Process Parameters Affecting Listeria innocua Inactivation by Pulsed Light

The impact of several processing parameters on the effectiveness of pulsed light (PL) technology was investigated. The distribution of the light on the surface of the PL treatment chamber and the fluence striking on the sample under different treatment conditions were evaluated. Higher pulse fluences were registered when input voltage increased and the distance from the sample to the light source decreased. Distance from the sample to the xenon lamp also played a major role in the light distribution onto the surface of the treatment zone. Whereas quartz shelf placed close to the xenon lamp resulted in a non-uniform distribution of fluence, a homogeneous light distribution was found when increasing the distance to the flash lamp. Moreover, the impact of several PL processing parameters on microbial inactivation was investigated. Neither pulse voltage nor total voltage determined the antimicrobial effectiveness of PL technology in the operating range tested. Reduction in Listeria innocua counts increased with the number of pulses and the pulse fluence, being total fluence reaching the samples the most relevant process factor affecting microbial inactivation by PL. Considering the importance of this processing parameter, an accurate quantification of the fluence striking on the sample will be needed in order to design effective PL treatments.     

Inactivation of Listeria innocua on Frankfurters by Ultraviolet Light and Flash Pasteurization

ABSTRACT:  Listeria monocytogenes , a psychrotrophic foodborne pathogen, is a recurring postprocess contaminant on ready-to-eat meat (RTE) products including frankfurters. Flash (Steam) Pasteurization (FP) and ultraviolet light (254 nm-UVC) has been shown to reduce levels of L. monocytogenes and L. innocua on frankfurters. In this study, the use of UVC light followed by FP to inactivate L. innocua, a nonpathogenic surrogate for L.monocytogenes , on frankfurters that contained sodium diacetate and potassium lactate (SDA/PL) in a pilot-plant setting was investigated. Application of UVC (1.0 J/cm²), followed by FP (0.75 s steam/121 °C) resulted in inactivation of 3.19 log L. innocua , while application of UVC (4.0 J/cm²), followed by FP (3 s steam/121 °C) resulted in inactivation of 3.89 log of L. innocua . A refrigerated storage study (8 °C) of frankfurters that contained SDA/PL that were treated with UVC followed by FP revealed the growth of L. innocua was inhibited for approximately 8 wk following application of the interventions. The use of UVC in combination with FP had little effect on frankfurter color and texture. The combination of UVC, FP, and SDA/PL was found to be an effective hurdle process for decontamination of frankfurter surfaces.     

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.     

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.     

Impact of Process Parameters on Listeria innocua Inactivation Kinetics by Pulsed Light Technology

The impact of several pulsed light (PL) processing parameters on microbial inactivation was evaluated in buffered water systems using Listeria innocua as test microorganism. Reduction in L. innocua population increased directly with pulse energy, pulse fluence and the number of light pulses, and inversely with the distance between samples and a xenon lamp. Overall, the higher the amount of light received by the target microorganism by both direct and reflected light, the larger the loss of cell counts. Total fluence striking on the samples per area unit was shown to be the most relevant process factor affecting L. innocua inactivation by PL. Microbial population decreased with total fluence, obtaining more than 7 log reductions after 0.4 J?cm^?2. The inactivation kinetics was clearly sigmoidal, showing an initial shoulder in the inactivation curve. No significant reduction (<1 log) in L. innocua counts was induced at fluences lower than 0.04 J?cm^?2. From this threshold total fluence, L. innocua inactivation increased exponentially to the maximum detectable level. Since total fluence is the most relevant process factor affecting microbial inactivation by PL, this parameter must be reported to describe PL processing conditions.     

