Reduction and survival of Listeria monocytogenes in ready-to-eat meats after irradiation

A five-strain Listeria monocytogenes culture was inoculated onto six different types of ready-to-eat (RTE) meats (frankfurters, ham, roast beef, bologna, smoked turkey with lactate, and smoked turkey without lactate). The meats were vacuum packed and stored at 4 degrees C for 24 h prior to irradiation. Populations of L. monocytogenes were recovered by surface plating on nonselective and selective media. The margins of safety studied include 3-log (3D) and 5-log (5D) reduction of pathogenic bacteria to achieve an optimal level of reduction while retaining organoleptic qualities of the meats. A 3-log reduction of L. monocytogenes was obtained at 1.5 kGy when nonselective plating medium was used. The dosages for 3-log reduction were 1.5 kGy for bologna, roast beef, and both types of turkey and 2.0 kGy for frankfurters and ham on the basis of use of selective medium. The D10-values ranged from 0.42 to 0.44 kGy. A 5-log reduction of L. monocytogenes was obtained at 2.5 kGy with nonselective medium. With selective medium, the dosages were 2.5 kGy for bologna, roast beef, and both types of turkey and 3.0 kGy for frankfurters and ham. Survival of L. monocytogenes in the same RTE meat types after irradiation was also studied. Meats were inoculated with 5 log L. monocytogenes per g and irradiated at doses of 2.0 and 4.0 kGy. Recovery of the surviving organisms was observed during storage at temperatures of 4 and 10 degrees C for 12 weeks. Preliminary results showed no growth in meats irradiated at 4.0 kGy. Survivors were observed for irradiated meats at 2.0 kGy stored at 10 degrees C after the second week. No growth was observed in samples irradiated at 2.0 kGy stored at 4 degrees C until the fifth week.     

Impact of dilution ratios on Listeria monocytogenes growth during University of Vermont medium enrichment of deli meats

In the U.S. Department of Agriculture (USDA) method for Listeria detection, a 25-g composite food sample is enriched in 225 ml of University of Vermont medium (UVM), giving a detection limit of 0.04 CFU/g. However, in a recent large-scale four-state deli meat survey for L. monocytogenes, 125-g samples enriched in 1,125 ml of UVM were requested to increase the detection limit to 0.008 CFU/g. To circumvent problems associated with large volumes of UVM, the impact on L. monocytogenes growth of lower dilution ratios used for enrichment and most-probable-number (MPN) detection was compared with the results obtained using the conventional 1:10 dilution. In this study, 125-g samples of cured turkey, uncured turkey, ham, and roast beef were inoculated with a six-strain L. monocytogenes cocktail to contain approximately 1 x 10(3) CFU/g. This cocktail was then diluted 1:3, 1:5, or 1:10 in UVM, homogenized, enriched at 30 degrees C, and periodically plated on modified Oxford agar to determine generation times during 24 h of incubation. The same enrichment protocol was also assessed in a three-tube MPN assay using 125-g samples inoculated with L. monocytogenes to contain approximately 1 CFU/g. The effects of two homogenization methods, stomaching and pulsifying, on Listeria growth were compared using oven-roasted turkey breast diluted 1:3, 1:5, and 1:10 in UVM. Overall, the growth rates, generation times, and MPN values for each of the four selected deli meats were similar (P > 0.05) using UVM enrichment ratios of 1:3, 1:5, and 1:10, with no significant (P > 0.05) differences in L. monocytogenes growth rate or generation time between experiments using pulsifying and stomaching. These findings indicate that lower volumes of UVM can be used in the USDA procedure when examining deli meats without compromising Listeria recovery.     

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.     

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.     

