Response of Listeria monocytogenes and Vibrio parahaemolyticus to High Hydrostatic Pressure

Listeria monocytogenes Scott A and CA, were subjected at 23°C to hydrostatic pressures ranging from 2,380 to 3,400 atm and Vibrio parahaemolyticus T-3765-1 from 680 to 1,700 atm. For L. monocytogenes Scott A, pressurization in ultra-high temperature-processed (UHT) milk and raw milk appeared to provide a protective effect and lessened cell death as compared to pressurization in phosphate-buffered saline (100 mM, pH 7.0). A population of about 10⁶ CFU/mL L. monocytogenes was killed by exposure to 3,400 atm within 80 min at 23°C in UHT milk. A population of about 10⁶ CFU/mL V. parahaemolyticus was killed by exposure to 1,700 atm within 10 min at 23°C in clam juice.     

The fate of Listeria monocytogenes during the manufacture of Camembert-type cheese: A comparison between raw milk and milk treated with high hydrostatic pressure

Camembert-type cheese was produced from: raw bovine milk; raw milk inoculated with 2 or 4 log CFU/ml Listeria monocytogenes; raw milk inoculated with L. monocytogenes and subsequently pressure-treated at 500 MPa for 10 min at 20 °C; or uninoculated raw milk pressure-treated under these conditions. Cheeses produced from both pressure-treated milk and untreated milk had the typical composition, appearance and aroma of Camembert. Curd and cheese made from inoculated, untreated milk contained large numbers of L. monocytogenes throughout production. An initial inoculum of 1.95 log CFU/ml in milk increased to 4.52 log CFU/g in the curd and remained at a high level during ripening, with 3.85 log CFU/g in the final cheese. Pressure treatment inactivated L. monocytogenes in the raw milk at both inoculum levels and the pathogen was not detected in any of the final cheeses produced from pressure-treated milk. Therefore high pressure may be useful to inactivate L. monocytogenes in raw milk that is to be used for the production of soft, mould-ripened cheese.

Industrial relevance

This paper demonstrates the potential of high pressure (HP) for treatment of raw milk to be used in the manufacture of soft cheeses. HP treatment significantly reduced the level of Listeria monocytogenes in the raw milk and so allowed the production of safer non-thermally processed camembert-like soft cheese.     

Inactivation of Listeria innocua in milk and orange juice by ultrahigh-pressure homogenization

The aim of this work was to evaluate the bactericidal efficacy of ultrahigh-pressure homogenization (UHPH) against Listeria innocua ATCC 33090 inoculated into milk and orange juice. We also intended to study the effect of inlet temperature on the lethality and production of sublethal injuries in this microorganism and its ability to survive, repair, and grow in refrigerated storage after UHPH treatment. Samples of ultrahigh-temperature whole milk and ultrahigh-temperature orange juice inoculated at a concentration of approximately 7.0 log (CFU per milliliter) were immediately pressurized at 300 MPa on the primary homogenizing valve and at 30 MPa on the secondary valve, with inlet temperatures of 6.0 +/- 1.0 degrees C and 20 +/- 1.0 degrees C. L. innocua viable counts and injured cells were measured 2 h after UHPH treatment and after 3, 6, and 9 days of storage at 4 degrees C for milk and after 3, 6, 9, 12, 15, 18, and 21 days of storage at 4 degrees C for orange juice. Both the inlet temperature and the food matrix influenced significantly (P < 0.05) the inactivation of L. innocua, which was higher in whole milk at the 20 degrees C inlet temperature. The UHPH treatment caused few or no sublethal injuries in L. innocua. During storage at 4 degrees C after treatments, counts increased by approximately 2 logarithmic units from day 0 to 9 in whole milk, whereas in orange juice counts diminished by approximately 2.5 logarithmic units from day 0 to 18.     

Development of a rapid, simple paddle-style dipstick dye immunoassay specific for Listeria monocytogenes

We have developed two types of new paddle-style dipstick dye immunoassays. The first is genus Listeria specific and the second is specific to Listeria monocytogenes. They are based respectively, on antisera raised against heat-killed L. monocytogenes cells and against internalin B crude extract, a virulence protein found only in the pathogenic L. monocytogenes. The minimum detectable level for L. monocytogenes is 2×10⁷ CFU ml⁻¹ for strain number 88/049 in pure culture. Detection is unaffected by the presence of high numbers (approximately log 8.0 CFU/ml) of the other microorganisms tested. When the dipsticks were applied to milk samples inoculated with L. monocytogenes reference material (ALM92), there was a strong response to the enrichment cultures. The new assay may prove useful in detection of L. monocytogenes in enrichment cultures of milk and ice cream food samples.     

