Ionizing radiation sensitivity of Listeria monocytogenes ATCC 49594 and Listeria innocua ATCC 51742 inoculated on endive (Cichorium endiva)

Ionizing radiation inactivates the pathogenic bacteria that can contaminate leafy green vegetables. Leaf pieces and leaf homogenate of endive (Cichorium endiva) were inoculated with the pathogen Listeria monocytogenes (ATCC 49594) or Listeria innocua (ATCC 51742), a nonpathogenic surrogate bacterium. The radiation sensitivity of the two strains was similar, although L. innocua was more sensitive to the type of suspending leaf preparation. During refrigerated storage after irradiation, the population of L. monocytogenes on inoculated endive was briefly suppressed by 0.42 kilogray (kGy), a dose calibrated to achieve a 99% reduction. However, the pathogen regrew after 5 days until it exceeded the bacterial levels on the control after 19 days in storage. Treatment with 0.84 kGy, equivalent to a 99.99% reduction, suppressed L. monocytogenes throughout refrigerated storage. Doses up to 1.0 kGy had no significant effect on the color of endive leaf material, regardless of whether taken from the leaf edge or the leaf midrib. The texture of leaf edge material was unaffected by doses up to 1.0 kGy, whereas the maximum dose tolerated by leaf midrib material was 0.8 kGy. These results show that endive leaves may be treated with doses sufficient to achieve at least a 99.99% reduction of L. monocytogenes with little or no impact on the product's texture or color.     

The effect of homogenization of whole milk, skim milk and milk fat on nisin activity against Listeria innocua

Whole milk, skim milk and an emulsion of milk fat in water, inoculated with approx. 10(5) cfu/ml of Listeria innocua, were treated at 30 degrees C with 100 IU/ml of nisin, homogenization at 200 bar or both procedures. Nisin activity and survival of L. innocua after treatments were determined. Recovery of nisin activity from non-homogenized whole milk treated with 100 IU/ml of nisin was complete, whereas a loss of 18 to 28% of activity was detected in non-homogenized fat-in-water emulsion. Loss in nisin activity due to homogenization represented up to 64% in whole milk and 62% in fat-in-water emulsion. Nisin addition by itself achieved a reduction in L. innocua counts of 3.7-3.8 log units in whole milk and 3.6 log units in fat-in-water emulsion compared to numbers in untreated samples. When nisin-containing whole milk and fat-in-water emulsion were homogenized, L. innocua counts were only reduced by 2.6-2.9 log units and 2.5 log units, respectively, compared to numbers in untreated samples. Homogenization of nisin-containing skim milk resulted in a loss of nisin activity of 20% but achieved a reduction of 3.0 log units in L. innocua counts.     

Effect of thermosonication,pulsed electric field and their combination on inactivation of Listeria innocua in milk

The impact of thermosonication (TS) and pulsed electric field (PEF), individually and combined, on the survival of Listeria innocua 11288 (NCTC) in milk was investigated. TS (400 W, 160 s) without pre-heating reduced L. innocua by 1.2 log₁₀ cfu mL^?1, while shorter treatment times produced negligible inactivation, suggesting TS to be a hurdle rather than an effective standalone treatment. PEF (30 and 40 kV cm^?1, 50 μs) at 10 °C caused a reduction of L. innocua of 1.1 and 3.3 log cycles, respectively. The highest field strength (40 kV cm^?1) combined with TS (80 s) led to 6.8 log₁₀ cfu mL^?1 inactivation. Milk pre-heated to 55 °C (over 60 s) prior to TS followed by PEF (30 and 40 kV cm^?1) showed inactivation between 4.5 and 6.9 log₁₀ cfu mL^?1, the latter being comparable (P > 0.05) with thermal pasteurisation. The data indicate that TS followed by PEF represents a valid alternative for L. innocua inactivation in milk.     