Effects of Lactic Acid on the Growth Characteristics of Listeria monocytogenes on Cooked Ham Surfaces

The surfaces of ready-to-eat meats are susceptible to postprocessing contamination by Listeria monocytogenes. This study examined and modeled the growth characteristics of L. monocytogenes on cooked ham treated with lactic acid solutions (LA). Cooked ham was inoculated with L. monocytogenes (ca. 10(3) CFU/g), immersed in 0, 0.5, 0.75, 1.0, 1.25, 1.5, and 2.0% LA for 30 min, vacuum packaged, and stored at 4, 8, 12, and 16°C. LA immersion resulted in <0.7 log CFU/g immediate reduction of L. monocytogenes on ham surfaces, indicating the immersion alone was not sufficient for reducing L. monocytogenes. During storage, no growth of L. monocytogenes occurred on ham treated with 1.5% LA at 4 and 8°C and with 2% LA at all storage temperatures. LA treatments extended the lag-phase duration (LPD) of L. monocytogenes and reduced the growth rate (GR) from 0.21 log CFU/day in untreated ham to 0.13 to 0.06 log CFU/day on ham treated with 0.5 to 1.25% LA at 4°C, whereas the GR was reduced from 0.57 log CFU/day to 0.40 to 0.12 log CFU/day at 8°C. A significant extension of the LPD and reduction of the GR of L. monocytogenes occurred on ham treated with >1.25% LA. The LPD and GR as a function of LA concentration and storage temperature can be satisfactorily described by a polynomial or expanded square-root model. Results from this study indicate that immersion treatments with >1.5% LA for 30 min may be used to control the growth of L. monocytogenes on cooked meat, and the models would be useful for selecting LA immersion treatments for meat products to achieve desired product safety.     

Controlling Listeria monocytogenes on sliced ham and turkey products using benzoate, propionate, and sorbate

The objective of this study was to identify concentrations of sorbate, benzoate, and propionate that prevent the growth of Listeria monocytogenes on sliced, cooked, uncured turkey breast and cured ham. Sixteen test formulations plus a control formulation for each product type were manufactured to include potassium sorbate, sodium benzoate, or sodium propionate, used alone and combined (up to 0.3% [wt/wt]), or with sodium lactate-sodium diacetate combinations. Products were inoculated with L. monocytogenes (5 log CFU/100-g package) and stored at 4, 7, or 10 degrees C for up to 12 weeks, and triplicate samples per treatment were assayed biweekly by plating on modified Oxford agar. Data showed that 0.1% benzoate, 0.2% propionate, 0.3% sorbate, or a combination of 1.6% lactate with 0.1% diacetate prevented the growth of L. monocytogenes on ham stored at 4 degrees C for 12 weeks, compared with greater than a 1-log increase at 4 weeks for the control ham without antimicrobials. When no nitrite was included in the formulation, 0.2% propionate used alone, a combination of 0.1% propionate with 0.1% sorbate, or a combination of 3.2% lactate with 0.2% diacetate was required to prevent listerial growth on the product stored at 4 degrees C for 12 weeks. Inhibition was less pronounced when formulations were stored at abuse temperatures. When stored at 7 degrees C, select treatments delayed listerial growth for 4 weeks but supported significant growth at 8 weeks. All treatments supported more than a 1-log increase in listerial populations when stored at 10 degrees C for 4 weeks. These results verify that antimycotic agents inhibit the growth of L. monocytogenes on ready-to-eat meats but aremore effective when used in combination with nitrite.     

Modeling the lag phase and growth rate of Listeria monocytogenes in ground ham containing sodium lactate and sodium diacetate at various storage temperatures

