Ozone treatment for reduction of Escherichia coli 0157:H7 and Salmonella serotype typhimurium on beef carcass surfaces

The effectiveness of an aqueous ozone treatment in reducing Escherichia coli O157:H7 and Salmonella serotype Typhimurium on hot carcass surfaces was determined with the use of a model carcass spray cabinet. Carcass surface regions were removed from carcasses and inoculated with feces containing 10(6) to 10(7) CFU each of E. coli O157:H7 and Salmonella Typhimurium per g and were then exposed to a water wash or to a water wash followed by a sanitizing ozone treatment. Water washes were applied at 28 degrees C beginning at a pressure of 10 lb/in2 and gradually increasing to 400 lb/in2. Ozone treatment was carried out by spraying surfaces with an aqueous ozone solution (80 lb/in2 at 28 degrees C) containing 95 mg of ozone per liter. Pathogen reductions achieved with ozone treatment were not significantly different from those achieved with a water wash alone. In addition, ozone treatment did not reduce E. coli O157:H7 or Salmonella Typhimurium contamination that was spread over the carcass surface as a result of the water wash. Under the conditions of this study, the aqueous ozone treatment applied resulted in no significant improvement over a water wash in reducing pathogens on beef carcass surfaces.     

Lactic acid sprays reduce bacterial pathogens on cold beef carcass surfaces and in subsequently produced ground beef

Organic acids have been shown to be effective in reducing the presence of pathogenic bacteria on hot beef carcass surfaces; however, application for decontaminating chilled carcasses has not been fully evaluated. In this study, a postchill, 30-s lactic acid spray (500 ml of 4% L-lactic acid, 55 degrees C) was applied onto outside rounds that had been contaminated with Escherichia coli O157:H7 and Salmonella Typhimurium, subsequent to prechill hot carcass treatments consisting of water wash alone or water wash followed by a 15-s lactic acid spray (250 ml of 2% L-lactic acid, 55 degrees C). The prechill treatments reduced both pathogens by 3.3 to 3.4 log cycles (water wash alone) to 5.2 log cycles (water wash and lactic acid). In all cases, the postchill acid treatment produced an additional reduction in E. coli O157:H7 of 2.0 to 2.4 log cycles and of 1.6 to 1.9 log cycles for Salmonella Typhimurium. The counts of both pathogens remained significantly lower in ground beef produced from the outside rounds that received prechill and postchill acid spray than from those that received a postchill spray only. These data indicate that organic acid sprays may be successfully applied for pathogen reduction in beef carcass processing after the cooler, especially when combined with prechill treatments.     

Efficacy of washing meat surfaces with 2% levulinic, acetic, or lactic acid for pathogen decontamination and residual growth inhibition

We compared spray washing at 55.4 °C with 2% levulinic acid to that with lactic or acetic acid for decontamination of pathogenic bacteria inoculated onto meat surfaces, and their residual protection against later growth of pathogenic bacteria. The model systems included Escherichia coli O157:H7 on beef plate, Salmonella on chicken skin and pork belly, and Listeria monocytogenes on turkey roll. In the decontamination studies, acid washes lowered recoverable numbers of pathogens by 0.6 to 1 log/cm(2) as compared to no-wash controls, and only lactic acid lowered the number of pathogens recovered as compared to the water wash. Washing with levulinic acid at 68.3 or 76.7 °C did not result in additional decontamination of E. coli. Acetic acid prevented residual growth of E. coli and L. monocytogenes, and it reduced numbers of Salmonella on chicken skin to below recoverable levels. Overall, levulinic acid did not provide as effective decontamination as lactic acid nor residual protection as acetic acid.     

