No Change in Risk for Antibiotic-Resistant Salmonellosis from Beef, United States, 2002-2010.

Restricting antibiotic use in food production animals is a target for reducing antimicrobial drug-resistant infections in humans. We used US surveillance data to estimate the probability of antibiotic-resistant nontyphoidal salmonellosis per meal made with beef during 2002-2010. Applying data for nontyphoidal Salmonella in raised-without-antibiotics cattle, we tested the effect of removing antibiotic use from all beef cattle production. We found an average of 1.2 (95% credible interval 0.6-4.2) antibiotic-resistant nontyphoidal salmonellosis cases per 1 million beef meals made with beef initially contaminated with antibiotic-resistant nontyphoidal Salmonella at slaughter or retail and 0.031 (95% credible interval 0.00018-0.14) cases per 1 million meals irrespective of beef contamination status. Neither outcome showed sustained change except for increases in 2003 and 2009 (>98% confidence) when larger or more outbreaks occurred. Switching all beef production to a raised-without-antibiotics system may not have a significant effect on antibiotic-resistant nontyphoidal salmonellosis (94.3% confidence).

Seasonal Prevalence of Shiga Toxin-Producing Escherichia coli on Pork Carcasses for Three Steps of the Harvest Process at Two Commercial Processing Plants in the United States.

Shiga toxin-producing Escherichia coli (STEC) is a foodborne pathogen that has a significant impact on public health, with strains possessing the attachment factor intimin referred to as enterohemorrhagic E. coli (EHEC) and associated with life-threatening illnesses. Cattle and beef are considered typical sources of STEC, but their presence in pork products is a growing concern. Therefore, carcasses (n = 1,536) at two U.S. pork processors were sampled once per season at three stages of harvest (poststunning skins, postscald carcasses, and chilled carcasses) and then examined using PCR for Shiga toxin genes (stx), intimin genes (eae), aerobic plate count (APC), and Enterobacteriaceae counts (EBC). The prevalence of stx on skins, postscald, and chilled carcasses was 85.3, 17.5, and 5.4%, respectively, with 82.3, 7.8, and 1.7% of swabs, respectively, having stx and eae present. All stx-positive samples were subjected to culture isolation that resulted in 368 STEC and 46 EHEC isolates. The most frequently identified STEC were serogroups O121, O8, and O91 (63, 6.7, and 6.0% of total STEC, respectively). The most frequently isolated EHEC was serotype O157:H7 (63% of total EHEC). Results showed that scalding significantly reduced (P < 0.05) carcass APC and EBC by 3.00- and 2.50-log10 CFU/100 cm2, respectively. A seasonal effect was observed, with STEC prevalence lower (P < 0.05) in winter. The data from this study show significant (P < 0.05) reduction in the incidence of STEC (stx) from 85.3% to 5.4% and of EHEC (stx plus eae) from 82.3% to 1.7% within the slaughter-to-chilling continuum, respectively, and that potential EHEC can be confirmed present throughout using culture isolation.IMPORTANCE Seven serogroups of STEC are responsible for most (>75%) cases of severe illnesses caused by STEC and are considered adulterants of beef. However, some STEC outbreaks have been attributed to pork products, although the same E. coli are not considered adulterants in pork because little is known of their prevalence along the pork chain. The significance of the work presented here is that it identifies disease-causing STEC, EHEC, demonstrating that these same organisms are a food safety hazard in pork as well as beef. The results show that most STEC isolated from pork are not likely to cause severe disease in humans and that processes used in pork harvest, such as scalding, offer a significant control point to reduce contamination. The results will assist the pork processing industry and regulatory agencies to optimize interventions to improve the safety of pork products.

Impact of "Raised without Antibiotics" Beef Cattle Production Practices on Occurrences of Antimicrobial Resistance.

