Industrial application of an antilisterial strain of Lactobacillus sakei as a protective culture and its effect on the sensory acceptability of cooked, sliced, vacuum-packaged meats.

The application of a protective lactic acid bacterium (LAB) during the commercial production of cooked meat products is described. The LAB, a strain of Lactobacillus sakei, was previously isolated from cooked ham and inhibited growth of Listeria monocytogenes and Escherichia coli O157:H7 in this product. L. sakei was applied to the cooked products at a concentration of 10(5)-10(6) cfu/g immediately before slicing and vacuum-packaging using a hand-operated spraying bottle. The LAB strain inhibited growth of 10(3) cfu/g of a cocktail of three rifampicin resistant mutant L. monocytogenes strains both at 8 degrees C and 4 degrees C. Consumer acceptance tests of cooked ham and of servelat sausage, a Norwegian non-fermented cooked meat sausage, showed that control and inoculated products were equally acceptable. The products were still acceptable after storage for 28 days at 4 degrees C and, after opening the packages, for a further 5 days at 4 degrees C. The findings presented here confirm that the L. sakei strain is suitable for use as a protective culture and may technically easily be implemented in the commercial production of cooked meat products.     

Pathogen testing of ready-to-eat meat and poultry products collected at federally inspected establishments in the United States, 1990 to 1999

The Food Safety and Inspection Service (FSIS) conducted microbiological testing programs for ready-to-eat (RTE) meat and poultry products produced at approximately 1,800 federally inspected establishments. All samples were collected at production facilities and not at retail. We report results here for the years 1990 through 1999. Prevalence data for Salmonella, Listeria monocytogenes, Escherichia coli O157:H7, or staphylococcal enterotoxins in nine different categories of RTE meat and poultry products are presented and discussed. The prevalence data have certain limitations that restrict statistical inferences, because these RTE product-testing programs are strictly regulatory in nature and not statistically designed. The cumulative 10-year Salmonella prevalences were as follows: jerky, 0.31%; cooked, uncured poultry products, 0.10%; large-diameter cooked sausages, 0.07%; small-diameter cooked sausages, 0.20%; cooked beef, roast beef, and cooked corned beef, 0.22%; salads, spreads, and patés, 0.05%; and sliced ham and luncheon meat, 0.22%. The cumulative 3-year Salmonella prevalence for dry and semidry fermented sausages was 1.43%. The cumulative 10-year L. monocytogenes prevalences were as follows: jerky, 0.52%; cooked, uncured poultry products, 2.12%; large-diameter cooked sausages, 1.31%; small-diameter cooked sausages, 3.56%; cooked beef, roast beef, and cooked corned beef, 3.09%; salads, spreads, and patés, 3.03%; and sliced ham and luncheon meat, 5.16%. The cumulative 3-year L. monocytogenes prevalence for dry and semidry fermented sausages was 3.25%. None of the RTE products tested for E. coli O157:H7 or staphylococcal enterotoxins was positive. Although FSIS and the industry have made progress in reducing pathogens in these products, additional efforts are ongoing to continually improve the safety of all RTE meat and poultry products manufactured in federally inspected establishments in the United States.     

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.     

Effect of acid resistance of Escherichia coli O157:H7 on efficacy of buffered lactic acid to decontaminate chilled beef tissue and effect of modified atmosphere packaging on survival of Escherichia coli O157:H7 on red meat.

The present study examined the effect of pH-independent acid resistance of Escherichia coli O157:H7 on efficacy of buffered lactic acid to decontaminate chilled beef tissue. A varied level of acid resistance was observed among the 14 strains tested. Eight strains were categorized as acid resistant, four strains as acid sensitive, and two strains demonstrated acid-inducible acid resistance. The survival of an acid-resistant (II/45/4) and acid-sensitive (IX/8/16) E. coli O157:H7 strain on chilled beef tissue treated with 1 and 2% buffered lactic acid, sterile water, or no treatment (control) was followed. A gradual reduction of E. coli O157:H7 was noticed during the 10 days of storage at 4 degrees C for each of the treatments. Decontamination with 1 and 2% buffered lactic acid did not appreciably affect the pathogen. Differences in the pH-independent acid resistance of the strains had no effect on the efficacy of decontamination. The effect of modified atmosphere packaging (MAP) on survival of E. coli O157:H7 in red meat was also studied. MAP (40% CO2/60% N2) or vacuum did not significantly influence survival of E. coli O157:H7 on inoculated sliced beef (retail cuts) meat compared to packing in air. The relative small outgrowth of lactic acid bacteria during storage under vacuum for 28 days did not affect survival of E. coli O157:H7. Neither lactic acid decontamination nor vacuum or MAP packaging could enhance reduction of E. coli O157:H7 on beef, thus underlining the need for preventive measures to control the public health risk of E. coli O157:H7.     