Inactivation of Listeria innocua in liquid whole egg by pulsed electric fields and nisin

Consumer demand for fresh-like products with little or no degradation of nutritional and organoleptic properties has led to the study of new technologies in food preservation. Pulsed electric fields (PEF) is a nonthermal preservation method used to inactivate microorganisms mainly in liquid foods. Microorganisms in the presence of PEF suffer cell membrane damage. Nisin is a natural antimicrobial known to disrupt cell membrane integrity. Thus the combination of PEF and nisin represents a hurdle for the survival of Listeria innocua in liquid whole egg (LWE). L. innocua suspended in LWE was subjected to two different treatments: PEF and PEF followed by exposure to nisin. The selected frequency and pulse duration for PEF was 3.5 Hz and 2 micros, respectively. Electric field intensities of 30, 40 and 50 kV/cm were used. The number of pulses applied to the LWE was 10.6, 21.3 and 32. The highest extent of microbial inactivation with PEF was 3.5 log cycles (U) for an electric field intensity of 50 kV/cm and 32 pulses. Treatment of LWE by PEF was conducted at low temperatures, 36 degrees C being the highest. Exposure of L. innocua to nisin following the PEF treatment exhibited an additive effect on the inactivation of the microorganism. Moreover, a synergistic effect was observed as the electric field intensity, number of pulses and nisin concentration increased. L. innocua exposed to 10 IU nisin/ml after PEF exhibited a decrease in population of 4.1 U for an electric field intensity of 50 kV/cm and 32 pulses. Exposure of L. innocua to 100 IU nisin/ml following PEF resulted in 5.5 U for an electric field intensity of 50 kV/cm and 32 pulses. The model developed for the inactivation of L. innocua by PEF and followed by exposure to nisin proved to be accurate (p = 0.05) when used to model the inactivation of the microorganism by PEF in LWE with 1.2 or 37 IU nisin/ml. The presence of 37 IU nisin/ml in LWE during the PEF treatment for an electric field intensity of 50 kV/cm and 32 pulses resulted in a decrease in the population of L. innocua of 4.4 U.     

Inactivation of Escherichia coli and Listeria innocua in kiwifruit and pineapple juices by high hydrostatic pressure

Escherichia coli and Listeria innocua in kiwifruit and pineapple juices were exposed to high hydrostatic pressure (HHP) at 300 MPa for 5 min. Both bacteria showed equal resistance to HHP. Using low (0 degrees C) or sub-zero (-10 degrees C) temperatures instead of room temperature (20 degrees C) during pressurization did not change the effectiveness of HHP treatment on both bacteria in studied juices. Pulse pressure treatment (multiple pulses for a total holding time of 5 min at 300 MPa) instead of continuous (single pulse) treatment had no significant (p>0.05) effect on the microbial inactivation in kiwifruit juice; however, in pineapple juice pulse treatment, especially after 5 pulses, increased the inactivation significantly (p<0.05) for both bacteria. Following storage of pressure-treated (350 MPa, 20 degrees C for 60 s x 5 pulses) juices at 4, 20 and 37 degrees C up to 3 weeks, the level of microbial inactivation further increased and no injury recovery of the bacteria were detected. This work has shown that HHP treatment can be used to inactivate E. coli and L. innocua in kiwifruit and pineapple juices at lower pressure values at room temperature than the conditions used in commercial applications (>400 MPa). However, storage period and temperature should carefully be optimized to increase the safety of HHP treated fruit juices.     

Inactivation of Listeria innocua on frankfurters that contain potassium lactate and sodium diacetate by flash pasteurization

ABSTRACT:  Listeria monocytogenes , a psychrotrophic foodborne pathogen, is a recurring postprocess contaminant on ready-to-eat meat (RTE) products, including frankfurters. Potassium lactate (PL) and sodium diacetate (SDA) are FDA-approved antimicrobials that inhibit the growth of L. monocytogenes when incorporated into the formulation of fine emulsion sausage. Flash (steam) pasteurization (FP) has been shown to reduce levels of L. monocytogenes , and its surrogate L. innocua , on frankfurter surfaces. The ability of FP to inactivate and prevent the growth of the L. monocytogenes surrogate L. innocua in a pilot plant setting was investigated. FP treatment (1.5 s, 121 °C) of single layers of frankfurters that were surface-inoculated with either 5, 4, or 3 log CFU/g of L. innocua immediately before FP (1.5 s, 121 °C) resulted in log reductions of 1.97 (± 0.11), 2.03 (± 0.10), or 2.07 (± 0.14), respectively. Inoculum level had no effect on the inactivation of L. innocua . Following 8 wk of refrigerated storage (4 °C), L. innocua levels decreased by 0.5 log in non-FP-treated frankfurter packs, while the 2 log reduction of L. innocua was maintained for FP-treated frankfurters. FP (1.5 s, 121 °C) had no effect on frankfurter color or texture. Because the numbers of L. monocytogenes associated with contaminations of ready-to-eat meats are typically very low, the use of FP in combination with PL and SDA has the potential to reduce the number of frankfurter recalls and foodborne illness outbreaks.     

