Efficacy of antimicrobials for the disinfection of pathogen contaminated green bell pepper and of consumer cleaning methods for the decontamination of knives

While there is strong focus on eliminating pathogens from produce at a commercial level, consumers can employ simple methods to achieve additional pathogen reductions in the domestic kitchen. To determine the ability of antimicrobials to decontaminate peppers, samples of green bell pepper were inoculated with Salmonella enterica and Escherichia coli O157:H7 and then immersed in 3% (v/v) hydrogen peroxide (H?O?), 2.5% (v/v) acetic acid (AA), 70% (v/v) ethyl alcohol (EtOH), or sterile distilled water (SDW). The potential for transfer of pathogens from contaminated peppers to other non-contaminated produce items, and the effect of knife disinfection in preventing this cross contamination, were also tested. Knife disinfection procedures were evaluated by chopping inoculated peppers into 1 cm2 pieces with kitchen knives. Experimental knives were then treated by either no treatment (control), wiping with a dry sterile cotton towel, rinsing under running warm water for 5 or 10s, or applying a 1% (v/v) lauryl sulfate-based detergent solution followed by rinsing with warm running water for 10s. Following disinfection treatment, knives were used to slice cucumbers. Exposure to H?O? for 5 min and EtOH for 1 min resulted in reductions of 1.3±0.3 log?? CFU/cm2 for both pathogens. A 5 min exposure to AA resulted in a reduction of S. enterica of 1.0±0.7 log?? CFU/cm2 and E. coli of 0.7±0.8 log?? CFU/cm2. No differences (p ≥ 0.05) were found between numbers of pathogens on knives and numbers of pathogens transferred to cucumber slices, suggesting that organisms remaining on knife surfaces were transferred to cucumbers during slicing. Findings suggest that EtOH and H?O? may be effective antimicrobials for in-home decontamination of peppers, and that use of detergent and warm water is effective for decontamination of implements used during meal preparation.     

Decontamination of knives used in the meat industry: effect of different water temperature and treatment time combinations on the reduction of bacterial numbers on knife surfaces

Previous regulations in Australia and internationally required that knives used during the slaughter and dressing of carcasses be sanitized by brief submersion in water at 82 degrees C. Many current international regulations allow science-based equivalent alternative procedures to be used. However, limited time-temperature data are available on the response of bacteria to hot-water treatment on knives. The present study was undertaken to determine the effect of combinations of time and temperature ranging from 1 to 60 s and 60 to 82 degrees C on the disinfection of knives artificially contaminated with Escherichia coli and Listeria monocytogenes. In addition, the effect of a prerinse at 40 degrees C on the disinfection of artificially contaminated knives treated under the same controlled conditions as above was established. The experiments, which were carried out with knives in a meat matrix at each of 42 time and temperature combinations, with and without the prerinse, were performed in a laboratory water bath. Bacterial reductions were established by plate counts from the knife blade before and after immersion. Mean log reductions were subjected to statistical analysis, and basic models were generated from the results. The results demonstrated that dipping knives in water for shorter times at higher temperatures, for example, 82 degrees C for 1 s, or longer times at lower temperatures can produce equivalent inactivation of bacteria. Prerinsing knives at 40 degrees C increases the performance of the subsequent dipping step. Models produced from the data in this study can be used to predict suitable combinations of time and temperature to achieve a desired bacterial reduction.     

Transfer of Escherichia coli O157:H7 to romaine lettuce due to contact water from melting ice

Ice can be used to chill romaine lettuce and maintain relative humidity during transportation. Escherichia coli O157:H7 may contaminate water used for ice. The objective of this study was to determine the potential for E. coli O157:H7 contamination of romaine lettuce from either ice contaminated with the pathogen or by transfer from lettuce surfaces via melting ice. In experiment 1, lettuce was spot inoculated with E. coli O157:H7 and chilled with ice prepared from uncontaminated tap water. In experiment 2, water inoculated with this pathogen was frozen and used to ice lettuce. Three heads of lettuce were stacked in each container and stored at 4 or 20 degrees C. After the ice melted, E. coli O157:H7 attachment to and recovery from the lettuce leaves were determined. For experiment 1, the population of E. coli O157:H7 attached to inoculated sites averaged 3.8 and 5.5 CFU/cm2 at 4 and 20 degrees C, respectively. Most of the uninoculated sites became contaminated with the pathogen due to ice melt. For experiment 2, 3.5 to 3.8 log CFU E. coli O157:H7 per cm2 was attached to the top leaf on the first head. After rinsing with chlorinated water (200 microg/ml), E. coli O157:H7 remained on the surface of the top head (1.8 to 2.0 log CFU/cm2). There was no difference in numbers of E. coli O157:H7 recovered from each sampling site at 4 and 20 degrees C. Results show that E. coli O157:H7 can be transferred onto other produce layers in shipping containers from melted ice made of contaminated water and from contaminated to uncontaminated leaf surfaces.     