The specific antimicrobial resistance (AMR) decreases that can be expected from reducing antimicrobial (AM) use in U.S. beef production have not been defined. To address this data gap, feces were recovered from 36 lots of "raised without antibiotics" (RWA) and 36 lots of "conventional" (CONV) beef cattle. Samples (n = 719) were collected during harvest and distributed over a year. AMR was assessed by (i) the culture of six AM-resistant bacteria (ARB), (ii) quantitative PCR (qPCR) for 10 AMR genes (ARGs), (iii) a qPCR array of 84 ARGs, and (iv) metagenomic sequencing. Generally, AMR levels were similar, but some were higher in CONV beef cattle. The prevalence of third-generation cephalosporin-resistant (3GCr) Escherichia coli was marginally different between production systems (CONV, 47.5%; RWA, 34.8%; P = 0.04), but the seasonal effect (summer, 92.8%; winter, 48.3%; P < 0.01) was greater. Erythromycin-resistant (ERYr) Enterococcus sp. concentrations significantly differed between production systems (CONV, 1.91 log10 CFU/g; RWA, 0.73 log10 CFU/g; P < 0.01). Levels of aadA1, ant(6)-I, bla ACI, erm(A), erm(B), erm(C), erm(F), erm(Q), tet(A), tet(B), tet(M), and tet(X) ARGs were higher (P < 0.05) in the CONV system. Aggregate abundances of all 43 ARGs detected by metagenomic sequencing and the aggregate abundances of ARGs in the aminoglycoside, ß-lactam, macrolide-lincosamide-streptogramin B (MLS), and tetracycline AM classes did not differ (log2 fold change < 1.0) between CONV and RWA systems. These results suggest that further reductions of AM use in U.S. beef cattle production may not yield significant AMR reductions beyond MLS and tetracycline resistance.IMPORTANCE The majority of antimicrobial (AM) use in the United States is for food-animal production, leading to concerns that typical AM use patterns during "conventional" (CONV) beef cattle production in the United States contribute broadly to antimicrobial resistance (AMR) occurrence. In the present study, levels of AMR were generally similar between CONV and "raised without antibiotics" (RWA) cattle. Only a limited number of modest AMR increases was observed in CONV cattle, primarily involving macrolide-lincosamide-streptogramin B (MLS) and tetracycline resistance. Macrolides (tylosin) and tetracyclines (chlortetracycline) are administered in-feed for relatively long durations to reduce liver abscesses. To ensure judicious AM use, the animal health, economic, and AMR impacts of shorter duration in-feed administration of these AMs should be examined. However, given the modest AMR reductions observed, further reductions of AM use in U.S. beef cattle production may not yield significant AMR reductions beyond MLS and tetracycline resistance.

Evaluation of Bacteriophage Application to Cattle in Lairage at Beef Processing Plants to Reduce Escherichia coli O157:H7 Prevalence on Hides and Carcasses.

Escherichia coli O157:H7 is a major food safety concern for the beef industry. Several studies have provided evidence that cattle hides are the main source of beef carcass contamination during processing and that reductions in the E. coli O157:H7 load on the hides of cattle entering processing facilities will lead to reductions in carcass contamination. Bacteriophages have been proposed as a novel preharvest antimicrobial intervention to reduce the levels of E. coli O157:H7 on cattle hides. The objective of this study was to evaluate a commercialized phage application administered in the lairage area of commercial beef processing plants for the ability to reduce E. coli O157:H7 contamination of cattle hides and carcasses. Cattle lots either received phage spray treatment (n = 289) or did not (n = 301), as they entered the lairage environments in two separate experiments at two different commercial beef processing plants. Hide and carcass samples were collected and analyzed for E. coli O157:H7 prevalence and concentration. Cattle hides receiving phage treatment had an E. coli O157:H7 prevalence of 51.8%, whereas untreated hides had a prevalence of 57.6%. For carcass samples, the E. coli O157 prevalence in treated and untreated samples was 17.1% and 17.6%, respectively. The results obtained from these experiments demonstrated that the treatment of cattle hides with bacteriophages before processing did not produce a significant reduction of E. coli O157:H7 on cattle hides or beef carcasses during processing.