Influence of treatment temperature on the inactivation of Listeria innocua by pulsed electric fields

The effect of treatment temperature on the bactericidal effectiveness of pulsed electric fields (PEF) applied on Listeria innocua suspended in McIllvaine buffer was investigated. Electric field intensity and number of applied pulses were applied in the ranges of 31–40 kV/cm and 5–35 pulses, respectively. Studied treatment temperatures were sustained for 10 s, and ranged between 19°C and 59°C depending on the amount of energy delivered by the PEF treatment. The application of PEF at higher temperatures proved to be more effective than either PEF at low temperatures or the applied thermal treatments by themselves. A maximum bacterial inactivation of 6-log cycles was obtained by applying either: 20 pulses of 40 kV/cm at 65°C, 25 pulses of 36 kV/cm at 61°C, or 31 pulses of 31 kV/cm at 56°C. On the other hand, a thermal treatment of 66°C sustained for 30 s reduced the bacterial population on its own by only 5-log cycles, and the application of 60 pulses of 31 kV/cm at 30°C caused only 3-log cycles of bacterial inactivation. The findings in this study suggest that PEF technology may be effectively used as an enhanced mild thermal preservation method.     

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.     

Effect of the pressurizing ramp on the inactivation of Listeria innocua considering thermofluiddynamical processes

The effect of the pressurization ramp on the inactivation kinetics of Listeria innocua suspended in Tris buffer has been studied experimentally in an optical pressure cell. Additionally, the thermofluiddynamical processes occurring at the different pressure ramps have been considered. The results obtained at room temperature show that the inactivation effect of high pressure remains the same in case of a fast pressurization ramp (500 MPa/min) and a slow depressurization ramp (100 MPa/min) in comparison to a slow pressurization ramp (100 MPa/min) and a fast depressurization ramp (500 MPa/min). At the chosen test conditions, the fluiddynamical and thermal processes of the two variations differ only slightly from each other. This has been ascertained by measuring the velocity and temperature distributions occurring at the pressure processes. The conclusion is that the pressurization and depressurization rate, respectively, do not effect per se the inactivation kinetics of Listeria innocua.     

Inactivation of Listeria innocua in skim milk by pulsed electric fields and nisin

Pulsed electric fields (PEF) is an emerging nonthermal processing technology used to inactivate microorganisms in liquid foods such as milk. PEF results in loss of cell membrane functionality that leads to inactivation of the microorganism. There are many processes that aid in the stability and safety of foods. The combination of different preservation factors, such as nisin and PEF, to control microorganisms should be explored. The objective of this research was to study the inactivation of Listeria innocua suspended in skim milk by PEF as well as the sensitization of PEF treated L. innocua to nisin. The selected electric field intensity was 30, 40 and 50 kV/cm and the number of pulses applied was 10.6, 21.3 and 32. The sensitization exhibited by PEF treated L. innocua to nisin was assessed for 10 or 100 IU nisin/ml. A progressive decrease in the population of L. innocua was observed for the selected field intensities, with the greatest reduction being 2 1/2 log cycles (U). The exposure of L. innocua to nisin after PEF had an additive effect on the inactivation of the microorganism as that exhibited by the PEF alone. As the electric field, number of pulses and nisin concentration increased, synergism was observed in the inactivation of L. innocua as a result of exposure to nisin after PEF. The reduction of L. innocua accomplished by exposure to 10 IU nisin/ml after 32 pulsed electric fields was 2, 2.7, and 3.4 U for an electric field intensity of 30, 40, and 50 kV/cm, respectively. Population of L. innocua subjected to 100 IU nisin/ml after PEF was 2.8-3.8 U for the selected electric field intensities and 32 pulses. The designed model for the inactivation of L. innocua as a result of the PEF followed by exposure to nisin proved to be accurate in the prediction of the inactivation of L. innocua in skim milk containing 1.2 or 37 IU nisin/ml. Inactivation of L. innocua in skim milk containing 37 IU nisin/ml resulted in a decrease in population of 3.7 U.     

Baroprotection of vegetative bacteria by milk constituents: A study of Listeria innocua

The baroprotective effect of milk constituents on Listeria innocua 4202 treated at 350 or 500 MPa for 5 min was examined. High-pressure (HP) treatment of L. innocua 4202 (1×10⁹ cfu mL⁻¹) resulted in complete inactivation in simulated milk ultra-filtrate (SMUF), a ∼5.0 log reduction in phosphate-buffered saline and a ∼2.9 log reduction in milk. The addition of micellar casein to SMUF increased survival of the bacterium by 3 logs, compared with SMUF alone, but the protective effect was negated if the minerals associated with the casein micelles were removed. The colloidal minerals calcium (30 mm), magnesium (5 mm), citrate (10 mm) and phosphate (20 mm), suspended in SMUF increased survival by ∼3.3, ∼1.7, ∼3.3 and ∼3.5 logs, respectively. The buffering capacity of the suspending medium was found to be a key factor in microbial baroresistance. Buffering by phosphate and citrate in milk may protect microorganisms against changes in pH during HP treatment, whereas the divalent cations calcium and magnesium may protect cell membranes against HP.     

