Growth of Listeria monocytogenes in different retail delicatessen meats during simulated home storage

Delicatessen meats are reported to be the leading vehicle of foodborne listeriosis in the United States. Listeria monocytogenes can reach high numbers in these products during storage, and the growth rate is largely dictated by product formulation and storage temperature. To assess the impact of product age on Listeria growth, five commercial brands each of cured and uncured turkey breast, ham, and roast beef (three lots per brand) were sliced (approximately 25 g per slice) at the beginning of the shelf life, the midpoint, and the last allowable day of sale, surface inoculated with an eight-strain cocktail of L. monocytogenes (approximately 40 CFU/g), and then quantitatively examined for Listeria, lactic acid bacteria, and mesophilic aerobic bacteria during aerobic storage at 4, 7, or 10°C. As expected, L. monocytogenes grew faster in deli meats without rather than with Listeria inhibitors (lactate and/or diacetate) and at the highest storage temperature (10°C). Lag-phase durations for L. monocytogenes in deli meats with and without Listeria inhibitors were 9.21, 6.96, and 5.00 and 6.35, 3.30, and 2.19 days at 4, 7, and 10°C, respectively. Generation times for L. monocytogenes in deli meats with and without Listeria inhibitors were 1.59, 1.53, and 0.85 and 0.94, 0.50, and 0.36 at 4, 7, and 10°C, respectively. Maximum population densities for L. monocytogenes in deli meats with and without Listeria inhibitors were 5.26, 5.92, and 5.97 and 8.47, 8.96 and 9.34 log CFU/g at 4, 7, and 10°C, respectively. Although lactate and diacetate suppressed L. monocytogenes growth, the extent of inhibition differed, ranging from total inhibition in roast beef to only partial inhibition in ham and cured turkey. Listeria growth was also impacted by lot-to-lot variation in the concentrations of Listeria inhibitors, product pH, and background microflora. These data will be useful for developing recommendations for "best consumed by" dating for deli meats using a risk-based approach.     

Cross-contamination between processing equipment and deli meats by Listeria monocytogenes

Contamination of luncheon meats by Listeria monocytogenes has resulted in outbreaks of listeriosis and major product recalls. Listeriae can survive on processing equipment such as meat slicers which serve as a potential contamination source. This study was conducted to determine (i) the dynamics of cross-contamination of L. monocytogenes from a commercial slicer and associated equipment onto sliced meat products, (ii) the influence of sample size on the efficacy of the BAX-PCR and U.S. Department of Agriculture-Food Safety and Inspection Service enrichment culture assays to detect L. monocytogenes on deli meat, and (iii) the fate of L. monocytogenes on sliced deli meats of different types during refrigerated storage. Three types of deli meats, uncured oven-roasted turkey, salami, and bologna containing sodium diacetate and potassium lactate, were tested. A five-strain mixture of L. monocytogenes was inoculated at ca.10(3) CFU onto the blade of a commercial slicer. Five consecutive meat slices were packed per package, then vacuum sealed, stored at 4 degrees C, and sampled at 1 and 30 days postslicing. Two sample sizes, 25 g and contents of the entire package of meat, were assayed. Total numbers of L. monocytogenes-positive samples, including the two sample sizes and two sampling times, were 80, 9, and 3 for turkey, salami, and bologna, respectively. A higher percentage of turkey meat samples were L. monocytogenes positive when contents of the entire package were assayed than when the 25-g sample was assayed (12.5 and 7.5%, respectively). Lower inoculum populations of ca. 10(1) or 10(2) CFU of L. monocytogenes on the slicer blade were used for an additional evaluation of oven-roasted turkey using two additional sampling times of 60 and 90 days postslicing. L. monocytogenes-positive samples were not detected until 60 days postslicing, and more positive samples were detected at 90 days than at 60 days postslicing. When BAX-PCR and enrichment culture assays were compared, 12, 8, and 2 L. monocytogenes-positive samples were detected by both the enrichment culture and BAX-PCR, BAX-PCR only, and enrichment culture only assays, respectively. The number of L. monocytogenes-positive samples and L. monocytogenes counts increased during storage of turkey meat but decreased for salami and bologna. Significantly more turkey samples were L. monocytogenes positive when the contents of the entire package were sampled than when 25 g was sampled. Our results indicate that L. monocytogenes can be transferred from a contaminated slicer onto meats and can survive or grow better on uncured oven-roasted turkey than on salami or bologna with preservatives. Higher L. monocytogenes cell numbers inoculated on the slicer blade resulted in more L. monocytogenes-positive sliced meat samples. In addition, the BAX-PCR assay was better than the enrichment culture assay at detecting L. monocytogenes on turkey meat (P < 0.05).     