ABSTRACT:  Refrigerated ready-to-eat (RTE) meats contaminated with Listeria monocytogenes were implicated in several listeriosis outbreaks. Lactate and diacetate have been shown to control L. monocytogenes in RTE meats. The objective of this study was to examine and model the effect of lactate (1.0% to 4.2%) and diacetate (0.05% to 0.2%) in ground ham on the lag phase duration (LPD, h) and growth rate (GR, log CFU/h) of L. monocytogenes at a range of temperatures (0 to 45 °C). A 6-strain mixture of L. monocytogenes was inoculated into ground ham containing lactate and diacetate, and stored at various temperatures. The LPD and GR of L. monocytogenes in ham as affected by lactate, diacetate, and storage temperature were analyzed and accurately represented with mathematical equations. Resulting LPD and GR equations for storage temperatures within the range of 0 to 36 °C significantly represented the experimental data with a regression coefficient of 0.97 and 0.96, respectively. Significant factors ( P < 0.05) that affected the LPD were temperature, lactate, diacetate, and the interactions of all three, whereas only temperature and the interactions between temperature and lactate and diacetate had a significant effect on GR. At suboptimal growth temperatures (≤12 °C) the increase of lactate and diacetate concentrations, individually or in combination, extended the LPD. The effect of higher concentrations of both additives on reducing the GR was observed only at temperatures that were more suitable for growth of L. monocytogenes , that is, 15 to 35 °C. These data may be used to assist in determining concentrations of lactate and diacetate in cooked ham products to control the growth of L. monocytogenes over a wide range of temperatures during manufacturing, distribution, and storage.     

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.     

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).     

Acidified sodium chlorite treatment for inhibition of Listeria monocytogenes growth on the surface of cooked roast beef

The effects of acidified sodium chlorite (ASC) against Listeria monocytogenes on the surface of cooked roast beef were investigated. L. monocytogenes, strain V7, serotype 1/2a, was inoculated at numbers of 6.0 log CFU/g onto 5-g cubes of cooked regular or spicy roast beef. The samples were allowed to air dry for 1 h. The cooked roast beef samples were dipped into ASC or sprayed with ASC solutions of 250, 500, 750, or 1,000 ppm, then placed in bags with or without a vacuum and refrigerated at 4 degrees C. L. monocytogenes counts were determined after 0, 7, 14, 21, and 28 days of storage by spread plating roast beef samples onto Oxford agar plates that were incubated at 37 degrees C for 48 h. At day 28, the number of L. monocytogenes on the > or = 500 ppm ASC-treated spicy roast beef samples had count reductions that were >4.0 log CFU/g, whereas the same concentrations of ASC-treated regular roast beef samples had approximately a 2.5 log CFU/g reduction in L. monocytogenes counts when compared with the untreated samples. No significant differences (P > 0.05) were observed in L. monocytogenes counts between the vacuum- or nonvacuum-packaged ASC-treated cooked roast beef samples. Sensory evaluation showed no significant differences (P > 0.05) between ASC-treated and untreated roast beef. ASC can be used as a processing aid in the form of a dip or spray treatment to control L. monocytogenes on the surface of cooked roast beef.     

Effect of packaging and storage time on survival of Listeria monocytogenes on kippered beef steak and turkey tenders

The objective of our study was to determine effect of packaging method and storage time on reducing Listeria monocytogenes in shelf-stable meat snacks. Commercially available kippered beef steak strips and turkey tenders were dipped into a 5-strain L. monocytogenes cocktail, and dried at 23 °C until a water activity of 0.80 was achieved. Inoculated samples were packaged with 4 treatments: (1) vacuum, (2) nitrogen flushed with oxygen scavenger, (3) heat sealed with oxygen scavenger, and (4) heat sealed without oxygen scavenger. Samples were stored at 23 °C and evaluated for L. monocytogenes levels at 0, 24, 48, and 72 h. Initial levels (time 0) of L. monocytogenes were approximately 5.7 log CFU/cm2 for steak and tenders. After 24 h of storage time, a 1 log CFU/cm2 reduction of L. monocytogenes was observed for turkey tenders for all packaging treatments. After 48 h, turkey tenders showed >1 log CFU/cm2 reduction of L. monocytogenes for all packaging treatments except for vacuum, where only 0.9 log CFU/cm2 reduction was observed. After 72 h, reductions for all packaging treatments for turkey tenders ranged from 1.5 to 2.4 log CFU/cm2. For kippered beef steak, there was no interaction between the packaging treatments and all storage times (P > 0.05) whereas, time was different (P <0.05). For kippered beef steak, there was 1 log reduction of L. monocytogenes at 24 and 48 h of storage times at 23 °C for all packaging treatments and a 2.1 log CFU/ cm2 L. monocytogenes reduction at 72 h of storage time. PRACTICAL APPLICATIONS: Processors of kippered beef steak and turkey tenders could use a combination of vacuum or nitrogen-flushing or heat sealed with an oxygen scavenger packaging methods and a holding time of 24 h prior to shipping to reduce potential L. monocytogenes numbers by ≥1 log. However, processors should be encouraged to hold packaged product a minimum of 72 h to enhance the margin of safety for L. monocytogenes control.     