The advantages of solid fat content determination in cocoa butter and cocoa butter equivalents by the Karlshamns method

This paper describes the preparation of samples and the evaluation techniques applied to determine the Solid Fat Content (SFC) by means of pulsed nuclear magnetic resonance (NMR) in confectionery fats characterized by distinct polymorphism (cocoa butter, cocoa butter equivalents and their mixtures). Due to the deviation of SFC values in certain softer cocoa butter equivalents at the temperature of 20 °C, an internal method devised at Karlshamns Oils & Fats AB is applied as well. When compared to the official IUPAC 2.150 (b) method, Karlshamns' method involves a lower temperature and a prolonged period of time at the stage of the final SFC stabilization in the pre-treatment of fat samples. The calculation of SFC in the analysed samples for the purposes of this paper was carried out by employing both methods (the indirect method and a serial procedure). The results are presented graphically. Differences were reflected in the SFC of the fats and their mixtures that were under observation depending on the method and temperatures applied. The obtained results indicated a considerable advantage of employing the Karlshamns method since the results yielded by this method are more lucid. Moreover, the behaviour of fats in the production process can be more easily predicted on the basis of these results.     

Performance of selected milk fat fractions in cold-spreadable butter.

Based on solid fat content profiles, milk fat fractions produced by fractional crystallization procedures employing melted milk fat and milk fat dissolved in acetone were selected for incorporation into soft butter samples. Butter samples made from low melting liquid fractions or a combination of primarily low melting liquid fractions and a small amount of high melting solid fractions exhibited good spreadability at refrigerator temperature (4 degrees C) but were almost melted at room temperature (21 degrees C). Butters made with a high proportion of low melting liquid fraction, a small proportion of high melting solid, and a small proportion of very high melting solid fractions were still spreadable at refrigerator temperature and maintained their physical form at room temperature. Very high melting fractions, which provided key structural functionality in spreadable butter, were obtained from acetone fractionation. Because the use of acetone in processing may hinder or prevent commercialization of these fractions, other means to obtain very high melting fractions from milk fat should be pursued.     

Effect of spectral range in surface inactivation of Listeria innocua using broad-spectrum pulsed light

Pulsed light (PL) treatment is an alternative to traditional thermal treatment that has the potential to achieve several log-cycle reductions in the concentration of microorganisms. One issue that is still debated is related to what specifically causes cell death after PL treatments. The main objective of this work was to elucidate which portions of the PL range are responsible for bacterial inactivation. Stainless steel coupons with controlled surface properties were inoculated with a known concentration of Listeria innocua in the stationary growth phase and treated with 1 to 12 pulses of light at a pulse rate of 3 pulses per s and a pulse width of 360 micros. The effects of the full spectrum (lambda = 180 to 1,100 nm) were compared with the effects obtained when only certain regions of UV, visible, and near-infrared light were used. The effectiveness of the treatments was determined in parallel by the standard plate count and most-probable-number techniques. At a fluence of about 6 J/cm(2), the full-spectrum PL treatment resulted in a 4.08-log reduction of L. innocua on a Mill finish surface, the removal of lambda < 200 nm diminished the reduction to only 1.64 log, and total elimination of UV light resulted in no lethal effects on L. innocua. Overwhelmingly, the portions of the PL spectrum responsible for bacterial death are the UV-B and UV-C spectral ranges (X < 300 nm), with some death taking place during exposure to UV-A radiation (300 < lambda < 400 nm) and no observable death upon exposure to visible and near-infrared light (lambda > 400 nm). This work provides additional supporting evidence that cell death in PL treatment is due to exposure to UV light. Additionally, it was shown that even a minor modification of the light path or the UV light spectrum in PL treatments can have a significant negative impact on the treatment intensity and effectiveness.     