Effective cleaning and sanitizing of polysulfone ultrafiltration membrane systems.

Polysulfone UF membranes that were soiled by Cheddar cheese whey were successfully cleaned in place. This cleaning procedure was completed in about 1 h. Most cleaning chemicals used were common and inexpensive. The cleaning procedure consisted of rinsing the membrane system for 2 min with water initially and after each cleaning solution. Sodium hydroxide at pH 11.0, with .1% of a nonionic surfactant added, was circulated for 20 min. After a 2-min rinse with water, a 1:1 mixture of nitric and phosphoric acids at pH 2 was circulated for 20 min and rinsed again with water. Finally, sodium hydroxide at pH 11.0, with 200 ppm of sodium hypochlorite added, was circulated for 20 min and rinsed. All cleaning solutions and all rinse waters were at 54 degrees C. Membranes cleaned by this procedure were found to be free from whey residue under examination by scanning electron microscopy. The cleaning process did not damage the membranes even when it was used continuously for 300 h. Microbial populations on the membrane were estimated by incubating small (4-cm2) sections of membrane in screw-cap vials filled with trypticase soy broth. From the portion of vials showing growth after 72 h at 32 degrees C, a most probable microbial population was calculated. Santizing cleaned polysulfone UF membranes with 100 ppm of sodium hypochlorite or 100 ppm of dichloroisocyanurate at 54 degrees C resulted in membranes free from viable microorganisms. When dichloroisocyanurate was used at 10 degrees C and 200 ppm, a most probable microbial population of 290/m2 was found. No microbial growth was detected when cleaned and sanitized membranes were stored in tap water for 24 h. This technique for cleaning UF membranes does not require the use of a holding solution containing santizers to control the growth of residual microorganisms.     

Hot water and organic acid interventions to control microbiological contamination on hog carcasses during processing

Pork skin and muscle tissue were washed with water at temperatures from 25 to 80 degrees C. Water temperatures of 65 and 80 degrees C resulted in greater population reductions of Enterobacteriaceae on pork muscle tissue than lower water temperatures. There was no observable effect of water temperature on population reductions of Enterobacteriaceae on pork skin. Water temperatures of 55, 65, and 80 degrees C reduced the populations of Enterobacteriaceae on inoculated scalded carcasses processed in a university abattoir by 1 to 1.5 log/cm2. Following the water wash with an organic acid rinse resulted in further numerical reductions in populations, although these were not statistically different from the water wash alone. The jowls of both scalded and skinned carcasses processed in a commercial establishment were directly inoculated with a fecal material slurry and then processed with organic acid rinsing only, hot water washing only, or a combination of hot water washing followed by organic acid rinsing. The hot water and acid treatment reduced the populations of mesophilic aerobic bacteria and Escherichia coli by approximately 2 log cycles on both scalded and skinned hog carcasses. The combined treatment resulted in 60% of the scalded carcasses and 40% of the skinned carcasses with undetectable levels of E. coli after direct fecal inoculation of the carcasses. Hot water washing followed by organic acid rinsing can significantly improve the microbiological quality of pork carcasses.     

Identification of inadequately cleaned equipment used in a sheep carcass-breaking process.