Antimicrobial activity of buckwheat starch films containing zinc oxide nanoparticles against Listeria monocytogenes on mushrooms

Buckwheat starch (BS) films containing zinc oxide nanoparticles (ZnO‐N; 0%, 1.5%, 3% and 4.5%) were prepared, and their physical, optical and antimicrobial properties were examined. As ZnO‐N content increased from 0% to 4.5%, TS increased from 14.99 to 19.09 MPa and E decreased from 25.60% to 20.65%. In addition, L* and a* values decreased, whereas b*, ΔE and opacity increased. Regarding antimicrobial activity, the BS/ZnO‐N films had the reductions of 2.96–3.74 log CFU mL^?1 against Listeria monocytogenes after 8 h based on viable cell count assay. The BS film containing 3% ZnO‐N, an optimal concentration chosen in this study, was applied to fresh‐cut mushroom packaging, and the film exhibited antimicrobial activity against L. monocytogenes, resulting in a reduction of 0.86 log CFU g^?1 after 6 days of storage. Thus, these results indicate that the BS/ZnO‐N film can be used as a biodegradable packaging material.     

Inhibitor‐Assisted High‐Pressure Inactivation of Bacteria in Skim Milk

The combined inactivation effects of high hydrostatic pressure (HHP) and antimicrobial compounds (potassium sorbate and ε‐polylysine [ε‐PL]) on 4 different bacterial strains present in skim milk and the effect of these treatments on milk quality were investigated in this study. HHP treatment at 500 MPa for 5 min reduced the populations of Escherichia coli, Salmonella enterica Typhimurium, Listeria monocytogenes, and Staphylococcus aureus from 6.5 log colony‐forming units (CFUs) or higher to less than 1 log CFU/mL. Compared to HHP alone, HHP with potassium or ε‐PL resulted in significantly higher reductions in the bacterial counts. After 5 min of treatment with HHP (500 MPa) and ε‐PL (2 mg/mL), no growth of E. coli, S. enterica Typhimurium, or L. monocytogenes in skim milk was observed during 15 d of refrigerated storage (4 ± 1 °C). Scanning electron microscopy analysis revealed that the synergistic treatments caused more serious damage to the bacterial cell walls. Quality assessments of the treated samples indicated that the combined treatments did not influence the color, the turbidity, the concentrations of –SH group of the proteins, or the in vitro digestion patterns of the milk. This study demonstrates that HHP with potassium or ε‐PL may be useful in the processing of milk or milk‐containing foods.     

Searching potential candidates for development of protective cultures: Evaluation of two Lactobacillus strains to reduce Listeria monocytogenes in artificially contaminated milk

The aims of this work were (a) to select the minimum concentrations of two bacteriocin-producing lactobacilli strains to reduce Listeria monocytogenes growth in whole milk, (b) to evaluate the individual and combined application of these chosen lactobacilli concentrations as protective culture in this food and, (c) to estimate if lactobacilli growth caused undesirable changes in the visible characteristics of milk. The selected minimum lactobacilli concentration with antilisterial activity was 10⁶ CFU ml⁻¹ for both lactobacilli. Although a high initial concentration of L. monocytogenes (10⁴ CFU ml⁻¹) was added to milk, a notable reduction on listerial counts (2–4 log CFU ml⁻¹) was achieved in comparison to the control. These lactobacilli showed a good adaptability in milk reaching optimal counts and not causing undesirable visible changes. In conclusion, the use of these lactobacilli could be considered as a bio-strategy potentially effective to limit the contamination by L. monocytogenes in certain milk-derived products.     