Impact of bacterial stress and biofilm-forming ability on transfer of surface-dried Listeria monocytogenes during slicing of delicatessen meats

Listeria monocytogenes contamination of delicatessen slicer blades can lead to cross-contamination of luncheon meats. A cocktail of 3 strong or 3 weak biofilm-forming strains of L. monocytogenes suspended in turkey slurry was used to inoculate stainless steel delicatessen slicer blades at a level of 6 log CFU/blade. The cocktails were used with or without injury (cold-shocked at 4 degrees C for 2 h, or chlorine-injured at 100 ppm for 1 min). Inoculated blades were held at 22 degrees C/78+/-2% relative humidity for 6 and 24 h, before being used to generate 30 slices from chubs of roast turkey breast or Genoa salami. Slices (25 g) were diluted 1:5 in University of Vermont Medium, homogenized by stomaching and then pour-plated using tryptose phosphate agar supplemented with esculin and ferric ammonium citrate. Greater cumulative transfer to the 30 slices was seen for the strong (3.62 log CFU) as opposed to weak biofilm-forming cocktails (3.12 log CFU) with transfer also significantly greater to turkey (3.61 log CFU) than to salami (3.12 log CFU). Among the three treatments, cold-shock significantly increased subsequent L. monocytogenes transfer (3.69 log CFU) compared to the uninjured control (3.30 log CFU) and chlorine-injury (3.12 log CFU). Significantly greater transfer was also seen for blades used after 6 as opposed to 24 h of incubation. Differences in product composition and survival of L. monocytogenes, as seen via viability staining, are likely reasons for these observed differences in transfer.     

Postpackage pasteurization of ready-to-eat deli meats by submersion heating for reduction of Listeria monocytogenes

A mixed cocktail of four strains of Listeria monocytogenes was resuspended in product purge and added to a variety of ready-to-eat (RTE) meat products, including turkey, ham, and roast beef. All products were vacuum sealed in shrink-wrap packaging bags, massaged to ensure inoculum distribution, and processed by submersion heating in a precision-controlled steam-injected water bath. Products were run in pairs at various time-temperature combinations in either duplicate or triplicate replications. On various L. monocytogenes-inoculated RTE deli meats, we were able to achieve 2- to 4-log cycle reductions when processed at 195 degrees F (90.6 degrees C), 200 degrees F (93.3 degrees C), or 205 degrees F (96.1 degrees C) when heated from 2 to 10 min. High-level inoculation with L. monocytogenes (approximately 10(7) CFU/ml) ensured that cells infiltrated the least processed surface areas, such as surface cuts, folds, grooves, and skin. D- and z-value determinations were made for the Listeria cocktail resuspended in product purge of each of the three meat categories. However, reduction of L. monocytogenes in product challenge studies showed much less reduction than was observed during the decimal reduction assays and was attributed to a combination of surface phenomena, including surface imperfections, that may shield bacteria from the heat and the migration of chilled purge to the product surface. The current data indicate that minimal heating regimens of 2 min at 195 to 205 degrees F can readily provide 2-log reductions in most RTE deli meats we processed and suggest that this process may be an effective microbial intervention against L. monocytogenes on RTE deli-style meats.     