Interventions for the reduction of Salmonella Typhimurium DT 104 and non-O157:H7 enterohemorrhagic Escherichia coli on beef surfaces

A study was conducted to determine if slaughter interventions currently used by the meat industry are effective against Salmonella Typhimurium definitive type 104 (DT 104) and two non-O157:H7 enterohemorrhagic Escherichia coli (EHEC). Three separate experiments were conducted by inoculating prerigor beef surfaces with a bovine fecal slurry containing Salmonella Typhimurium and Salmonella Typhimurium DT 104 (experiment 1), E. coli O157:H7 and E. coli O111:H8 (experiment 2), or E. coli O157:H7 and E. coli O26:H11 (experiment 3) and spray washing with water, hot water (72 degrees C), 2% acetic acid, 2% lactic acid, or 10% trisodium phosphate (15 s, 125 +/- 5 psi, 35 +/- 2 degrees C). Remaining bacterial populations were determined immediately after treatments (day 0), after 2 days of aerobic storage at 4 degrees C, and after 7, 21, and 35 days of vacuum-packaged storage at 4 degrees C. In addition to enumeration, confirmation of pathogen serotypes was performed for all treatments on all days. Of the interventions investigated, spray treatments with trisodium phosphate were the most effective, resulting in pathogen reductions of >3 log10 CFU/cm2, followed by 2% lactic acid and 2% acetic acid (>2 log10 CFU/cm2). Results also indicated that interventions used to reduce Salmonella Typhimurium on beef surfaces were equally effective against Salmonella Typhimurium DT 104 immediately after treatment and again after long-term, refrigerated, vacuum-packaged storage. Similarly, E. coli O111:H8 and E. coli O26:H11 associated with beef surfaces were reduced by the interventions to approximately the same extent as E. coli O157:H7 immediately after treatment and again after long-term, refrigerated, vacuum-packaged storage. It was also demonstrated that phenotypic characterization may not be sufficient to identify EHECs and that the organisms should be further confirmed with antibody- or genetic-based techniques. Based on these findings, interventions used by the meat industry to reduce Salmonella spp. and E. coli O157:H7 appear to be effective against DT 104 and other EHEC.     

Reduction of Escherichia coli O157:H7 and Salmonella typhimurium on beef carcass surfaces using acidified sodium chlorite.

The efficacy of a phosphoric acid-activated acidified sodium chloride (PASC) spray and a citric acid-activated acidified sodium chlorite (CASC) spray applied at room temperature (22.4 to 24.7 degrees C) in combination with a water wash was compared with that of a water wash only treatment for reduction of Escherichia coli O157:H7 and Salmonella Typhimurium inoculated onto various hot-boned individual beef carcass surface regions (inside round, outside round, brisket, flank, and clod). Initial counts of 5.5 and 5.4 log CFU/cm2 were obtained after inoculation with E. coli O157:H7 and Salmonella Typhimurium, respectively. Initial numbers for both pathogens were reduced by 3.8 to 3.9 log cycles by water wash followed by PASC spray and by 4.5 to 4.6 log cycles by water wash followed by CASC spray. The sprays consisted of applying 140 ml of the appropriate sanitizing solution for 10 s at 69 kPa. Corresponding reduction values obtained by water wash alone were 2.3 log. The performance of CASC appeared to be consistently better than that of PASC. In general, no effect of the carcass surface region was observed on the log reductions for either pathogen, except for the inside round, which consistently had lower reductions. Both PASC and CASC were capable of effectively reducing pathogens spread to areas beyond the initial contaminated area of the cuts to levels close to or below the counting method detection limit (0.5 log CFU/cm2). However, 30 to 50% of the carcasses treated by these antimicrobial solutions still yielded countable colonies. Results of this study indicate that acidified sodium chlorite sprays are effective for decontaminating beef carcass surfaces.     

Effect of xylitol on adhesion of Salmonella Typhimurium and Escherichia coli O157:H7 to beef carcass surfaces

