Postprocessing antimicrobial treatments to control Listeria monocytogenes in commercial vacuum-packaged bologna and ham stored at 10 degrees C

The antilisterial effect of chemical dipping solutions on commercial bologna and ham slices, inoculated (3 to 4 log CFU/ cm2) after processing, was evaluated during storage in vacuum packages at 10 degrees C. Samples were inoculated with a 10-strain composite of Listeria monocytogenes and subsequently immersed (25+/-2 degrees C) for 2 min in 2.5% acetic acid (AA), 2.5% lactic acid (LA), 5% potassium benzoate (PB), or 0.5% Nisaplin (commercial form of nisin, equivalent to 5,000 IU/ml of nisin) solutions, either singly or sequentially (Nisaplin plus AA, Nisaplin plus LA, or Nisaplin plus PB), and then vacuum packaged and stored at 10 degrees C for 48 days. In addition to microbiological analysis, sensory evaluations were performed on uninoculated samples treated with AA, LA, or PB. Initial reductions (day 0) of the pathogen, compared with the controls, on bologna and ham samples treated with AA, LA, or PB ranged from 0.4 to 0.7 log CFU/cm2. Higher (P < 0.05) initial reductions (2.4 to 2.9 log CFU/cm2) were obtained for samples treated with Nisaplin alone and when followed by AA, LA, or PB. L. monocytogenes populations on control bologna and ham samples increased from 3.4 log CFU/cm2 (day 0) to 7.4 and 7.8 log CFU/ cm2, respectively, in 8 days at 10 degrees C. Listericidal effects were observed for all treatments tested, except for Nisaplin applied on its own, during storage at 10 degrees C. The sequential treatment of Nisaplin plus LA reduced L. monocytogenes to undetectable levels in both products at the end of storage. The sequential treatments were also found to inhibit growth of spoilage microorganisms. Sensory evaluations indicated that dipping (2 min) of ham samples in AA (2.5%), LA (2.5%), or PB (5%) led to lower sensory scores. However, since results of this study indicated that these treatments caused extensive listericidal effects, there is possibly a potential to reduce the levels of chemicals applied and still achieve adequate antilisterial activity without major negative effects on product quality.     

Survival of Listeria monocytogenes in a simulated dynamic gastrointestinal model during storage of inoculated bologna and salami slices in vacuum packages

Listeria monocytogenes counts were determined during storage (82 days, 4 degrees C) in vacuum packages of inoculated bologna and salami slices and after exposure to a simulated dynamic model of the stomach and small intestine. Variables controlled in the model included gastric emptying and gastrointestinal fluid secretion rates, gradual gastric acidification, and intestinal pH maintenance. L. monocytogenes populations increased on bologna and decreased on salami, reaching 8.7 and 1.4 log CFU/g, respectively, on day 82. Inactivation rates (IR) during gastric exposure of bologna and salami ranged from 0.079 (day 14) to 0.158 (day 57) log CFU/g/min and from 0.013 (day 42) to 0.051 (day 1) log CFU/g/min, respectively. On corresponding days, gastric IR for cells on salami were lower than on bologna, suggesting potential protective effects of the former product. However, it is also possible that the low initial L. monocytogenes levels reached with storage of salami (< or = 2.5 log CFU/g after day 27) may have resulted in slower reductions than in the high levels on bologna. Gradual decline of gastric pH allowed survival in the gastric compartment during the initial stages, which resulted in a large fraction of the cells being delivered into the intestinal compartment. Intestinal IR ranged from 0.003 to 0.048 (bologna) and from 0.002 to 0.056 (salami) log CFU/g/min throughout storage. Although findings indicated potential effects of salami against gastric killing of L. monocytogenes, any effects of the food matrix per se on the gastrointestinal survival of the pathogen were overwhelmed by the high and low contamination levels reached on bologna and salami, respectively, during storage.     

