Sensitivity analysis of Salmonella enteritidis levels in contaminated shell eggs using a biphasic growth model

Salmonella enteritidis (SE) is a common foodbome pathogen, the transmission of which is primarily associated with the consumption of contaminated Grade A shell eggs. In order to estimate the level of SE present in raw shell eggs, it is necessary to consider the protective effects of the egg albumin, which effectively inhibits SE growth in a time- and temperature-dependent manner. In this study, a SE growth model was produced by combining two mathematical equations that described both the extended lag phase of SE growth (food component) and a SE growth model (pathogen component). This biphasic growth model was then applied to various egg handling scenarios based on the farm-to-table continuum, including in-line and off-line processing facilities with consideration of key events in production, processing, transportation, and storage. Seasonal effects were also studied. Monte Carlo simulation was used to characterize variability in temperature and time parameter values influencing the level of SE to which individuals are exposed. The total level of SE consumed was estimated under best, most likely, and time-temperature abusive handling scenarios. The model estimated that, in most cases, there was no SE growth in contaminated eggs handled under most likely practices, because 10-70% of the yolk membrane remained intact. Under abusive handling scenarios, complete loss of yolk membrane integrity frequently occurred by the time eggs reach the distribution phase, followed by subsequent SE growth, which was often quite rapid. In general, the effect of season and processing method (in-line vs. off-line) was minimal. Further sensitivity analysis demonstrated that the initial SE contamination level significantly influenced the final exposure levels only under no-abuse or mildly abusive conditions. The results of our study suggest that, for maximum reduction of SE exposure level, cooling strategies should not only focus on the on-farm or processing phases, but should emphasize the importance of cooling strategies at the distribution and consumer phases of the farm-to-fork continuum.     

Correlation of eggshell strength and Salmonella enteritidis contamination of commercial shell eggs

Shell quality has been identified as a heritable trait that can be manipulated by genetic selection. Previous research has concluded that many methods of determining shell quality produce variable results. With the development of newer, more precise measuring technologies, shell strength can now be assessed in a consistent, objective fashion. A research project was conducted to determine what role shell strength might play in affecting external Salmonella Enteritidis contamination of egg contents. Visibly clean eggs were collected from an in-line shell egg-processing facility at the accumulator. Eggs were inoculated by dipping in a concentrated suspension of nalidixic acid-resistant Salmonella Enteritidis. After storage, eggs were assessed for shell strength and both external and internal Salmonella Enteritidis contamination. In the first study, there was a significant difference (P < 0.05) in shell strength among the three replicates. No differences between treatments were found for shell strength or Salmonella Enteritidis contamination of contents. In the second study, there were no replicate differences for any of the monitored factors. When rinsate and content samples were enriched, 100% of the rinsates were positive for Salmonella Enteritidis. No content samples were shown to be contaminated with Salmonella Enteritidis during direct plating, but 3 to 5% of the samples from each replicate were positive after enrichment. Correlation analysis of the results from each study found only weak correlations between shell strength and Salmonella Enteritidis contamination on eggshell surface or contents. Within the range of shell strengths recorded in this study, the correlation analysis suggests that shell strength does not play a major role in Salmonella Enteritidis contamination. Further work with eggs that represent a greater range of shell strengths could provide a clearer indication of the interaction of shell strength and Salmonella Enteritidis contamination.     

Efficacy of electrolyzed water in inactivating Salmonella enteritidis and Listeria monocytogenes on shell eggs

The efficacy of acidic electrolyzed (EO) water produced at three levels of total available chlorine (16, 41, and 77 mg/ liter) and chlorinated water with 45 and 200 mg/liter of residual chlorine was investigated for inactivating Salmonella Enteritidis and Listeria monocytogenes on shell eggs. An increasing reduction in Listeria population was observed with increasing chlorine concentration from 16 to 77 mg/liter and treatment time from 1 to 5 min, resulting in a maximal reduction of 3.70 log CFU per shell egg compared with a deionized water wash for 5 min. There was no significant difference in antibacterial activities against Salmonella and Listeria at the same treatment time between 45 mg/liter of chlorinated water and 14-A acidic EO water treatment (P > or = 0.05). Chlorinated water (200 mg/liter) wash for 3 and 5 min was the most effective treatment; it reduced mean populations of Listeria and Salmonella on inoculated eggs by 4.89 and 3.83 log CFU/shell egg, respectively. However, reductions (log CFU/shell egg) of Listeria (4.39) and Salmonella (3.66) by 1-min alkaline EO water treatment followed by another 1 min of 14-A acidic EO water (41 mg/liter chlorine) treatment had a similar reduction to the 1-min 200 mg/liter chlorinated water treatment for Listeria (4.01) and Salmonella (3.81). This study demonstrated that a combination of alkaline and acidic EO water wash is equivalent to 200 mg/liter of chlorinated water wash for reducing populations of Salmonella Enteritidis and L. monocytogenes on shell eggs.     