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.

Validation of a New Method of Sampling Beef Manufacturing Trimmings for Pathogen Testing Using a Manual Sampling Mitt Approach.

The USDA Food Safety Inspection Service has declared Escherichia coli O157:H7, and six additional Shiga toxin-producing E. coli (STEC) are adulterants for nonintact raw beef products. The U. S. beef processing industry has implemented several antimicrobial intervention technologies throughout the carcass dressing process to remove or destroy foodborne pathogens present on beef carcasses. Despite these efforts, STEC have been shown to cause finished product contamination, albeit at prevalences typically <0.5%. Recent work described the development and validation of improved methods for collecting samples from raw beef trimmings. One of the methods, the Manual Sampling Device (MSD) method, uses the manual implementation of the MicroTally® Swab (MT-Swab) to vigorously scrub the surface of raw beef manufacturing trimmings for pathogen detection. The work described herein reports the data from an evaluation of a novel MSD method using the MicroTally® Mitt (MT-Mitt). The MT-Mitt provides a more user-friendly option for sample collection than the MT-Swab. A series of trials were conducted with a total of 360 matched samples comparing manual sampling of raw beef manufacturing trimmings using the MT-Swab, N60-excision, or N60-plus methods to a novel method using the MT-Mitt. The results of these trials collectively demonstrate that manual sampling of raw beef manufacturing trimmings using the MT-Mitt provides organism recovery that is not significantly different from that of the MT-Swab, N60-excision, and N60-plus methods. Thus, the MT-Mitt method provides an alternative sampling method with organism recovery that is not significantly different from previous methods for sampling beef manufacturing trimmings for pathogen detection and some implementation advantages pertaining to labor and ease of use.

Evaluation of Peracetic Acid Treatment on Beef Trimmings and Subprimals Against Salmonella and E. Coli O157:H7 Within Regulatory Retained Water Limitations.

The application of antimicrobial treatments to beef trimmings prior to grinding for the reduction of microbial contamination in ground beef has increased recently. However, raw single-ingredient meat products are not permitted by Food Safety and Inspection Services (FSIS) to retain more than 0.49% water resulting from postevisceration processing. The effectiveness of antimicrobials with the limited water retention is not well documented. The objective of this study was to determine the effectiveness of peracetic acid at varied concentrations against E. coli O157:H7 and Salmonella on the surface of beef trimmings and beef subprimals that was applied at industry operating parameters within the retained water requirement. One hundred and forty-four each of beef trimmings and subprimals were used to evaluate the effect of different concentrations of peracetic acid solution on reducing E. coli O157:H7 and Salmonella on surfaces of fresh beef within the FSIS requirement of ≤0.49% retained water from antimicrobial spray treatments using a conveyor system. A ten-strain cocktail mixture was inoculated on surfaces of fresh beef and subjected to water or four different concentrations of peracetic acid (130, 150, 200, and 400 ppm). Spray treatments with 130, 150, and 200 ppm peracetic acid reduced (P ≤ 0.05) E. coli O157:H7 and Salmonella at least 0.2 log on surfaces of beef trimmings and subprimals. Spray treatment with 400 ppm peracetic acid resulted in approximately 0.5 and 0.3 log reduction of E. coli O157:H7 and Salmonella, respectively. Results indicate that all concentrations (130-400 ppm) of peracetic acid significantly reduced E. coli O157:H7 and Salmonella on beef trimmings and subprimals compared to untreated controls. Thus, a range from 130 to 400 ppm of peracetic acid can be used during beef processing to improve the safety of beef trimmings and subprimals when weight gain is limited to ≤0.49% to meet regulatory requirements.

Identifying a list of Salmonella serotypes of concern to target for reducing risk of salmonellosis.