COMPARISON OF SELECTIVE AND NONSELECTIVE ENRICHMENT MEDIA FOR THE ISOLATION OF LISTERIA SPECIES FROM RETAIL FOODS

Enrichment in a nonselective medium, Buffered Peptone Water (BPW) was compared with selective enrichment in University of Vermont Medium (UVMI and UVMII) for the isolation of Listeria spp. from foods. The selectivity of the 2 types of media for the pathogenic strain, Listeria monocytogenes, was also compared. In total, 221 food samples including beef burgers, ham, turkey, lettuce, broccoli, carrots, coleslaw, salads, fish, and ice cream, were purchased from local retail outlets and examined for the presence of Listeria species and L. monocytogenes using both enrichment media Listeria species were detected in 57 (25.8%) samples using UVM, and 56 (25.3%) using BPW. L. monocytogenes was present in 33(14.9%) samples enriched in UVM and in 29(13.1%) samples enriched in BPW. The advantages and disadvantages of selective and nonselective enrichment for detection of Listeria species from a range of foods are discussed.     

Growth of L. monocytogenes strain F2365 on ready-to-eat turkey meat does not enhance gastrointestinal listeriosis in intragastrically inoculated A/J mice

There have been significant outbreaks of listeriosis associated with consumption of contaminated ready-to-eat (RTE) turkey meat products. In this study, we investigated whether growth on RTE deli turkey meat sends environmental signals to listerial cells that makes them more virulent in the gastrointestinal tract of mice. L. Listeria monocytogenes strain F2365 grew from a starting inoculum of 10(3) CFU/mL to final numbers of 10(8)-10(9) CFU/mL (within 12 days at 10 degrees C) when inoculated onto sliced processed, or whole muscle, turkey breast, or into emulsified whole turkey breast. We did not observe any difference in the numbers of CFU recovered from the spleens and livers of A/J mice inoculated intragastrically with L. monocytogenes grown on sliced turkey meat, in emulsified turkey meat, or in brain heart infusion broth. These results suggest that growth on RTE sliced deli turkey, or in RTE emulsified deli turkey, does not enhance the ability of L. monocytogenes F2365 to cause gastrointestinal listeriosis in intragastrically challenged A/J mice.     

Transfer of Listeria monocytogenes during slicing of turkey breast, bologna, and salami with simulated kitchen knives

In response to continued concerns regarding Listeria cross-contamination during the slicing of deli meats, a series of specially prepared grade 304 and 316 stainless steel kitchen knife blades was inoculated with a six-strain Listeria monocytogenes cocktail (10(8), 10(5), and 10(3) CFU per blade) composed of two weak, two medium, and two strong biofilm-forming strains. The blades were then attached to an Instron 5565 electromechanical compression analyzer and used to slice whole chubs of delicatessen turkey breast, bologna, and salami to entirety (30 slices) at a cutting speed of 8.3 mm/s. Homogenates of the slices in University of Vermont Medium were surface or pour plated with modified Oxford agar and then enriched. Listeria transfer from knife blades inoculated at 10(8) CFU per blade was logarithmic, with a 2-log decrease seen after 8 to 12 slices and direct counts obtained thereafter out to 30 slices. However, blades containing 10(5) and 10(3) CFU per blade typically yielded direct counts out to only 20 and 5 slices, respectively. Normalizing data on a log scale for the first 10 slices resulted in significantly greater Listeria transfer and "tailing" from grade 304 as opposed to grade 316 stainless (P < 0.05) for all three products. After 1 year of use, surface roughness values as determined by surface profilometry were significantly greater (P < 0.001) for grade 304 than for grade 316 stainless blades. Cutting force and blade sharpness were not significantly different (P > 0.05) within stainless steel grade (P < 0.05) for each product. However, significant differences in cutting force were seen between salami and turkey (P < 0.05) for grades 304 and 316 stainless, respectively. In addition to compositional differences in the deli meats and knife blades, wear and scoring on the blade likely affected Listeria transfer during slicing.     