Effect of a post-packaging pasteurization process on inactivation of a Listeria innocua surrogate in meat products

Effects of surface pasteurization on inactivation of Listeria innocua were investigated. Surface temperature, monitored during post-packaging pasteurization, was used to predict the lethality of L. monocytogenes. Temperatures reached 70°C for lean and fat sausages within 9 min of treatment. An inoculation study validated the efficacy of post-processing and the thermal lethality of L. monocytogenes. Pre-cooked sausage and ham, inoculated with approximately 10⁷ CFU/cm² of L. innocua, were heated to a surface temperature of 70°C. Numbers of L. innocua were reduced by 7 log on surface-inoculated sausage. Guidelines for safe, ready-to-eat meat products are provided for small scale meat processors.     

Transmission electron microscopy of Listeria innocua treated by pulsed electric fields and nisin in skimmed milk

Pulsed electric fields (PEF) is a nonthermal food preservation process where organoleptic and nutritional properties of the food are maintained. PEF is known to inactivate microorganisms by causing dielectric breakdown of the cell membrane, thus altering the functionality of the membrane as a semipermeable barrier. The extent of damage of the cell membrane, whether visible in the form of a pore or as loss of membrane functionality leads to the inactivation of the microorganism. The objective of this study was to investigate under transmission electron microscopy (TEM) the morphological changes on Listerit innocua as a result of PEF treatment in skimmed milk containing nisin. L. innocua was subjected to PEF at selected electric field intensities of 30, 40, and 50 kV/cm. L. innocua was treated by PEF in both skimmed milk with and without 37 IU nisin/ml. L. innocua treated by PEF in skimmed milk exhibited an increase in the cell wall roughness. cytoplasmic clumping, leakage of cellular material, and rupture of the cell walls and cell membranes. L. innocua subjected to PEF in skimmed milk containing 37 IU nisin/ml exhibited an increased cell wall width. At the highest electric field intensity, 50 kV/cm, elongation of the cell length was observed. There were no morphological differences between cells treated by PEF in skimmed milk with or without nisin. The combination of PEF and nisin exhibit an additive effect in the morphological damage observed on L. innocua. Pore formation was observed on L. innocua for an electric field intensity of 40 kV/cm. The inactivation of L. innocua was a consequence of rupture of the cell membrane and loss of cell membrane functionality.     

Effects of steam and lactic acid treatments on inactivation of Listeria innocua surface-inoculated on chicken skins

Effectiveness of combined steam (10 and 60 s, 70 degrees C and 98 degrees C) and chemical treatments, using concentrated solutions of lactic acid (1 and 30 min, 5% and 10% lactic acid), on the inactivation of Listeria innocua inoculated on the surface of chicken skins have been studied. Surviving bacteria on the skin were enumerated immediately after treatment, and after 7 days of storage at 4 degrees C. The most effective treatment was the combination of steam of 98 degrees C and 10% lactic acid with its immediate efficacy being mainly attributed to the applied heat treatment. However, after 7-day storage, the treatment's effectiveness was mainly due to the applied acid treatment, which prevented growth of the bacteria that survived the heat treatment. Milder treatments (70 degrees C steam, 5% lactic acid) revealed a genuine synergy between the heat and acid treatments, paving the way for an effective means of reducing bacterial load on the surface of poultry without affecting the product's "raw" appearance.     

A predictive model for the inactivation of Listeria innocua in cooked poultry products during postpackage pasteurization

Contamination of Listeria monocytogenes in ready-to-eat poultry products poses potential risk of listeriosis to the public. To control the level of Listeria contamination, attention has been focused on the postpackage pasteurization of fully cooked poultry products. In this study, we sought to develop a model to predict the thermal inactivation of L. monocytogenes in chicken drumettes during postpackage hot water pasteurization. Fully cooked chicken drumettes were inoculated with Listeria innocua as a surrogate microorganism for Listeria monocytogenes, vacuum packaged, and treated in hot water baths at 60, 70, 80, and 90°C for different heating times. Experimental results showed that a 7-log CFU/g reduction of L. innocua occurred at 54, 28, 18, and 10 min at 60, 70, 80, and 90°C, respectively. The Weibull model was used to fit the survival curves of L. innocua at each heating temperature. The root mean square errors and residual plots indicated good agreements between the predicted and observed values. The predictive model was further validated by predicting a new data set generated in the pilot-plant tests. Model performance was evaluated by the acceptable prediction zone method, and the results indicated that the percentages of acceptable prediction errors were 100, 100, 82.4, and 87.5% at 60, 70, 80 and 90°C, respectively, which were all greater than the threshold acceptable value of 70% , indicating good performance of the model. The developed predictive model can be used as a tool to predict thermal inactivation behaviors of L. monocytogenes in ready-to-eat chicken drumettes products.     