Quantitative Evaluation of Thermal Inactivation Kinetics of Free-Floating Versus Surface-Attached Listeria innocua Cells

Surface pasteurization is one of the decontamination treatments that can contribute to better preservation of meat products retaining most of their quality characteristics relatively intact if compared with the raw products. The current research compares the kinetics of free-floating and surface attached Listeria innocua cells by using integrated microbial and heat transfer modelling approaches. Surface pasteurization treatments are applied on a (abiotic) Teflon® model system in a novel steam surface decontamination rig. The experimental set-up prevented following four technological aspects to occur, (1) cold purge migration to the surface during the heating process, (2) inactivation kinetics of a cocktail of microbes, (3) protective effect of food components, and (4) physical distribution of bacteria throughout the depth of the product skin. Microbial load predictions are performed based on the inactivation parameters obtained during free-floating cell experiments. These predictions, when compared with the microbial data of the surface treatments, prove that the surface attached cells were much more heat resistant, despite the experimental set-up preventing the aforementioned (technological) events to occur. Indeed, surface attached cells can have different physiological/phenotypical/genetical characteristics, such as cell aggregations, colony formations, presence of flagella. In a final step, three techniques are implemented to evaluate mathematically the kinetics of the surface attached cells. Overall, this research’s significance is lying in the quantitative assessment of microbial heat resistance. The technological reasons underlying the increased microbial heat resistance on biotic and abiotic surfaces should be reevaluated, taking into account possible physiological/phenotypical/genetical characteristics.     

Aerosol studies with Listeria innocua and Listeria monocytogenes

Aerosol studies of Listeria monocytogenes in food processing plants have been limited by lack of a suitable surrogate microorganism. The objective of this study was to investigate the potential of using green fluorescent protein-labeled strains of Listeria innocua as a surrogate for L. monocytogenes for aerosol studies. These studies were conducted in a laboratory bioaerosol chamber and a pilot food-processing facility. Four strains of L. innocua and five strains of L. monocytogenes were used. In the laboratory chamber study, Listeria cells were released into the environment at two different cell numbers and under two airflow conditions. Trypticase soy agar (TSA) plates and oven-roasted breasts of chicken and turkey were placed in the chamber to monitor Listeria cell numbers deposited from aerosols. A similar experimental design was used in the pilot plant study; however, only L. innocua was used. Results showed that L. monocytogenes and L. innocua survived equally well on chicken and turkey breast meats and TSA plates. No-fan and continuous fan applications, which affected airflow, had no significant effect on settling rates of aerosolized L. monocytogenes and L. innocua in the bioaerosol chamber or L. innocua in the pilot plant study. Listeriae cell numbers in the air decreased rapidly during the first 1.5 h following release, with few to no listeriae detected in the air at 3 h. Aerosol particles with diameters of 1 and 2 microM correlated directly with the number of Listeria cells in the aerosol but not with particles that were 0.3, 0.5, and 5 microM in diameter. Results indicate that L. innocua can be used as a surrogate for L. monocytogenes in an aerosol study.     

Inactivation kinetics and factors of variability in the pulsed light treatment of Listeria innocua cells