Survival of Listeria monocytogenes in a simulated recirculating brine chiller system

Contamination by Listeria monocytogenes of processed meats after cooking presents a significant food safety risk. The purpose of this study was to determine the survival of L. monocytogenes in a simulated recirculating brine chiller system using pH values of 5, 6, and 7 with free chlorine concentrations of 0, 3, 5, and 10 ppm in 20% salt brine at -12 degrees C. At pH values of 5, 6, and 7 with chlorine concentrations of 2 and 3 ppm, using 10(8) CFU in a test tube system, an immediate drop of 0.28 log CFU/ml with no significance between treatments (P > 0.05), followed by a steady survival phase with a slope close to 0, was observed. In brine at a pH of 5 with 5 and 10 ppm of chlorine, an initial drop of 0.8 log CFU/ml was observed, which was followed by a steady survival phase with a destruction slope close to zero. At an inoculation concentration of 10(2) CFU in a test tube system (pH values of 5 and 7 with 0 and 10 ppm of chlorine), the average initial drop for all treatments was 0.1 log CFU/ml, which was followed by a steady survival phase. In a recirculating system, very few cells were destroyed during the brine chilling process, but only low numbers of L. monocytogenes were recovered from the brine and uninoculated hot dogs. Although little destruction of L. monocytogenes was noted, the dilution effect observed during the study indicates that environmental contamination of a brine chiller system poses little danger of postcooking contamination for processed meats if the system is regularly cleaned and sanitized.     

Mathematical model of Listeria monocytogenes cross-contamination in a fish processing plant

Listeriosis is a foodborne disease caused by the bacterium Listeria monocytogenes. The food industry and government agencies devote considerable resources to reducing contamination of ready-to-eat foods with L. monocytogenes. Because inactivation treatments can effectively eliminate L. monocytogenes present on raw materials, postprocessing cross-contamination from the processing plant environment appears to be responsible for most L. monocytogenes food contamination events. An improved understanding of cross-contamination pathways is critical to preventing L. monocytogenes contamination. Therefore, a plant-specific mathematical model of L. monocytogenes cross-contamination was developed, which described the transmission of L. monocytogenes contamination among food, food contact surfaces, employees' gloves, and the environment. A smoked fish processing plant was used as a model system. The model estimated that 10.7% (5th and 95th percentile, 0.05% and 22.3%, respectively) of food products in a lot are likely to be contaminated with L. monocytogenes. Sensitivity analysis identified the most significant input parameters as the frequency with which employees' gloves contact food and food contact surfaces, and the frequency of changing gloves. Scenario analysis indicated that the greatest reduction of the within-lot prevalence of contaminated food products can be achieved if the raw material entering the plant is free of contamination. Zero contamination of food products in a lot was possible but rare. This model could be used in a risk assessment to quantify the potential public health benefits of in-plant control strategies to reduce cross-contamination.     

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.     

Efficacy of chlorine and a peroxyacetic acid sanitizer in killing Listeria monocytogenes on iceberg and Romaine lettuce using simulated commercial processing conditions

The efficacy of chlorine (100 microg/ml) and a peroxyacetic acid sanitizer (80 microg/ml; Tsunami 100) in killing Listeria monocytogenes inoculated at populations of 1 to 2, 2 to 3, and 4 to 5 log CFU/g of iceberg lettuce pieces, shredded iceberg lettuce, and Romaine lettuce pieces was determined by treatment conditions simulating those used by a commercial fresh-cut lettuce processor. The lettuce/treatment solution ratio was 1:100 (wt/vol), treatment temperature was 4 degrees C, and total treatment time was 30 s. Compared with washing in water, treatment of iceberg lettuce pieces containing all levels of inoculum and shredded iceberg lettuce containing 2 to 3 or 4 to 5 log CFU/g with chlorine or Tsunami resulted in significant reductions (P < or = 0.05) of pathogen populations. Populations recovered from Romaine lettuce pieces treated with chlorine or Tsunami were not significantly different from populations recovered from pieces washed with water, regardless of the inoculum level. Within lettuce type and inoculum level, in no instance was the number of L. monocytogenes recovered from lettuce treated with chlorine or Tsunami significantly different. The rate of decrease in free chlorine concentration in treatment solution as affected by the weight/volume ratio (1:100, 1:10, 2:10, and 4:10) of lettuce and solution was determined. The rate of reduction increased as the ratio decreased. The overall order of magnitude of reduction was shredded iceberg lettuce > iceberg pieces > Romaine pieces. The highest reductions in free chlorine concentration in solutions used to treat shredded lettuce are attributed to the release of tissue juices, which increases the concentration of soluble organic materials available for reaction with chlorine.     