There is an increasing awareness in the field of Salmonella epidemiology that focusing control efforts on those serotypes which cause severe human health outcomes, as opposed to broadly targeting all Salmonella, will likely lead to the greatest advances in decreasing the incidence of salmonellosis. Yet, little guidance exists to support validated, scientific selection of target serotypes. The goal of this perspective is to develop an approach to identifying serotypes of greater concern and present a case study using meat- and poultry-attributed outbreaks to examine challenges in developing a standardized framework for defining target serotypes.

Proportions and Serogroups of Enterohemorrhagic Shiga Toxin-producing Escherichia coli in Feces of Fed and Cull Beef and Cull Dairy Cattle at Harvest.

Cattle are considered a primary reservoir of Shiga toxin (stx)-producing Escherichia coli that cause enterohemorrhagic disease (EHEC), and contaminated beef products are one vehicle of transmission to humans. However, animals entering the beef harvest process originate from differing production systems: feedlots, dairies, and beef breeding herds. The objective of this study was to determine if fed cattle, cull dairy, and or cull beef cattle carry differing proportions and serogroups of EHEC at harvest. Feces were collected via rectoanal mucosal swabs (RAMSs) from 1,039 fed cattle, 1,058 cull dairy cattle, and 1,018 cull beef cattle at harvest plants in seven U.S. states (CA, GA, NE, PA, TX, WA, and WI). The proportion of the stx gene in feces of fed cattle (99.04%) was not significantly different (P > 0.05) than in the feces of cull dairy (92.06%) and cull beef (91.85%) cattle. When two additional factors predictive of EHEC (intimin and ecf1 genes) were considered, EHEC was significantly greater (P < 0.05) in fed cattle (77.29%) than in cull dairy (47.54%) and cull beef (38.51%) cattle. The presence of E. coli O157:H7 and five common non-O157 EHEC of serogroups O26, O103, O111, O121, and O145 was determined using molecular analysis for single nucleotide polymorphisms (SNPs) followed by culture isolation. SNP analysis identified 23.48%, 17.67%, and 10.81% and culture isolation confirmed 2.98%, 3.31%, and 3.00% of fed, cull dairy, and cull beef cattle feces to contain one of these EHEC, respectively. The most common serogroups confirmed by culture isolation were O157, O103, and O26. Potential EHEC of fourteen other serogroups were isolated as well, from 4.86%, 2.46%, and 2.01% of fed, cull dairy, and cull beef cattle feces, respectively; with the most common being serogroups O177, O74, O98, and O84. The identification of particular EHEC serogroups in different types of cattle at harvest may offer opportunities to improve food safety risk management.

Cross-sectional study examining Salmonella enterica carriage in subiliac lymph nodes of cull and feedlot cattle at harvest.

Bovine peripheral lymph nodes (LNs), including subiliac LNs, have been identified as a potential source of human exposure to Salmonella enterica, when adipose trim containing these nodes is incorporated into ground beef. In order to gain a better understanding of the burden of S. enterica in peripheral LNs of feedlot and cull cattle, a cross-sectional study was undertaken in which 3327 subiliac LNs were collected from cattle at harvest in seven plants, located in three geographically distinct regions of the United States. Samples were collected in three seasons: Fall 2010, Winter/Spring 2011, and Summer/Fall 2011. A convenience sample of 76 LNs per day, 2 days per season (approximately 1 month apart), was collected per plant, from carcasses held in the cooler for no less than 24 h. Every 10(th) carcass half on a rail was sampled, in an attempt to avoid oversampling any single cohort of cattle. Median point estimates of S. enterica contamination were generally low (1.3%); however, median Salmonella prevalence was found to be greater in subiliac LNs of feedlot cattle (11.8%) compared to those of cull cattle (0.65%). Enumeration analysis of a subset of 618 feedlot cattle LNs showed that 67% of those harboring S. enterica (97 of 144) did so at concentrations ranging from <0.1 to 1.8 log10 CFU/g, while 33% carried a higher burden of S. enterica, with levels ranging from 1.9 to >3.8 log10 CFU/g. Serotyping of S. enterica isolated identified 24 serotypes, with the majority being Montevideo (44.0%) and Anatum (24.8%). Antimicrobial susceptibility phenotypes were determined for all isolates, and the majority (86.1%) were pansusceptible; however, multidrug-resistant isolates (8.3%) were also occasionally observed. As Salmonella contained within LNs are protected from carcass interventions, research is needed to define opportunities for mitigating the risk of Salmonella contamination in LNs of apparently healthy cattle.

