Efficacy of pulsed light for shelf-life extension and inactivation of Listeria monocytogenes on ready-to-eat cooked meat products

Pulsed light (PL) was tested for its utility to improve the microbial quality and safety of ready-to-eat cooked meat products. Vacuum-packaged ham and bologna slices were superficially inoculated with Listeria monocytogenes and treated with 0.7, 2.1, 4.2 and 8.4 J/cm². PL treatment at 8.4 J/cm² reduced L. monocytogenes by 1.78 cfu/cm² in cooked ham and by 1.11 cfu/cm² in bologna. The effect of PL on lipid oxidation and sensory properties was also investigated. The 2-thiobarbituric acid values were very low and chromaticity parameters were within the normal values reported for cooked meat products. PL at 8.4 J/cm² did not affect the sensory quality of cooked ham, while treatments above 2.1 J/cm² negatively influenced the sensory properties of bologna. The combination of PL and vacuum packaging provided ham with an additional shelf-life extension of 30 days compared with only vacuum packaging. The shelf-life of bologna was not extended by PL.

Industrial relevance

The efficacy of pulsed light for the decontamination of surfaces offers excellent possibilities to ensure food safety and to extend shelf-life of ready-to-eat (RTE) products. The results of this study indicate that Listeria monocytogenes can be reduced by approximately 2 log cfu/cm² in RTE cooked ham and 1 log cfu/cm² in bologna using a fluence of 8.4 J/cm². This dose does not affect the sensory properties of ham and triples its shelf-life when compared with conventional RTE products. On the contrary, fluences above 2.1 J/cm² are not suitable for the treatment of bologna since sensory quality is modified.     

Control of food spoiling bacteria in cooked meat products with nisin,lacticin 3147, and a lacticin 3147-producing starter culture

Nisin, in the form of the commercial product Nisaplin, and lacticin 3147 in whey powdered form were added to minced pork-meat in amounts of 0.15% (w/w) and 1.5% (w/w), respectively. The meat was cooked and inoculated with a Staphylococcus aureus strain of meat origin and a Listeria innocua strain at a level of 10⁷ or 10⁵ CFU g^–1. The batches were stored vacuum-packaged for 21 days at 8 °C. Nisin and lacticin 3147 immediately reduced the L. innocua population at the time of inoculation. Nisin showed higher inhibitory activity than lacticin 3147. During the storage period, a slight L. innocua growth was observed in the batches inoculated with the larger inoculum, and a bacteriostatic effect was observed against Listeria in the batches inoculated with 10⁵ CFU g^–1. Nisin maintained a constant S. aureus population in the cooked batch inoculated with 10⁷ CFU g^–1, although the bacteriocin was capable of reducing the amount of S. aureus by 90% in the batch inoculated with 10⁵ CFU g^–1. On the other hand, lacticin 3147 did not show an inhibitory effect against S. aureus in the cooked meat. The starter culture Lactococcus lactis DPC 303-T4 (containing the conjugative plasmid encoding production of lacticin 3147) was inoculated in a portion of a Longissimus dorsi pork muscle with brine. L. lactis DPC 303-T4 performed a good fermentation, but lacticin 3147 production was not found after 7 days at 12 °C of storage.     

Safety and quality of ready-to-eat dry fermented sausages subjected to E-beam radiation

The inactivation kinetics in the death of Listeria innocua NTC 11288 (more radioresistant than five different strains of Listeria monocytogenes) and Salmonella Enterica serovar Enteritidis and S. enterica serovar Typhimurium by E-beam irradiation has been studied in two types of vacuum-packed RTE dry fermented sausages (“salchichon” and “chorizo”) in order to optimize the sanitation treatment of these products. A treatment of 1.29 kGy was calculated to reach the food safety objective (FSO) according to the “zero tolerance” criterion for the three strains. No irradiation treatment was necessary to meet the 10² c.f.u./g microbiological criterion for L. monocytogenes. Dry fermented sausages treated with 2 kGy had negligible sensory (appearance, odour and taste) modifications. Therefore, this treatment produces safe dry fermented sausages with similar sensory properties to the non-irradiated product.     