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.     

Inhibition of Listeria monocytogenes by sodium diacetate and sodium lactate on wieners and cooked bratwurst.

The inhibition of Listeria monocytogenes by sodium lactate and sodium diacetate was evaluated for wieners containing pork, turkey, and beef and for cooked bratwurst containing beef and pork. Both products were supplied by commercial manufacturers. Treated products were surface-inoculated with 10(5) CFU of L. monocytogenes per package and vacuum-packed in gas-impermeable pouches. Wieners were stored for 60 days at 4.5 degrees C, and bratwurst were stored for 84 days at 3 and 7degrees C. A surface treatment that consisted of dipping wieners into solutions containing < or = 6% lactate and < or = 3% diacetate for 5 s did not delay pathogen growth compared with that for untreated wieners. In additional trials, the antilisterial activity of lactate and diacetate in wiener and bratwurst formulations was evaluated. Lactate levels ranged from 1.32 to 3.4%, and diacetate was evaluated at 0.1 and 0.25%. The growth of L. monocytogenes was delayed for 4 and 12 weeks at 7 and 3 degrees C, respectively, on uncured, unsmoked bratwurst formulated with 3.4% lactate/0.1% diacetate, compared with 1 and 2 weeks, respectively, for the formulation containing 2% lactate. L. monocytogenes grew by > or = 1 log unit after 4 weeks' storage at 3 or 7 degrees C on cured, smoked bratwurst without lactate or diacetate, but growth was inhibited for 12 weeks on cured, smoked bratwurst formulated with 3.4% lactate and 0.1% diacetate. Sodium lactate levels of > or = 3% and combinations of > or = 1% lactate plus > or = 0.1% diacetate prevented listerial growth on wieners stored for 60 days at 4.5 degrees C. These results indicate that dipping wieners in lactate-diacetate solutions is not an efficient way to apply these antimicrobial agents to wieners. However, the inclusion of combinations of sodium lactate and sodium diacetate in wiener or bratwurst formulations inhibits the growth of L monocytogenes at < or = 7 degrees C, and an additional margin of safety was observed for products that are cured and smoked.     

Prepackage surface pasteurization of ready-to-eat meats with a radiant heat oven for reduction of Listeria monocytogenes

In this paper, a thermal process for the surface pasteurization of ready-to-eat (RTE) meat products for the reduction of Listeria monocytogenes on such products (turkey bologna, roast beef, corned beef, and ham) is described. The process involves the passage of products through a "tunnel" of heated coils on a stainless steel conveyor belt at various treatment times relevant to the manufacture of processed meat for the surface pasteurization of RTE meat products. Two inoculation procedures, dip and contact inoculation, were examined with the use of a four-strain cocktail of L. monocytogenes prior to heat processing. With the use of radiant heat prepackage surface pasteurization, 1.25 to 3.5-log reductions of L. monocytogenes were achieved with treatment times of 60 to 120 s and air temperatures of 475 to 750 degrees F (246 to 399 degrees C) for these various RTE meats. Reduction levels differed depending on the type of inoculation method used, the type of product used, the treatment temperature, and the treatment time. Prepackage pasteurization (60 s) was also combined with postpackage submerged water pasteurization for formed ham (60 or 90 s), turkey bologna (45 or 60 s), and roast beef (60 or 90 s), resulting in reductions of 3.2 to 3.9. 2.7 to 4.3, and 2.0 to 3.75 log cycles, respectively. These findings demonstrate that prepackage pasteurization, either alone or in combination with postpackage pasteurization, is an effective tool for controlling L. monocytogenes surface contamination that may result from in-house handling.     

