James O. Reagan

Microbial Populations on Animal Hides and Beef Carcasses at Different Stages of Slaughter in Plants Employing Multiple-Sequential Interventions for Decontamination

Multiple-sequential interventions were applied commercially to reduce beef carcass contamination in eight packing plants. The study evaluated microbial populations on animal hides and changes in carcass microbial populations at various stages in the slaughtering process. Sponge swab samples yielded mean (log CFU/100 cm2) total plate counts (TPC), total coliform counts (TCC), and Escherichia coli counts (ECC) on the exterior hide in the ranges of 8.2 to 12.5, 6.0 to 7.9, and 5.5 to 7.5, respectively, while corresponding contamination levels on carcass surfaces, after hide removal but before application of any decontamination intervention, were in the ranges of 6.1 to 9.1, 3.0 to 6.0, and 2.6 to 5.3, respectively. Following the slaughtering process and application of multiple-sequential decontamination interventions that included steam vacuuming, pre-evisceration carcass washing, pre-evisceration organic acid solution rinsing, hot water carcass washing, postevisceration final carcass washing, and postevisceration organic acid solution rinsing, mean TPC, TCC, and ECC on carcass surfaces were 3.8 to 7.1, 1.5 to 3.7, and 1.0 to 3.0, respectively, while corresponding populations following a 24 to 36 h chilling period were 2.3 to 5.3, 0.9 to 1.3, and 0.9, respectively. The results support the concept of using sequential decontamination processes in beef packing plants as a means of improving the microbiological quality of beef carcasses.

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

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

Effect of chemical dehairing on the prevalence of Escherichia coli O157:H7 and the levels of aerobic bacteria and enterobacteriaceae on carcasses in a commercial beef processing plant.

The objective of this experiment was to test the hypothesis that cleaning cattle hides by removing hair and extraneous matter before hide removal would result in improved microbiological quality of carcasses in commercial beef processing plants. To test this hypothesis, we examined the effect of chemical dehairing of cattle hides on the prevalence of Escherichia coli O157:H7 and the levels of aerobic bacteria and Enterobacteriaceae on carcasses. Samples from 240 control (conventionally processed) and 240 treated (chemically dehaired before hide removal) hides (immediately after stunning but before treatment) and preevisceration carcasses (immediately after hide removal) were obtained from four visits to a commercial beef processing plant. Total aerobic plate counts (APC) and Enterobacteriaceae counts (EBC) were not (P > 0.05) different between cattle designated for chemical dehairing (8.1 and 5.9 log CFU/100 cm2 for APC and EBC, respectively) and cattle designated for conventional processing (8.0 and 5.7 log CFU/100 cm2 for APC and EBC, respectively). However, E. coli O157:H7 hide prevalence was higher (P < 0.05) for the control group than for the treated group (67% versus 88%). In contrast to hides, the bacterial levels were lower (P < 0.05) on the treated (3.5 and 1.4 log CFU/100 cm2 for APC and EBC) than the control (5.5 and 3.2 log CFU/100 cm2 for APC and EBC) preevisceration carcasses. Prevalence of E. coli O157:H7 was lower (P > 0.05) on treated than on control preevisceration carcasses (1% versus 50%). These data indicate that chemical dehairing of cattle hides is an effective intervention to reduce the incidence of hide-to-carcass contamination with pathogens. The data also imply that any effective hide intervention process incorporated into beef processing procedures would significantly reduce carcass contamination by E. coli O157:H7.

Trimming and Washing of Beef Carcasses as a Method of Improving the Microbiological Quality of Meat