Effects of 10% xylitol (a five-carbon sugar alcohol) on adhesion of Escherichia coli O157:H7 and Salmonella Typhimurium to meat surfaces were examined with three approaches. First, beef outside round was inoculated with rifampin-resistant E. coli O157:H7 and Salmonella Typhimurium dispersed in xylitol or peptone solution. Samples were rinsed with water or not rinsed in a 2 x 2 factorial arrangement. No interaction existed between inoculum and rinsing treatments (P > 0.84). Incubation in xylitol had minimal impact on pathogen adhesion (P > 0.76); however, rinsing reduced pathogen cell counts (P < 0.01). Second, meat samples were treated with water, xylitol, or no rinse; inoculated with pathogens dispersed in peptone solution (8.6 log CFU/ml for each pathogen); and then treated with water, xylitol, or no rinse in a 3 x 3 factorial arrangement. No interactions were observed (P > 0.50). Postinoculation rinsing reduced pathogen loads (P < 0.01) without difference between water and xylitol (P > 0.64). Third, carcass surfaces inoculated with pathogens (5.5 log CFU/cm2) were treated with 35 degrees C water wash, 2.5% L-lactic acid spray, 10% xylitol spray, lactic acid plus xylitol, or hot water plus xylitol. Lactic acid treatments reduced Salmonella Typhimurium at 0 h (P < 0.01) and 24 h (P < 0.02). Hot water treatments tended to reduce Salmonella Typhimurium at 0 h (P < 0.07). Xylitol did not reduce pathogens (P > 0.62) or increase effectiveness of other treatments. Xylitol does not influence E. coli O157:H7 and Salmonella Typhimurium adhesion to meat surfaces.     

Evaluation of various antimicrobial interventions for the reduction of Escherichia coli O157:H7 on bovine heads during processing

The effectiveness of electrolyzed oxidizing water, FreshFx, hot water, DL-lactic acid, and ozonated water was determined using a model carcass spray-washing cabinet. A total of 140 beef heads obtained from a commercial processing line were inoculated with Escherichia coli O157:H7 on the cheek areas. Each head was exposed to a simulated preevisceration wash and then had antimicrobial wash treatments. Hot water, lactic acid, and FreshFx treatments reduced E. coli O157:H7 on inoculated beef heads by 1.72, 1.52, and 1.06 log CFU/cm2, respectively, relative to the simulated preevisceration wash. Electrolyzed oxidizing water and ozonated water reduced E. coli O157:H7 less than 0.50 log CFU/cm2. Hot water, lactic acid, and FreshFx could be used as decontamination washes for the reduction of E. coli O157:H7 on bovine head and cheek meat.     

Efficacy of a peroxyacetic acid formulation as an antimicrobial intervention to reduce levels of inoculated Escherichia coli O157:H7 on external carcass surfaces of hot-boned beef and veal

The efficacy of a peroxyacetic acid formulation (POAA) at reducing Escherichia coli O157:H7 contamination on external carcass surfaces of hot-boned beef and veal with a commercial spray apparatus was determined. Hot-boned external carcass surfaces were inoculated with either a high dose (10(6) CFU/cm2) in fresh bovine feces or with a low dose (10(3) CFU/cm2) in diluent of laboratory-cultured E. coli O157:H7. Treatments included a water wash, a POAA (180 ppm) wash, or a water plus POAA wash. Samples were extracted from the external carcass surface with a cork borer to determine the numbers of viable E. coli O157:H7 remaining on the carcass surface after treatment. Although a water wash alone resulted in a 1.25 (94.4%) and a 1.31 (95.1%) mean log reduction on veal and beef inoculated with a high dose of E. coli O157:H7, the POAA treatment resulted in a substantially greater mean log reduction of 3.56 and 3.59 (>99.9%). The water wash only resulted in a 33.9% reduction on veal and 62.8% on beef inoculated with a low dose of E. coli O157:H7, whereas POAA treatment greatly improved pathogen reduction to 98.9 and 97.4% on veal and beef, respectively. The combination of a water wash followed by a POAA treatment resulted in a similar E. coli O157:H7 reduction to that achieved by POAA treatment alone. In conclusion, POAA treatment significantly reduced viable E. coli O157:H7 numbers on experimentally contaminated beef and veal carcasses, which justifies its use as a chemical intervention for the removal of this human pathogen.     