Fate of Listeria monocytogenes inoculated onto the surface of model Turkey frankfurter pieces treated with zein coatings containing nisin, sodium diacetate, and sodium lactate at 4 degrees C

The antimicrobial effects of zein coatings containing nisin, sodium lactate, and sodium diacetate against Listeria monocytogenes on turkey frankfurters at 4 degrees C were determined. Our objectives were to determine whether zein, nisin, lactate, and diacetate alone or in combination could control the growth of L. monocytogenes on full-fat turkey frankfurters at 4 degrees C and to determine whether lactate or diacetate had any synergistic effect on the activity of nisin. Turkey frankfurter pieces surface inoculated with L. monocytogenes strain V7 were treated with zein-ethanol-glycerol (ZEG), zein-propylene-glycol (ZPR), ethanol-glycerol (EG), propylene glycol (PR), nisin (N), sodium lactate (L), or sodium diacetate (D) alone or in combination. Over 28 days, treatment with N or D alone reduced L. monocytogenes counts on frankfurters by 6.6 or 6.3 log CFU/g, respectively. N-D treatment reduced L. monocytogenes by 6 log CFU/g. The zein solvents EG and PR reduced L. monocytogenes by about 5.6 and 5.2 log CFU/g, respectively, similar to the results obtained with ZEG and ZPR, which suggests that zein powder per se had no antimicrobial activity. After 28 days, ZEG-N-D, ZEG-N-D-L, ZPR-N-D, and ZPR-N-D-L yielded no detectable CFU. L alone was ineffective. No synergies were observed. N and D when used singly and the combinations of N-D, ZEG-N-D, ZEG-N-D-L, ZPR-N-D, ZPR-N-D-L, EG, and PR were effective as inhibitors of the growth of recontaminating L. monocytogenes cells on full-fat turkey frankfurters.     

Attachment of Listeria monocytogenes on ready-to-eat meats

Five individual strains of Listeria monocytogenes and a mixed cocktail of all five were studied for attachment on frankfurters, ham, bologna, and roast beef relative to their cell surface characteristics. The ratio of strongly attached (sessile) L. monocytogenes cells compared with total (sessile and planktonic) attached cells on ready-to-eat meats was also determined. Because bacterial cell surfaces were characterized by net negative charge and hydrophobicity, electrostatic interaction chromatography and cationized ferritin methods were chosen to study net negative charge distribution on the bacterial cell surface, whereas hydrophobic interaction chromatography and contact angle measurement were used to examine the cell surface hydrophobicity. No differences (P > 0.05) were observed in cell surface charge or cell surface hydrophobicity among strains. Approximately 84 to 87% L. monocytogenes were found to attach strongly to ready-to-eat meats within 5 min. No differences (P > 0.05) were found among strains or among meats. Micrographs observed from scanning electron microscopy showed no differences among the strains but showed a difference in age of cells (mixed culture) in terms of surface negative charge distribution. More surface negatively charged sites were observed at 0 and 7 days and much fewer at 3 days during storage of washed, harvested cells in buffer at 4 degrees C (aged cells under cold and nutrient deprivation), indicating a possible change in cell surface properties. Because no difference in strains was observed, the contact angle measurement study was carried out with the five-strain mixed culture. The surface hydrophobicity increased in frankfurters, decreased in roast beef, and was unchanged in ham and bologna as a result of inoculation.     

