Efficacy of a Blend of Sulfuric Acid and Sodium Sulfate against Shiga Toxin-Producing Escherichia coli, Salmonella, and Nonpathogenic Escherichia coli Biotype I on Inoculated Prerigor Beef Surface Tissue.

A study was conducted to investigate the efficacy of a sulfuric acid-sodium sulfate blend (SSS) against Escherichia coli O157:H7, non-O157 Shiga toxin-producing E. coli (STEC), Salmonella, and nonpathogenic E. coli biotype I on prerigor beef surface tissue. The suitability of using the nonpathogenic E. coli as a surrogate for in-plant validation studies was also determined by comparing the data obtained for the nonpathogenic inoculum with those for the pathogenic inocula. Prerigor beef tissue samples (10 by 10 cm) were inoculated (ca. 6 log CFU/cm2) on the adipose side in a laboratory-scale spray cabinet with multistrain mixtures of E. coli O157:H7 (5 strains), non-O157 STEC (12 strains), Salmonella (6 strains), or E. coli biotype I (5 strains). Treatment parameters evaluated were two SSS pH values (1.5 and 1.0) and two spray application pressures (13 and 22 lb/in2). Untreated inoculated beef tissue samples served as controls for initial bacterial populations. Overall, the SSS treatments lowered inoculated (6.1 to 6.4 log CFU/cm2) bacterial populations by 0.6 to 1.5 log CFU/cm2 (P < 0.05), depending on inoculum type and recovery medium. There were no main effects (P ≥ 0.05) of solution pH or spray application pressure when SSS was applied to samples inoculated with any of the tested E. coli inocula; however, solution pH did have a significant effect (P < 0.05) when SSS was applied to samples inoculated with Salmonella. Results indicated that the response of the nonpathogenic E. coli inoculum to the SSS treatments was similar (P ≥ 0.05) to that of the pathogenic inocula tested, making the E. coli biotype I strains viable surrogate organisms for in-plant validation of SSS efficacy on beef. The application of SSS at the tested parameters to prerigor beef surface tissue may be an effective intervention for controlling pathogens in a commercial beef harvest process.

Antimicrobial Efficacy of a Sulfuric Acid and Sodium Sulfate Blend, Peroxyacetic Acid, and Cetylpyridinium Chloride against Salmonella on Inoculated Chicken Wings.

Studies were conducted to evaluate the efficacy of a commercial blend of sulfuric acid and sodium sulfate (SSS) in reducing Salmonella on inoculated whole chilled chicken wings and to compare its efficacy to peroxyacetic acid (PAA) and cetylpyridinium chloride (CPC). Wings were spot inoculated (5 to 6 log CFU/ml of sample rinsate) with a five-strain mixture of novobiocin- and nalidixic acid-resistant Salmonella and then left untreated (control) or treated by immersing individual wings in 350 ml of antimicrobial solution. An initial study evaluated two treatment immersion times, 10 and 20 s, of SSS (pH 1.1) and compared cell recoveries following rinsing of treated samples with buffered peptone water or Dey/Engley neutralizing broth. In a second study, inoculated wings were treated with SSS (pH 1.1; 20 s), PAA (700 ppm, 20 s), or CPC (4,000 ppm, 10 s) and analyzed for survivors immediately after treatment (0 h) and after 24 h of aerobic storage at 4°C. Color and pH analyses were also conducted in the latter study. Recovery of Salmonella survivors following treatment with SSS (10 or 20 s) was not (P ≥ 0.05) affected by the type of cell recovery rinse solution (buffered peptone water or Dey/Engley neutralizing broth), but there was an effect (P < 0.05) of SSS treatment time. Immersion of samples for 10 or 20 s in SSS resulted in pathogen reductions of 0.8 to 0.9 and 1.1 to 1.2 log CFU/ml, respectively. Results of the second study showed that there was an interaction (P < 0.05) between antimicrobial type and storage time. Efficacy against Salmonella at 0 h increased in the order CPC , SSS , PAA; however, after 24 h of aerobic storage, pathogen counts of SSS- and PAA-treated wings did not differ (P ≥ 0.05). Overall, the results indicated that SSS applied at pH 1.1 for 20 s was an effective antimicrobial intervention to reduce Salmonella contamination on chicken wings.

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.

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.

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.

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 shiftt.

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.

Identification of midgut and salivary glands as specific and distinct barriers to efficient tick-borne transmission of Anaplasma marginale.

Understanding the determinants of efficient tick-borne microbial transmission is needed to better predict the emergence of highly transmissible pathogen strains and disease outbreaks. Although the basic developmental cycle of Anaplasma and Ehrlichia spp. within the tick has been delineated, there are marked differences in the ability of specific strains to be efficiently tick transmitted. Using the highly transmissible St. Maries strain of Anaplasma marginale in Dermacentor andersoni as a positive control and two unrelated nontransmissible strains, we identified distinct barriers to efficient transmission within the tick. The Mississippi strain was unable to establish infection at the level of the midgut epithelium despite successful ingestion of infected blood following acquisition feeding on a bacteremic animal host. This inability to colonize the midgut epithelium prevented subsequent development within the salivary glands and transmission. In contrast, A. marginale subsp. centrale colonized the midgut and then the salivary glands, replicating to a titer indistinguishable from that of the highly transmissible St. Maries strain and at least 100 times greater than that previously associated with successful transmission. Nonetheless, A. marginale subsp. centrale was not transmitted, even when a large number of infected ticks was used for transmission feeding. These results establish that there are at least two specific barriers to efficient tick-borne transmission, the midgut and salivary glands, and highlight the complexity of the pathogen-tick interaction.

Effects of Meat Type, Storage Time and Temperature on Various Physical, Chemical and Microbiological Characteristics of Ground Pork.

Whole-hog sausage was prepared from hot- and cold-boned pork raw materials to determine the effects of meat type, storage temperature and length of storage on various processing and bacteriological characteristics. Samples were stored at -1 and 4°C for 0, 28 and 56 d. Various physical, chemical and microbiological properties of the sausage were evaluated. Thiobarbituric acid (TBA) values were not affected by meat type (pre or postrigor). Hunter-Color values varied significantly among the meat types and storage temperatures. Total bacterial counts varied significantly among the hot- and cold-boned pork sausage samples (day 0). Cold-boned sausage stored at -1°C had lower plate counts of the various treatments for days 28 and 56. Pseudomonas was the predominant organism found in hotand cold-boned sausage samples. Hot-boned sausage exhibited a more diverse bacterial population than did cold-boned sausage. More gram-positive organisms were found in hot-boned sausage samples. Cold-boned sausage had a lower total bacterial count at day 0 and maintained lower counts and therefore a longer shelf life throughout the study when held at -1°C.

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.

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.