Survival of Listeria monocytogenes in commercial cheese brines.

The survival of Listeria monocytogenes was determined in commercial cheese brines collected from cheese factories in Wisconsin and northern Illinois. Survival of L. monocytogenes inoculated into commercial cheese brines ranged from < 7 d to over 259 d. Survival did not correlate with pH, salt content, nitrogen content, mineral content, or inherent microbial populations but was negatively associated with addition of sodium hypochlorite at the dairy plant. The L. monocytogenes generally survived longer in brines held at 4 degrees C than at 12 degrees C. Sodium hypochlorite or hydrogen peroxide inactivated L. monocytogenes when added to commercial brines in the lab at 10 to 100 ppm or 0.001% to 0.02%, respectively. Addition of 1% potassium sorbate or 1% sodium benzoate also decreased survival of L. monocytogenes. Laboratory filtration of commercial brines had a negative effect on survival in one of three brines tested. The L. monocytogenes did not grow during incubation in any of the commercial brine samples tested.     

Comparison of Listeria monocytogenes virulence in a mouse model

Listeriosis results from exposure to the foodborne pathogen Listeria monocytogenes. Although many different strains of L. monocytogenes are isolated from food, no definitive tests currently predict which isolates are most virulent. The objectives of this study were to address two major data gaps for risk assessors, variability among L. monocytogenes strains in pathogenicity and virulence. Strains used in our monkey clinical trial or additional food isolates were evaluated for their virulence and infectivity in mice. All strains were equally pathogenic to immunocompromised mice, causing deaths to 50% of the population 3 days after exposure to doses ranging from 2 to 3 log CFU. Doses resulting in 50% deaths on the fifth day after administration were 1 to 2 log lower than those on the third day, indicating that the full course of pathogenicity exceeds the 3-day endpoint in immunocompromised mice. Three strains were chosen for further testing for their virulence and infectivity in liver and spleen in normal (immunocompetent) mice. Virulence was not significantly different (P > 0.05) among the three strains, all resulting in deaths to 50% of mice at 5 to 7 log CFU by 5 days after administration. All strains were equally infective in liver or spleen, with higher numbers of L. monocytogenes directly correlated with higher doses of administration. In addition, there was no preference of organs by any strains. The lack of strain differences may reflect the limitation of the mouse model and suggests the importance of using various models to evaluate the pathogenicity and virulence of L. monocytogenes strains.     

Effect of acid adaptation on growth during storage at 10 degrees C and resistance to simulated gastric fluid of Listeria monocytogenes inoculated onto bologna formulated with or without antimicrobials

The fate of acid-adapted and nonadapted Listeria monocytogenes inoculated onto bologna slices (formulated with or without antimicrobials) was examined during storage and after exposure to in vitro gastric challenge. Bologna slices formulated with no antimicrobials (control), 3% sodium lactate (SL), or 1.8% SL plus 0.25% sodium diacetate (SD) were inoculated (2 log CFU/cm2) with a 10-strain composite of acid-adapted or nonadapted L. monocytogenes strains. Growth or survival of the two inocula on bologna was evaluated during vacuum-packaged storage (10 degrees C) for up to 36 days. Survival of previously acid-adapted or nonadapted L. monocytogenes on stored bologna exposed to simulated gastric fluid (adjusted to pH 1.0 with HCl) for 20, 40, and 60 min also was determined. As expected, inclusion of antimicrobials in the product formulation inhibited growth of L. monocytogenes during storage of vacuum-packaged bologna compared with growth on control samples. Acid adaptation of L. monocytogenes prior to product inoculation did not affect subsequent survival or growth on bologna or resistance to simulated gastric fluid (P > 0.05). Survival of L. monocytogenes exposed to simulated gastric fluid during storage increased with product age, growth phase of the cells, and possibly age of the cells, particularly for control samples (no antimicrobials), in which the pathogen grew uninhibited to approximately 6 log CFU/cm2 by day 8 of storage. Inhibition of L. monocytogenes growth on product formulated with antimicrobials was associated with only sporadic and small numbers of survivors following exposure of these samples to simulated gastric fluid, especially in samples stored longer. However, cell numbers in these treatment groups before the gastric challenge did not exceed 3.8 log CFU/cm2. Inhibition of growth on product with antimicrobials precluded detection of survivors resistant to the effects of simulated gastric fluid.     