Heat transfer models for predicting Salmonella enteritidis in shell eggs through supply chain distribution

ABSTRACT:  Egg and egg preparations are important vehicles for Salmonella enteritidis infections. The influence of time–temperature becomes important when the presence of this organism is found in commercial shell eggs. A computer-aided mathematical model was validated to estimate surface and interior temperature of shell eggs under variable ambient and refrigerated storage temperature. A risk assessment of S. enteritidis based on the use of this model, coupled with S. enteritidis kinetics, has already been reported in a companion paper published earlier in JFS . The model considered the actual geometry and composition of shell eggs and was solved by numerical techniques (finite differences and finite elements). Parameters of interest such as local ( h ) and global ( U ) heat transfer coefficient, thermal conductivity, and apparent volumetric specific heat were estimated by an inverse procedure from experimental temperature measurement. In order to assess the error in predicting microbial population growth, theoretical and experimental temperatures were applied to a S. enteritidis growth model taken from the literature. Errors between values of microbial population growth calculated from model predicted compared with experimentally measured temperatures were satisfactorily low: 1.1% and 0.8% for the finite difference and finite element model, respectively.     

Preenrichment versus direct selective agar plating for the detection of Salmonella Enteritidis in shell eggs

The relative effectiveness of two methods for the recovery of Salmonella Enteritidis (SE) from jumbo and medium shell eggs was compared. The first method used in the comparison consisted of a preenrichment of the sample, and the second method was developed by the U.S. Department of Agriculture's Animal and Plant Health Inspection Service (APHIS). Three bulk lots of blended, pooled eggs, each containing 220 liquid whole eggs that were thoroughly mixed manually were artificially inoculated with different levels of SE cells between approximately 10(0) and 10(3) CFU/ml. Twenty samples containing the contents of approximately 10 eggs each (by weight) were withdrawn from each of the inoculated bulk lots and incubated for 4 days at room temperature (ca. 23 degrees C). For the APHIS method, each sample was cultured by direct plating onto brilliant green (BG), brilliant green with novobiocin (BGN), xylose lysine desoxycholate (XLD), and xylose lysine agar Tergitol 4 (XLT4) agars. For the preenrichment method, 25-g portions from each pool were enriched in modified tryptic soy broth with 30 mg/liter of FeSO4. After 24 h of incubation, the preenrichments were subcultured to tetrathionate and Rappaport-Vassiliadis broths, and streaked to BG, BGN, bismuth sulfite, XLD, and XLT4 agar plates. SE isolates were confirmed biochemically and serologically. In all of the experiments, the preenrichment method recovered significantly more SE isolates (P < 0.05) of all the phage types and inoculum levels than did the APHIS method. From a total of 539 jumbo egg test portions analyzed, 381 (71%) were SE-positive by the preenrichment method and 232 (43%) were positive by the APHIS method. From a total of 360 medium egg test portions analyzed, 223 (62%) were SE-positive by the preenrichment method and 174 (48%) were positive by the APHIS method. The preenrichment method provided greater sensitivity for the isolation of SE in contaminated egg slurries than did the APHIS method.     

Inactivation of Salmonella enteritidis during boiling of eggs

A series of inactivation curves for Salmonella enteritidis were determined for boiling eggs using different conditions of time and temperature. No significant influence of egg weight could be found on the temperature evolution in the yolk. The inactivation curves consistently showed an initial slow decline in bacterial number at lower temperatures, after which a very rapid inactivation took place. It was not possible to reproduce this behavior using a traditional inactivation model. A pragmatic model existing in two parts was therefore constructed. When the temperature is below a certain threshold, the inactivation follows a second order temperature dependence. Above the temperature threshold, standard Bigelow inactivation kinetics are assumed.This model could describe the data reasonably well, provided that the decimal reduction time in the Bigelow model was assumed to be different for a fast or slow heating process, respectively. The results suggest that the bacteria are more resistant towards a slower heating process, which is confirmed by analyzing the raw data. A fail-safe model can be obtained by using the parameters associated with the slow heating process. The statistical properties of the calibrated model are satisfactory, and a cross-validation shows that it can be used for egg boiling conditions outside its calibration range.     