SURVIVAL OF SALMONELLA TYPHIMURIUM, ESCHERICHIA COLI O157:H7 AND LISTERIA MONOCYTOGENES DURING STORAGE ON BEEF SANITIZED WITH ORGANIC ACIDS

Sterile beef tissue was inoculated with either Salmonella typhimurium, Escherichia coli O157:H7 or Listeria monocytogenes Scott A and washed with 23C distilled water, 1% lactic acid or 1% acetic acid. The washed tissue was subjected to simulated dry chilling or spray chilling followed by storage at 5C. The washed tissue was stored at 5C for up to 21 days at 26% relative humidity, and total bacterial populations were determined by plating on nonselective and selective agars. There was no significant difference in the surviving populations of S. typhimurium, Escherichia coli O157:H7, or L. monocytogenes after storage, irrespective of chilling method. The surviving populations of bacteria were significantly lower on acid washed adipose tissue, when compared to the comparable water washed tissue. These results indicate that although injury and recovery of pathogenic bacteria may occur as a result of organic acid carcass sanitizing treatments, there was no practical significance of this phenomenon after 3 days of storage.     

Evaluation of peroxyacetic acid as a post-chilling intervention for control of Escherichia coli O157:H7 and Salmonella Typhimurium on beef carcass surfaces

Four experiments were conducted to test the efficacy of peroxyacetic acid as a microbial intervention on beef carcass surfaces. In these experiments, beef carcass surfaces were inoculated with fecal material (no pathogens) or fecal material containing rifampicin-resistant Escherichia coli O157:H7 and Salmonella Typhimurium. Inoculated surfaces were subjected to a simulated carcass wash with and without 2% l-lactic acid treatment before chilling. In Experiments 1 and 2, the chilled carcass surfaces were sprayed with peroxyacetic acid (200 ppm; 43°) for 15 s. Peroxyacetic acid had no effect on microbial counts of any organism measured on these carcass surfaces. However, lactic acid reduced counts of E. coli Type I (1.9log(10) CFU/cm(2)), coliforms (3.0log(10) CFU/cm(2)), E. coli O157:H7 (2.7log(10) CFU/cm(2)), and S. Typhimurium (2.8log(10) CFU/cm(2)) entering the chilling cooler and prevented growth during the chilling period. In Experiment 3, peroxyacetic acid at different concentrations (200, 600, and 1000 ppm) and application temperatures (45 and 55 °C) were used to investigate its effectiveness in killing E. coli O157:H7 and S. Typhimurium compared to 4% l-lactic acid (55 °C). Application temperature did not affect the counts of either microorganism. Peroxyacetic acid concentrations up to 600 ppm had no effect on these microorganisms. Concentrations of 1000 ppm reduced E. coli O157:H7 and S. Typhimurium by up to 1.7 and 1.3log(10) CFU/cm(2), respectively. However, 4% lactic acid reduced these organisms by 2.7 and 3.4log(10) CFU/cm(2), respectively. In Experiment 4, peroxyacetic acid (200 ppm; 43 °C) was applied to hot carcass surfaces. This treatment caused a 0.7log(10) CFU/cm(2) reduction in both E. coli O157:H7 and S. Typhimurium. The collective results from these experiments indicate that peroxyacetic acid was not an effective intervention when applied to chilled inoculated carcass piece surfaces.     

Treatments using hot water instead of lactic acid reduce levels of aerobic bacteria and Enterobacteriaceae and reduce the prevalence of Escherichia coil O157:H7 on preevisceration beef carcasses

Lactic acid has become the most commonly used organic acid for treatment of postevisceration beef carcasses. Many processors have also implemented 2% lactic acid washes on preevisceration carcasses. We previously demonstrated that hot water washing and steam vacuuming are effective carcass interventions. Because of the effectiveness of hot water, we compared its use with that of lactic acid as a preevisceration wash in a commercial setting. A commercial hot water carcass wash cabinet applying 74 degrees C (165 degrees F) water for 5.5 s reduced both aerobic plate counts and Enterobacteriaceae counts by 2.7 log CFU/100 cm2 on preevisceration carcasses. A commercial lactic acid spray cabinet that applied 2% L-lactic acid at approximately 42 degrees C (105 to 110 degrees F) to preevisceration carcasses reduced aerobic plate counts by 1.6 log CFU/100 cm2 and Enterobacteriaceae counts by 1.0 log CFU/100 cm2. When the two cabinets were in use sequentially, i.e., hot water followed by lactic acid, aerobic plate counts were reduced by 2.2 log CFU/100 cm2 and Enterobacteriaceae counts were reduced by 2.5 log CFU/100 cm2. Hot water treatments reduced Escherichia coli O157:H7 prevalence by 81%, and lactic acid treatments reduced E. coli O157:H7 prevalence by 35%, but the two treatments in combination produced a 79% reduction in E. coli O157:H7, a result that was no better than that achieved with hot water alone. These results suggest that hot water would be more beneficial than lactic acid for decontamination of preevisceration beef carcasses.     