Efficacy of Short Thermal Treatment Time Against Escherichia coli O157:H7 and Salmonella on the Surface of Fresh Beef.

Thermal treatment interventions consistently provide effective pathogen reductions. However, the cost of maintaining high temperature of 95°C in order to raise the surface temperature of carcasses to 82°C is very expensive. Therefore, beef processors need to identify thermal application times and temperatures that optimize the treatment effects with less maintenance cost. The objectives of this study were to determine the efficacy of hot water or steam at 71°C for 6 s and cascade e-ion plasma treatment for 2 s in reducing pathogens on the surface of fresh beef compared to the thermal treatment at 82°C for 15 s. Hot water at 71°C for 6 s reduced Escherichia coli O157:H7 and Salmonella by 2.38 and 2.48 log CFU/cm2, while steam treatment at 71°C for 6 s reduced E. coli O157:H7 and Salmonella by 2.94 and 3.06 log CFU/cm2, respectively. Cascade e-ion plasma treatment for 2 s reduced E. coli O157:H7 on surface of fresh beef by 1.89 log CFU/cm2. The findings indicate that short treatment time with appropriate temperature could serve as an effective carcass intervention to improve the safety of fresh beef.

Determination of Verification Parameters for Using the Manual Sampling Device for Fresh Raw Beef Trim.

Multifaceted food safety systems are used by the beef processing industry to minimize risk of bacterial contamination of the finished product. These systems are comprised of several parts including the conditional release of product requiring a sample to produce a negative result on a pathogen test prior to sending the product into the food supply. The methods of sample collection require verification activities that ensure the sampling protocols are performed adequately. The research described herein was done to determine the parameters for use in verifying adequate beef trim sampling for the Manual Sampling Device (MSD) method. In addition, the efficacy of repeated sampling via multiple applications of the MSD procedure on a fresh raw beef trim combo was investigated. The results show that MSD sample collection thatcoversless than the entire combo surface, but at least one-halfof the combo surface and is collected for a minimum of 90 s, is adequate for the recovery of organisms and prevalence targets from fresh raw beef trim. In addition, the evidence that MSD sample collection thatoccurs forless than the recommended time, butnot less than 30 s per side of the swab, is adequate for the recovery of organisms and prevalence targets from raw beef trim. Finally, results show that in a scenario where an in-plant MSD sample and a regulatory MSD sample are required from the same combo, two MSD samples can be collected from the same combo bin with similar test results for both samples. While the recommended MSD protocol specifications will not be changed, the data presented herein provide support for some flexibility in accepting test results when verification activities indicate that sampling did not occur as specified in the recommended procedure.

Fate of Shiga Toxin-Producing Escherichia coli (STEC) and Salmonella during Kosher Processing of Fresh Beef.

