U.S. beef production chain is low.

Disinfectant and Antibiotic Susceptibility Profiles of Escherichia coli O157:H7 Strains from Cattle Carcasses, Feces, and Hides and Ground Beef from the United States

The disinfectant and antibiotic susceptibility profiles of 344 Escherichia coli O157:H7 strains from cattle carcasses, feces, and hides and ground beef from the United States were determined. A low prevalence of antibiotic resistance was observed (14%). The highest prevalences of resistance were to sulfisoxazole (10.5%), tetracycline (9.9%), streptomycin (7%), and chloramphenicol (4.9%). Four strains were resistant to eight antibiotics (two strains from ground beef and one strain each from hide and preevisceration carcass swabs of cull cattle at harvest). Pulsed-field gel electrophoresis analysis of the E. coli O157:H7 strains revealed two major groups (designated 1 and 2) composed of 17 and 20 clusters, respectively. Clusters 1A, 1B, 1C, and 1G.1 were associated with multidrug-resistant strains. There was no observed correlation between disinfectant resistance and antibiotic resistance. Sixty-nine (20%) of the 344 strains were resistant to chlorhexidine or benzalkonium chloride or the MICs of benzyldimethyldodecylammonium chloride were elevated. Inducible resistance was observed at elevated concentrations of antibiotics (1.4%) and disinfectants (6.1%). The highest rate of disinfectant inducible resistance was to OdoBan, quaternary ammonium chlorides, and the surface disinfectants F25, FS512, and MG, which are used in dairies, restaurants, and food processing plants. High MICs (1,024 to 4,096 m g/ml) of acetic, lactic, and citric acids were found. The decreasing order of acid potency based on molar MICs (MICs(molar)) was acetic, citric, and lactic acid. The correlation of the concentration of dissociated organic acids and MICs(molar) strongly suggests that the observed inhibition of E. coli O157:H7 was primarily due to dissociated forms of the acids.

Prevalence, enumeration, serotypes, and antimicrobial resistance phenotypes of salmonella enterica isolates from carcasses at two large United States pork processing plants.

The objective of this study was to characterize Salmonella enterica contamination on carcasses in two large U.S. commercial pork processing plants. The carcasses were sampled at three points, before scalding (prescald), after dehairing/polishing but before evisceration (preevisceration), and after chilling (chilled final). The overall prevalences of Salmonella on carcasses at these three sampling points, prescald, preevisceration, and after chilling, were 91.2%, 19.1%, and 3.7%, respectively. At one of the two plants, the prevalence of Salmonella was significantly higher (P < 0.01) for each of the carcass sampling points. The prevalences of carcasses with enumerable Salmonella at prescald, preevisceration, and after chilling were 37.7%, 4.8%, and 0.6%, respectively. A total of 294 prescald carcasses had Salmonella loads of >1.9 log CFU/100 cm(2), but these carcasses were not equally distributed between the two plants, as 234 occurred at the plant with higher Salmonella prevalences. Forty-one serotypes were identified on prescald carcasses with Salmonella enterica serotypes Derby, Typhimurium, and Anatum predominating. S. enterica serotypes Typhimurium and London were the most common of the 24 serotypes isolated from preevisceration carcasses. The Salmonella serotypes Johannesburg and Typhimurium were the most frequently isolated serotypes of the 9 serotypes identified from chilled final carcasses. Antimicrobial susceptibility was determined for selected isolates from each carcass sampling point. Multiple drug resistance (MDR), defined as resistance to three or more classes of antimicrobial agents, was identified for 71.2%, 47.8%, and 77.5% of the tested isolates from prescald, preevisceration, and chilled final carcasses, respectively. The results of this study indicate that the interventions used by pork processing plants greatly reduce the prevalence of Salmonella on carcasses, but MDR Salmonella was isolated from 3.2% of the final carcasses sampled.