Survey of Listeria monocytogenes in ready-to-eat foods

The purpose of this study was to develop data on the risk of listeriosis to support a science-based strategy for addressing Listeria monocytogenes in foods in the United States. Eight categories of ready-to-eat foods were collected over 14 to 23 months from retail markets at Maryland and northern California FoodNet sites. The product categories included luncheon meats, deli salads, fresh soft "Hispanic-style" cheeses, bagged salads, blue-veined and soft mold-ripened cheeses, smoked seafood, and seafood salads. The presence and levels of L. monocytogenes in the samples were determined by rapid DNA-based assays in combination with culture methods. Of 31,705 samples tested, 577 were positive. The overall prevalence was 1.82%. with prevalences ranging from 0.17 to 4.7% among the product categories. L. monocytogenes levels in the positive samples varied from <0.3 MPN (most probable number) per g to 1.5 x 10(5) CFU/g, with 402 samples having levels of <0.3 MPN/g, 21 samples having levels of >10(2) CFU/g, and the rest of the samples having intermediate levels. No obvious trends with respect to seasonality were observed. Significant differences (P < 0.05) between the sampling sites were found, with higher prevalences for threes categories in northern California and for two categories in Maryland. Significantly (P < 0.001) higher prevalences were found for in-store-packaged samples than for manufacturer-packaged samples of luncheon meats, deli salads, and seafood salads, while 16 of the 21 samples with higher counts were manufacturer packaged. The data collected in this study help to fill gaps in the knowledge about the occurrence of L. monocytogenes in foods, and this new information should be useful in the assessment of the risk posed by L. monocytogenes to consumers.     

Effect of inhibitory extracts derived from liquid smoke combined with postprocess pasteurization for control of Listeria monocytogenes on ready-to-eat meats

Surface pasteurization was examined in combination with low-phenolic antimicrobial extracts derived from liquid smoke to inhibit and prevent the growth of Listeria monocytogenes during the shelf life of ready-to-eat meats. In preliminary trials with retail frankfurters, one smoke derivative (2-min dip) produced a 0.3-log reduction of L. monocytogenes and a 1-min in-bag pasteurization (73.9 degrees C) produced a 2.9-log reduction, whereas a combination of the two treatments produced a 5.3-log reduction that resulted in no detectable Listeria by week 3 under accelerated shelf-life conditions (10 degrees C). In trials with frankfurters manufactured without lactate or diacetate that were treated with a shortened 1-s dip, this smoke extract and one with reduced smoke flavor and color both produced a > 4.5-log reduction of L. monocytogenes on frankfurters when heated at 73.9 degrees C for 1 min, with no recoverable Listeria detected for 10 weeks when stored at 6.1 degrees C. When deli turkey breast chubs manufactured without lactate, diacetate, or nitrite were treated with a 1-s dip in combination with radiant-heat pasteurization (270 degrees C), growth of L. monocytogenes was retarded but not prevented. However, in a similar study in which smoke extract treatment of deli turkey breast was combined with in-bag postpackage pasteurization (water submersion at 93.3 degrees C), a 60-, 45-, or even 30-s heat treatment resulted in a 2- to 3-log reduction of L. monocytogenes, with no growth on the meat during 10 weeks of storage at 6.1 degrees C. These findings indicate that reduced-acid low-phenolic antimicrobial liquid smoke derivatives combined with surface pasteurization are capable of reducing or preventing growth of L. monocytogenes to meet the criteria for the U.S. Department of Agriculture Food Safety and Inspection Service Alternative 1 process for ready-to-eat deli meat products manufactured without lactate or diacetate.     

Effect of prepackage and postpackage pasteurization on postprocess elimination of Listeria monocytogenes on deli turkey products

Surface pasteurization for inactivation of Listeria monocytogenes was evaluated for radiant heat prepackage pasteurization, submersed water postpackage pasteurization, and combinations of the two techniques on various types of ready-to-eat deli turkey products obtained from at least four different manufacturers. Products were inoculated either by in-package liquid inoculum or surface sponge-contact with approximately 10(9) CFU of L. monocytogenes. Additional testing of radiant heat pasteurization was performed with low-level inoculation of product undersides with approximately 100 CFU of L. monocytogenes followed by enrichment recovery after pasteurization. Prepackage pasteurization provided 2.0 to 2.8 log reductions when processed for 60 s and 2.8 to 3.8 log reductions when processed for 75 s. An improved radiant oven provided 3.53 (60 s) and 4.76 (75 s) log reductions of L. monocytogenes. No positive samples were detected after enrichment when 40 samples of deli turkey (4 to 4.5 kg) undersides were inoculated at low levels and processed for 75 s. Submersed water postpackage pasteurization provided 1.95 to 3.0 log reductions when processed for 2, 3, 4, or 5 min, and combinations of the two processes gave 3.0 to 4.0 log inactivation of L. monocytogenes using either 60 + 60 s or 60 + 90 s for the prepackage and postpackage pasteurization processes, respectively. These processes, either individually or in combination, can provide postprocess elimination of bacteria for the manufacture of safe ready-to-eat deli meats.     