Efficacy of bacteriophage LISTEXP100 combined with chemical antimicrobials in reducing Listeria monocytogenes in cooked turkey and roast beef

The aim of this study was to verify the effectiveness of the commercially available anti-Listeria phage preparation LISTEX^™P100 in reducing Listeria monocytogenes on ready-to-eat (RTE) roast beef and cooked turkey in the presence or absence of the chemical antimicrobials potassium lactate (PL) and sodium diacetate (SD). Sliced RTE meat cores at 4 and 10 °C were inoculated with cold-adapted L. monocytogenes to result in a surface contamination level of 10³ CFU/cm². LISTEX^TMP100 was applied at 10⁷ PFU/cm² and samples taken at regular time intervals during the RTE product's shelf life to enumerate viable L. monocytogenes. LISTEX^™P100 was effective during incubation at 4 °C with initial reductions of L. monocytogenes of 2.1 log₁₀ CFU/cm² and 1.7 log₁₀ CFU/cm², respectively, for cooked turkey and roast beef without chemical antimicrobials (there was no significant difference to the initial L. monocytogenes reductions in the presence of LISTEX^TMP100 for cooked turkey containing PL and roast beef containing SD-PL). In the samples containing no chemical antimicrobials, the presence of phage resulted in lower L. monocytogenes numbers, relative to the untreated control, of about 2 log CFU/cm² over a 28-day storage period at 4 °C. An initial L. monocytogenes cell reduction of 1.5 log₁₀ CFU/cm² and 1.7 log₁₀ CFU/cm², respectively, for cooked turkey and roast beef containing no chemical antimicrobials was achieved by the phage at 10 °C (abusive temperature). At this temperature, the L. monocytogenes cell numbers of samples treated with LISTEX™ P100 remained below those of the untreated control only during the first 14 days of the experiment for roast beef samples with and without antimicrobials. On day 28, the L. monocytogenes numbers on samples containing chemical antimicrobials and treated with LISTEX^TMP100 stored at 4 and 10 °C were 4.5 log₁₀ CFU/cm² and 7.5 log₁₀ CFU/cm², respectively, for cooked turkey, and 1.2 log₁₀ CFU/cm² and 7.2 log₁₀ CFU/cm², respectively, for roast beef. In both cooked turkey samples with and without chemical antimicrobials stored at 10 °C, the phage-treated samples had significantly lower numbers of L. monocytogenes when compared to the untreated controls throughout the 28-day storage period (P < 0.0001). For roast beef and cooked turkey containing chemical antimicrobials treated with LISTEX^TMP100 and stored at 4 °C, no more than a 2 log CFU/cm² increase of L. monocytogenes was observed throughout the stated shelf life of the product. This study shows that LISTEX^™P100 causes an initial reduction of L. monocytogenes numbers and can serve as an additional hurdle to enhance the safety of RTE meats when used in combination with chemical antimicrobials.     

Control of Listeria monocytogenes on turkey frankfurters by generally-recognized-as-safe preservatives

Generally-recognized-as-safe chemicals applied to the surfaces of turkey frankfurters were evaluated for their ability to reduce populations of or inhibit the growth of Listeria monocytogenes. Frankfurters were treated prior to inoculation by dipping for 1 min in a solution of one of four preservatives (sodium benzoate, sodium propionate, potassium sorbate, and sodium diacetate) at three different concentrations (15, 20, and 25% [wt/vol]), with < 0.3% of the preservative being present for each frankfurter. Subsequently, 0.1 ml of a five-strain mixture of L. monocytogenes (10(6) CFU/ml) was used to surface inoculate each frankfurter separately in a sterile stomacher bag. Inoculated frankfurter bags were held at 4, 13, and 22 degrees C, and L. monocytogenes cells were enumerated at 0, 3, 7, 10, and 14 days of storage. The results of this study revealed that at all three concentrations of all four preservatives, the initial populations of L. monocytogenes decreased immediately by 1 to 2 log10 CFU/g. After 14 days of storage at 4 degrees C, L. monocytogenes counts for all treated frankfurters were 3 to 4 log10 CFU/g less than those for the untreated frankfurters. After 14 days of storage at 13 degrees C, L. monocytogenes counts for frankfurters treated with 25% sodium benzoate or 25% sodium diacetate were 3.5 to 4.5 log10 CFU/g less than those for untreated frankfurters, and those for frankfurters treated with 25% sodium propionate or 25% potassium sorbate were 2.5 log10 CFU/g less than those for untreated frankfurters. In all instances, the degree of growth inhibition was directly proportional to the concentration of the preservative. Only frankfurters treated with 25% sodium diacetate or sodium benzoate were significantly inhibitory to L. monocytogenes when held at 22 degrees C for 7 days or longer. Interestingly, the untreated frankfurters held at 22 degrees C were spoiled within 7 days, with copious slime formation, whereas there was no evidence of slime on any treated frankfurters after 14 days of storage.     