Factors influencing inactivation of Salmonella enteritidis in hard-cooked eggs

The inside of a hen's egg, once considered sterile, is now known to occasionally harbor Salmonella Enteritidis. At least two recent outbreaks of salmonellosis in which Salmonella Enteritidis PT34 was involved have been associated with hard-cooked eggs. This study was undertaken to compare D56 degrees C values of Salmonella Senftenberg 775W and six strains of Salmonella Enteritidis isolated from outbreaks associated with eggs. D56 degrees C values for Salmonella Enteritidis in liquid egg yolk ranged from 5.14 to 7.39 min; the D56 degrees C value for Salmonella Senftenberg was 19.96 min. The two PT34 strains from outbreaks associated with hard-cooked eggs did not exhibit significantly higher resistance to heat compared with two PT4 strains and one strain each of PT8 and PT13a. A PT4 strain and a PT34 strain of Salmonella Enteritidis were separately inoculated (10(7) to 10(8) CFU) into the yolk of medium and extra large shell eggs at 10 and 21 degrees C, and survival was monitored using two cooking methods: (i) placing eggs in water at 23 degrees C, heating to 100 degrees C, removing from heat, and holding for 15 min (American Egg Board method) and (ii) placing eggs in water at 100 degrees C, then holding for 15 min at this temperature. Within the 15-min holding periods, inactivation was more rapid using the method recommended by the American Egg Board compared with method 2. Within each cooking method, inactivation was most rapid in medium eggs initially at 21 degrees C. The PT4 strain survived in yolk of extra large eggs initially at 10 degrees C when eggs were held in boiling water 9 min using method 2. The final temperature of the yolk in these eggs was 62.3 +/- 2 degrees C. Of the two methods evaluated for hard cooking eggs, the American Egg Board method is clearly most effective in killing Salmonella Enteritidis in the yolk.     

Efficacy of disinfection of shell eggs externally contaminated with Salmonella enteritidis. Implications for egg testing.

Experimental contamination of the surface of shell eggs by dipping in a culture of Salmonella enteritidis resulted in the presence of Salmonella enteritidis in/on the shells as well as shell membranes but not in the egg content. Disinfection with Lugol's solution, chlorhexidine, ethanol, quarternary ammonium solutions or flaming after dipping in ethanol failed to achieve complete decontamination of the shell and membranes with resulting false positives when eggs were broken for culturing of the content. Dipping eggs for three seconds in boiling water resulted in complete destruction of Salmonella enteritidis in shells and membranes but sometimes caused the eggs to crack. A method of aseptically opening eggs without risk of contaminating the content from the shell or membrane was developed. Salmonella enteritidis deposited in/on the shell and membranes did not multiply during storage of the eggs at 20 degrees C for four weeks, the counts seemed to decrease. No Salmonella enteritidis was detected in the contents of any contaminated eggs.     

Simple and rapid methods for detecting Salmonella enteritidis in raw eggs

The Centers for Disease Control and Prevention estimates there were 300,000 cases of Salmonella enteritidis (SE) in 1997. Egg products were associated with many of the cases. To address this problem, many producers implemented flock surveillance of the SE situation at their facilities. A rapid and simple method for detecting SE from poultry samples is critical for the effective implementation of such testing strategies. A lateral flow device for the detection of SE utilized in this study was manufactured by Neogen, Lansing, MI. The test panel is a presumptive qualitative test system that detects only members of Group D1 Salmonella species. A series of studies were conducted to optimize the test procedure for raw eggs with different sample preparations. A novel antigen extraction method was developed for use with the test panel kit. The detection limit of the test panel kit was increased approximately tenfold when the extraction method was used. Detection of SE was 100% in raw egg pools inoculated with 10 SE cells per ml of egg and incubated at a 1:10 ratio in buffered peptone water (BPW) or tetrathionate brilliant green broth (TBG) for 24 h at 37 degrees C. The developed lateral flow test kit could provide a simple, rapid, and inexpensive method for egg producers and processors to test specifically for Salmonella group D1 serovars, such as SE, in egg samples.     