Investigating the control of Listeria monocytogenes on alternatively-cured frankfurters using natural antimicrobial ingredients or post-lethality interventions

The objective of this study was to investigate natural antimicrobials including cranberry powder, dried vinegar and lemon juice/vinegar concentrate, and post-lethality interventions (lauric arginate, octanoic acid, thermal treatment and high hydrostatic pressure) for the control of Listeria monocytogenes on alternatively-cured frankfurters. Lauric arginate, octanoic acid, and high hydrostatic pressure (400 MPa) reduced L. monocytogenes populations by 2.28, 2.03, and 1.88 log10 CFU per g compared to the control. L. monocytogenes grew in all post-lethality intervention treatments, except after a 600 MPa high hydrostatic pressure treatment for 4 min. Cranberry powder did not inhibit the growth of L. monocytogenes, while a dried vinegar and a vinegar/lemon juice concentrate did. This study demonstrated the bactericidal properties of high hydrostatic pressure, octanoic acid and lauric arginate, and the bacteriostatic potential of natural antimicrobial ingredients such as dried vinegar and vinegar/lemon juice concentrate against L. monocytogenes.     

Wound healing effect of cuttlebone extract in burn injury of rat

This study was designed to evaluate the morphological and physiological characteristics of high pressure (HP)-treated Listeria monocytogenes treated at different pHs. L. monocytogenes in phosphate buffered saline (PBS, pH 7.2) adjusted with HCl or lactic acid to pH 4.0, 5.0, and 6.0 was subjected to 300 MPa for 5 min. The numbers of HP-treated L. monocytogenes at pH 4.0 adjusted with HCl or lactic acid were reduced by more than 6 log CFU/mL, while the numbers of non-HP-treated L. monocytogenes were not significantly reduced at pH 4.0. The propidium iodide (PI) fluorescence intensity of HP-treated cells adjusted with lactic acid was increased from 65 (pH 6.0) to 78% (pH 4.0). The HP-induced inactivation of L. monocytogenes in low pH was not directly associated with the membrane disruption. The results provide valuable information for understanding the HP-induced inactivation mechanisms and enhancing microbial lethality in acidic food systems.     

Antilisterial Properties of Marinades during Refrigerated Storage and Microwave Oven Reheating against Post‐Cooking Inoculated Chicken Breast Meat

This study evaluated growth of Listeria monocytogenes inoculated on cooked chicken meat with different marinades and survival of the pathogen as affected by microwave oven reheating. During aerobic storage at 7 °C, on days 0, 1, 2, 4, and 7, samples were reheated by microwave oven (1100 W) for 45 or 90 s and analyzed microbiologically. L. monocytogenes counts on nonmarinated (control) samples increased (P < 0.05) from 2.7 ± 0.1 (day‐0) to 6.9 ± 0.1 (day‐7) log CFU/g during storage. Initial (day‐0) pathogen counts of marinated samples were <0.5 log CFU/g lower than those of the control, irrespective of marinating treatment. At 7 d of storage, pathogen levels on samples marinated with tomato juice were not different (P ≥ 0.05; 6.9 ± 0.1 log CFU/g) from those of the control, whereas for samples treated with the remaining marinades, pathogen counts were 0.7 (soy sauce) to 2.0 (lemon juice) log CFU/g lower (P < 0.05) than those of the control. Microwave oven reheating reduced L. monocytogenes counts by 1.9 to 4.1 (45 s) and >2.4 to 5.0 (90 s) log CFU/g. With similar trends across different marinates, the high levels of L. monocytogenes survivors found after microwave reheating, especially after storage for more than 2 d, indicate that length of storage and reheating time need to be considered for safe consumption of leftover cooked chicken.     

Effect of carbon dioxide pressure on Listeria monocytogenes in physiological saline and foods

The effect of 15,30 and 60 atm CO₂pressure at 25,35 and 45°C on Listeria monocytogenes was studied. A biphasic curve was observed in the CO₂treatments. A primary curve was characterized with very little killing and a secondary curve was showed straight-line inactivation kinetics. An increase of pressure and/or temperature enhanced the antimicrobial effects of CO₂. Listeria monocytogenes suspended in physiological saline containing 1% brain–heart infusion broth was completely inactivated under 60 atm CO₂treatment in 115,75 and 60 min at 25,35 and 45°C, respectively. A minimum D -value was obtained under 60 atm CO₂pressure at 45°C. High pressure CO₂treatment technique can possibly be applied to reduce microbiol load in whole and skim milk; and carrot, orange, peach, and apple juices.     