Evaluation of meat born lactic acid bacteria as protective cultures for the biopreservation of cooked meat products

In this study, 91 strains, originating from meat products, were subjected to a step-by-step screening and characterisation to search for potential protective cultures to be used in the cooked cured meat industry. Strains were first tested on their homofermentative and psychrotrophic character and salt tolerance. Secondly, the antibacterial capacities towards Listeria monocytogenes, Leuconostoc mesenteroides, Leuconostoc carnosum and Brochotrix thermosphacta were determined in an agar spot test. In total, 38% of the tested strains were inhibitory towards all indicator strains. However, 91%, 88% and 74% of the strains could inhibit, respectively, L. monocytogenes, B. thermosphacta and Leuc. mesenteroides. Finally, 12 strains, with the highest antibacterial capacities, were evaluated on their competitive nature by comparing their growth rate, acidifying character and lactic acid production at 7 degrees C under anaerobic conditions in a liquid broth. All 12 strains, except for a bacteriocin producing Lactobacillus plantarum strain and the lactocin S producing Lactobacillus sakei 148, combined a fast growth rate with a deep and rapid acidification caused by the production of high levels of lactic acid. The 12 selected strains were then further investigated for their growth capacity on a model cooked ham product to establish whether the presence of these cultures on the ham did not negatively influence the sensory properties of the ham. All strains grew in 6 days at 7 degrees C from a level of 10(5)-10(6) to 10(7)-10(8) cfu/g and again the bacteriocin producing L. plantarum strain was the slowest growing strain. As the glucose level of the model cooked ham product was low (0.09+/-0.03%), growth of the putative protective cultures resulted in glucose depletion and a limited lactic acid production and accompanying pH decrease. Cooked ham inoculated with isolates 13E, 10A, 14A (all three identified as L. sakei subsp. carnosus by SDS-PAGE) and with strains L. sakei 148 (LS5) and L. sakei subsp. carnosus SAGA 777 (LS8) were not rejected by the sensory panel at the 34th day of the vacuum packaged storage at 7 degrees C. Therefore, these strains could have potential for the use as protective culture in cooked meat products.     

Determination of 5-log pathogen reduction times for heat-processed, acidified vegetable brines

Recent outbreaks of acid-resistant food pathogens in acid foods, including apple cider and orange juice, have raised concerns about the safety of acidified vegetable products. We determined pasteurization times and temperatures needed to assure a 5-log reduction in the numbers of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella strains in acidified cucumber pickle brines. Cocktails of five strains of each pathogen were (separately) used for heat-inactivation studies between 50 and 60 degrees C in brines that had an equilibrated pH value of 4.1. Salmonella strains were found to be less heat resistant than E. coli O157:H7 or L. monocytogenes strains. The nonlinear killing curves generated during these studies were modeled using a Weibull function. We found no significant difference in the heat-killing data for E. coli O157:H7 and L. monocytogenes (P = 0.9709). The predicted 5-log reduction times for E. coli O157:H7 and L. monocytogenes were found to fit an exponential decay function. These data were used to estimate minimum pasteurization times and temperatures needed to ensure safe processing of acidified pickle products and show that current industry pasteurization practices offer a significant margin of safety.     