Individual-based modelling of growth and migration of Salmonella enteritidis in hens' eggs

An individual-based model (IbM) was developed to describe the growth and migration of Salmonella enteritidis in hens' eggs. The Bacteria Simulator (BacSim) environment was used to implement the model; the bacteria are represented by spheres that grow by nutrient uptake and divide in two daughter cells upon exceeding a certain threshold volume. Motility of the Salmonella bacteria was described by a run and tumble mechanism. For the sake of simplicity, the bacteria were assumed to grow exponentially, an appropriate assumption for the initial phase of growth relevant for shelf-life predictions. Both albumen and yolk were assumed to be homogeneous. The impact of several model parameters (chemotaxis, growth rate, initial contamination numbers and bacterial swimming speed) was assessed by a sensitivity analysis. The results show that chemotaxis towards the yolk would have a strong effect on the time needed to reach the vitelline membrane, an aspect that future research should focus on. The contamination position had less impact on the time to reach the vitelline membrane. The simulation results illustrate the need for more detailed knowledge on the subject of bacterial migration in hens' eggs. Our model can easily incorporate this knowledge when it becomes available.     

Growth of Salmonella enteritidis in yolk from eggs laid by immunized hens

After hyperimmunization of laying hens with Salmonella enteritidis, antibodies can be found in egg yolks. This study was conducted to ascertain whether the growth of S. enteritidis would be suppressed in the presence of antibodies contained in egg yolk. Specifically pathogen-free (SPF)-laying hens were immunized with S. enteritidis; eggs were collected, the yolk was separated and the concentration of S. enteritidis antibodies was determined quantitatively by using the enzyme-linked immunosorbent assay (ELISA), the radial immunodiffusion and the bicinchoninic acid protein assay. Then, the yolk was inoculated with approximately 10, 100 or 1000 S. enteritidis cells/ml and incubated at 15, 20 and 30 degrees C for 0, 2, 6 and 24 h. The growth of organisms in each yolk was examined, and the generation times were calculated. The egg yolk from nonimmunized hens served as negative control. The highest level of antibody concentration was found in the hyperimmunized group. There was no difference in the generation times of S. enteritidis between the antibody-positive yolk and the negative yolk at the three different incubation temperatures. The results suggest that antibodies in the yolk do not influence the growth of S. enteritidis, even if the hens are highly immunized.     

肠炎沙门氏菌的LAMP检测方法的建立

建立肠炎沙门氏菌(Salmonella enteritidis)的环介导等温扩增(Loop-mediated isothermal amplification,LAMP)方法,实现对肠炎沙门氏菌的快速检测.通过针对肠炎沙门氏菌血清型特异性基因lygD设计LAMP内外引物对,优化LAMP扩增反应条件,采用包括肠炎沙门氏菌在内的10种不同菌株进行LAMP引物特异性检测;通过系列梯度稀释肠炎沙门氏菌菌液进行LAMP扩增,计算检出限;并对鲜鸡蛋模拟样本进行LAMP检测.结果表明LAMP法可快速特异地检测出肠炎沙门氏菌;细菌培养液检出限为2.33×101 cfu/mL,鲜鸡蛋模拟样品为2.67×l01cfu/mL.该方法反应灵敏度高,可用于食品中肠炎沙门氏菌的快速检测.     

Detection of Salmonella enteritidis and Salmonella typhimurium in foods using a rapid,multiplex real-time recombinase polymerase amplification assay

Salmonella has been recognized as a major foodborne pathogen for humans and animals. In this study, a multiplex real-time recombinase polymerase amplification (RPA) was developed for simultaneous detection of Salmonella enterica serovars, Salmonella enteritidis and Salmonella typhimurium, from chicken, eggs, lettuce, and papaya. The reaction was performed for 20 min at 35°C, and the detection limit of the assay was 10² CFU/ml for pure culture. In food application, the limit of detection (LOD) of S. enteritidis and S. typhimurium using multiplex real-time RPA without enrichment procedure was 10² CFU/25 g, respectively. After enrichment, the LOD of S. enteritidis and S. typhimurium was 10 CFU/25 g. Moreover, the result for Salmonella spp. was not significantly different from those obtained using a culture-based method. Additionally, the assay has a lower cross-reactivity with other pathogenic microorganisms and a good stability performance. Thus, the developed multiplex RPA assay could be used as a rapid tool for the detection of S. enteritidis and S. typhimurium in food.     