A study to compare procedures and interventions for removing physical and bacterial contamination from beef carcasses was conducted in six carcass conversion operations that were representative of modern, high-volume plants and located in five different states. Treatment procedures included trimming, washing, and the current industry practice of trimming followed by washing. In addition, hot (74 to 87.8°C at the pipe) water washing and rinsing with ozone (0.3 to 2.3 ppm) or hydrogen peroxide (5%) were applied as intervention treatments. Beef carcasses were deliberately contaminated with bovine fecal material at >4.0 log colony-forming units (CFU)/cm2 in order to be better able to observe the decontaminating effects of the treatments. Carcasses were visually scored by 2 to 3 trained personnel for the level of gross contamination before and after treatment. Samples (10 by 15 cm, 0.3 to 0.5 cm thick) for microbiological testing were excised as controls or after application of each procedure or intervention and analyzed for aerobic mesophilic plate counts, Escherichia coli Biotype I counts, and presence or absence of Listeria spp., Salmonella spp., and Escherichia coli O157:H7. Average reductions in aerobic plate counts were 1.85 and 2.00 log CFU/cm2 for the treatments of trimming-washing and hot-water washing, respectively. Hydrogen peroxide and ozone reduced aerobic plate counts by 1.14 and 1.30 log CFU/cm2, respectively. In general, trimming and washing of beef carcasses consistently resulted in low bacterial populations and scores for visible contamination. However, the data also indicated that hot- (74 to 87.8°C at the pipe) water washing was an effective intervention that reduced bacterial and fecal contamination in a consistent manner.

Prevalence of Escherichia coli O157 and Levels of Aerobic Bacteria and Enterobacteriaceae Are Reduced When Hides Are Washed and Treated with Cetylpyridinium Chloride at a Commercial Beef Processing Plant

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

Sources and Extent of Microbiological Contamination of Beef Carcasses in Seven United States Slaughtering Plants

This study determined microbiological loads of beef carcasses at different stages during the slaughtering to chilling process in seven (four steer/heifer and three cow/bull) plants. Potential sources of contamination (feces, air, lymph nodes) were also tested. Each facility was visited twice, once in November through January (wet season) and again in May through June (dry season). Carcasses were sampled by aseptic excision of surface tissue (100 cm2) from the brisket, flank, and rump (30 samples each) after hide removal (pre-evisceration), after final carcass washing, and after 24-h carcass chilling. The samples were analyzed individually by standard procedures for aerobic plate counts (APC), total coliform counts (TCC), Escherichia coli biotype I counts (ECC), and presence of Salmonella. Incidence of Salmonella was higher on dry feces of older compared to younger animals, fresh feces of younger compared to older animals, and on cow/bull carcasses compared to steer/heifer carcasses. Most factors and their interactions had significant (P < or = 0.05) effects on the bacterial counts obtained. Depending on plant and season, APC, TCC, and ECC were < or =10(4), < or =10(2), and < or =10(1) CFU/cm2 in 46.7 to 93.3, 50.0 to 100.0, and 74.7 to 100.0% of the samples, respectively. TCC exceeded 10(3) CFU/cm2 in 2.5% (wet season) and 1.5% (dry season) of the samples. ECC exceeded 10(2) CFU/cm2 in 8.7%, 0.3%, and 1.5% of the pre-evisceration, final carcass-washing, and 24-h carcass-chilling samples, respectively, during the wet season; the corresponding numbers during the dry season were 3.5%, 2.2%, and 3.0%, respectively. These data should serve as a baseline for future comparisons in measuring the microbiological status of beef carcasses, as the new inspection requirements are implemented.

Steam vacuuming as a pre-evisceration intervention to decontaminate beef carcasses

One steam-vacuuming unit (Unit A) was evaluated for removal of visible contamination and reduction of bacterial counts on beef carcass surfaces in five processing plants; a second steam-vacuuming unit (Unit B) was evaluated in two of those same plants at a later date. Experimental treatments included appropriate Controls: steam vacuuming carcass surfaces with or without visible contamination, and knife trimming surfaces with visible contamination. Depending on the processing plant, carcasses were tested on the midline or on the round. Each treatment was applied to a 103-cm2 area of the carcass surface, which was scored for visible contamination and analyzed for aerobic plate counts (APC) at 25°C and for total coliform counts (TCC). Average reductions in APC of 0.57 (Unit A) and 0.72 (Unit B) log CFU/cm2 and in TCC of 0.33 (Unit A) and 0.26 (Unit B) log CFU/cm2 were obtained by steam-vacuuming carcass surfaces which had no visible fecal contamination. Steam vacuuming and knife trimming effectively (P < 0.05) cleaned soiled carcass surfaces and reduced microbial counts. Knife trimming reduced APC and TCC by 1.38 and 1.61 log CFU/cm2 in the Unit A experiment and by 1.64 and 1.72 log CFU/cm2 in the Unit B experiment, respectively. Steam vacuuming carcass surfaces soiled with visible contamination reduced APC and TCC by 1.73 and 1.67 log CFU/cm2 (Unit-A) and by 2.03 and 2.13 log CFU/cm2 (Unit B), respectively. The results of this study suggest that both steam-vacuuming systems available at the time of the study were at least as effective as knife trimming in decontaminating beef carcasses with areas of visible contamination 2.54 cm in the greatest dimension.