Comparison of different enrichment broths and background flora for detection of heat-injured Listeria monocytogenes in whole milk

Various primary enrichment broths, including University of Vermont medium (UVM), Listeria enrichment broth (LEB), modified LEB, and aerobic and anaerobic L-PALCAMY, were compared with aerobic and anaerobic Pennsylvania State University (PSU) broths for the detection of severely heat-injured (62.8 degrees C for 5, 10, or 15 min; no colony appearance after heat injury on aerobic Trypticase soy agar containing 0.6% yeast extract and modified Oxford medium) Listeria monocytogenes Scott A. Anaerobic conditions were produced by adding L-cysteine and then purging the headspace with N2. The effect of uninjured background flora (10(3) CFU/ml of Enterococcus faecium) on frequency of detection was examined. Anaerobic PSU broth resulted in the lowest false-positive rate and the highest frequency of detection of severely heat-injured L. monocytogenes compared with UVM, LEB, and modified LEB (P < 0.05). The presence of E. faecium significantly enhanced the detection of heat-injured (10 min at 62.8 degrees C) L. monocytogenes in aerobic and anaerobic PSU and aerobic and anaerobic L-PALCAMY broths (P < 0.05). The highest concentration of uninjured E. faecium (>10(6) CFU/ml) inhibited the detection of heat-injured L. monocytogenes (P < 0.05). A heat-resistant, LiCl-tolerant Lactobacillus isolate from raw milk increased the rate of both false-positive and false-negative reactions.     

Transfer of Listeria monocytogenes during mechanical slicing of turkey breast, bologna, and salami

A commercial delicatessen slicer was used as the vector for sequential quantitative transfer of Listeria monocytogenes (i) from an inoculated slicer blade (approximately 10(8), 10(5), or 10(3) CFU per blade) to 30 slices of uninoculated delicatessen turkey, bologna, and salami, (ii) from inoculated product (approximately 10(8) CFU/cm2) to the slicer, and (iii) from inoculated product (10(8), 10(5), or 10(3) CFU/cm2) to 30 slices of uninoculated product via the slicer blade. Cutting force and product composition also were assessed for their impact on L. monocytogenes transfer. Five product contact areas on the slicer, which were identified from residue of product bathed in Glow-Germ, were also sampled using a 1-ply composite tissue technique after inoculated product had been sliced. After being sliced with inoculated blades, each product slice was surface or pour plated on modified Oxford agar and/ or enriched in University of Vermont medium. Greater transfer (P < 0.05) occurred from inoculated turkey (10(8) CFU/cm2) to the five slicer contact areas from an application force of 4.5 kg as compared with 0 kg. On uninoculated product sliced with blades inoculated at 10(8) CFU per blade, L. monocytogenes populations decreased logarithmically to 10(2) CFU per slice after 30 slices. Findings for the inoculated slicer blade and product (10(5) CFU per blade or cm2) were similar; L. monocytogenes concentrations were 102 CFU per slice after 5 slices and enriched samples were generally negative for L. monocytogenes after 27 slices. For uninoculated product sliced with blades inoculated at 10(3) CFU per blade, the first 5 slices typically produced L. monocytogenes at approximately 10 CFU per slice by direct plating, and enrichments were negative for L. monocytogenes after 15 slices. The higher fat and lower moisture content of salami compared with turkey and bologna resulted in a visible fat layer on the blade that likely prolonged L. monocytogenes transfer. As a result of cross-contamination, those delicatessen-sliced meats that allow growth of L. monocytogenes during prolonged refrigerated storage likely pose an increased public health risk for certain consumers.     