Biochemical and genetic evidence for production of enterocins A and B by Enterococcus faecium WHE 81

Enterococcus faecium WHE 81, isolated from cheese, has been reported to produce a bacteriocin called “enterocin 81” [J. Appl. Microbiol. 85 (1998) 521.]. Purification of “enterocin 81” was carried out using ammonium sulfate precipitation, desalting on ODP-90 reverse-phase column, and purification through SP Sepharose HP cation exchange and C₂/C₁₈ reverse-phase chromatographies. The antimicrobial was eluted from the C₂/C₁₈ column as four individually active fractions, designated A81, B81, C81 and D81. The purification procedure used proved particularly efficient for the bacteriocin in fraction D81, with a yield of 46%, while only 3.8% the bacteriocin in fraction B81 could be collected. MALDI-TOF mass spectrometry of the bacteriocins in fractions B81 and D81 showed respective masses of 4833.0 and 5462.2 Da. Amino acid sequencing of the two peptides revealed two class-II bacteriocins whose sequences were similar to those of enterocin A and enterocin B, respectively. Using proper primers, chromosomal fragments of 212 and 216 bp enclosing bacteriocin structural genes were PCR-amplified. Cloning of the amplicons and their sequencing revealed two genes with sequences identical to the structural genes of enterocins A and B, respectively. It was therefore clearly established that E. faecium WHE 81 produces bacteriocins respectively identical to enterocins A and B. Our results, combined with data from previous reports, suggest that the two bacteriocins may be widespread among enterococcal strains and may play an important role in controlling the growth of pathogens and other undesirable bacteria in certain fermented food products.     

Effects of curing sodium nitrite additive and natural meat fat on growth control of Listeria monocytogenes by the bacteriocin-producing Lactobacillus curvatus strain CWBI-B28

In realistic model meat systems, the separate and combined effects of fat content and sodium nitrite on the antilisterial activity of the bacteriocin of Lactobacillus curvatus CWBI-B28 were studied. In laboratory fermentations where Listeria monocytogenes was co-cultured at 4 °C with bacteriocin-producing CWBI-B28 in lean pork meat (fat content: 13%) without added nitrite, a strong antilisterial effect was observed after one week. The effect was maintained for an additional week, after which a slight and very gradual rebound was observed. Both added nitrite (20 ppm) and a high-fat content (43%) were found to antagonise this antilisterial effect, the Listeria cfu count reached after six weeks being 200 times as high in high-fat meat with added nitrite than in lean meat without nitrite. This antagonism could not be attributed to slower growth of the bacteriocin-producing strain, since CWBI-B28 grew optimally in fat-rich meat with 20 ppm sodium nitrite. Bacteriocin activity was also measured in the samples. The observed activity levels are discussed in relation to the degree of antilisterial protection conferred.     

Lack of growth of Listeria monocytogenes and Staphylococcus aureus in temperature abuse of E-beam treated ready-to-eat (RTE) cooked ham

The behaviour of Listeria monocytogenes and Staphylococcus aureus in vacuum-packed cooked ham slices treated with an electron beam and stored at 4, 7 and 10 °C was investigated. Cooked ham slices were inoculated with L. monocytogenes and S. aureus and electron beam treated at 2 and 3 kGy. After treatment, a long temperature-dependent death phase was observed, followed by growth at a slower rate than in untreated samples. Assuming a hypothetical load of 10 cells/g or cm² of L. monocytogenes and S. aureus as an original contamination in an industrial situation, an E-beam treatment of vacuum-packed cooked ham slices at 2 kGy guarantees the microbiological safety of the product along its shelf life, even if a noticeable temperature (10 °C) abuse occur during its storage period. Likewise, the E-beam treatment gave rise to a substantial increase of the RTE cooked ham shelf life off-sensory features associated to the spoilage only were detected in non-treated samples (controls) after 8 and 18 days of storage at 10 °C and 7 °C, respectively.     

