Michael N. Guerini

Post-harvest interventions to reduce/eliminate pathogens in beef.

In 1999 the foodborne pathogens Salmonella, Listeria, Campylobacter, and Escherichia coli (both O157 and non-O157) were estimated to cause more than 6 million illnesses and approximately 9000 deaths each year. However, the most recent Centers for Disease Control and Prevention report on the sources and incidence of foodborne disease, released in 2004, has shown a dramatic decrease in E. coli O157:H7 infections. Since raw beef products are the most frequently foodborne sources of these pathogens, the results of this report demonstrate that the microbiological quality of raw beef has improved greatly. During the intervening years, post-harvest interventions have continually improved, with new attention to hide decontamination and innovative treatments of carcasses. In addition, a system to hold and test beef trim or ground beef for E. coli O157:H7 before its release into commerce has provided an even greater level of safety. In this paper, we review the latest information on the prevalence of E. coli O157:H7 and other pathogens on beef, the evidence identifying the hide as the primary source of pathogens on beef carcasses, the efficacy of various hide and carcass interventions, and other developments that have led or have the potential to lead to even greater improvements in the microbial quality of beef.

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.

Salmonella and Escherichia coli O157:H7 Contamination on Hides and Carcasses of Cull Cattle Presented for Slaughter in the United States: an Evaluation of Prevalence and Bacterial Loads by Immunomagnetic Separation and Direct Plating Methods

ABSTRACT The hide and carcass hygiene of cull cattle at slaughter in four geographically distant regions of the United States was examined from July 2005 to April 2006 by measuring the aerobic plate counts (APC) and the prevalences and loads of Salmonella and Escherichia coli O157:H7. The geometric mean log10 APC CFU/100 cm2 levels on hides and preevisceration and postintervention carcasses ranged from 6.17 to 8.19, 4.24 to 6.47, and 1.46 to 1.96, respectively, and were highest in the summer (P < 0.0001). The average prevalences of Salmonella on hides and preevisceration and postintervention carcasses were 89.6% (95% confidence interval [CI], 85.1 to 94.0), 50.2% (95% CI, 40.9 to 59.5), and 0.8% (95% CI, 0.18 to 1.42), respectively. The prevalences of E. coli O157:H7 were 46.9% (95% CI, 37.3 to 56.6) and 16.7% (95% CI, 9.8 to 23.6) on hides and preevisceration carcasses, respectively. Examination of the concomitant incidence of Salmonella and E. coli O157:H7 showed that, on average, 33.3% (95% CI, 15.9 to 69.8) of cattle hide and 4.1% (95% CI, 0.98 to 17.3) of preevisceration carcass samples were contaminated with both pathogens. The pathogen prevalence on hides and carcasses was not significantly affected by the season; however, significant differences were observed between plants with respect to the incoming pathogen load and the ability to mitigate hide-to-carcass transfer. In spite of these differences, postintervention carcass contamination was significantly reduced (P < 0.001), likely as a result of the use of one or more of the processing interventions employed at each of the four processing plants examined.

Prevalence and Characterization of Salmonellae in Commercial Ground Beef in the United States

ABSTRACT Commercially produced ground beef samples (n = 4,136) were collected from seven regions of the United States over a 24-month period (July 2005 to June 2007) and analyzed for the presence of Salmonella enterica by using methods that concurrently provided total prevalence and enumerable levels. The overall prevalence of Salmonella strains was 4.2%. Enumeration showed that 94.2% were present at levels below 2 CFU/g. Regional monthly prevalences of Salmonella strains varied from 1.8% to 6.5% but were not statistically different (P > 0.05). All Salmonella isolates were serotyped and their antibiotic susceptibilities determined and analyzed by pulsed-field gel electrophoresis (PFGE). The most common serotypes identified were Salmonella enterica serotypes Montevideo, Anatum, Muenster, and Mbandaka, with these accounting for one-half of the isolates obtained. The prevalence of multidrug-resistant (MDR) Salmonella was determined to be 0.6%. The most common MDR serotypes were Salmonella enterica serotypes Dublin, Reading, and Typhimurium. MDR strains had resistance to between 2 and 10 antibiotics. There were no regional differences in prevalence of MDR Salmonella. PFGE analysis revealed that indistinguishable XbaI and AvrII restriction digest patterns (RDPs) could be observed in isolates of the same serotype found in different regions and months of sampling. The RDPs of 19 Salmonella strains were compared to RDPs in the PulseNet USA database. Thirteen were indistinguishable from existing patterns, and the number of records for each ranged from 1 to 478. These data show that Salmonella prevalence in commercial ground beef is low and suggest that attempts to identify sources contributing to Salmonella in ground beef by serotype, antibiogram, and PFGE cannot be made without additional documented evidence.

Enumeration of Salmonella and Escherichia coli O157:H7 in ground beef, cattle carcass, hide and faecal samples using direct plating methods†

Aim:  To develop and validate high throughput methods for the direct enumeration of viable and culturable Salmonella and Escherichia coli O157:H7 in ground beef, carcass, hide and faecal (GCHF) samples from cattle.

Prevalence and level of Escherichia coli O157:H7 in feces and on hides of feedlot steers fed diets with or without wet distillers grains with solubles.