Aeromonads deposited on pork during a carcass-breaking process were recovered on hydrophobic grid membrane filters placed on ampicillin-dextrin-ethanol agar. Isolates from 85 honey-yellow colonies from filters on that medium were Aeromonas hydrophila (95%) or Vibrio sp. (5%). Presumptive aeromonads, coliforms, and Escherichia coli in swab samples from product passing through a sheep carcass-breaking process were enumerated by hydrophobic grid membrane filtration techniques with a detection level of 1 CFU/100 cm2. Total aerobic counts were determined by a spread plate procedure with a detection level of 1 CFU/cm2 The numbers of aerobes, coliforms, and E. coli in the product were apparently unaffected by the carcass-breaking process, although coliforms and E. coli appeared to be redistributed from shoulder to loin and leg portions. However, the numbers of aeromonads on product increased by about two orders of magnitude as a result of the process. Few bacteria were recovered from most cleaned, large items of equipment, but aerobes, coliforms, and aeromonads were recovered at log total numbers of 5.25, 3.96, and 3.26, respectively, from most of 25 samples from bars supporting a conveyor belt. Also, aerobes, coliforms, E. coli, and aeromonads were recovered from 25 supposedly cleaned steel mesh gloves at log total numbers of 10.14, 5.54, 4.73, and 8.30, respectively. Those findings indicate that inspection of cleaned equipment and microbiological sampling of only food-contacting surfaces, as is the current practice at meat plants, cannot provide assurance that the cleaning of carcass-breaking equipment is adequate. Instead, enumeration of indicator organisms on product passing through a process seems to be required as well, with subsequent sampling of equipment to identify sources of contaminants if increases in numbers during processing are observed. For surety of adequate cleaning, enumeration of several types of indicator organism may be necessary, as increases during processing in the numbers of organisms that are present in relatively large numbers on product entering a process may be difficult to detect.     

Efficacy of ozonated and electrolyzed oxidative waters to decontaminate hides of cattle before slaughter

The hides of cattle are the primary source of pathogens such as Escherichia coli O157:H7 that contaminate preevisceration carcasses during commercial beef processing. A number of interventions that reduce hide contamination and subsequent carcass contamination are currently being developed. The objective of this study was to determine the efficacy of ozonated and electrolyzed oxidizing (EO) waters to decontaminate beef hides and to compare these treatments with similar washing in water without the active antimicrobial compounds. Cattle hides draped over barrels were used as the model system. Ozonated water (2 ppm) was applied at 4,800 kPa (700 lb in2) and 15 degrees C for 10 s. Alkaline EO water and acidic EO water were sequentially applied at 60 degrees C for 10 s at 4,800 and 1,700 kPa (250 lb in2), respectively. Treatment using ozonated water reduced hide aerobic plate counts by 2.1 log CFU/100 cm2 and reduced Enterobacteriaceae counts by 3.4 log CFU/100 cm2. EO water treatment reduced aerobic plate counts by 3.5 log CFU/100 cm2 and reduced Enterobacteriaceae counts by 4.3 log CFU/100 cm2. Water controls that matched the wash conditions of the ozonated and EO treatments reduced aerobic plate counts by only 0.5 and 1.0 log CFU/100 cm2, respectively, and each reduced Enterobacteriaceae counts by 0.9 log CFU/100 cm2. The prevalence of E. coli O157 on hides was reduced from 89 to 31% following treatment with ozonated water and from 82 to 35% following EO water treatment. Control wash treatments had no significant effect on the prevalence of E. coli O157:H7. These results demonstrate that ozonated and EO waters can be used to decontaminate hides during processing and may be viable treatments for significantly reducing pathogen loads on beef hides, thereby reducing pathogens on beef carcasses.     

Evaluation of hot-water and sanitizer dip treatments of knives contaminated with bacteria and meat residue

Hot water (HW; 82.2 degrees C, 180 degreesF) is used for sanitation of meat cutting implements in most slaughter facilities, but validation of actual practices against meat-borne bacterial pathogens and spoilage flora is lacking. Observed implement immersions in HW in two large pork processing plants were found to typically be < or = 1 s. Impact of these practices on bacteria on metal surfaces was assessed in the laboratory, and alternative treatments were investigated. Knives were inoculated with raw pork residues and Escherichia coli O157:H7, Salmonella Typhimurium DT104, Clostridium perfringens, and Lactobacillus spp. and were sampled before and after 1- or 15-s dips of blades in HW, warm water (48.9 degrees C), or warm sanitizers (neutral or acid quaternary ammonium compounds [QAC] at 400 ppm, or peroxyacetic acid at 700 ppm H2O2 and 165 ppm peroxyacetic acid). Simultaneous scrubbing and 15-s dipping in HW or acid QAC was also evaluated. Reductions on knives dipped for 1 s were usually < 1 log and were not significantly different (P > 0.05) between treatments. Reductions of E. coli O157:H7 after 15 s in HW, neutral QAC, acid QAC, or peroxyacetic acid were 3.02, 2.38, 3.04, and 1.52 log, respectively. Reductions of other bacteria due to HW were not significantly different from sanitizers and were significantly greater than warm water for all bacteria except C. perfringens. Combined scrubbing and 15-s dipping in HW resulted in a 2.91- and 2.25-log reduction of E. coli O157:H7 and Salmonella Typhimurium DT104, respectively, whereas reduction caused by acid QAC was significantly less at about 1.7 log each. Brief dip treatments of contaminated knives have limited efficacy, but longer immersions cause greater reductions that were not enhanced by scrubbing. QAC is a suitable alternative to HW in this application.     