Pulsed light (PL) treatment can effectively reduce microbial populations in clear substrates and on surfaces, but its effectiveness varies as a function of substrate or treatment-related factors. For PL to be successfully adopted by the food industry, all factors of influence, as well as the inactivation kinetics for the microorganisms of concern, must be elucidated. In this study, the inactivation kinetics of Listeria innocua and the effect of inoculum size on PL inactivation were investigated. Stainless steel coupons (50.8 by 101.6 mm) of defined surface properties and transparent glass chamber slides (25.4 by 50.8 by 10 mm) were each inoculated with 1 ml of aqueous suspensions of L. innocua containing inoculum populations of up to 10(9) CFU. The thickness of the liquid layer in the glass slides was 1.16 mm. The inoculated substrates were exposed to PL treatment of up to 17 J/cm2 in a static PL chamber equipped with a pulsed Xenon lamp. Survivors were recovered and enumerated by both standard plate counting and most-probable-number procedures. The data indicated that in clear liquids, PL resulted in more than a 6-log reduction of L. innocua after a 12-J/cm2 treatment, regardless of the initial inoculum size. For the stainless steel surfaces, less than a 4-log reduction after a 12-J/cm2 treatment and a noticeable effect of substrate characteristics and inoculum size on inactivation were observed. The survivor curves showed pronounced tailing for all substrates used in the study. The Weibull model accurately predicted the survivor ratios for the PL treatment of L. innocua in clear liquids, with a shape and scale parameter of 0.33 and 3.01, respectively. The Weibull model resulted in significant overestimation of PL effectiveness for the stainless steel substrates, where the influence of various substrate properties and inoculum level on inactivation was significant.     

Inactivation Kinetics of Listeria innocua in Skim Milk in a Continuous Flow Processing System

Thermal inactivation of Listeria innocua in raw skim milk determined under continuous flow conditions was compared to results from the capillary tube method at 65, 68, and 70°C. A laboratory scale pasteurizer (LSP) was used to generate kinetic data under isothermal, continuous-flow conditions. Inactivation was monitored by sampling at various locations along the hold tubes. D_(657°C)-, D_(65°C)- and D_(70°C)- values for L. innocua M1 were 11.5, 3.5, and 1.6 sec, respectively, when determined by the LSP system and 16.5, 3.9,1.5 sec, respectively, when determined by the capillary tube method. Decimal reduction times for the two methods were predicted to be coincident at 69°C. Z_D-values for the capillary tube and LSP systems were significantly different. The data suggest that the mechanism of thermal destruction may be different between batch and continuous flow processes.     

Recovery of heat-injured Listeria innocua

Listeria innocua was subjected to thermal inactivation and the extent of heat-injured cells was quantified. Cultures were heated in liquid medium for different times, using temperatures in the range of 52.5 to 65.0 degrees C, and plated on Tryptic Soy Agar with 0.6% yeast extract (TSAYE) used as non-selective medium and on TSAYE plus 5% NaCl (TSAYE+NaCl) and Palcam agar with selective supplement (Palcam agar) as selective media. The difference observed in counts in non-selective and in selective media gave an indication of cell injury during the heat treatment. D- and z- values were calculated for all conditions considered. For each temperature, D-values obtained using non-selective recovery procedures were higher than the ones obtained using the two selective media. When comparing the selective media, it can be concluded that Palcam agar allowed recovery and growth of thermally injured cells and so it was less inhibitor than TSAYE+NaCl. Another important result was the influence of temperature on the degree of cellular injury. As temperature increases, the degree of heat-injured cells also increases, and consequently concern has to be taken with the temperature and the counting medium used in food processing studies. The results of this work clearly demonstrated that selective media used for Listeria monocytogenes enumeration/detection might not be suitable for the recovery of heat-injured cells, which can dangerously underestimate the presence of this foodborne pathogen.     

Inactivation of Listeria innocua and Escherichia coli in carrot juice by combining high pressure processing,nisin, and mild thermal treatments

This study explored the effectiveness of high pressure (200–500 MPa) alone or in combination with mild thermal treatments (35 and 50 °C) and nisin (25 and 50-ppm) on the inactivation of L. innocua and E. coli in carrot juice. Processing at 500 MPa at 20 °C for 2 min without nisin resulted in 4- and 5-log CFU/mL reduction of L. innocua and E. coli, respectively while incorporating 25-ppm nisin at same pressure and temperature rendered 7-log CFU/mL reduction. There was synergism between high pressure, nisin, and heat in all treatments to inactivate both microorganisms. After a 28-d of refrigerated storage, total plate counts were <2-log CFU/mL in carrot juice treated with combination of 300 MPa and 25-ppm nisin at 35 °C. All combinations resulted in less intense use of pressure, i.e. more energy efficient, cost effective processes while attaining high quality juices. The results of this study suggest that by using selected combinations of high pressure, nisin and mild temperatures, safe, clean-label, high-quality juices can be produced.Industrial relevanceThe results from this study show a synergistic effect on the inactivation of L. innocua and E. coli in carrot juice from the combined application of HPP, nisin, and mild temperatures. By replacing the use of HPP alone by these combinations will allow the use of reduced pressures over shorter period of times to process low-acid juices, lowering energy requirements and increasing throughput. This study will aid the beverage processing industry in the development of clean label juice products with fresh-like quality attributes and using considerable less energy to conventional processing.     