Evaluation of the sensitivity of microbiological criteria for Listeria monocytogenes in detecting unsafe food according to the prevalence of the pathogen and the shelf-life of the food

The sensitivity of criteria in detecting unsafe food was studied for a hypothetical ready-to-eat food supporting growth of Listeria monocytogenes during storage. The effects of the prevalence of the pathogen and of the duration of the shelf-life on the probability for servings to be unsafe and on the probabilities of detecting unsafe food were evaluated. The results obtained in this example show that the probability for servings to be unsafe increases with the prevalence and the shelf-life while the probabilities of detecting unsafe food depend above all on the prevalence of the pathogen. The current criteria used for L. monocytogenes can then sometimes be relatively ineffective to prevent the consumption of unsafe food with regard to the shelf-life of foods. The exposure assessment approach described in this study could be used to establish shelf-lives consistent with a defined acceptable risk.     

Effects of high hydrostatic pressure and varying concentrations of sodium nitrite from traditional and vegetable-based sources on the growth of Listeria monocytogenes on ready-to-eat (RTE) sliced ham

The objective of this study was to determine the effect the source of added nitrite and high hydrostatic pressure (HHP) had on the growth of Listeria monocytogenes on ready-to-eat (RTE) sliced ham. Use of 600 MPa HHP for 3 min resulted in an immediate 3.9–4.3 log CFU/g reduction in L. monocytogenes numbers, while use of 400 MPa HHP (3 min) provided less than 1 log CFU/g reduction. With the 600 MPa HHP treatment, sliced ham with a conventional concentration of sodium nitrite (200 ppm) was not different in L. monocytogenes growth from use with 50 or 100 ppm of sodium nitrite in pre-converted celery powder. Instrumental color values as well as residual nitrite and residual nitrate concentrations for cured (sodium nitrite and nitrite from celery powder) and uncured ham formulations are discussed.     

Microbial heat resistance of Listeria monocytogenes and the impact on ready-to-eat meat quality after post-package pasteurization

Several methods using bactericides, hydrostatic pressure, and post-package pasteurization technologies to control Listeria monocytogenes (LM) in ready-to-eat meats have been attempted. In addition to controlling LM contamination, any newly developed technology must have minimal effects on organoleptic properties. The objectives of this study were to: (1) determine the heat resistance of LM in two brands (A and B) of bologna differing in formulations, and, (2) evaluate the effects of post-package pasteurization on product quality. Fat content did not affect LM heat resistance in bologna at 55, 60, and 65 °C; however, Brand B bologna had a numerically lower inactivation rate. Microbial heat resistance differed (P < 0.05) with changes in pasteurization temperature. Time and temperature affected (P < 0.05) cook-loss and L^∗ Hunter color value for both bologna brands. These data show that post-package pasteurization is effective but suggest that meat formulations may need modification to prevent development of negative quality characteristics.     

Inactivation of Listeria monocytogenes on agar and processed meat surfaces by atmospheric pressure plasma jets

An apparatus for generating atmospheric pressure plasma (APP) jet was used to investigate the inactivation of Listeria monocytogenes on the surface of agar plates and slices of cooked chicken breast and ham. He, N₂ (both 7 L/min), and mixtures of each with O₂ (0.07 L/min) were used to produce the plasma jets. After treatment for 2 min with APP jets of He, He + O₂, N₂, or N₂ + O₂, the numbers of L. monocytogenes on agar plates were reduced by 0.87, 4.19, 4.26, and 7.59 log units, respectively. Similar treatments reduced the L. monocytogenes inoculated onto sliced chicken breast and ham by 1.37 to 4.73 and 1.94 to 6.52 log units, respectively, according to the input gas used with the N₂ + O₂ mixture being the most effective. Most APP jets reduced the numbers of aerobic bacteria on the meat surfaces to <10² CFU/g, and the numbers remained below that level of detection after storage at 10 °C for 7 days. The results indicate that APP jets are effective for the inactivation of L. monocytogenes on sliced meats and for prolonging the shelf-life of such foods.     