Occurrence of Campylobacter and Listeria monocytogenes in a poultry processing plant

The occurrence of Campylobacter and Listeria monocytogenes was studied in 645 samples from surfaces, water, and poultry products (chicken carcasses, chicken parts, viscera, and spoils) in a poultry processing plant in southern Brazil. The automated mini-VIDAS system was used to detect the presence of Campylobacter and L. monocytogenes on the samples. The positive samples were confirmed by conventional methods. Campylobacter and L. monocytogenes were found in 16.6 and 35.6% of the analyzed samples, respectively. The sampling points with the highest Campylobacter incidence were intestine (63.3%, 19 of 30 samples), gallbladder (46.7%, 14 of 30), carcasses before evisceration (33.33%, 10 of 30), and carcasses after plucking (30%, 9 of 30). For L. monocytogenes, the majority of positive samples were from frozen breast (100%, 15 of 15 samples), frozen wing (93.3%, 14 of 15), fresh breast (83.3%, 25 of 30), fresh wing (80%, 24 of 30), skin of breast and leg (76.7%, 23 of 30), frozen leg (60%, 9 of 15), and fresh leg (50%, 15 of 30).     

Bactericidal effect of enterocin 4 on Listeria monocytogenes in a model dairy system.

Enterococcus faecalis INIA 4 produced the bacteriocin enterocin 4 during growth in raw ewe's milk at 30 degrees C. Enterocin activity reached 2,200 to 3,600 AU/ml after 8 h, with a 1 to 8% (vol/vol) level of inoculum from an 18-h culture. An enterocin activity of 500 AU/ml significantly decreased counts of Listeria monocytogenes Ohio when incubated for 6 h in a model system consisting of filtrates from cultures of E. faecalis INIA 4 in raw ewe's milk, at pH 6.0 and 30 degrees C. However, an enterocin activity of 2,400 AU/ml was needed in the same conditions to significantly decrease counts of L. monocytogenes Scott A. All 22 wild L. monocytogenes strains isolated from ewe's milk and tested were inhibited by a filtrate containing 400 AU/ml of enterocin 4. Incubation in the filtrate for 6 h significantly lowered counts of 16 L. monocytogenes strains, and incubation for 24 h, counts of 21 strains.     

Heat inactivation of Listeria monocytogenes and Salmonella enterica serovar Typhi in a typical bologna matrix during an industrial cooking-cooling cycle

The heat resistance of Salmonella enterica serovar Typhi PF-724 and Listeria monocytogenes 2812 was determined in a commercial bologna batter. The heat inactivation of the two bacterial species was also studied in a semiautomatic pilot smokehouse under cooking conditions that reproduced an industrial bologna process. S. enterica serovar Typhi PF-724 was less heat resistant than L. monocytogenes 2812. The D-values (times required to reduce the population by 1 logarithmic cycle) for S. enterica serovar Typhi PF-724 ranged from 10.11 to 0.04 min for temperatures of 50 to 70 degrees C, while for L. monocytogenes 2812, the D-values were 2.5-, 4.9-, 3.8-, 3.3-, and 2-fold higher at 50, 55, 60, 65, and 70 degrees C, respectively, than for S. enterica serovar Typhi PF-724. However, the z-value (temperature required to reduce log D by 1 logarithmic cycle) for S. enterica serovar Typhi PF-724 (5.72 degrees C) was not significantly different from the z-value for L. monocytogenes 2812 (7.04 degrees C), indicating that a given increase in temperature would have a similar effect on the decimal reduction time for both bacterial species in that meat emulsion. Our data on experimentally inoculated batter also showed that processing bologna at a cooking-cooling cycle commonly used in the industry resulted in a minimum 5-log reduction for both S. enterica serovar Typhi PF-724 and L. monocytogenes 2812.     