Real-time PCR detection of 16S rRNA genes speeds most-probable-number enumeration of foodborne Listeria monocytogenes

Quantifying foodborne pathogens at concentrations of 0.1 to 1,000 CFU/g of food generally involves most-probable-number (MPN) enumeration, which takes at least 4 days. A real-time PCR assay (RTi-PCR) was developed to accelerate MPN enumeration of foodborne Listeria monocytogenes. Foods were spiked from 70 to 110 CFU/g, and triplicate subportions from 0.0001 to 1 g were selectively enriched for 48 h at 30 degrees C. For standard MPN enumeration, the enrichments were subcultured on Oxford agar (48 h at 35 degrees C) to isolate Listeria. For RTi-PCR MPN, the L. monocytogenes cells from the same enrichments were washed and resuspended in 2 ml of sterile water. DNA was extracted by boiling for 10 min. The DNA in the extract's supernatant was targeted with published oligonucleotide primers for amplifying an Lmo-specific sequence of 16S rRNA genes. Amplification was continuously monitored with SYBR Green. The resulting amplicon was characterized by its melting temperature. The L. monocytogenes specificity of the primers was confirmed by testing L. monocytogenes (15 strains), Listeria innocua (11 strains), and Listeria welshimeri, Listeria seeligeri, Listeria ivanovii, and Listeria grayi (1 strain each). Quantitatively spiked milk, lettuce, smoked salmon, Brie cheese, ice cream, pork paté, salami, ready-to-eat shrimp, raw ground beef, and fresh soft cheese were enumerated by both the standard and the PCR MPN method. The paired results from the two MPN methods agreed well, except for the fresh cheese. For some foods, l-g samples required a decimal dilution for a positive test result, suggesting concentration-dependent food ingredient interference with the RTi-PCR. This RTi-PCR method reduced the time necessary for the MPN enumeration of foodborne L. monocytogenes from 4 to 2 days.     

Comparison of public health impact of Listeria monocytogenes product-to-product and environment-to-product contamination of deli meats at retail

This study compared the relative public health impact in deli meats at retail contaminated with Listeria monocytogenes by either (i) other products or (ii) the retail environment. Modeling was performed using the risk of listeriosis-associated deaths as a public health outcome of interest and using two deli meat products (i.e., ham and turkey, both formulated without growth inhibitors) as model systems. Based on reported data, deli meats coming to retail were assumed to be contaminated at a frequency of 0.4%. Three contamination scenarios were investigated: (i) a baseline scenario, in which no additional cross-contamination occurred at retail, (ii) a scenario in which an additional 2.3% of products were cross-contaminated at retail due to transfer of L. monocytogenes cells from already contaminated ready-to-eat deli meats, and (iii) a scenario in which an additional 2.3% of products were contaminated as a result of cross-contamination from a contaminated retail environment. By using a previously reported L. monocytogenes risk assessment model that uses product-specific growth kinetic parameters, cross-contamination of deli ham and turkey was estimated to increase the relative risk of listeriosis-associated deaths by 5.9- and 6.1-fold, respectively, for contamination from other products and by 4.9- and 5.8-fold, respectively, for contamination from the retail environment. Sensitivity and scenario analyses indicated that the frequency of cross-contamination at retail from any source (other food products or environment) was the most important factor affecting the relative risk of listeriosis-associated deaths. Overall, our data indicate that retail-level cross-contamination of ready-to-eat deli meats with L. monocytogenes has the potential to considerably increase the risk of human listeriosis cases and deaths, and thus precise estimates of cross-contamination frequency are critical for accurate risk assessments.     