High pressure treatments on the inactivation of Salmonella Enteritidis and the physicochemical, rheological and color characteristics of sliced vacuum-packaged dry-cured ham

The effect of high pressure (HP) on Salmonella Enteritidis in sliced dry-cured ham stored under temperature abuse (8°C) during 60d was investigated. After treatment, reductions of S. Enteritidis were 1.06, 2.54 and 4.32 log units in ham treated at 400, 500 and 600MPa for 5min at 12°C, compared to non-pressurized samples. After 60d, counts of S. Enteritidis in ham treated at 400 and 500MPa were 2.56 and 2.66 log units lower than in non-treated ham, whereas the pathogen was only detected after enrichment in ham treated at 600MPa. Lipid oxidation increased with storage and pressurization, whereas total free amino acid contents were similar in HP and control samples after 60d. Dry-cured ham treated at the highest pressures exhibited lower shear resistance, whereas the maximum force to compress the sample was slightly changed. Color (L*, a* and b*) varied with pressurization and storage. Changes induced by HP in dry-cured ham were attenuated during storage.     

Modeling the growth rate and lag time of different strains of Salmonella enterica and Listeria monocytogenes in ready-to-eat lettuce

The growth parameters (growth rate, μ and lag time, λ) of three different strains each of Salmonella enterica and Listeria monocytogenes in minimally processed lettuce (MPL) and their changes as a function of temperature were modeled. MPL were packed under modified atmosphere (5% O₂, 15% CO₂ and 80% N₂), stored at 7–30 °C and samples collected at different time intervals were enumerated for S. enterica and L. monocytogenes. Growth curves and equations describing the relationship between μ and λ as a function of temperature were constructed using the DMFit Excel add-in and through linear regression, respectively. The predicted growth parameters for the pathogens observed in this study were compared to ComBase, Pathogen modeling program (PMP) and data from the literature. High R² values (0.97 and 0.93) were observed for average growth curves of different strains of pathogens grown on MPL. Secondary models of μ and λ for both pathogens followed a linear trend with high R² values (>0.90). Root mean square error (RMSE) showed that the models obtained are accurate and suitable for modeling the growth of S. enterica and L. monocytogenes in MP lettuce. The current study provides growth models for these foodborne pathogens that can be used in microbial risk assessment.     

Variation of microbial load and visual quality of ready-to-eat salads by vegetable type,season, processor and retailer

Microbial components and visual quality were determined on 1158 consumer units of ready-to-eat salads from several processors, two per each of 579 process lots, with residual shelf-life varying around a mode of five days, collected over 19 months in the years 2006–2008 from retail stores of two Italian cities close to a major producing and processing area. The salads were mainly baby leaf of single species (lettuce, arugula, spinach, lamb’s lettuce), with approximately 10% of the lots made up by mixes of 2–4 species. One unit per lot was analyzed on the day of collection and the other at the consume-by date. No Salmonella or Listeria monocytogenes was found (detection limit: presence in 25 g). Escherichia coli was detected in 27% of the lots (detection limit: 5 cfu/g), with probability of occurrence and counts highest in Autumn and for lettuce and arugula. Average visual quality was higher and other components of the microbial load were lower in Winter and Spring compared to Summer and Autumn (−0.6 log cfu/g of total aerobic counts, −1.3 log cfu/g of coliforms, −0.6 log cfu/g of yeasts and moulds). Lactic acid bacteria were detected more frequently in Spring and Summer (up to 50% of the lots). The rate of increase of microbial populations during shelf life was not affected by the level of initial contamination. At the consume-by date total aerobic count exceeded 7.2 log cfu/g for 50% of the lots and 7.7 log cfu/g for 25%. Salads from the biggest processor and retailer showed slightly higher visual quality scores, lower odds of E. coli occurrence and lower microbial loads. Visual quality scores showed significant negative relationships with the levels of lactic acid bacteria, coliforms and total viable counts.     

Evaluation of the effect of defrosting practices of ground beef on the heat tolerance of Listeria monocytogenes and Salmonella Enteritidis

The effect of common defrosting practices of ground beef, including (i) defrosting in the refrigerator (5 °C for 15 h), (ii) defrosting at room temperature (25 °C for 12 h) and (iii) defrosting in the microwave, on the heat tolerance of artificially inoculated Listeria monocytogenes and Salmonella Enteritidis, was studied. The thermal inactivation of S. Enteritidis was not, overall, affected by defrosting practices. In contrast, defrosting at room temperature resulted, overall, in an increased heat tolerance of L. monocytogenes compared to the rest tested defrosting practices. Inactivation kinetics of the two pathogens for the different defrosting practices were determined by fitting the data to the Weibull model. The δ parameter of the Weibull model (heat challenge time (min) required for the first 1-log reduction) for S. Enteritidis and for defrosting at 25 °C, microwave defrosting, defrosting at 5 °C and for the control (fresh ground beef inoculated with the pathogens just before the heat challenge trials) was 1.13, 1.62, 1.60 and 0.96, respectively, while the corresponding values for L. monocytogenes were 20.13, 10.82, 9.95 and 9.47, respectively. The findings of this study should be useful in risk assessments and in developing food handling guidelines for the consumers.     