Surface application of lysozyme, nisin, and EDTA to inhibit spoilage and pathogenic bacteria on ham and bologna

A study was conducted to determine if the effectiveness of an antimicrobial treatment for cooked ham and bologna would be increased or maintained when applied in a surface coating. Cooked 10-g disks of ham and bologna sausage received one of three treatments: no coating (control), coating with 0.2 g of 7% (wt/vol) gelatin gel (gel-control), or coating with 0.2 g of 7% gelatin gel containing 25.5 g/liter of lysozyme-nisin (1:3) plus 25.5 g/liter of EDTA (gel-treated). The samples were then inoculated with one of six test organisms: Brochothrix thermosphacta, Escherichia coli O157:H7, Lactobacillus sakei, Leuconostoc mesenteroides, Listeria monocytogenes, or Salmonella Typhimurium. Inoculated samples were vacuum packed and stored at 8 degrees C for 4 weeks. The antimicrobial gel treatment had an immediate bactericidal effect up to 4 log CFU/cm2 on the four gram-positive organisms tested (B. thermosphacta, Lactobacillus sakei, Leuconostoc mesenteroides, and Listeria monocytogenes) and inhibited the growth of these organisms during the 4 weeks of storage. The antimicrobial gel treatment also had a bactericidal effect on the growth of Salmonella Typhimurium during storage. The numbers of E. coli O157:H7 on ham were reduced by 2 log CFU/cm2 following treatment with both antimicrobial-containing and non-antimicrobial-containing gels during the 4-week storage period. No effect was observed on the growth of E. coli O157:H7 on bologna.     

Bactericidal effects of Lactobacillus reuteri and allyl isothiocyanate on Escherichia coli O157:H7 in refrigerated ground beef

Two naturally occurring antimicrobial agents were tested in packages of refrigerated ground beef for their ability to reduce the viability of Escherichia coli O157:H7 during storage. Allyl isothiocyanate (AITC) and Lactobacillus reuteri were tested separately and together for their action against a cocktail of five strains of E. coli O157:H7 in ground beef held at 4 degrees C for 25 days. Ground beef prepared from whole, raw inside round beef roasts was inoculated with low (3 log CFU/g) or high (6 log CFU/g) levels of the E. coli O157:H7 mixture. The beef was treated with AITC (about 1,300 ppm), L. reuteri, or both, along with 250 mM of glycerol per kg of meat at two levels (3 and 6 log CFU/g) and according to a design that yielded 8 controls plus 10 different treatments. Samples were analyzed for E. coli O157:H7 survivors, numbers of total bacteria, and lactic acid bacteria on days 0 to 25 at 5-day intervals. L. reuteri at both input levels with glycerol killed E. coli O157:H7 at both inoculated levels before day 20. AITC completely eliminated E. coli O157:H7 at the low-inoculum level (3 log CFU/g) and reduced viability >4.5 log CFU/g at the high-inoculum level (6 log CFU/g) by the end of the storage period. The combination of L. reuteri and AITC did not yield an additive effect against E. coli O157:H7 viability. L. reuteri in the presence of glycerol was highly effective against E. coli O157:H7 in ground beef during refrigerated storage (4 degrees C) in modified atmosphere packages. Sensory testing is planned to evaluate effects of treatments.     

Inactivation of Escherichia coli O157:H7, Salmonella enterica serotype enteritidis,and Listeria monocytogenes on lettuce by hydrogen peroxide and lactic acid and by hydrogen peroxide with mild heat