Tracking verocytotoxigenic Escherichia coli O157, O26, O111, O103 and O145 in Irish cattle

The purpose of this study was to investigate carriage and transfer of verocytotoxigenic Escherichia coli (VTEC) O157, O26, O111, O103 and O145 from faeces and hide to dressed carcasses of Irish cattle as well as establishing the virulence potential of VTEC carried by these cattle. Individual cattle was tracked and faecal samples, hide and carcass (pre-evisceration and post-wash) swabs were analysed for verotoxin (vt1 and vt2) genes using a duplex real-time PCR assay. Positive samples were screened for the five serogroups of interest by real-time PCR. Isolates were recovered from PCR positive samples using immunomagnetic separation and confirmed by latex agglutination and PCR. Isolates were subject to a virulence screen (vt1, vt2, eaeA and hlyA) by PCR. Isolates carrying vt genes were examined by Pulsed-Field Gel Electrophoresis (PFGE). Of the VTEC isolated, E. coli O157 was the most frequently recovered from hide (17.6%), faeces (2.3%) and pre-evisceration/post-wash carcass (0.7%) samples. VTEC O26 was isolated from 0.2% of hide swabs and 1.5% of faeces samples. VTEC O145 was isolated from 0.7% of faeces samples. VTEC O26 and VTEC O145 were not recovered from carcass swabs. Non-VTEC O103 was recovered from all sample types (27.1% hide, 8.5% faeces, 5.5% pre-evisceration carcass, 2.2% post-wash carcass), with 0.2% of hide swabs and 1.0% of faeces samples found to be positive for VTEC O103 isolates. E. coli O111 was not detected in any samples. For the four serogroups recovered, the direct transfer from hide to carcass was not observed. This study shows that while VTEC O157 are being carried by cattle presented for slaughter in Ireland, a number of other verotoxin producing strains are beginning to emerge.

P27. Clinical utility of uric acid in the detection of preeclampsia in hypertensive pregnancy – Preliminary results

In order to investigate the prevalence, O serogroup, virulence genes and antibiotic resistance of Shiga toxin-producing Escherichia coli (STEC) in two beef plants in China, a total of 600 samples collected from 6 sites (feces, hide, pre-evisceration carcasses, post-washing carcasses, chilled carcasses and meat, 50 samples per site in each plant) were screened for the existence of Shiga toxin-encoding genes by PCR. STEC strains in positives were isolated and characterized for serogroup and antibiotic sensitivity. The PCR prevalence rate in each site was 45.0%, 31.0%, 14.0%, 13.0%, 9.0% and 18.0%, respectively. Sixteen O serogroups including O157, O146 and O76 which are associated with disease were identified. The existence of both stx1 and stx2 genes was the most common among the isolated strains (42.3%). Among the overall 26 isolates, seven and three were resistant to at least three and ten antibiotics, indicating a high antibiotic resistance in STEC strains isolated from the study.

Diversity of Multidrug-Resistant Salmonella enterica Strains Associated with Cattle at Harvest in the United States

The prevalence and diversity of multidrug-resistant (MDR) Salmonella enterica strains associated with cattle at harvest in the United States were examined. Hides and carcasses of cattle were sampled at processing plants (n = 6) located in four geographically distant regions from July 2005 to April 2006. The mean prevalences of Salmonella on hides, preevisceration carcasses (immediately after hide removal), and postintervention carcasses (in the chiller and after the full complement of interventions) were 89.6%, 50.2%, and 0.8%, respectively. The values for MDR Salmonella enterica strains (defined as those resistant to two or more antimicrobials) as percentages of Salmonella prevalence were 16.7% (95% confidence interval [CI], 8.3 to 25.1%; median percent prevalence, 6.9%), 11.7% (95% CI, 4.4 to 19.0%; median, 4.8%), and 0.33% (95% CI, -0.3 to 0.70%; median, 0%), respectively. In this study, 16,218 Salmonella hide and carcass isolates were screened for antimicrobial resistance. Of these, 978 (6.0%) unique MDR S. enterica isolates were identified and serotyped and their XbaI pulsed-field gel electrophoresis (PFGE) profiles determined. The predominant MDR S. enterica serotypes observed were Newport (53.1%), Typhimurium (16.6%), and Uganda (10.9%). Differences in MDR S. enterica prevalence were detected, and PFGE analysis revealed both epidemic clusters (profiles found in plants in multiple regions/seasons) and endemic clusters (profiles observed in plants in limited regions/seasons) within several of the MDR serotypes examined. Despite these differences, multiple-hurdle processing interventions employed at all plants were found to be quite effective and decreased Salmonella carcass contamination by 98.4% (95% CI, 97.6 to 99.7%).

Animal- and truckload-level associations between Escherichia coli O157:H7 in feces and on hides at harvest and contamination of preevisceration beef carcasses.

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