Contamination patterns of Listeria monocytogenes in cold-smoked pork processing

Contamination patterns of Listeria monocytogenes were studied in a cold-smoked pork processing plant to identify the sources and possible reasons for the contamination. Environmental sampling combined with pulsed-field gel electrophoresis (PFGE) subtyping and serotyping were applied to investigate the genetic diversity of L. monocytogenes in the plant environment and ready-to-eat (RTE) cold-smoked pork products. A total of 183 samples were collected for contamination analyses, including samples of the product at different stages during manufacture (n = 136) and environmental samples (n = 47) in 2009. L. monocytogenes isolates, previously recovered from 73 RTE cold-smoked pork samples and collected from the same meat processing plant in 2004, were included in this study. The brining machine and personnel working with brining procedures were the most contaminated places with L. monocytogenes. The overall prevalence of L. monocytogenes in raw pork (18%) increased to 60% after the brining injections. The brining machine harbored six different PFGE types belonging to serotypes 1/2a, 1/2c, 4b, and 4d, which were found on the feeding teeth, smooth surfaces, and spaces of the machine, thus potentially facilitating dissemination of L. monocytogenes contamination. Two PFGE types (2 and 8) belonging to serotypes 1/2a and 1/2c were recovered from RTE cold-smoked pork collected in 2004, and from surfaces of the brining machine sampled in 2009, and may indicate the presence of persistent L. monocytogenes strains in the plant. Due to poor hygiene design, removal of the brining machine from the production of cold-smoked meat products should be considered to reduce L. monocytogenes contamination in the finished products.     

Tracking of Listeria monocytogenes in smoked fish processing plants

Four smoked fish processing plants were used as a model system to characterize Listeria monocytogenes contamination patterns in ready-to-eat food production environments. Each of the four plants was sampled monthly for approximately 1 year. At each sampling, four to six raw fish and four to six finished product samples were collected from corresponding lots. In addition, 12 to 14 environmental sponge samples were collected several hours after the start of production at sites selected as being likely contamination sources. A total of 234 raw fish, 233 finished products, and 553 environmental samples were tested. Presumptive Listeria spp. were isolated from 16.7% of the raw fish samples, 9.0% of the finished product samples, and 27.3% of the environmental samples. L. monocytogenes was isolated from 3.8% of the raw fish samples (0 to 10%, depending on the plant), 1.3% of the finished product samples (0 to 3.3%), and 12.8% of the environmental samples (0 to 29.8%). Among the environmental samples, L. monocytogenes was found in 23.7% of the samples taken from drains, 4.8% of the samples taken from food contact surfaces, 10.4% of the samples taken from employee contact surfaces (aprons, hands, and door handles), and 12.3% of the samples taken from other nonfood contact surfaces. Listeria spp. were isolated from environmental samples in each of the four plants, whereas L. monocytogenes was not found in any of the environmental samples from one plant. Overall, the L. monocytogenes prevalence in the plant environment showed a statistically significant (P < 0.0001) positive relationship with the prevalence of this organism in finished product samples. Automated EcoRI ribotyping differentiated 15 ribotypes among the 83 L. monocytogenes isolates. For each of the three plants with L. monocytogenes-positive environmental samples, one or two ribotypes seemed to persist in the plant environment during the study period. In one plant, a specific L. monocytogenes ribotype represented 44% of the L. monocytogenes-positive environmental samples and was also responsible for one of the two finished product positives found in this plant. In another plant, a specific L. monocytogenes ribotype persisted in the raw fish handling area. However, this ribotype was never isolated from the finished product area in this plant, indicating that this operation has achieved effective separation of raw and finished product areas. Molecular subtyping methods can help identify plant-specific L. monocytogenes contamination routes and thus provide the knowledge needed to implement improved L. monocytogenes control strategies.     