Inactivation of Listeria monocytogenes in Skim Milk and Liquid Egg White by Antimicrobial Bottle Coating with Polylactic Acid and Nisin

ABSTRACT: This study was to develop an antimicrobial bottle coating method to reduce the risk of outbreaks of human listeriosis caused by contaminated liquid foods. Liquid egg white and skim milk were inoculated with Listeria monocytogenes Scott A and stored in glass jars that were coated with a mixture of polylactic acid (PLA) polymer and nisin. The efficacy of PLA per nisin coating in inactivating L. monocytogenes was investigated at 10 and 4 °C. The pathogen grew well in skim milk without PLA/nisin coating treatments, reaching 8 log CFU/mL after 10 d and then remained constant up to 42 d at 10 °C. The growth of Listeria at 4 °C was slower than that at 10 °C, taking 21 d to obtain 8 log CFU/mL. At both storage temperatures, the PLA coating with 250 mg nisin completely inactivated the cells of L. monocytogenes after 3 d and throughout the 42-d storage period. In liquid egg white, Listeria cells in control and PLA coating without nisin samples declined 1 log CFU/mL during the first 6 d at 10 °C and during 28 d at 4 °C, and then increased to 8 or 5.5 log CFU/mL. The treatment of PLA coating with 250 mg nisin rapidly reduced the cell numbers of Listeria in liquid egg white to undetectable levels after 1 d, then remained undetectable throughout the 48 d storage period at 10 °C and the 70 d storage period at 4 °C. These data suggested that the PLA/nisin coating treatments effectively inactivated the cells of L. monocytogenes in liquid egg white and skim milk samples at both 10 and 4 °C. This study demonstrated the commercial potential of applying the antimicrobial bottle coating method to milk, liquid eggs, and possibly other fluid products.     

Inactivation of foodborne pathogens in milk using dynamic high pressure

Improving the microbiological safety of perishable foods is currently a major preoccupation in the food industry. The aim of this study was to investigate the inactivation of three major food pathogens (Listeria monocytogenes [LSD 105-1], Escherichia coli O157:H7 [ATCC 35150], and Salmonella enterica serotype Enteritidis ATCC [13047]) by dynamic high pressure (DHP) in order to evaluate its potential as a new alternative for the cold pasteurization of milk. The effectiveness of DHP treatment against L. monocYtogenes, E. coli O157:H7, and Salmonella Enteritidis was first evaluated in 0.01 M phosphate-buffered saline (PBS) at pH 7.2 as a function of applied pressure (100, 200, and 300 MPa) and of the number of passes (1, 3, and 5) at 25 degrees C. A single pass at 100 MPa produced no significant inactivation of the three pathogens, while increasing the pressure up to 300 MPa or the number of passes to five increased inactivation. From an initial count of 8.3 log CFU/ml, complete inactivation of viable L. monocytogenes was achieved after three successive passes at 300 MPa, while 200-MPa treatments with three and five passes completely eliminated viable Salmonella Enteritidis and E. coli O157:H7, respectively. The effectiveness of DHP for the inactivation of these pathogens was compared to that of hydrostatic high pressure (HHP) using the same pressure (200 MPa, single pass at 25 degrees C). In general, two additional log reductions in viable count were obtained with DHP DHP was less effective against L. monocytogenes and E. coli O157:H7 in raw milk than in PBS. After five passes at 200 MPa, an 8.3-log reduction was obtained for E. coli O157:H7, while a reduction of about 5.8 log CFU/ml was obtained for L. monocytogenes exposed to 300 MPa for five passes. Exposing milk or buffer samples to mild heating (45 to 60 degrees C) prior to dynamic pressurization enhanced the lethal effect of DHP The inactivation of pathogens also depended on the initial bacterial concentration. The highest reduction was obtained when the bacterial load did not exceed 10(5) CFU/ml. In conclusion, DHP was shown to be very effective for the destruction of the tested pathogens. It offers a promising alternative for the cold pasteurization of milk and possibly other liquid foods.     