Effect of Packaging-Film Thicknesses on Thermal Inactivation of Salmonella and Listeria innocua in Fully Cooked Chicken Breast Meat

ABSTRACT: The study was conducted to determine the effect of packaging-film thicknesses on thermal inactivation of Salmonella and Listeria innocua in cooked chicken breast meat. The meat was inoculated to contain 107 CFU/g of Salmonella or L. innocua and vacuum-packaged in a thickness of 0.0762- or 0.2032-mm film. The packaged meat was heat-treated in a water bath at 68°C for 10 to 120 s and then cooled in an ice-water bath. The survivors of Salmonella or L. innocua were enumerated. This study revealed that packaging-film thicknesses affected thermal inactivation of Salmonella and L. innocua . The results are useful for surface pasteurization of fully cooked and vacuum-packaged meat and poultry products.     

Characterization of Listeria monocytogenes and Listeria innocua from a vegetable processing plant by RAPD and REA

The incidence of Listeria monocytogenes in a vegetable processing plant was investigated over a 23-month period. Frozen ready-to-eat vegetable samples, well as the plant environment, were sampled. The molecular subtyping techniques, Random Amplified Polymorphic DNA (RAPD) and Restriction Endonuclease Analyses (REA), were performed to help investigate the origin and routes of Listeria dissemination.

The low and sporadic incidence of L. monocytogenes made it impossible to establish an epidemiological sequence in the processing plant, though a case of cross-contamination between tomato and ratatouille was detected. Listeria innocua subtyping, however, allowed us to determine the prevalence of several strains in vegetables, and their presence on machinery samples suggested the possibility of cross-contamination during processing.

The low incidence of L. monocytogenes indicated that the risk of listeriosis transmission by vegetable consumption is low. On the other hand, the isolation of the same strain of L. innocua in several surveys pointed out the risk of colonisation on surfaces and machinery. The persistence of Listeria spp. is a cause for concern as can lead to future contamination of vegetables processed in the plant and to a possible increased risk for health. Therefore, periodic controls for the presence of Listeria spp. and a further review of the cleaning and disinfection procedures used in frozen vegetable plants are recommended.     

Inactivation of Listeria innocua in nisin-treated salmon (Oncorhynchus keta) and sturgeon (Acipenser transmontanus) caviar heated by radio frequency

Recent regulatory concerns about the presence of the pathogen Listeria monocytogenes in ready-to-eat aquatic foods such as caviar has prompted the development of postpackaging pasteurization processes. However, caviar is heat labile, and conventional pasteurization processes affect the texture, color, and flavor of these foods negatively. In this study, chum salmon (Oncorhynchus keta, 2.5% total salt) caviar or ikura and sturgeon (Acipenser transmontanus, 3.5% total salt) caviar were inoculated with three strains of Listeria innocua in stationary phase at a level of more than 10(7) CFU/g. L innocua strains were used because they exhibit an equivalent response to L monocytogenes for many physicochemical processing treatments, including heat treatment. The products were treated by immersion in 500 IU/ml nisin solution and heat processed (an 8-D process without nisin or a 4-D process with 500 IU/ml nisin) in a newly developed radio frequency (RF; 27 MHz) heating method at 60, 63, and 65 degrees C. RF heating along with nisin acted synergistically to inactivate L. innocua cells and total mesophilic microorganisms. In the RF-nisin treatment at 65 degrees C, no surviving L. innocua microbes were recovered in sturgeon caviar or ikura. The come-up times in the RF-heated product were significantly lower compared with the water bath-heated caviar at all treatment temperatures. The visual quality of the caviar products treated by RF with or without nisin was comparable to the untreated control.     

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.