Effect of Zataria multiflora Boiss. essential oil and nisin on Salmonella typhimurium and Staphylococcus aureus in a food model system and on the bacterial cell membranes

The effects of different concentrations of Zataria multiflora Boiss. essential oil (EO: 0, 5, 15 and 30 μl 100 ml⁻¹) and nisin (N: 0, 0.25 and 0.5 μg ml⁻¹), temperatures (T: 25 and 8 °C), and storage times (up to 21 days) on growth of Salmonella typhimurium and Staphylococcus aureus in a commercial barley soup were evaluated in a factorial design study. The growth of S. typhimurium was significantly (P < 0.05) decreased by EO concentrations and their combinations with N concentrations at 8 °C. For S. aureus, the viable count was significantly (P < 0.05) inhibited by EO and N concentrations and their combinations, incubated at both storage temperatures. The mechanism of the antimicrobial action of EO, N, and their combinations against cell membranes of the tested organisms were also studied by measurement of the release of cell constituents and by the electronic microscopy observations of the cells. The significant increase of the cell constituents’ release of both organisms was observed as a result of treatments with EO and EO in combination with N. Electronic microscopy observations revealed that the cell membranes of S. typhimurium treated by EO and EO in combination with N were significantly damaged, while cells treated with only N looked similar to untreated cells. The electron micrographs of treated cells of S. aureus with EO, N, and their combination also showed important morphological damages and disrupted membranes.     

The microbiological safety of ready-to-eat specialty meats from markets and specialty food shops: A UK wide study with a focus on Salmonella and Listeria monocytogenes

From 2359 specialty meats (continental sausages, cured/fermented, dried meats) sampled from markets and specialty food shops, 98.9% of samples were of satisfactory or acceptable microbiological quality. However, 16 (0.7%) were unsatisfactory as a result of Escherichia coli, Staphylococcus aureus or Listeria spp. contamination (≥10² CFU/g), and nine (0.4%) were unacceptable due to presence of Salmonella spp. or Listeria monocytogenes (>10² CFU/g). Meats with unacceptable levels of L. monocytogenes were within shelf life (range: 8–143 days remaining). Nine different subtypes of L. monocytogenes were detected with sero/AFLP type 1/2c VII predominating (37%), although this subtype was not overrepresented in any particular meat type (P > 0.05). Ninety-six percent of continental sausages and cured/fermented products were stored at <8 °C at premises, including seven of the nine unacceptable samples. These nine meats were all pre-packed prior to supply to retail premises (OR = 0.1 P = 0.003) indicating that contamination with bacterial pathogens occurred earlier in the production chain. Most samples (72.7%, 8/11) with unsatisfactory levels of E. coli were sliced on request, suggesting cross-contamination at point of sale. This study highlights the importance of ensuring that products do not become contaminated before final packaging, that storage conditions are controlled, and that durability dates are an accurate indication of the shelf life of the product so as to minimise the potential for L. monocytogenes to be present at levels hazardous to health at the point of sale.     

Combined effect of enterocin AS-48 and high hydrostatic pressure to control food-borne pathogens inoculated in low acid fermented sausages

The single and combined effects of enterocin AS-48 and high hydrostatic pressure (HHP) on Listeria monocytogenes, Salmonellaenterica, and Staphylococcus aureus was investigated in fuet (a low acid fermented sausage) during ripening and storage at 7 °C or at room temperature. AS-48 (148 AU g⁻¹) caused a drastic 5.5 log cfu g⁻¹ decrease in L. monocytogenes (P < 0.001) and a significant (P < 0.01) inhibition (1.79 logs) for Salmonella at the end of ripening (10 d). After pressurization (400 MPa) and storage Listeria counts remained below 5 cfu g⁻¹ in all fuets containing AS-48 (pressurized or not). HHP alone had no anti-Listeria effect. HHP treatment significantly reduced Salmonella counts, with lowest levels in pressurized fuets with AS-48. S. aureus showed similar growth for all treatments and storage conditions. These results indicate that AS-48 can be applied alone to control L. monocytogenes and combined with HHP treatment to control Salmonella in fuets.     