Inactivation of Listeria monocytogenes in raw and hot smoked trout fillets by high hydrostatic pressure processing combined with liquid smoke and freezing

High hydrostatic pressure (HHP; 200 MPa for 15 min), liquid smoke (0.50%, v/v) and freezing (−80 °C, overnight) was used to eliminate Listeria monocytogenes in BHI broth, raw and smoked trout. The bactericidal effect of liquid smoke solutions (L9 and G6), HHP and their combinations was evaluated against L. monocytogenes LO28, EGD-e and 10403S and further continued with the most resistant strain (10403S) to the combined treatment. For first time, a synergistic effect of liquid smoke and HHP was observed and was further enhanced by freezing prior to HHP. The effect of HHP and liquid smoke, prior to freezing was highest in BHI compared to raw and smoked trout. A major synergistic effect of HHP, liquid smoke and freezing was observed, reaching a 5.48 or 1.93 log CFU/g reduction when smoked or raw trout was used respectively. Furthermore, high injury levels occurred, among treatments reaching up to 55.98%.Industrial relevanceThis paper illustrates for first time, the possibility of using a very low pressure in combination with liquid smoke and freezing to eliminate L. monocytogenes. It was demonstrated that treatment of trout samples with liquid smoke followed by freezing prior to pressurization at 200 MPa for 15 min reduced the number of L. monocytogenes by more than 5-log CFU/g. Such a remarkable bacterial inactivation at a very low pressure (compared to common industrial practices) is a significant achievement that could allow production of safer and novel products by HHP at an affordable price, as the cost of equipment manufacture as well as the maintenance and running costs could be reduced substantially at lower operation pressures.     

Multiplex PCR identification of Listeria monocytogenes isolates from milk and milk‐processing environments

A multiplex PCR procedure based on genes iap (coding for the invasion‐associated 60 kDa protein or p60) and hly (coding for listeriolysin O) of Listeria monocytogenes was to used to investigate the status of its contamination along the major milk‐processing environments. Duplex PCR amplified fragments of the iap gene at about 1.45 kb from all strains of major Listeria spp. tested and a 420 bp fragment of hly from L. monocytogenes reference strains. With triplex PCR, all L. monocytogenes strains exhibited a 420 bp fragment of hly as well as a 700 bp fragment of iap instead of its 1.45kb PCR product. The tentative L. monocytogenes isolates (n = 27) out of 566 samples of milk and milk‐processing environments in the PALCAM agar from cultures of buffered listeria enrichment broth were further subjected to both API and triplex PCR identification. Both methods identified the same 20 isolates as L. monocytogenes. The triplex PCR procedure detected as low as 3.2 × 10¹ cfu mL^?1 of listerial cells even in the presence of L. innocua (10⁸ cfu mL^?1). It could detect 1.4 × 10² listerial cells mL^?1 directly from milk artificially contaminated with the bacteria. Lower levels of L. monocytogenes cells in milk (1.45 × 10¹ to 1.45 × 10⁰ cfu mL^?1) could be detectable if the inoculated milk samples were cultured for 3–6 h. The bacterium was found not only in raw milk, sewage water and vessel surfaces but also in pasteurized milk. Copyright © 2005 Society of Chemical Industry     

Inactivation of Escherichia coli in orange juice using ozone

This research investigated the efficacy of gaseous ozone for the inactivation of Escherichia coli ATCC 25922 and NCTC 12900 strains in orange juice. Orange juice inoculated with E. coli (10⁶ CFU mL^− 1) as a challenge microorganism was treated with ozone at 75–78 µg mL^− 1 for different time periods (0–18 min). The efficacy of ozone for inactivation of both strains of E. coli was evaluated as a function of different juice types: model orange juice, fresh unfiltered juice, juice without pulp, and juice filtered through 500 µm or 1 mm sieves. Fast inactivation rates for total reduction of E. coli were achieved in model orange juice (60 s) and in juice with low pulp content (6 min). However, in unfiltered juice inactivation was achieved after 15–18 min. This indicated that juice organic matter interferes with antibacterial activity of gaseous ozone. The effect of prior acid (pH 5.0) exposure of E. coli strains on the inactivation efficacy of ozone treatment was also investigated. There was a strain effect observed, where prior acid exposure resulted in higher inactivation times in some cases by comparison with the control cells. However, the overarching influence on inactivation efficacy of ozone was related to the pulp content. Generally, the applied gaseous ozone treatment of orange juice resulted in a population reduction of 5 log cycles.

Industrial relevance

To facilitate the preservation of unstable nutrients many juice processors have investigated alternatives to thermal pasteurisation, including un-pasteurised short shelf life juices with high retail value. This trend has continued within the European Union. However within the US recent regulations by the FDA have required processors to achieve a 5-log reduction in the numbers of the most resistant pathogens in their finished products. Pathogenic E. coli may survive in acid environments such as fruit juices for long periods. This study demonstrates that the use of ozone as a non-thermal technology is effective for inactivation of E. coli and acid exposed E. coli in orange juice. Information on the design of the ozone treatment for inactivation of E. coli which results into safe juice products is also among the main outputs of this work. Ozone auto-decomposition makes this technology safe for fruit juice processing.     