Combined application of electrolysed water and ultrasound to improve the sanitation of knives in the meat industry

This study investigated the effect of the combination of basic electrolysed water (BEW) and slightly acid electrolysed water (SAEW) with ultrasound (US) for cleaning and sanitation of the knives used in slaughtering processes. The knives were sonicated in a US bath using two modes of operation (normal and sweep) in two steps: (i) 5 min with BEW and (ii) 10 min with SAEW at 35 °C. The microbiological counts and the possible changes in the physical structure of the knives were evaluated. The association BEW + SAEW + US, in the sweep mode, provided lower mesophilic, enterobacterial, Staphylococcus aureus and yeast counts when compared to the values recommended by the international legislation. In addition, these conditions removed all organic residues from the knife blades and promoted the highest migration rate of the residues to the US water bath (12.35 mg/L·min), without modifications in the knife blades. Thus, cleaning and sanitation of knives was feasible with the association of BEW + SAEW + US, which could be a useful alternative for the meat industry.     

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.     

Lactic acid and trisodium phosphate treatment of lamb breast to reduce bacterial contamination

Lactic acid and trisodium phosphate (TSP) were evaluated for the ability to reduce Escherichia coli and aerobic plate counts (APCs) on lamb breasts that were inoculated with a lamb fecal paste. A 90-s water rinse was applied followed by either a 9-s (55 degrees C) 2% lactic acid spray, a 60-s (55 degrees C) 12% TSP dip, or a combined treatment of both lactic acid and TSP treatments. Lactic acid reduced E. coli and APCs by 1.6 log10/cm2, and TSP caused a 1.8-log10/cm2 reduction in E. coli and a 0.7-log10/cm2 reduction in APCs. Combined reductions by the lactic acid spray followed by the TSP dip were 1.8 and 1.5 log10/cm2 for E. coli and APCs, respectively. Lactic acid and trisodium phosphate, used alone or in combination, were effective in reducing numbers of E. coli and could be useful as pathogen intervention steps in lamb slaughter processing.     

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.     

Sanitation and design of lettuce coring knives for minimizing Escherichia coli O157:H7 contamination

This study was undertaken to examine the effect of ultrasound in combination with chlorine on the reduction of Escherichia coli O157:H7 populations on lettuce coring knives. Two new coring devices designed to mitigate pathogen attachment were fabricated and evaluated. The coring rings of the knives were dip inoculated with soil slurry containing 10? E. coli cells and treated with chlorinated water with and without ultrasonication for 30, 60, and 120 s. The rough welding joints on currently used in-field lettuce coring knives provided a site conducive to bacterial attachment and resistant to cell removal during sanitation treatment. The two modified coring knives harbored significantly fewer E. coli cells than did the currently used commercial model, and the efficacy of the disinfection treatment was high (P < 0.05). Ultrasound treatment reduced the E. coli O157:H7 counts to below the detection limit of 1.10 log CFU/cm2 at both the coring ring blade and welding joint within 30 s in 1 ppm of chlorinated water. The redesigned coring knives and an ultrasound plus chlorine combination treatment may provide practical options for minimizing the microbial safety hazards of lettuce processed by core-in-field operations.     