Traditional kosher meat processing involves the following steps after slaughtering: soaking with water to remove blood, salting to help draw out more blood, and rinsing to remove salt. However, the impact of the salt used on foodborne pathogens and beef quality is not well understood. The objectives of the current study were to determine the effectiveness of salt in reducing pathogens in a pure culture model, on surfaces of inoculated fresh beef during kosher processing, and the effect of salt on beef quality. The pure culture studies indicated that the reduction of E. coli O157:H7, non-O157 STEC, and Salmonella increased with increasing salt concentrations. With salt concentrations from 3 to 13%, salt reduced E. coli O157:H7, non-O157 STEC, and Salmonella ranging from 0.49 to 1.61 log CFU/mL. For kosher processing, the water-soaking step did not reduce pathogenic and other bacteria on the surface of fresh beef. Salting and rinsing steps reduced non-O157 STEC, E. coli O157:H7, and Salmonella ranging from 0.83 to 1.42 log CFU/cm2, and reduced Enterobacteriaceae, coliforms, and aerobic bacteria by 1.04, 0.95, and 0.70 log CFU/cm2, respectively. The salting process for kosher beef resulted in reducing pathogens on the surface of fresh beef, color changes, increased salt residues, and increased lipid oxidation on the final products.

Genetic and phenotypic associations of mitochondrial DNA copy number, SNP, and haplogroups with growth and carcass traits in beef cattle.

Mitochondrial DNA copy number (mtDNA CN) is heritable and easily obtained from low-pass sequencing (LPS). This study investigated the genetic correlation of mtDNA CN with growth and carcass traits in a multi-breed and crossbred beef cattle population. Blood, leucocyte, and semen samples were obtained from 2,371 animals and subjected to LPS that resulted in nuclear DNA (nuDNA) and mtDNA sequence reads. Mitochondrial DNA CN was estimated as the ratio of mtDNA to nuDNA coverages. Variant calling was performed from mtDNA, and 11 single nucleotide polymorphisms (SNP) were identified in the population. Samples were classified in taurine haplogroups. Haplogroup and mtDNA type were further classified based on the 11 segregating SNP. Growth and carcass traits were available for between 7,249 and 60,989 individuals. Associations of mtDNA CN, mtDNA haplogroups, mtDNA types, and mtDNA SNP with growth and carcass traits were estimated with univariate animal models, and genetic correlations were estimated with a bivariate animal model based on pedigree. Mitochondrial DNA CN tended (P-value ≤0.08) to be associated with birth weight and weaning weight. There was no association (P-value >0.10) between mtDNA SNP, haplogroups, or types with growth and carcass traits. Genetic correlation estimates of mtDNA CN were -0.30 ± 0.16 with birth weight, -0.31 ± 0.16 with weaning weight, -0.15 ± 0.14 with post-weaning gain, -0.11 ± 0.19 with average daily dry-matter intake, -0.04 ± 0.22 with average daily gain, -0.29 ± 0.13 with mature cow weight, -0.11 ± 0.13 with slaughter weight, -0.14 ± 0.13 with carcass weight, -0.07 ± 0.14 with carcass backfat, 0.14 ± 0.14 with carcass marbling, and -0.06 ± 0.14 with ribeye area. In conclusion, mtDNA CN was negatively correlated with most traits investigated, and the genetic correlation was stronger with growth traits than with carcass traits.

This study investigated mitochondrial DNA copy number (mtDNA CN) as a potential genetic indicator of growth and carcass traits in a composite beef cattle population. Mitochondrial DNA CN was previously shown to be under genetic control. The current study estimated the genetic relationship of mtDNA CN with growth and carcass traits. Blood, leucocyte, and semen samples were obtained from 2,371 animals and subjected to whole-genome sequencing at a low depth that resulted in nuclear DNA and mtDNA sequence reads. Mitochondrial DNA CN was estimated as the ratio of mtDNA to nuclear DNA coverages. Growth and carcass traits were available for between 7,249 and 60,989 individuals. Genetic parameters were estimated from an animal model based on pedigree. Genetic correlation estimates of mtDNA CN with growth and carcass traits were low to moderate and mostly negative. These indicate that selection for lower mtDNA would be associated with an increase in birth weight, weaning weight, post-weaning gain, average daily dry-matter intake, mature cow weight, slaughter weight, and carcass weight. Therefore, the by-product of whole-genome sequencing at a low depth could be used as an indicator trait for growth and carcass traits in genetic evaluations, but the genetic relationships are not likely strong enough to prioritize mtDNA ahead of routinely used indicator traits.