Validation of a stochastic modelling approach for Listeria monocytogenes growth in refrigerated foods

A stochastic modelling approach was developed to describe the distribution of Listeria monocytogenes contamination in foods throughout their shelf life. This model was designed to include the main sources of variability leading to a scattering of natural contaminations observed in food portions: the variability of the initial contamination, the variability of the biological parameters such as cardinal values and growth parameters, the variability of individual cell behaviours, the variability of pH and water activity of food as well as portion size, and the variability of storage temperatures. Simulated distributions of contamination were compared to observed distributions obtained on 5 day-old and 11 day-old cheese curd surfaces artificially contaminated with between 10 and 80 stressed cells and stored at 14 °C, to a distribution observed in cold smoked salmon artificially contaminated with approximately 13 stressed cells and stored at 8 °C, and to contaminations observed in naturally contaminated batches of smoked salmon processed by 10 manufacturers and stored for 10 days a 4 °C and then for 20 days at 8 °C. The variability of simulated contaminations was close to that observed for artificially and naturally contaminated foods leading to simulated statistical distributions properly describing the observed distributions. This model seems relevant to take into consideration the natural variability of processes governing the microbial behaviour in foods and is an effective approach to assess, for instance, the probability to exceed a critical threshold during the storage of foods like the limit of 100 CFU/g in the case of L. monocytogenes.     

Survival of Listeria monocytogenes strains in a dry sausage model

The survival of five inoculated Listeria monocytogenes strains (DCS 31, DCS 184, AT3E, HT4E, and HR5E) was studied in dry fermented sausages prepared using two different starter cultures (starter A and B) with or without a protective Lactobacillus plantarum DDEN 2205 strain. L. monocytogenes was detected throughout ripening in every sausage sample in which the L. plantarum DDEN 2205 strain had not been used. The use of either starter A, with a high concentration of protective culture, or starter B, with a low concentration of protective culture, resulted in L. monocytogenes-negative sausages after 17 days of ripening. Differential survival was noted among the L. monocytogenes strains during fermentation. Strains AT3E and DCS 31 survived in sausages with protective cultures more often than did the other strains, whereas HT4E and HR5E were inhibited during ripening by all starter and protective cultures used. Protective cultures such as L. plantarum may be used as part of a hurdle strategy in dry sausage processing, but variations in susceptibility of different L. monocytogenes strains can create problems if other hurdles are not included.     

Distribution of Listeria monocytogenes subtypes within a poultry further processing plant

Samples from environmental sites and raw product in a chicken further processing plant were collected every 6 weeks for 12 months. Each sample site was examined before and after a complete production shift. All samples were examined for the presence of Listeria monocytogenes, which was detected in floor drains on the raw product side of the plant preoperation and in drains on both raw and cooked sides following 8 h of processing operation. L. monocytogenes also was detected in raw product and once in fully cooked product but never on cooked product contact surfaces. One hundred sixty-one isolates were collected from 75 positive samples. All isolates were subtyped using a sequence-based method, and 14 unique subtypes were detected through the course of the study. Four of these types were found repeatedly and appeared to be resident in the plant. Three of the four resident strains were detected on raw product at some point during the year-long study, suggesting that raw product may be one source of L. monocytogenes in the processing plant environment. These data highlight the need for research to investigate why some types of L. monocytogenes persist in a processing plant environment but others do not.     