Survival of Escherichia coli O157:H7 and Salmonella in apple cider and orange juice as affected by ozone and treatment temperature

Inactivation of Escherichia coli O157:H7 and Salmonella in apple cider and orange juice treated with ozone was evaluated. A five-strain mixture of E. coli O157:H7 or a five-serovar mixture of Salmonella was inoculated (7 log CFU/ml) into apple cider and orange juice. Ozone (0.9 g/h) was pumped into juices maintained at 4 degrees C, ambient temperature (approximately 20 degrees C), and 50 degrees C for up to 240 min, depending on organism, juice, and treatment temperature. Samples were withdrawn, diluted in 0.1% peptone water, and surface plated onto recovery media. Recovery of E. coli O157:H7 was compared on tryptic soy agar (TSA), sorbitol MacConkey agar, hemorrhagic coli agar, and modified eosin methylene blue agar; recovery of Salmonella was compared on TSA, bismuth sulfite agar, and xylose lysine tergitol 4 (XLT4) agar. After treatment at 50 degrees C, E. coli O157:H7 populations were undetectable (limit of 1.0 log CFU/ml; a minimum 6.0-log CFU/ml reduction) after 45 min in apple cider and 75 min in orange juice. At 50 degrees C, Salmonella was reduced by 4.8 log CFU/ml (apple cider) and was undetectable in orange juice after 15 min. E. coli O157:H7 at 4 degrees C was reduced by 4.8 log CFU/ml in apple cider and by 5.4 log CFU/ml in orange juice. Salmonella was reduced by 4.5 log CFU/ml (apple cider) and 4.2 log CFU/ml (orange juice) at 4 degrees C. Treatment at ambient temperature resulted in population reductions of less than 5.0 log CFU/ml. Recovery of E. coli O157:H7 and Salmonella on selective media was substantially lower than recovery on TSA, indicating development of sublethal injury. Ozone treatment of apple cider and orange juice at 4 degrees C or in combination with mild heating (50 degrees C) may provide an alternative to thermal pasteurization for reduction of E. coli O157:H7 and Salmonella in apple cider and orange juice.     

Recovery of Escherichia coli Biotype I and Enterococcus spp. during refrigerated storage of beef carcasses inoculated with a fecal slurry.

Three beef front quarters/carcasses were inoculated with a slurry of cattle manure. During storage at 4 degrees C, two sponge samples from each of three sites (i.e., 100 cm2 from each of two fat surfaces and 100 cm2 from a lean surface) were taken from each of the three carcasses on days 0, 1, 3, 7, and 10 after inoculation. The initial numbers of Escherichia coli averaged 2.0 log10 CFU/cm2 (1.21 to 2.47 log10 CFU/cm2) using the Petrifilm method and 2.09 log10 most probable number (MPN)/cm2 (0.88 to 2.96 log10 MPN/cm2) using the MPN method. The initial numbers of enterococci averaged 3.34 log10 CFU/cm2 (3.07 to 3.79 log10 CFU/cm2) using kanamycin esculin azide agar. In general, an appreciable reduction in the numbers of E. coli occurred during the first 24 h of storage; for the Petrifilm method an average reduction of 1.37 log10 CFU/cm2 (0.69 to 1.71 log10 CFU/cm2) was observed, and for the MPN method an average reduction of 1.52 log10 MPN/cm2 (0.47 to 2.08 log10 MPN/cm2) was observed. E. coli were not detected (<-0.12 log10 CFU/cm2) using Petrifilm on day 7 of the storage period on two (initial counts of 1.21 and 2.29 log10 CFU/cm2) of the three carcasses. However, viable E. coli cells were recovered from these two carcasses after a 24-h enrichment at 37 degrees C in EC broth. Viable E. coli cells were detected at levels of -0.10 log10 CFU/cm2 on the third carcass (initial count of 2.47 log10 CFU/cm2) after 7 days at 4 degrees C. No significant difference in recovery of viable cells was observed between the MPN and Petrifilm methods on days 0, 1, and 3 (P > 0.05). However, viable E. coli cells were recovered from all three carcasses by the MPN method on day 7 at an average of -0.29 log10 MPN/ cm2 (-0.6 to -0.1 log10 MPN/cm2). On day 10, viable cells were recovered by the MPN method from two of the three carcasses at -0.63 and -0.48 log10 MPN/cm2 but were not recovered from the remaining carcass (<-0.8 log10 MPN/cm2). Similar to E. coli, the greatest reduction (average of 1.26 log10 CFU/cm2, range = 1.06 to 1.45 log10 CFU/cm2) in the numbers of enterococci occurred during the first 24 h of storage. Because of higher initial numbers and a slightly slower rate of decrease, the numbers of Enterococcus spp. were significantly higher (P < 0.017) than the numbers of E. coli Biotype I after 3, 7, and 10 days of storage. These results suggest that enterococci may be useful as an indicator of fecal contamination of beef carcasses.     