Impedimetric Characterization of Adsorption of Listeria monocytogenes on the Surface of an Aluminum‐Based Immunosensor

The impedimetric characteristics of an immunosensor depend on the electrical properties of an immunosensor substrate. The impedimetric characteristics of an immunosensor compared with adsorption of Listeria monocytogenes were investigated on an aluminum surface insulated with an electrically resistive aluminum oxide layer. Antibody for L. monocytogenes (anti‐L. monocytogenes) was immobilized on an aluminum surface that was insulated with a native air‐formed aluminum oxide layer. The resistance of impedance (R) value of an aluminum‐based immunosensor decreased, especially at 10⁴ to 10⁶ Hz, where the effect of the reactance of impedance (X) was minimal when L. monocytogenes was adsorbed on the immunosensor surface. The R value of the immunosensor at 81 kHz decreased proportionally to the concentration of L. monocytogenes from 1.3 to 4.3 log CFU mL^?1. The adsorption of L. monocytogenes produced local protrusions on the immunosensor surface, causing physicochemical changes in the ionic layer formed on the immunosensor surface by a sinusoidal electrical signal input, which might help electrical current to flow and cause the R value to decrease.     

Recoverability of Listeria monocytogenes after coculture with Tetrahymena pyriformis

Bactivory by protozoa is a major factor that limits the number of bacteria in nature and may control the presence of Listeria monocytogenes. The effectiveness of Tetrahymena pyriformis destruction of L. monocytogenes was measured. Within 1 hr, 35–40 T. pyriformis cells ingested an average of 1,219 CFU of L. monocytogenes. Gentamicin was then added to kill un-ingested Listeria. In 24 hr, the recoverable bacteria were reduced at an exponential rate to undetectable levels (<1 per culture). A genetically diverse set of L. monocytogenes cultures all reduced Listeria recovery by the same degree. In assays without addition of gentamicin, numbers of attached L. monocytogenes cells were lessened from an average of log 6.5 CFU/2 ml culture to log 4.7 CFU/2 ml culture. T. pyriformis was capable of lowering numbers of both free-swimming and attached L. monocytogenes. This technology may have applications to control L. monocytogenes in food processing environments.     

Inactivation of foodborne pathogens in ready-to-eat salad using UV-C irradiation

To examine the applicability of ultraviolet (UV)-C irradiation on the inactivation of foodborne pathogen in ready-to-eat salad, it was inoculated with Escherichia coli O157:H7 and Listeria monocytogenes and then irradiated with UV-C light. Radiation dose required for 90% reduction (d _R) values of E. coli O157:H7 and L. monocytogenes were determined to be 0.21 and 2.48 J/m², respectively. Foodborne pathogen populations significantly (p<0.05) decreased with increasing UV-C irradiation. UV-C irradiation at 8,000 J/m² reduced the populations of E. coli O157:H7 and L. monocytogenes on ready-to-eat salad by 2.16 and 2.57 log CFU/g, respectively.     

Effect of high hydrostatic pressure on Listeria innocua 910 CECT inoculated into Ewe's milk.

Ewe's milk standardized to 6% fat was inoculated with Listeria innocua 910 CECT at a concentration of 10(7)CFU/ml and treated by high hydrostatic pressure. Treatments consisted of combinations of pressure (200, 300, 350, 400, 450, and 500 MPa), temperature (2, 10, 25, and 50 degrees C), and time (5, 10, and 15 min). To determine numbers of L. innocua, listeria selective agar base with listeria selective supplement and plate count agar was used. Low-temperature (2 degrees C) pressurizations produced higher L. innocua inactivation than treatments at room temperatures (25 degrees C). Pressures between 450 and 500 MPa for 10 to 15 min were needed to achieve reductions of 7 to 8 log units. The kinetics of destruction of L. innocua were first order with D-values of 3.12 min at 2 degrees C and 400 MPa and 4 min at 25 degrees C and 400 MPa. A baroprotective effect of ewe's milk (6% fat) on L. innocua was observed in comparison with other studies using different media and similar pressurization conditions.