Investigation of chemical rinses suitable for very small meat plants to reduce pathogens on beef surfaces

Numerous antimicrobial interventions are capable of reducing the prevalence of harmful bacteria on raw meat products. There is a need to identify effective and inexpensive antimicrobial interventions that could, in practice, be used in very small meat plants because of limited financial, space, and labor resources. Eight antimicrobial compounds (acetic acid, citric acid, lactic acid, peroxyacetic acid, acidified sodium chlorite, chlorine dioxide, sodium hypochlorite, and aqueous ozone) were applied at various concentrations with small, hand-held spraying equipment, and bactericidal effectiveness was examined. Beef plate pieces were inoculated with fecal slurry containing a pathogen cocktail (Escherichia coli O157:H7, Salmonella Typhimurium, Campylobacter coli, and Campylobacter jejuni) and natural populations of aerobic plate counts, coliforms, and E. coli. Antimicrobial solutions were applied to beef surfaces via a portable, pressurized hand-held spray tank, and treated surfaces were subjected to appropriate methods for the enumeration and isolation of pathogens and hygiene indicators. Relative antimicrobial effectiveness was determined (from greatest to least): (i) organic acids, (ii) peroxyacetic acid, (iii) chlorinated compounds, and (iv) aqueous ozone. Using the equipment described, a 2% lactic acid rinse provided 3.5- to 6.4-log CFU/cm(2) reductions across all bacterial populations studied. Conversely, aqueous ozone yielded 0.02- to 2.9-log CFU/cm(2) reductions in pathogens and hygiene indicators, and did not differ significantly from a control tap water rinse (P = 0.055 to 0.731). This 2% lactic acid rinse will be subsequently combined with a previously described water wash to create a multistep antimicrobial intervention that will be examined under laboratory conditions and validated in very small meat plants.     

Antimicrobial effect of herb extracts against Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella typhimurium associated with beef

The effects of plant extracts against pathogenic bacteria in vitro are well known, yet few studies have addressed the effects of these compounds against pathogens associated with muscle foods. A series of experiments was conducted to determine the effectiveness of a commercially available, generally recognized as safe, herb extract dispersed in sodium citrate (Protecta One) or sodium chloride (Protecta Two) against Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes associated with beef. In the first experiment, E. coli O157:H7, Salmonella typhimurium, and L. monocytogenes inoculated onto beef and subjected to surface spray treatments with 2.5% solutions of Protecta One or Protecta Two were not affected by immediate application (day 0) of the herbal extracts. However, after 7 days of storage at 4 degrees C, E. coli O157:H7 was reduced by >1.3 log10 CFU/cm2 by Protecta Two; L. monocytogenes was reduced by 1.8 and 1.9 log10 CFU/cm2 by Protecta One and Protecta Two, respectively; Salmonella typhimurium was not reduced >0.3 log10 CFU/cm2 by either extract by day 7. In the second experiment, 2.5% Protecta Two (wt/vol or wt/wt) added to inoculated lean and adipose beef trim, processed, and packaged as ground beef chubs (80% lean, 20% adipose), did not reduce pathogen populations >0.5 log10 CFU/g up to 14 days at 4 degrees C. In the third experiment, surface spray treatments of beef with 2.5% lactic acid or 2.5% solutions of Protecta One or Protecta Two, vacuum packaged, and stored up to 35 days at 4 degrees C did reduce E. coli O157:H7, L. monocytogenes, and Salmonella Typhimurium slightly. These studies suggest that the use of herb extracts may afford some reductions of pathogens on beef surfaces; however, the antimicrobial activity may be diminished in ground beef by adipose components.     