Iceberg lettuce is a major component in vegetable salad and has been associated with many outbreaks of foodborne illnesses. In this study, several combinations of lactic acid and hydrogen peroxide were tested to obtain effective antibacterial activity without adverse effects on sensory characteristics. A five-strain mixture of Escherichia coli O157:H7, Salmonella enterica serotype Enteritidis, and Listeria monocytogenes was inoculated separately onto fresh-cut lettuce leaves, which were later treated with 1.5% lactic acid plus 1.5% hydrogen peroxide (H2O2) at 40 degrees C for 15 min, 1.5% lactic acid plus 2% H2O2 at 22 degrees C for 5 min, and 2% H2O2 at 50 degrees C for 60 or 90 s. Control lettuce leaves were treated with deionized water under the same conditions. A 4-log reduction was obtained for lettuce treated with the combinations of lactic acid and H2O2 for E. coli O157:H7 and Salmonella Enteritidis, and a 3-log reduction was obtained for L. monocytogenes. However, the sensory characteristics of lettuce were compromised by these treatments. The treatment of lettuce leaves with 2% H2O2 at 50 degrees C was effective not only in reducing pathogenic bacteria but also in maintaining good sensory quality for up to 15 days. A < or = 4-log reduction of E. coli O157:H7 and Salmonella Enteritidis was achieved with the 2% H2O2 treatment, whereas a 3-log reduction of L. monocytogenes was obtained. There was no significant difference (P > 0.05) between pathogen population reductions obtained with 2% H2O2 with 60- and 90-s exposure times. Hydrogen peroxide residue was undetectable (the minimum level of sensitivity was 2 ppm) on lettuce surfaces after the treated lettuce was rinsed with cold water and centrifuged with a salad spinner. Hence, the treatment of lettuce with 2% H2O2 at 50 degrees C for 60 s is effective in initially reducing substantial populations of foodborne pathogens and maintaining high product quality.     

Competitive inhibition of Listeria monocytogenes in ready-to-eat meat products by lactic acid bacteria

Forty-nine strains of lactic acid bacteria (LAB), isolated from commercially available ready-to-eat (RTE) meat products, were screened for their ability to inhibit the growth of Listeria monocytogenes at refrigeration (5 degrees C) temperatures on agar spot tests. The three most inhibitory strains were identified as Pediococcus acidilactici, Lactobacillus casei, and Lactobacillus paracasei by 16S rDNA sequence analysis. Their antilisterial activity was quantified in associative cultures in deMan Rogosa Sharpe (MRS) broth at 5 degrees C for 28 days, resulting in a pathogen reduction of 3.5 log10 cycles compared to its initial level. A combined culture of these strains was added to frankfurters and cooked ham coinoculated with L. monocytogenes, vacuum packaged, and stored at 5 degrees C for 28 days. Bacteriostatic activity was observed in cooked ham, whereas bactericidal activity was observed in frankfurters. Numbers of L. monocytogenes were 4.2 to 4.7 log10 and 2.6 log10 cycles lower than controls in frankfurters and cooked ham, respectively, after the 28-day refrigerated storage. In all cases, numbers of LAB increased by only 1 log10 cycle. The strain identified as P. acidilactici was possibly a bacteriocin producer, whereas the antilisterial activity of the other two strains was due to the production of organic acids. There was no significant difference (P > 0.05) in the antilisterial activity detected in frankfurters whether the LAB strains were used individually or as combined cultures. Further studies over a 56-day period indicated no impact on the quality of the product. This method represents a potential antilisterial intervention in RTE meats, because it inhibited the growth of the pathogen at refrigeration temperatures without causing sensory changes.     

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 a challenge testing protocol to assess the stability of ready-to-eat cooked meat products against growth of Listeria monocytogenes

Challenge testing of ready-to-eat (RTE) foods with Listeria monocytogenes is recommended to assess the potential for growth. The present study was undertaken to evaluate a protocol for challenge testing applied to RTE cooked meat products. In order to choose L. monocytogenes strains with a representative behaviour, initially, the variability of the response of multiple L. monocytogenes strains of human and food origin to different stress and growth conditions was established. The strains were not inhibited in their growth at moderate acid pH (5.25) and the four strains tested in particular showed a similar acid-adaptive response. Growth of the various strains under four different combined stress conditions indicated that no L. monocytogenes strain had consistently significant longer or shorter lag phase or higher or lower maximum specific growth rates. The effect of choice of strain and history (pre-incubation temperature 7 or 30 degrees C) on growth of L. monocytogenes under optimum conditions (Brain Heart Infusion, BHI) and modified BHI simulating conditions of cooked ham and paté was studied. In general, all four L. monocytogenes strains behaved similarly. In BHI, no difference in lag phase was observed for the cold-adapted and standard inoculum, whereas in BHI adjusted to ham and paté conditions, a ca. 40-h reduction of the lag phase was noted for the cold-adapted inoculum. Subsequently, microbial challenge testing of L. monocytogenes in modified atmosphere packaged sliced cooked ham and paté was performed. A mixed inoculum of four L. monocytogenes strains and an inoculum level of ca. 1-10 cfu/g was used. On vacuum packed sliced cooked ham, the concentration of 100 cfu/g, the safety limit considered as low risk for causing listeriosis, was exceeded after 5 days whereas ca. 10(5) cfu/g were obtained after 14 days when also LAB spoilers reached unacceptable numbers (ca. 10(7) cfu/g) whether standard or cold-adapted inoculum was used. The concentration of sodium lactate determined the opportunities for growth of L. monocytogenes in paté. If growth of L. monocytogenes in paté was noticed, the threshold of 100 cfu/ml was crossed earlier for the cold-adapted inoculum compared to the standard inoculum.     