Evaluation of atmospheric pressure plasma to improve the safety of sliced cheese and ham inoculated by 3-strain cocktail Listeria monocytogenes

The objective of this study was to evaluate the efficacy of atmospheric pressure plasma (APP), which is capable of operating at atmospheric pressure in air, in sliced cheese and ham inoculated by 3-strain cocktail of Listeria monocytogenes (ATCC 19114, 19115, and 19111, LMC). The process parameters considered were input power (75, 100, 125, and 150 W) and plasma exposure time (60, 90, and 120 s). Microbial log reduction increased with increases of input power and plasma exposure time. After 120 s APP treatments at 75, 100, and 125 W, the viable cells of LMC were reduced by 1.70, 2.78, and 5.82 log in sliced cheese, respectively. More than 8 log reductions can be achieved in 120 s at 150 W. In contrast, reductions after 120 s ranged from 0.25 to 1.73 log CFU/g in sliced ham. Calculated D values, the exposure time required to inactivate 90% of a population, from the survival curves of 75, 100, 125, and 150 W of APP treatments were 71.43, 62.50, 19.65, and 17.27 s for LMC in sliced cheese, respectively, and those in sliced ham were 476.19, 87.72, 70.92, and 63.69 s. No viable cells were detected at 125 and 150 W of APP treatment in sliced cheese, irrespective of plasma exposure time, after 1 week at a detection limit of 10¹ CFU/g. These results indicate that the inactivation effects of APP on L. monocytogenes are strongly dependent on the type of food.     

Evaluation of hygiene practices and microbiological quality of cooked meat products during slicing and handling at retail

Cooked meat ready-to-eat products are recognized to be contaminated during slicing which, in the last years, has been associated with several outbreaks. This work aimed to find out possible relation between the hygiene practice taking place at retail point during slicing of cooked meat products in small and medium-sized establishments (SMEs) and large-sized establishments (LEs) and the microbiological quality of sliced cooked meat products. For that, a checklist was drawn up and filled in based on scoring handling practice during slicing in different establishments in Cordoba (Southern Spain). In addition, sliced cooked meats were analyzed for different microbiological indicators and investigated for the presence of Listeria spp. and Listeria monocytogenes. Results indicated that SMEs showed a more deficient handling practices compared to LEs. In spite of these differences, microbiological counts indicated similar microbiological quality in cooked meat samples for both types of establishments. On the other hand, Listeria monocytogenes and Listeria inocua were isolated from 7.35% (5/68) and 8.82% (6/68) of analyzed samples, respectively. Positive samples for Listeria spp. were found in establishments which showed acceptable hygiene levels, though contamination could be associated to the lack of exclusiveness of slicers at retail points. Moreover, Listeria spp presence could not be statistically linked to any microbiological parameters; however, it was observed that seasonality influenced significantly (P < 0.05) L. monocytogenes presence, being all samples found during warm season (5/5). As a conclusion, results suggested that more effort should be made to adequately educate handlers in food hygiene practices, focused specially on SMEs.     

Role of quantity and quality of fat in meat models inoculated with Listeria innocua or Salmonella Typhimurium treated by high pressure and refrigerated stored

Several variables can influence the effects of high hydrostatic pressure processing (HPP), but the role of fat in the treated sample is still uncertain. We designed a model by which controlling the known variables we could elucidate that role. We applied 400 MPa for 2 min to minced chicken samples inoculated with Listeria innocua and Salmonella Typhimurium mixed with 10% and 20% of three fat types with different fatty acid composition. Microbial counts were performed during 60 days of refrigerated storage either at 2 °C or 8 °C.Immediately after HPP bacterial growth was independent of the type and percentage of fat content, but a possible effect of type of fat could be observed after 60 days of cold storage.