Inactivation of Escherichia coli O157:H7 and Listeria monocytogenes on plastic kitchen cutting boards by electrolyzed oxidizing water.

One milliliter of culture containing a five-strain mixture of Escherichia coli O157:H7 (approximately 10(10) CFU) was inoculated on a 100-cm2 area marked on unscarred cutting boards. Following inoculation, the boards were air-dried under a laminar flow hood for 1 h, immersed in 2 liters of electrolyzed oxidizing water or sterile deionized water at 23 degrees C or 35 degrees C for 10 or 20 min; 45 degrees C for 5 or 10 min; or 55 degrees C for 5 min. After each temperature-time combination, the surviving population of the pathogen on cutting boards and in soaking water was determined. Soaking of inoculated cutting boards in electrolyzed oxidizing water reduced E. coli O157:H7 populations by > or = 5.0 log CFU/100 cm2 on cutting boards. However, immersion of cutting boards in deionized water decreased the pathogen count only by 1.0 to 1.5 log CFU/100 cm2. Treatment of cutting boards inoculated with Listeria monocytogenes in electrolyzed oxidizing water at selected temperature-time combinations (23 degrees C for 20 min, 35 degrees C for 10 min, and 45 degrees C for 10 min) substantially reduced the populations of L. monocytogenes in comparison to the counts recovered from the boards immersed in deionized water. E. coli O157:H7 and L. monocytogenes were not detected in electrolyzed oxidizing water after soaking treatment, whereas the pathogens survived in the deionized water used for soaking the cutting boards. This study revealed that immersion of kitchen cutting boards in electrolyzed oxidizing water could be used as an effective method for inactivating foodborne pathogens on smooth, plastic cutting boards.     

Application of reuterin produced by Lactobacillus reuteri 12002 for meat decontamination and preservation

Lactobacillus reuteri strain 12002 was used for reuterin production in the two-step fermentation process. A batch culture fermentation was used to produce a maximum biomass of L. reuteri. Then cells were harvested, resuspended in a glycerol-water solution, and anaerobically incubated to produce reuterin. The lyophilized supernatants (approximately 4000 activity units (AU) of reuterin per ml) were diluted in distilled water for decontamination and preservation trials. The MIC values of reuterin for Escherichia coli O157:H7 and Listeria monocytogenes were 4 and 8 AU/ml, respectively. In meat decontamination experiments, the surface of cooked pork was inoculated with either L. monocytogenes or E. coli O157:H7 at a level of approximately log10 5 CFU/cm2, incubated for 30 min at 7 degrees C, and decontaminated by exposure to reuterin (500 AU/ml). The bactericidal effect of reuterin was analyzed 15 s and 24 h after exposure at 7 degrees C. After 15 s of exposure to reuterin, viable numbers decreased by 0.45 and 0.3 log10 CFU/cm2 for E. coli O157:H7 and L. monocytogenes, respectively. After 24 h the numbers decreased by 2.7 log10 CFU/cm2 for E. coli O157:H7 and by 0.63 log10 CFU/cm2 for L. monocytogenes. In the same experiment, the combined effect of reuterin and lactic acid was also investigated. Adding lactic acid (5%, vol/vol) to reuterin significantly enhanced (P < or = 0.05) the efficacy of reuterin. No additional effect (P < or = 0.05) was found when ethanol (40%) was added to the mixture of reuterin and lactic acid. To evaluate the preservative effect of reuterin during meat storage, reuterin was added to raw ground pork contaminated with E. coli O157:H7 or L. monocytogenes. Reuterin at a concentration of 100 AU/g resulted in a 5.0-log10 reduction of the viability of E. coli O157:H7 after 1 day of storage at 7 degrees C. Reuterin at a concentration of 250 AU/g reduced the number of the viable cells of L. monocytogenes by log10 3.0 cycles after 1 week of storage at 7 degrees C.     