Inactivation of Escherichia coli O157:H7, Salmonella typhimurium DT104, and Listeria monocytogenes on inoculated alfalfa seeds with a fatty acid-based sanitizer

Alfalfa seeds were inoculated with a three-strain cocktail of Escherichia coli O157:H7, Salmonella enterica subsp. enterica serovar Typhimurium DT104, or Listeria monocytogenes by immersion to contain approximately 6 to 8 log CFU/g and then treated with a fatty acid-based sanitizer containing 250 ppm of peroxyacid, 1,000 ppm of caprylic and capric acids (Emery 658), 1,000 ppm of lactic acid, and 500 ppm of glycerol monolaurate at a reference concentration of 1X. Inoculated seeds were immersed at sanitizer concentrations of 5X, 10X, and 15X for 1, 3, 5, and 10 min and then assessed for pathogen survivors by direct plating. The lowest concentration that decreased all three pathogens by >5 log was 15. After a 3-min exposure to the 15X concentration, populations of E. coli O157:H7, Salmonella Typhimurium DT104, and L. monocytogenes decreased by >5.45, >5.62, and >6.92 log, respectively, with no sublethal injury and no significant loss in seed germination rate or final sprout yield. The components of this 15x concentration (treatment A) were assessed independently and in various combinations to optimize antimicrobial activity. With inoculated seeds, treatment C (15,000 ppm of Emery 658, 15,000 ppm of lactic acid, and 7,500 ppm of glycerol monolaurate) decreased Salmonella Typhimurium, E. coli O157:H7, and L. monocytogenes by 6.23 and 5.57 log, 4.77 and 6.29 log, and 3.86 and 4.21 log after 3 and 5 min of exposure, respectively. Treatment D (15,000 ppm of Emery 658 and 15,000 ppm of lactic acid) reduced Salmonella Typhimurium by >6.90 log regardless of exposure time and E. coli )157:H7 and L. monocytogenes by 4.60 and >5.18 log and 3.55 and 3.14 log after 3 and 5 min, respectively. No significant differences (P > 0.05) were found between treatments A, C, and D. Overall, treatment D, which contained Emery 658 and lactic acid as active ingredients, reduced E. coli O157:H7, Salmonella Typhimurium, and L. monocytogenes populations by 3.55 to >6.90 log and may provide a viable alternative to the recommended 20,000 ppm of chlorine for sanitizing alfalfa seeds.     

Multiplex PCR for simultaneous detection of Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7 in meat samples

A multiplex PCR method was developed for simultaneous detection of Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7 in meat samples. DNA detection sensitivity for this method was 10(3) CFU/ml for each pathogen. When this protocol was used for the detection of each of the above pathogenic bacteria in spiked pork samples, 1 cell per 25 g of inoculated sample could be detected within 30 h. In the samples of naturally contaminated meat, Salmonella spp., L. monocytogenes, and E. coli O157:H7 were detected over the same time period. Excellent agreement was obtained for the results of multiplex PCR and the conventional culture method, which suggests that the multiplex PCR is a reliable and useful method for rapid screening of meat products for Salmonella spp., L. monocytogenes, and E. coli O157:H7 contamination.     