Fate of Listeria monocytogenes in commercial ham, formulated with or without antimicrobials, under conditions simulating contamination in the processing or retail environment and during home storage

Commercial cured ham formulated with or without potassium lactate and sodium diacetate was inoculated with Listeria monocytogenes and stored to simulate conditions of processing, retail, and home storage. The ham was sliced, inoculated with a 10-strain composite of L. monocytogenes (1 to 2 log CFU/cm2), vacuum packaged, and stored at 4 degrees C to simulate contamination following lethality treatment at processing (first shelf life). After 10, 20, 35, and 60 days of storage, packages were opened, samples were tested, and bags with remaining slices were reclosed with rubber bands. At the same times, portions of original product (stored at 4 degrees C in original processing bags) were sliced, inoculated, and packaged in delicatessen bags to simulate contamination during slicing at retail (second shelf life). Aerobic storage of both sets of packages at 7 degrees C for 12 days was used to reflect domestic storage conditions (home storage). L. monocytogenes populations were lower (P < 0.05) during storage in ham formulated with lactate-diacetate than in product without antimicrobials under both contamination scenarios. Inoculation of ham without lactate-diacetate allowed prolific growth of L. monocytogenes in vacuum packages during the first shelf life and was the worst case contamination scenario with respect to pathogen numbers encountered during home storage. Under the second shelf life contamination scenario, mean growth rates of the organism during home storage ranged from 0.32 to 0.45 and from 0.18 to 0.25 log CFU/cm2/day for ham without and with lactate-diacetate, respectively, and significant increases in pathogen numbers (P < 0.05) were generally observed after 4 and 8 days of storage, respectively. Regardless of contamination scenario, 12-day home storage of product without lactate-diacetate resulted in similar pathogen populations (6.0 to 6.9 log CFU/cm2) (P > 0.05). In ham containing lactate-diacetate, similar counts were found during the home storage experiment under both contamination scenarios, and only in 60-day-old product did samples from the first shelf life have higher (P < 0.05) pathogen numbers than those found in samples from the second shelf life. These results should be useful in risk assessments and for the establishment of "sell by" and "consume by" date labels for refrigerated ready-to-eat meat products.     

The fate of two Listeria monocytogenes serotypes in "cig kofte" at different storage temperatures

Cig kofte is a traditional Turkish food prepared from minced beef, bulgur, onions, garlic and varieties of spices. It is generally consumed within a few hours. However, leftovers can be kept in refrigerator or in room temperature up to 24h until they are consumed. In this study, survival and growth of two Listeria monocytogenes serotypes were investigated in cig kofte during the storage. For this purpose, the prepared samples were separately contaminated with serotypes 1/2b or 4b of L. monocytogenes at the level of 10(4)CFU/g and stored at 4°C and 21°C. L. monocytogenes colonies were counted at the beginning, 3rd, 6th, 12th and 24th hours of the storage. At 4°C, L. monocytogenes 4b significantly increased (P<0.05) from 4.12 to 5.49log(10)CFU/g but L. monocytogenes 1/2b remained constant (P>0.05) during the storage period. At 21°C, both L. monocytogenes 1/2b and 4b increased significantly (P<0.05) from 4.56 to 5.16log(10)CFU/g and from 4.23 to 5.65log(10)CFU/g, respectively. The physicochemical and microbiological characteristics of the cig kofte did not inhibit the growths of L. monocytogenes serotypes during the storage. These results indicated that L. monocytogenes was able to survive and grow in cig kofte at the both storage temperatures of 4°C and 21°C and cig kofte seemed to be a suitable medium for this pathogen.