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.

Efficacy of Antimicrobial Interventions Used in Meat Processing Plants against Antimicrobial Tolerant Non-Antibiotic-Resistant and Antibiotic-Resistant Salmonella on Fresh Beef.

ABSTRACT: Salmonella is a common cause of foodborne illness in the United States, and several strains of Salmonella have been identified as resistant to antibiotics. It is not known whether strains that are antibiotic resistant (ABR) and that have some tolerance to antimicrobial compounds are also able to resist the inactivation effects of antimicrobial interventions used in fresh meat processing. Sixty-eight Salmonella isolates (non-ABR and ABR strains) were treated with half concentrations of lactic acid (LA), peracetic acid (PAA), and cetylpyridinium chloride (CPC), which are used in beef processing plants to screen for tolerant strains. Six strains each from non-ABR and ABR Salmonella that were most tolerant of LA (2%), PAA (200 ppm), and CPC (0.4%) were selected. Selected strains were inoculated on surfaces of fresh beef and subjected to spray wash treatment with 4% LA, 400 ppm PAA, or 0.8% CPC for the challenge study. Tissue samples were collected before and after each antimicrobial treatment for enumeration of survivors. Spray treatment with LA, PAA, or CPC significantly reduced non-ABR Salmonella and ABR Salmonella on surfaces of fresh beef by 1.95, 1.22, and 1.33 log CFU/cm2, and 2.14, 1.45, and 1.43 log CFU/cm2, respectively. The order of effectiveness was LA > PAA = CPC. The findings also indicated that LA, PAA, and CPC were equally (P ≤ 0.05) effective against non-ABR and ABR Salmonella on surfaces of fresh beef. These data contribute to the body of work that indicates that foodborne pathogens that have acquired both antibiotic resistance and antimicrobial tolerance are still equally susceptible to meat processing antimicrobial intervention treatments.

A Farm-to-Fork Quantitative Microbial Exposure Assessment of ß-Lactam-Resistant Escherichia coli among U.S. Beef Consumers.

Integrated quantitative descriptions of the transmission of ß-lactam-resistant Escherichia coli (BR-EC) from commercial beef products to consumers are not available. Here, a quantitative microbial exposure assessment model was established to simulate the fate of BR-EC in a farm-to-fork continuum and provide an estimate of BR-EC exposure among beef consumers in the U.S. The model compared the per-serving exposures from the consumption of intact beef cuts, non-intact beef cuts, and ground beef. Additionally, scenario analysis was performed to evaluate the relative contribution of antibiotic use during beef cattle production to the level of human exposure to BR-EC. The model predicted mean numbers of BR-EC of 1.7 × 10-4, 8.7 × 10-4, and 6.9 × 10-1 CFU/serving for intact beef cuts, non-intact beef cuts, and ground beef, respectively, at the time of consumption. Sensitivity analyses using the baseline model suggested that factors related to sectors along the supply chain, i.e., feedlots, processing plants, retailers, and consumers, were all important for controlling human exposure to BR-EC. Interventions at the processing and post-processing stages are expected to be most effective. Simulation results showed that a decrease in antibiotic use among beef cattle might be associated with a reduction in exposure to BR-EC from beef consumption. However, the absolute reduction was moderate, indicating that the effectiveness of restricting antibiotic use as a standalone strategy for mitigating human exposure to BR-EC through beef consumption is still uncertain. Good cooking and hygiene practices at home and advanced safety management practices in the beef processing and post-processing continuum are more powerful approaches for reducing human exposure to antibiotic-resistant bacteria in beef products.

Evaluating the Shelf Life and Sensory Properties of Beef Steaks from Cattle Raised on Different Grass Feeding Systems in the Western United States.