Validation of lactic acid bacteria, lactic acid, and acidified sodium chlorite as decontaminating interventions to control Escherichia coli O157:H7 and Salmonella Typhimurium DT 104 in mechanically tenderized and brine-enhanced (nonintact) beef at the purveyor

After three different outbreaks were linked to the consumption of nonintact meat products contaminated with Escherichia coli O157:H7, the U.S. Food Safety and Inspection Service published notice requiring establishments producing mechanically tenderized and moisture-enhanced beef products to reassess their respective hazard analysis and critical control point systems, due to potential risk to the consumers. The objective of this study was to validate the use of lactic acid bacteria (LAB), acidified sodium chlorite (ASC), and lactic acid (LA) sprays when applied under a simulated purveyor setting as effective interventions to control and reduce E. coli O157:H7 and Salmonella Typhimurium DT 104 in inoculated U.S. Department of Agriculture (USDA) Choice strip loins (longissimus lumborum muscles) pieces intended for either mechanical blade tenderization or injection enhancement with a brine solution after an aging period of 14 or 21 days at 4.4°C under vacuum. After the mechanical process, translocation of E. coli O157:H7 and Salmonella Typhimurium DT 104 from the surface into the internal muscles occurred at levels between 1.00 and 5.72 log CFU/g, compared with controls. LAB and LA reduced internal E. coli O157:H7 loads up to 3.0 log, while ASC reduced the pathogen 1.4 to 2.3 log more than the control (P < 0.05), respectively. Salmonella Typhimurium DT 104 was also reduced internally 1.3 to 2.8, 1.0 to 2.3, and 1.4 to 1.8 log after application of LAB, LA, and ASC, respectively. The application of antimicrobials by purveyors prior to mechanical tenderization or enhancement of steaks should increase the safety of these types of products.     

Thermal process validation for Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes in ground turkey and beef products

At 55 to 70 degrees C, thermal inactivation D-values for Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes were 19.05 to 0.038, 43.10 to 0.096, and 33.11 to 0.12 min, respectively, in ground turkey and 21.55 to 0.055, 37.04 to 0.066, and 36.90 to 0.063 min, respectively, in ground beef. The z-values were 5.73, 5.54, and 6.13 degrees C, respectively, in ground turkey and 5.43, 5.74, and 6.01 degrees C, respectively, in ground beef. In both ground turkey and beef, significant (P < 0.05) differences were found in the D-values between E. coli O157:H7 and Salmonella or between E. coli O157:H7 and L. monocytogenes. At 65 to 70 degrees C, D-values for E. coli O157:H7, Salmonella, and L. monocytogenes were also significantly (P < 0.05) different between turkey and beef. The obtained D- and z-values were used in predicting process lethality of the pathogens in ground turkey and beef patties cooked in an air impingement oven and confirmed by inoculation studies for a 7-log (CFU/g) reduction of E. coli O157:H7, Salmonella, and L. monocytogenes.     

Reduction of Escherichia coli O157:H7 and Salmonella in ground beef using lactic acid bacteria and the impact on sensory properties

Studies were conducted to determine whether four strains of lactic acid bacteria (LAB) inhibited Escherichia coli O157: H7 and Salmonella in ground beef at 5 degrees C and whether these bacteria had an impact on the sensory properties of the beef. The LAB consisted of frozen concentrated cultures of four Lactobacillus strains, and a cocktail mixture of streptomycin-resistant E. coli O157:H7 and Salmonella were used as pathogens. Individual LAB isolates at 10(7) CFU/ml were added to tryptic soy broth containing a pathogen concentration of 10(5) CFU/ml. Samples were stored at 5 degrees C, and pathogen populations were determined on days 0, 4, 8, and 12. After 4 days of storage, there were significant differences in numbers of both pathogens exposed to LAB isolates NP 35 and NP 3. After 8 and 12 days of storage, all LAB reduced populations of both pathogens by an average of 3 to 5 log cycles. A second study was conducted in vacuum-packaged fresh ground beef. The individual LAB isolates resulted in an average difference of 1.5 log cycles of E. coli O157:H7 after 12 days of storage, and Salmonella populations were reduced by an average of 3 log cycles. Following this study, a mixed concentrated culture was prepared from all four LAB and added to ground beef inoculated with pathogen at 10(8) CFU/g. After 3 days of storage, the mixed culture resulted in a 2.0-log reduction in E. coli O157:H7 compared with the control, whereas after 5 days of storage, a 3-log reduction was noted. Salmonella was reduced to nondetectable levels after day 5. Sensory studies on noninoculated samples that contained LAB indicated that there were no adverse effects of LAB on the sensory properties of the ground beef. This study indicates that adding LAB to raw ground beef stored at refrigeration temperatures may be an important intervention for controlling foodborne pathogens.     