Identification of Escherichia coli O157:H7 surrogate organisms to evaluate beef carcass intervention treatment efficacy

We compared the survival of potential pathogen surrogates-meat-hygiene indicators (non-Escherichia coli coliforms), biotype I E. coli, and lactic acid bacteria starter cultures-with survival of an E. coli O157:H7 (ECO157:H7) inoculum in beef carcass intervention trials. Survival of one lactic acid bacteria starter culture (Bactoferm LHP Dry [Pediococcus acidilactici and Pediococcus pentosaceus]), a five-isolate biotype I inoculum, and a five-isolate non-E. coli coliform inoculum, was compared with survival of a 12-isolate ECO157:H7 inoculum in interventions by using beef brisket (adipose and lean), cod fat membrane, or neck tissue. Treatments were grouped by abattoir size: small (6-day dry aging; 22°C acid treatment [2.5% acetic acid, 2% lactic acid, or Fresh Bloom], followed by 1-day dry aging; hot water) and large (warm acid treatment [5% acetic acid or 2% lactic acid] with or without a preceding hot water treatment). Reductions in pathogen and surrogate inocula were determined with excision sampling. A surrogate was considered a suitable replacement for ECO157:H7 if the intervention produced a reduction in surrogate levels that was not significantly greater (P≥0.05) than that observed for ECO157:H7. All three surrogate inocula were suitable as ECO157 surrogates for dry aging and acid spray plus dry-aging treatments used by small abattoirs. No one inoculum was suitable as an ECO157 surrogate across all intervention treatments used by large abattoirs. Effects seen on neck tissue were representative of other tissues, and the low value of the neck supports its use as the location for evaluating treatment efficacy in in-plant trials. Results support using nonpathogenic surrogate organisms to validate beef carcass intervention efficacy.     

Antimicrobial activity of cetylpyridinium chloride washes against pathogenic bacteria on beef surfaces

Cetylpyridinium chloride (CPC), a water-soluble, neutral pH, colorless compound, is widely used in oral hygiene products to inhibit bacteria responsible for plaque. Previously, researchers have demonstrated that CPC not only reduces Salmonella typhimurium on poultry but also prevents cross-contamination. To determine the effectiveness of CPC against pathogens associated with lean and adipose beef surfaces, several spray-washing experiments (862 kPa, 15 s, 35 degrees C) with 1% (wt/vol) CPC were conducted. On lean beef surfaces, CPC immediately reduced 5 to 6 log10 CFU/cm2 of Escherichia coli O157:H7 and Salmonella typhimurium to virtually undetectable levels (0 log10 CFU/cm2), as well as after 35 days of refrigerated (4 degrees C), vacuum-packaged storage. On adipose beef surfaces, 5 log10 CFU/cm2 Salmonella typhimurium and E. coli O157:H7 were reduced immediately (>2.5 log10 CFU/cm2) with 1% CPC; by day 35 the reduction was <1.3 log10 CFU/cm2. Further plate overlay analyses indicated that the effectiveness of CPC against pathogens on adipose surfaces was not hampered by the presence of meat components or fatty acids. Additional chemical and microbiological analyses of 1% CPC-treated beef surfaces subjected to a secondary water wash (following contact times of 0, 5, 10, 15, or 30 min) or grinding did reduce pathogenic bacteria and CPC levels. However, residual CPC levels following any of the treatments were considered excessive for human consumption. Despite the residual levels, this study is the first to demonstrate the effect of CPC on pathogenic bacteria associated with beef surfaces immediately after treatment and also after long-term, refrigerated, vacuum-packaged storage.     

Fate of Escherichia coli O157:H7, Salmonella typhimurium DT 104, and Listeria monocytogenes in fresh meat decontamination fluids at 4 and 10 degrees C.