Incidence of Salmonella on Beef Carcasses Relating to the U.S. Meat and Poultry Inspection Regulations

This article is part of a major study designed to collect baseline contamination data by sampling beef carcasses in seven slaughtering plants (four steer-heifer and three cow-bull plants) during both a dry season (November to January) and a wet season (May to June). Samples (n = 30) were excised from each of three carcass anatomical sites (brisket, flank, and rump) at each of three points in the slaughtering chain (pre-evisceration, following final carcass washing, after 24-h carcass chilling). A total of 3,780 samples (100 cm2 each) were analyzed for presence of Salmonella; aerobic plate counts, total coliform counts, and Escherichia coli counts were also made. After 24-h chilling, average incidence (expressed as a percentage) of Salmonella in the brisket, flank, and rump samples, respectively, for steer-heifer carcasses was 0.8+/-1.7, 0, and 2.5+/-5.0 for the wet season and 0.8+/-1.7, 0, and 0 for the dry season; the corresponding percentages for cowbull carcasses were 4.4+/-2.0, 2.2+/-3.9, and 1.1+/-1.9 for the wet season and 2.2+/-3.9, 1.1+/-1.9, and 0 for the dry season. Depending on plant and season, ranges of probabilities of chilled steer-heifer carcasses passing the U.S. regulatory requirements for Salmonella contamination were 0.24 to 1.0 for the brisket, 1.0 for the flank, and 0.002 to 1.0 for the rump; the corresponding ranges for the chilled cow-bull carcasses were 0.25 to 1.0, 0.25 to 1.0, and 0.70 to 1.0. When the number of positive brisket, flank, and rump samples were combined, the probabilities of passing the regulatory requirements were 0.242 to 1.0 and 0.772 to 1.0 for the wet and dry seasons, respectively, in steer-heifer plants and 0.368 to 0.974 and 0.865 to 1.0 in cow-bull plants. Correlation coefficients of aerobic plate counts, total coliform counts, and E. coli counts with Salmonella incidence were higher (P< or =0.05) for cow-bull samples that had increased incidence of the pathogen when compared to steer-heifer samples.

Protocol for Evaluating the Efficacy of Cetylpyridinium Chloride as a Beef Hide Intervention

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

Incidence of Escherichia coli O157:H7 in Frozen Beef Patties Produced over an 8-Hour Shift†

A ground beef patty processor detected Escherichia coli O157:H7 in five production lots during routine testing with polymerase chain reaction (PCR) technology. This finding stimulated research to determine the incidence and potential entry points of the pathogen during processing. One of these lots (53,960 kg) was divided into 71 pallets (760 kg each) of food service ground beef patties. Ten cartons (19 kg each) were removed from each pallet, for a total of 710 cartons. Four patties were taken from each carton and subdivided to provide comparable samples for E. coli O157:H7 analyses by three different laboratories. Two laboratories employed different immunoassay tests, and one used PCR to screen samples. One sample set was analyzed for aerobic plate, coliform, and E coli Biotype I counts to determine if any relationship existed between these microbial groups and the incidence of E. coli O157:H7. For 73 samples, presumptive positive results for E. coli O157:H7 were obtained by one or more methods. For 48 of these 73 samples, positive results for the pathogen were culture confirmed. The largest number (29) of culture-confirmed positive E. coli O157:H7 results were detected by PCR. Most positive results were obtained during a short segment of processing. All culture-confirmed E. coli O157:H7 strains were further characterized by two genetic subtyping techniques, resulting in two to four different patterns, depending on the subtyping procedure employed. For any sample tested, the aerobic plate count was < 3.0 log CFU/g, and coliform and E. coli Biotype I counts were < or = 1.00 log CFU/g. The results of this study suggest that most positive samples were associated with a contaminated batch of raw material introduced just before the 1725- to 1844-h processing segment. These results also indicate that more aggressive sampling plans and genetic screening technologies such as PCR may be used to better detect low levels of E. coli O157:H7 in ground beef products.