Acid tolerance of acid-adapted and nonacid-adapted Escherichia coli O157:H7 strains in beef decontamination runoff fluids or on beef tissue

This study assessed the acid tolerance response (ATR) of stationary phase, acid-adapted (tryptic soy broth [TSB]+1% glucose) or nonacid-adapted (glucose-free TSB) Escherichia coli O157:H7 strains (ATCC43889, ATCC43895, ATCC51658 and EO139), grown individually or in a mixed culture, prior to inoculation of beef or meat decontamination runoff (washings) fluids (acidic [pH 4.95] or nonacidic [pH 7.01]). The inoculated beef was left untreated or treated by dipping for 30s in hot water (75 degrees C) followed by 2% lactic acid (55 degrees C). Inoculated beef samples and washings were stored aerobically at 4 or 15 degrees C for 6d, and at set intervals (0, 2, and 6d) were exposed (for 0, 60, 120, and 180min) to pH 3.5 (adjusted with lactic acid) TSB plus 0.6% yeast extract. Overall, there were no significant (P0.05) differences in responses of cultures prepared as individual or mixed strains. Decontamination of meat did not affect the subsequent ATR of E. coli O157:H7 other than resulting in lower initial pathogen levels exposed to acidic conditions. In this study, E. coli O157:H7 appeared to become more tolerant to acid following incubation in acidic washings of sublethal pH (4.89-5.22) compared to nonacidic washings (pH 6.97-7.41) at 4 degrees C or in both types of washings incubated at 15 degrees C. The ATR of the pathogen inoculated into washings was enhanced when cells were previously acid-adapted and incubated at 4 degrees C. Similarly, the ATR on meat was increased by previous acid-adaptation of the inoculum in broth and enhanced by storage at 4 degrees C. Populations on treated meat were consistently lower than those on untreated meat during storage and following exposure to acid. Although on day-0 there were no significant (P0.05) differences in ATR between acid-adapted and nonacid-adapted populations on meat, acid-adapted cells displayed consistently higher resistance through day-6. This suggests that acid-adapted E. coli O157:H7 introduced on meat may become resistant to subsequent lactic acid exposure following storage at 4 degrees C.     

Disinfection of mung bean seed with gaseous acetic acid.

Mung bean seed inoculated with Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes (3 to 5 log CFU/g) was exposed to gaseous acetic acid in an aluminum fumigation chamber. Salmonella Typhimurium and E. coli O157:H7 were not detected by enrichment of seeds treated with 242 microl of acetic acid per liter of air for 12 h at 45 degrees C. L. monocytogenes was recovered by enrichment from two of 10 25-g seed samples treated in this manner. Fumigation with gaseous acetic acid was also lethal to indigenous bacteria and fungi on mung bean seed. The treatment did not significantly reduce seed germination rates, and no differences in surface microstructure were observed between treated and untreated seed viewed by scanning electron microscopy.     

Reduction of Listeria monocytogenes, Salmonella spp., and Escherichia coli O157:H7 During Processing of Country-cured Hams

ABSTRACT: The country-cured ham process, including curing, equalization, cold-smoked or nonsmoked, and aging up to 6 mo, was validated and showed its effectiveness in achieving a 6-log reduction of Listeria monocytogenes, Salmonella spp., and Escherichia coli O157:H7. The viable counts of L. monocytogenes populations decreased to below detection levels after 206 d, Salmonella populations required 122 d, and E. coli O157:H7 required 66 d. However, L. monocytogenes -inoculated hams were positive and Salmonella spp-inoculated and E. coli O157:H7-inoculated hams were negative following enrichment procedures at the end of the aging process. Therefore, the survival of L. monocytogenes on country-cured ham represents a risk.