Consumer interest in grass-fed beef has been steadily rising due to consumer perception of its potential benefits. This interest has led to a growing demand for niche market beef, particularly in the western United States. Therefore, the objective of this study was to assess the impact of feeding systems on the change in microbial counts, color, and lipid oxidation of steaks during retail display, and on their sensory attributes. The systems included: conventional grain-fed (CON), 20 months-grass-fed (20GF), 25-months-grass-fed (25GF) and 20-months-grass-fed + 45-day-grain-fed (45GR). The results indicate that steaks in the 20GF group displayed a darker lean and fat color, and a lower oxidation state than those in the 25GF group. However, the feeding system did not have an impact on pH or objective tenderness of beef steaks. In addition, consumers and trained panelist did not detect a difference in taste or flavor between the 20GF or 25GF steaks but expressed a preference for the CON and 45GR steaks, indicating that an increased grazing period may improve the color and oxidative stability of beef, while a short supplementation with grain may improve eating quality.

Twenty-Four-Month Longitudinal Study Suggests Little to No Horizontal Gene Transfer In Situ between Third-Generation Cephalosporin-Resistant Salmonella and Third-Generation Cephalosporin-Resistant Escherichia coli in a Beef Cattle Feedyard.

ABSTRACT: Third-generation cephalosporins (3GCs) are preferred treatments for serious human Salmonella enterica infections. Beef cattle are suspected to contribute to human 3GC-resistant Salmonella infections. Commensal 3GC-resistant Escherichia coli are thought to act as reservoirs of 3GC resistance because these strains are isolated more frequently than are 3GC-resistant Salmonella strains at beef cattle feedyards. During each of 24 consecutive months, four samples of pen surface material were obtained from five pens (N = 480) at a Nebraska feedyard to determine to the contribution of 3GC-resistant E. coli to the occurrence of 3GC-resistant Salmonella. Illumina whole genome sequencing was performed, and susceptibility to 14 antimicrobial agents was determined for 121 3GC-susceptible Salmonella, 121 3GC-resistant Salmonella, and 203 3GC-resistant E. coli isolates. 3GC-susceptible Salmonella isolates were predominantly from serotypes Muenchen (70.2%) and Montevideo clade 1 (23.1%). 3GC-resistant Salmonella isolates were predominantly from serotypes Montevideo clade 2 (84.3%). One bla gene type (blaCMY-2) and the IncC plasmid replicon were present in 100 and 97.5% of the 3GC-resistant Salmonella, respectively. Eleven bla gene types were detected in the 3GC-resistant E. coli, which were distributed across 42 multilocus sequence types. The blaCMY-2 gene and IncC plasmid replicon were present in 37.9 and 9.9% of the 3GC-resistant E. coli, respectively. These results suggest that 3GC resistance in Salmonella was primarily due the persistence of Salmonella Montevideo clade 2 with very minimal or no contribution from 3GC-resistant E. coli via horizontal gene transfer and that 3GC-resistant E. coli may not be a useful indicator for 3GC-resistant Salmonella in beef cattle production environments.

Meat tenderness: advances in biology, biochemistry, molecular mechanisms and new technologies.

Meat tenderness is an important quality trait critical to consumer acceptance, and determines satisfaction, repeat purchase and willingness-to-pay premium prices. Recent advances in tenderness research from a variety of perspectives are presented. Our understanding of molecular factors influencing tenderization are discussed in relation to glycolysis, calcium release, protease activation, apoptosis and heat shock proteins, the use of proteomic analysis for monitoring changes, proteomic biomarkers and oxidative/nitrosative stress. Each of these structural, metabolic and molecular determinants of meat tenderness are then discussed in greater detail in relation to animal variation, postmortem influences, and changes during cooking, with a focus on recent advances. Innovations in postmortem technologies and enzymes for meat tenderization are discussed including their potential commercial application. Continued success of the meat industry relies on ongoing advances in our understanding, and in industry innovation. The recent advances in fundamental and applied research on meat tenderness in relation to the various sectors of the supply chain will enable such innovation.