Bacterial pathogens may colonize meat plants and increase food safety risks following survival, stress hardening, or proliferation in meat decontamination fluids (washings). The objective of this study was to evaluate the ability of Escherichia coli O157:H7, Salmonella Typhimurium DT 104, and Listeria monocytogenes to survive or grow in spray-washing fluids from fresh beef top rounds sprayed with water (10 or 85 degrees C) or acid solutions (2% lactic or acetic acid, 55 degrees C) during storage of the washings at 4 or 10 degrees C in air to simulate plant conditions. Inoculated Salmonella Typhimurium DT 104 (5.4 +/- 0.1 log CFU/ml) died off in lactate (pH 2.4 +/- 0.1) and acetate (pH 3.1 +/- 0.2) washings by 2 days at either storage temperature. In contrast, inoculated E. coli O157:H7 (5.2 +/- 0.1 log CFU/ml) and L. monocytogenes (5.4 +/- 0.1 log CFU/ml) survived in lactate washings for at least 2 days and in acetate washings for at least 7 and 4 days, respectively; their survival was better in acidic washings stored at 4 degrees C than at 10 degrees C. All inoculated pathogens survived in nonacid (pH > 6.0) washings, but their fate was different. E. coli O157:H7 did not grow at either temperature in water washings, whereas Salmonella Typhimurium DT 104 failed to multiply at 4 degrees C but increased by approximately 2 logs at 10 degrees C. L. monocytogenes multiplied (0.6 to 1.3 logs) at both temperatures in water washings. These results indicated that bacterial pathogens may survive for several days in acidic, and proliferate in water, washings of meat, serving as potential cross-contamination sources, if pathogen niches are established in the plant. The responses of surviving pathogens in meat decontamination waste fluids to acid or other stresses need to be addressed to better evaluate potential food safety risks.     

Evaluation of peroxyacetic acid as a potential pre-grinding treatment for control of Escherichia coli O157:H7 and Salmonella Typhimurium on beef trimmings

Peroxyacetic acid was evaluated in four separate trials for ability to reduce populations of Escherichia coli O157:H7 and Salmonella serotype Typhimurium on fresh beef trim. Trial 1 examined the effectiveness of peroxyacetic acid on individual pieces of fresh beef trim. Trial 2 evaluated the efficacy of peroxyacetic acid at low levels of contamination on batches of fresh beef trim. Trial 3 studied a washing effect of water. Lastly, Trial 4 compared the effectiveness of peroxyacetic acid to lactic acid. At various inoculation levels, peroxyacetic acid reduced populations of both pathogens by approximately 1.0log(10)CFU/cm(2) on fresh beef trim. Trial 3 showed that approximately half of the reductions found in Trials 1 and 2 were due to a washing effect of the water dip. In addition, as shown in Trial 1, increases in concentrations (>200ppm) did not significantly increase log(10) reductions of both pathogens. Following a water dip in Trial 4, peroxyacetic acid caused a reduction of 0.7log(10)CFU/cm(2) in E. coli O157:H7 and 1.0log(10)CFU/cm(2) in Salmonella Typhimurium, whereas lactic acid caused a reduction of 1.3log(10)CFU/cm(2) in E. coli O157:H7 and 2.1log(10)CFU/cm(2) in S. Typhimurium following the water dip. These results show that peroxyacetic acid was not more effective than 2% l-lactic acid in reducing pathogens on fresh beef trim.     

Decontamination of cattle hides prior to slaughter using washes with and without antimicrobial agents

Two trials were conducted to determine the efficacy of cattle wash treatments in reducing pathogens on hides of cattle before slaughter. In trial I, live cattle (n = 120) were washed in an automated, commercial cattle wash system with one of four treatments (single water wash, double water wash, water wash with 0.5% L-lactic acid, or water wash with 50 ppm chlorine). Samples were collected at three locations (brisket, belly, and inside round) pre- and posttreatment to evaluate the effectiveness of treatments on the reduction of aerobic plate counts, coliforms, Escherichia coli and the incidence of Salmonella. For all three locations, bacterial numbers increased from 0.1 to 0.8 log CFU/cm2 posttreatment. In trial II, hide samples were inoculated in the laboratory with 6.0 log CFU/cm2 of rifampicin-resistant Salmonella serotype Typhimurium. Hide wash treatments included higher concentrations of chlorine (100, 200, and 400 ppm) and L-lactic acid (2, 4, and 6%), as well as other antimicrobial agents such as ethanol (70, 80, and 90%), acetic acid (2, 4, and 6%), and Oxy-Sept 333 (0.5, 2, and 4%). Spray wash treatments with ethanol and 4 to 6% concentrations of lactic acid had greater (P < 0.05) mean log reductions than 2% solutions of acetic or lactic acid, as well as 100, 200, and 400 ppm chlorine and the control water wash treatment. Spray wash treatments with Oxy-Sept 333 and 100, 200, or 400 ppm chlorine were not effective (P > 0.05) in reducing Salmonella Typhimurium compared to the (control) distilled water spray wash treatment. Several effective cattle hide interventions were identified in a controlled laboratory setting, but the high concentrations required for effectiveness would likely present problems from an animal welfare standpoint.     