Reduced detectability of Listeria monocytogenes in the presence of Listeria innocua

Recent foodborne crises have demonstrated the importance of monitoring food safety. In terms of microbiological criteria, food safety requires the reliable detection of pathogens such as Listeria monocytogenes along the food chain by appropriate analytical methods. However, indications exist that accompanying Listeria innocua strains suppress the growth of L. monocytogenes during selective enrichment, which may cause reduced or even inhibited detection. To study these effects, the limit of detection of L. monocytogenes was investigated in the presence of L. innocua using the International Organization for Standardization standard method ISO 11290-1 and the VIDAS LDUO system, an automated method based on enzyme-linked fluorescence technology. The challenge was to provide low initial Listeria concentrations at sufficient precision to quantify the influence on the probability of detection of L. monocytogenes. The application of reference materials appropriate for quantitative test methods and a standardized dilution procedure were necessary to ensure accurate CFU levels of defined proportions of mixtures of both Listeria species. During selective enrichment, overgrowth of L. monocytogenes by L. innocua could be confirmed, leading to high rates of false-negative results. Moreover, with both methods, a significant decrease in the detectability of L. monocytogenes could be quantified at ratios of 2:1 at very low concentrations representative of natural contamination levels often found in foods and environments. It is concluded that there is a need to improve existing procedures with respect to selective enrichment, as well as the detection techniques.     

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.     

Addition of olein from milk fat positively affects the firmness of butter

Butter is highly valued for its characteristic flavor and aroma; however, it has the disadvantage of unsatisfactory spreadability at low temperatures. The functional characteristics of butter can be modified by changing its composition or physical structure. The objective of this study was to evaluate the effect of olein on structure and composition of butter. Olein was obtained by two stage dry fractionation process of the anhydrous milk fat (AMF) and added to commercial cream prior to butter manufacture. The fractions were characterized for triacylglycerols composition, solid fat content, crystallization isotherm, and thermal behavior. Butter was manufactured using commercial cream or cream containing 50% olein. Butter samples were characterized for physicochemical composition, instrumental color, crystallization parameters, and firmness after 1 and 7 days of storage at 10 °C. The firmness of butter subjected to room temperature was also evaluated. Butter containing olein differed significantly from the control and had darker yellow color, higher crystallization time, and lower solid fat content after 120 min at 15 °C, and hence lower firmness after 1 and 7 days of refrigerated storage. Although lower firmness was observed over time for all samples at room temperature, butter containing olein exhibited lower firmness after both 1 and 7 days, thus suggesting changes in organization of solid fat crystal network in the liquid fat. The addition of olein to butter allowed obtaining a softer product, with more intense color and possible nutritional benefits due to the medium chain triglycerides and higher carotene levels.     

Effect of fat content on infection by Listeria monocytogenes in a mouse model

An estimated 2,500 cases of listeriosis occur annually in the United States. Listeriosis is particularly severe among pregnant women and immunocompromised individuals. Little is known regarding the effect of the food matrix on the ability of L. monocytogenes to survive in the gastrointestinal tract and cause systemic infection. Mice were inoculated with various doses of L. monocytogenes in skim milk, Half & Half, or whipping cream to determine whether differences in milk fat content influence the ability of L. monocytogenes to survive passage through the gut and infect the liver or spleen. The number of fecal samples positive for L. monocytogenes increased with increasing doses of L. monocytogenes for all three vehicles. The number of L. monocytogenes cells isolated from liver or spleen of mice dosed with L. monocytogenes was not significantly different among treatment vehicles. Dose-response models revealed that as the dosage of L. monocytogenes was increased in different milk vehicles, the number of L. monocytogenes cells in liver or spleen also increased. Although fat content of food had no dose-dependent effect on L. monocytogenes infection in the murine gastrointestinal tract, we cannot discount the possibility that it may be a factor in L. monocytogenes infections of humans because of differences in the physiology of gastrointestinal tracts of mice and humans.