Impact of nitrite on detection of Listeria monocytogenes in selected ready-to-eat (RTE) meat and seafood products

ABSTRACT:  The impact of sodium nitrite (NaNO₂) on detection and recovery of Listeria monocytogenes from select ready-to-eat (RTE) foods including smoked salmon, smoked ham, beef frankfurters, and beef bologna was assessed. Nitrite-containing (NC; 100 to 200 ppm NaNO₂) or nitrite-free (NF) foods were inoculated with a 5-strain cocktail of L. monocytogenes by immersion into Butterfield's buffer solution containing 5.4 to 7.4 × 10³ L. monocytogenes per milliliter. Inoculated products were vacuum-packaged and stored at 5 °C. A weekly comparative analysis was performed for presence of L. monocytogenes using 5 detection methods on products held at 5 °C for up to 8 wk. L. monocytogenes initially present at <100 CFU/g during the first 2 wk of storage increased throughout the study, attaining final populations of approximately 1 × 10⁴ to 1 × 10⁵ CFU/g. Lactic acid bacteria predominated throughout the study in all products. Exposure to NaNO₂ (100 to 200 ppm) resulted in 83% to 99% injury to the L. monocytogenes strains tested. The genetic-based BAX^® System (DuPont™ Qualicon, Wilmington, Del., U.S.A.) and modified USDA/FSIS methods detected 98% to 100% of Listeria -positive food samples and were consistently superior to and significantly different ( P < 0.05) from conventional cultural methods in recovering Listeria from NC samples. Data show that nitrite-induced injury adversely affects detection and recovery of L. monocytogenes from NC food, confirming earlier findings that nitrite-induced injury masks L. monocytogenes detection in NC RTE food products. Nitrite-injured Listeria can subsequently repair upon nitrite depletion and grow to high levels over extended refrigerated storage.     

Determination of thermal lethality of Listeria monocytogenes in fully cooked chicken breast fillets and strips during postcook in-package pasteurization

Fully cooked chicken breast fillets and strips were surface inoculated with a cocktail of Listeria monocytogenes culture. The inoculation level was 10(7) to 10(8) CFU/g meat. The inoculated products were vacuum packaged and pasteurized at 90 degrees C with a pilot-scale steam or hot water cooker. After heat treatment, the survivors of L. monocytogenes were enumerated. No significant difference was found on survivors of L. monocytogenes between steam- and hot water-treated products. To achieve a 7-log10 (CFU/g) reduction, approximately 5, 25, and 35 min were needed for single-packaged fillets, 227-g package strips, and 454-g strips, respectively. The results from this study were subsequently verified by a computer model that could predict the thermal lethality of pathogens in fully cooked meat and poultry products during postcook in-package pasteurization.     

Resistance of Listeria monocytogenes F2365 cells to synthetic gastric fluid is greater following growth on ready-to-eat deli turkey meat than in brain heart infusion broth

Ready-to-eat (RTE) deli meats have been categorized as high-risk foods for contraction of foodborne listeriosis. Several recent listeriosis outbreaks have been associated with the consumption of RTE deli turkey meat. In this study, we examined whether the growth of Listeria monocytogenes F2365 on commercially prepared RTE deli turkey meat causes listerial cells to become more resistant to inactivation by synthetic gastric fluid (SGF). Listerial cells grown on turkey meat to late logarithmic-early stationary phase were significantly more resistant to SGF at pH 7.0, 5.0, or 3.5 than listerial cells grown in brain heart infusion (BHI) broth. The pH was lower in the fluid in packages of turkey meat than in BHI broth (6.5 versus 7.5). However, listerial cells grown in BHI broth adjusted to a lower pH (6.0) did not exhibit enhanced resistance to SGF. The lesser resistance to SGF of listerial cells grown in BHI broth may be due, in part, to the presence of glucose (0.2%). This study indicates the environment presented by the growth of L. monocytogenes on deli turkey meat affects its ability to survive conditions it encounters in the gastrointestinal tract.     