Comparison of different washing treatments for reducing pathogens on orange surfaces and for preventing the transfer of bacterial pathogens to fresh-squeezed orange juice

The objectives of this study were to compare the effectiveness of various washing treatments for reducing Escherichia coli O157:H7, Salmonella sp., and Listeria monocytogenes populations on orange surfaces and to measure the effect of some of these treatments in preventing the transfer of pathogens during juice extraction. Orange surfaces inoculated with L. monocytogenes or a mixture of E. coli O157:H7 and Salmonella Typhimurium were washed by water spray and then sprayed with or dipped in water at 80°C for 1 min, 70% ethanol for 15, 30, or 45 s or 1, 2, or 4 min, 2 or 4% lactic acid solution at 55°C for 15, 30, or 45 s or 1, 2, or 4 min, or 200 mg/liter hypochlorite at pH 6.5 or 10 for 15 s. The surviving populations of these pathogens on the oranges were enumerated after each treatment. In a further stage, the ability of these pathogens to be transferred to the juice during extraction was tested. Juice was obtained from inoculated oranges that were subjected to selected treatments using chlorine, lactic acid, ethanol, and hot water as described above, and then bacterial counts in orange juice were determined. The application of these treatments reduced the populations of pathogens on orange surfaces by 1.9 to >4.9 log, 1.9 to >4.6 log, and 1.4 to 3.1 log cycles for E. coli O157:H7, Salmonella Typhimurium, and L. monocytogenes, respectively. The treatments using hot water or lactic acid showed greater reductions than other treatments. The time, antimicrobial concentration, and form of application affected the bacterial reduction. All treatments resulted in undetectable counts in the juice. Nevertheless, pathogens were recovered by the enrichment-plating method. Treatment of oranges before juice extraction may reduce the risk associated with consuming orange juice.     

Evaluation of Lactic Acid as an Initial and Secondary Subprimal Intervention for Escherichia coli O157:H7, Non-O157 Shiga Toxin-Producing E. coli, and a Nonpathogenic E. coli Surrogate for E. coli O157:H7

Lactic acid can reduce microbial contamination on beef carcass surfaces when used as a food safety intervention, but effectiveness when applied to the surface of chilled beef subprimal sections is not well documented. Studies characterizing bacterial reduction on subprimals after lactic acid treatment would be useful for validations of hazard analysis critical control point (HACCP) systems. The objective of this study was to validate initial use of lactic acid as a subprimal intervention during beef fabrication followed by a secondary application to vacuum-packaged product that was applied at industry operating parameters. Chilled beef subprimal sections (100 cm(2)) were either left uninoculated or were inoculated with 6 log CFU/cm(2) of a 5-strain mixture of Escherichia coli O157:H7, a 12-strain mixture of non-O157 Shiga toxin-producing E. coli (STEC), or a 5-strain mixture of nonpathogenic (biotype I) E. coli that are considered surrogates for E. coli O157:H7. Uninoculated and inoculated subprimal sections received only an initial or an initial and a second "rework" application of lactic acid in a custombuilt spray cabinet at 1 of 16 application parameters. After the initial spray, total inoculum counts were reduced from 6.0 log CFU/cm(2) to 3.6, 4.4, and 4.4 log CFU/cm(2) for the E. coli surrogates, E. coli O157:H7, and non-O157 STEC inoculation groups, respectively. After the second (rework) application, total inoculum counts were 2.6, 3.2, and 3.6 log CFU/cm(2) for the E. coli surrogates, E. coli O157:H7, and non-O157 STEC inoculation groups, respectively. Both the initial and secondary lactic acid treatments effectively reduced counts of pathogenic and nonpathogenic strains of E. coli and natural microflora on beef subprimals. These data will be useful to the meat industry as part of the HACCP validation process.