Loss of viability by Listeria monocytogenes in commercial bovine pepsin-rennet extract

Loss of viability by Listeria monocytogenes strains California, V7, and Scott A in commercial bovine pepsin-rennet extract was determined during storage for 56 d at 7 degrees C. Four levels (10(3) to 10(6)/ml) of L. monocytogenes were added to the coagulant, and McBride listeria agar was used to determine numbers of survivors. Selected colonies thought to be L. monocytogenes were confirmed biochemically. Samples also were tested during and after completion of cold enrichment (up to 8 wk at 4 degrees C). Coagulant inoculated with 10(3) to 10(4) L. monocytogenes/ml usually was free of viable cells of the pathogen after 28 d and sometimes after 14 d, as determined by direct plating and cold enrichment. When the inoculum was 10(5) to 10(6) cells/ml, samples of coagulant usually were free of viable L. monocytogenes after 42 d and sometimes after 28 d. The three strains of L. monocytogenes behaved similarly, although strain California was somewhat less hardy in the environment of the coagulant than were the other two strains. Bovine pepsin-rennet extract, before inoculation, was free of L. monocytogenes (direct plating and cold enrichment).     

Use of mild-heat treatment following high-pressure processing to prevent recovery of pressure-injured Listeria monocytogenes in milk

This study examined the inactivation of Listeria monocytogenes in milk by high-pressure processing (HPP) and bacterial recovery during storage after HPP. We developed a technique to inhibit the bacterial recovery during storage after HPP (550 MPa for 5 min) using a mild-heat treatment (30-50 degrees C). Various mild-heat treatments were conducted following HPP to investigate the condition on which the bacterial recovery was prevented. Immediately after HPP of 550 MPa at 25 degrees C for 5 min, no L. monocytogenes cells were detected in milk regardless of the inoculum levels (3, 5, and 7 log(10)CFU/ml). However, the number of L. monocytogenes cells increased by >8 log(10)CFU/ml regardless of the inoculum levels after 28 days of storage at 4 degrees C. Significant recovery was observed during storage at 25 degrees C; the bacterial number increased by >8 log(10)CFU/ml after 3 days of storage in the case of an initial inoculum level of 7 and 5 log(10)CFU/ml. Even in the case of an initial inoculum level of 3 log(10)CFU/ml, the bacterial number reached the level of 8 log(10)CFU/ml after 7 days of storage. No bacterial recovery was observed with storage at 37 degrees C for 28 days. Milk samples were treated by various mild-heat treatments (30-50 degrees C for 5-240 min) following HPP of 550 MPa at 25 degrees C for 5 min, and then stored at 25 degrees C for 70 days. The mild-heat treatment (e.g., 37 degrees C for 240 min or 50 degrees C for 10 min) inhibited the recovery of L. monocytogenes in milk after HPP. No recovery of L. monocytogenes in milk was observed during 70-day storage at 25 degrees C in samples that received mild-heat treatments such as mentioned above following HPP (550 MPa for 5 min). Moreover, the mild-heat treatment conditions (temperature and holding time) required to inhibit the recovery of L. monocytogenes in milk was modelled using a logistic regression procedure. The predicted interface of recovery/no recovery can be used to calculate the mild-heat treatment condition to control bacterial recovery during storage at 25 degrees C after HPP (550 MPa for 5 min). The results in this study would contribute to enhance the safety of high-pressure-processed milk.     

Incidence of Listeria monocytogenes in two milk processing environments, and assessment of Listeria monocytogenes blood agar for isolation

A year-long survey of two Northern Ireland milk processing plants for Listeria monocytogenes was carried out. Sample sites included the milk processing environment (walls, floors, drains, and steps), processing equipment, raw and pasteurised milk. The FDA listeria-selective enrichment procedure was used to process samples and an additional agar medium, L. monocytogenes Blood Agar (LMBA), was utilized as part of the isolation procedure in order to compare its performance to that of the recommended Oxford and Palcam agars. LMBA proved to be a very useful tool and was able to detect L. monocytogenes from 94.1% of sites compared to the 76.5% and 79.4% detection rate displayed by Oxford and Palcam agars, respectively. The overall incidence of listeria on equipment was 18.8% (6.3% L. monocytogenes), in the environment was 54.7% (40.6% L. monocytogenes) and in raw milk 44.4% (22.2% L. monocytogenes). On one occasion, L. welshimeri was isolated from pasteurised milk, probably demonstrating post-pasteurisation contamination of product. The main environmental sources of L. monocytogenes were considered to be a floor drain and stainless steel steps.