Elimination of Listeria monocytogenes from ready-to-eat turkey and cheese tortilla wraps using ionizing radiation

Listeria monocytogenes is a common postprocess contaminant on ready-to-eat foods including premade ready-to-eat sandwiches. One popular type of sandwich product is the tortilla wrap, which contains sliced luncheon meats and cheeses rolled within a flour tortilla. This study determined the radiation resistance of L. monocytogenes surface inoculated onto two types of commercially available wheat flour tortillas, processed cheese slices, and deli turkey meat. The D10-values for L. monocytogenes (the radiation dose required to inactivate 1 log of the pathogen) were 0.27 kGy when inoculated onto two flour tortilla types, 0.28 and 0.30 kGy when inoculated onto two types of sliced processed cheeses, and 0.58 and 0.65 kGy when inoculated onto two types of sliced deli turkey meat. When two types of tortilla wraps were assembled from the individual components and L. monocytogenes was inoculated into the interfaces between the individual components, the D10-values were 0.27 to 0.37 kGy in the tortilla and cheese interfaces, 0.33 to 0.41 kGy in the cheese and turkey interfaces, and 0.25 to 0.33 kGy in the turkey and tortilla interfaces. The ability of ionizing radiation to reduce pathogen levels on the complex tortilla, cheese, and luncheon meat product was limited by the higher radiation resistance of L. monocytogenes when inoculated onto the ready-to-eat turkey-meat component.     

Comparison of different most-probable-number methods for enumeration of Listeria in poultry

To estimate levels of Listeria spp. in poultry and to select the most appropriate enumeration method for routine analysis, 40 naturally contaminated retail chicken carcasses were tested in Ponferrada (León, N.W. Spain) using the direct plate count technique and various most-probable-number (MPN) designs (UVM I [University of Vermont modified Listeria enrichment broth], Fraser enrichment broth, or both were used in 3-, 5-, and 10-tube MPN techniques). MPN estimation was obtained from the number of tubes with Listeria confirmed (after streaking on PALCAM and modified Oxford agars: "true" MPN) and from the number of dark Fraser broth tubes ("predictive" MPN). Samples were analyzed in duplicate. Low levels of Listeria were found (< 110 CFU/g). The direct plate count technique was totally ineffective for enumerating Listeria in poultry. The single-step (UVM I) and the two-step (UVM I-Fraser) MPN methods gave comparable estimations and a low number of significantly discrepant predictions. Using a single-step method with Fraser broth, lower true MPNs were obtained. The number of tubes used (3, 5, or 10) did not have a substantial influence on the results. Similar estimations, highly correlated (r = 0.538 to 0.968; P < 0.001), were found with (true MPN) and without (predictive MPN) plating confirmation when using the two-step MPN method. The statistical evaluation of the differential character of Fraser broth as part of the two-step MPN method showed high sensitivity (87.5 to 92.5%), specificity (95.2 to 98.6%), efficiency (94.2 to 97.6%), and predictive values (73.6 to 89.9% for a positive test and 98.0 to 98.9% for a negative test). Taking into account these results, we suggest the convenience of using a 3- or 5-tube two-step (UVM I-Fraser) MPN method with estimations obtained from the number of tubes with darkening, without confirmation, in order to achieve great savings in time and money.     

Use of one-ply composite tissues in an automated optical assay for recovery of Listeria from food contact surfaces and poultry-processing environments

A novel one-ply composite tissue (CT) method using the Soleris (formerly BioSys) optical analysis system was compared with the conventional U.S. Department of Agriculture (USDA) environmental sponge enrichment method for recovery of Listeria from food contact surfaces and poultry-processing environments. Stainless steel and high-density polyethylene plates were inoculated to contain a six-strain L. monocytogenes cocktail at 10(4), 10(2), and 10 CFU per plate, whereas samples from naturally contaminated surfaces and floor drains from a poultry-processing facility were collected with CTs and environmental sponges. CT samples were transferred into Soleris system vials, and presumptive-positive samples were further confirmed. Sponge samples were processed for Listeria using the USDA culture method. L. monocytogenes recovery rates from inoculated stainless steel and polyethylene surfaces were then compared for the two methods in terms of sensitivity, specificity, and positive and negative predictive values. No significant differences (P > 0.05) were found between the two methods for recovery of L. monocytogenes from any of the inoculated stainless steel and polyethylene surfaces or environmental samples. Sensitivity, specificity, and overall accuracy of the CT-Soleris for recovery of Listeria from environmental samples were 83, 97, and 95%, respectively. Listeria was detected 2 to 3 days sooner with the CT-Soleris method than with the USDA culture method, thus supporting the increased efficacy of this new protocol for environmental sampling.