The objective of this study was to determine if wet distillers grains with solubles (WDGS) from corn in diets affected Escherichia coli O157:H7 in growing and finishing cattle; steers (n = 603) were randomly assigned to diets with or without WDGS. Hide and fecal samples were collected monthly (October through June) from each animal for enumeration and enrichment of E. coli O157:H7. In the growing phase (0 or 13.9% WDGS diets), fecal prevalence for E. coli O157:H7 in steers fed a diet with WDGS was twice that of the prevalence in control steers (P < 0.001). In the finishing phase (0 or 40% WDGS diets), the average prevalence in feces (P < 0.001) and on hides (P < 0.001) was higher for cattle fed WDGS. The average percentage of fecal E. coli O157:H7 enumerable samples during the finishing phase for cattle fed WDGS was 2.7% compared with 0.1% for control steers (P < 0.001). The average percentage of E. coli O157:H7 enumerable hide samples was not different between diets, but the cattle fed WDGS had higher levels (P < 0.05) of the pathogen. Animals fed WDGS had higher levels of E. coli (P < 0.001), higher pH values (P < 0.001), and lower concentrations of L-lactate (P < 0.001) in feces than those values of the control steers. These results indicate that feeding 40% WDGS could increase the level and prevalence of E. coli O157:H7 in and on feedlot cattle when E. coli O157:H7 is seasonally low.

Prevalence and Characterization of Salmonella in Bovine Lymph Nodes Potentially Destined for Use in Ground Beef

A potential source of pathogenic bacteria in ground beef is the lymphatic system, specifically the lymph nodes. Bacteria have been isolated from the lymph nodes of cattle at slaughter; however, most studies have dealt with mesenteric lymph nodes, which are not normally incorporated into ground beef. The objective of the current study was to determine the prevalence and multidrug-resistance status of Salmonella in bovine lymph nodes associated with lean and fat trimmings that might be utilized in ground beef production. Bovine lymph nodes (n = 1,140) were collected from commercial beef processing plants. Half of the lymph nodes sampled were obtained from cull cow and bull processing plants, and the remainder were obtained from fed beef processing plants. Lymph nodes located in chuck and flank adipose tissue were collected for this study. Salmonella prevalence in the lymph node samples was low, with an overall prevalence of 1.6% and a 95% confidence interval of 0.85 to 2.3%. Lymph nodes from cull cattle carcasses had a higher prevalence of Salmonella than did those from fed cattle carcasses. Lymph nodes from the flanks of cow and bull carcasses had the highest prevalence at 3.86%, whereas lymph nodes from the chuck region of fed cattle carcasses had the lowest prevalence at 0.35%. Three of the 18 Salmonella-positive lymph node samples contained multidrug-resistant Salmonella, and all 3 samples were from cull cattle.

Microbiological characterization of imported and domestic boneless beef trim used for ground beef.

The United States imports lean boneless beef trim from Australia (AUS), New Zealand (NZL), and Uruguay (URY) to meet demand for ground beef production. The reported incidence of and etiological agents responsible for foodborne diseases differ between these countries and the United States. Our objective was to determine whether current U.S. microbiological profiling adequately addresses the potential differences between foreign and domestic beef trim. We compared the hygienic status of imported and domestic (USA) beef trim by enumeration of aerobic bacteria, Enterobacteriaceae, coliforms, Escherichia coli, and Staphylococcus aureus. We also compared the prevalence of pathogens between imported and domestic samples by screening for the presence of Salmonella, Campylobacter spp., Listeria spp., and non-O157 Shiga toxin-producing E. coli (STEC). A total of 1,186 samples (487 USA, 220 AUS, 223 NZL, and 256 URY) of boneless beef trim were analyzed. Results of enumeration revealed significant differences between samples from all countries, with the lowest pathogen numbers in samples from AUS and the highest in samples from URY. Six Salmonella isolates (1 NZL, 1 URY, and 4 USA), 79 L. monocytogenes isolates (4 AUS, 5 NZL, 53 URY, and 17 USA), and 7 Campylobacter isolates (1 NZL, 1 URY, 5 USA) were found among the trim samples tested. Non-O157 STEC prevalence was 10% in NZL samples and about 30% in all of the other samples; 99 STEC strains were isolated. Serotyping of these isolates revealed that serotypes associated with hemolytic uremic syndrome were not different in prevalence between imported and domestic beef trim. Although it may be tempting to do so, these data cannot be used to compare the microbiological quality of beef trim between the countries examined. However, these results indicate that the current pathogen monitoring procedures in the United States are adequate for evaluation of imported beef trim.

Listeria prevalence and Listeria monocytogenes serovar diversity at cull cow and bull processing plants in the United States.

Listeria monocytogenes, the causative agent of epidemic and sporadic listeriosis, is routinely isolated from many sources, including cattle, yet information on the prevalence of Listeria in beef processing plants in the United States is minimal. From July 2005 through April 2006, four commercial cow and bull processing plants were sampled in the United States to determine the prevalence of Listeria and the serovar diversity of L. monocytogenes. Samples were collected during the summer, fall, winter, and spring. Listeria prevalence on hides was consistently higher during cooler weather (28 to 92% of samples) than during warmer weather (6 and 77% of samples). The Listeria prevalence data collected from preevisceration carcass ranged from undetectable in some warm season samples to as high as 71% during cooler weather. Listeria on postintervention carcasses in the chill cooler was normally undetectable, with the exception of summer and spring samples from one plant where > 19% of the carcasses were positive for Listeria. On hides, L. monocytogenes serovar 1/2a was the predominant serovar observed, with serovars 1/2b and 4b present 2.5 times less often and serovar 1/2c not detected on any hides sampled. L. monocytogenes serovars 1/2a, 1/2c, and 4b were found on postintervention carcasses. This prevalence study demonstrates that Listeria species are more prevalent on hides during the winter and spring and that interventions being used in cow and bull processing plants appear to be effective in reducing or eliminating Listeria contamination on carcasses.

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.