Decontamination of beef subprimal cuts intended for blade tenderization or moisture enhancement

The prevalence of Escherichia coli O157:H7 on beef subprimal cuts intended for mechanical tenderization was evaluated. This evaluation was followed by the assessment of five antimicrobial interventions at minimizing the risk of transferring E. coli O157:H7 to the interior of inoculated subprimal cuts during blade tenderization (BT) or moisture enhancement (ME). Prevalence of E. coli O157:H7 on 1,014 uninoculated beef subprimals collected from six packing facilities was 0.2%. Outside round pieces inoculated with E. coli O157:H7 at 10(4) CFU/100 cm2 were treated with (i) no intervention, (ii) surface trimming, (iii) hot water (82 degrees C), (iv) warm 2.5% lactic acid (55 degrees C), (v) warm 5.0% lactic acid (55 degrees C), or (vi) 2% activated lactoferrin followed by warm 5.0% lactic acid (55 degrees C) and then submitted to BT or ME. Prevalence (n=196) of internalized (BT and ME) E. coli O157:H7 was 99%. Enumeration of E. coli 0157:H7 (n=192) revealed mean surface reductions of 0.93 to 1.10 log CFU/100 cm2 for all antimicrobial interventions. E. coli O157:H7 was detected on 3 of the 76 internal BT samples and 73 of the 76 internal ME samples. Internal ME samples with no intervention had significantly higher mean E. coli O157:H7 populations than did those internal samples treated with an intervention, but there were no significant differences in E. coli O157:H7 populations among internal BT samples. Results of this study demonstrate that the incidence of E. coli O157:H7 on the surface of beef subprimal cuts is low and that interventions applied before mechanical tenderization can effectively reduce the transfer of low concentrations of E. coli O157:H7 to the interior of beef subprimal cuts.     

Studies to evaluate chemicals and conditions with low-pressure applications for reducing microbial counts on cattle hides

Studies were conducted to identify effective antimicrobials and application parameters that could be used as decontamination interventions to reduce microbial loads on cattle hides before removal from carcasses. In study I, hide swatches inoculated with Escherichia coli O157:H7 were sprayed with 10% acetic acid (at 23 and 55 degrees C), 10% lactic acid (at 23 and 55 degrees C), 3% sodium hydroxide (at 23 degrees C) or 4 and 5% sodium metasilicate (at 23 degrees C). All antimicrobials were evaluated independently after being applied alone, being applied after a water rinse, or being followed by a water rinse. Antimicrobial treatments followed by a water rinse lowered E. coli O157:H7 populations by 0.6 to 2.4 log CFU/cm2 and resulted in hides with a surface pH of 6.3 to 9.2. Treatments in which a water rinse was followed by antimicrobial application lowered E. coli O157:H7 populations by 1.5 to 5.1 log CFU/cm2 but resulted in hides with a surface pH of 3.9 to 10.5. In study II, whole hides were treated with one of four antimicrobials (acetic acid, lactic acid, sodium hydroxide, or sodium metasilicate) followed by a water rinse. Hides were evaluated for aerobic bacterial counts, total coliform counts, and E. coli counts. Generally, all antimicrobials resulted in greater reductions (P < 0.05) of E. coli counts when compared with the control; however, only acetic and lactic acids resulted in greater reductions (P < 0.05) of aerobic bacterial counts and total coliform counts compared with the controls. These antimicrobials could be used to reduce microbial contamination on hides, potentially reducing microbiological contamination transferred to carcasses or to the plant environment.     

Decontamination of beef carcass surface tissue by steam vacuuming alone and combined with hot water and lactic acid sprays

Hot beef carcass surface regions (outside round, brisket, and clod) contaminated with feces spread over a 5-cm2 (1-in2) area were cleaned using a steam-vacuum spot-cleaning system alone or combined with subsequent sanitizing treatments of hot water (95 degrees C at the nozzle), or warm (55 degrees C) 2% lactic acid spray, or combinations of these two sanitizing methods. These treatments were compared for effectiveness in reducing aerobic plate counts (APC) and counts of Enterobacteriaceae, total coliforms, thermotolerant coliforms, and Escherichia coli. All treatments significantly reduced the numbers of each group of bacteria on beef carcass surfaces. However, reductions obtained by steam vacuuming were significantly smaller than those obtained by a combination of steam vacuuming with any sanitizing treatment. No differences in bacterial reductions were observed between different carcass surface regions. Steam vacuuming reduced the number of different indicator organisms tested by ca. 3.0 log cycles but also spread the bacterial contamination to areas of the carcass surface adjacent to the contaminated sites. This relocated contamination after steam vacuuming was most effectively reduced by spraying with hot water and then lactic acid. This combined treatment consistently reduced the numbers of Enterobacteriaceae, total and thermotolerant coliforms, and E. coli to undetectable levels (<1.0 log10 CFU/cm2) on areas outside the initial 5-cm2 inoculated areas.