High voltage atmospheric cold plasma decontamination of Salmonella enteritidis on chicken eggs

Salmonella enteritidis (SE) accounts for more than 70% of Salmonella spp. infections in humans with a primary source being chicken eggs, that can result from post-lay SE cross-contamination of the shell from contaminated equipment or the environment. The objective of this study was to apply a HVACP treatment that can achieve a minimum 5-log reduction in SE on the surface of artificially inoculated shell eggs with an initial bacterial load of 10⁸ CFU/egg, after a previous disinfection. Optimized HVACP treatment conditions were an indirect treatment with air at 60% humidity at 100 kV for one minute treatment and six hours post-treatment or alternatively, five minutes of treatment and four hours post-treatment. Egg quality parameters of Haugh unit (HU), pH, color, and vitelline membrane and shell strength were tested under the optimized conditions and showed no significant difference (p > 0.05) between treated and untreated eggs.Industrial relevance: Missing information for a possible scale up of a cold plasma system for egg surface decontamination has been addressed by an optimization of HVACP treatment focused on treatment and post-treatment time, essential parameters to have into account in the food industry. These results demonstrate that HVACP is an effective decontamination method for SE on chicken shell eggs and provides a baseline for a future scale up of the process, showing that different combinations of treatment variables can achieve the desired decontamination without affecting to key quality parameters of the egg such as Haugh Unit or vitelline membrane strength.     

肠炎沙门氏菌EMA-PCR检测方法的建立

将荧光染料叠氮溴化乙锭（ethidium monoazide,EMA）与聚合酶链式反应（polymerase chain reaction,PCR）检测技术相结合,用于肠炎沙门氏菌活菌的检测。实验参数优化结果表明,当EMA终质量浓度50 μg/mL、曝光时间10 min时,可以抑制约107 CFU/mL肠炎沙门氏菌死菌DNA的扩增;活菌灵敏度检测结果显示,EMA-PCR方法检测限与单一PCR方法一样均为27.5 CFU/mL,说明EMA处理既不会影响活菌DNA的扩增也不会影响PCR方法的灵敏度。利用EMA-PCR方法检测死活混合菌液时发现,添加EMA的实验组DNA条带亮度会随着活菌比例的降低而变暗,且当样品中全是死菌时,没有目标条带出现;而不添加EMA的对照组DNA条带亮度没有变化,当样品中全是死菌时,目标条带依然清晰可见。说明添加EMA可以达到区分死活菌的目的,EMA-PCR方法只检测样品中的活菌,避免了死菌DNA造成假阳性的可能性。     

Detection of Salmonella in food over 30 h using enrichment and polymerase chain reaction

A 30-h method for the detection of Salmonella spp. in food was developed. The method involved preenrichment in buffered peptone water for 6–8 h, immunomagnetic separation (IMS) using Dynabeads^®anti-Salmonella , selective enrichment in Rappaport-Vassiliadis broth for 16–18 h, lysis of bacterial cells in sodium dodecylsulfate and NaOH solution at 95°C, and the polymerase chain reaction (PCR) using primers ST11 and ST15. The detection limit of the method was 10⁰cfu 25g⁻¹, as determined by the analysis of food samples artificially contaminated with S. enteritidis. When the reference material containing on average 5 cfu of S. panama was used for the artificial contamination, 3 out of 22 food samples were found to be false-negative. When the method was evaluated in comparison with the standard ISO 6579 method on 42 possibly naturally-contaminated food samples, one sample was found positive by the 30-h method, one sample was found positive by the ISO-method, and two samples were found positive by both methods. The developed method proved rapid, but produced a non-zero level of false-negative results.     

Estimating the annual fraction of eggs contaminated with Salmonella enteritidis in the United States

Using available data on the occurrence of Salmonella enteritidis (SE) in US layer flocks and eggs, and a probabilistic scenario tree method, an estimate of the fraction of SE-contaminated eggs produced annually is derived with attendant uncertainty. In lieu of a definitive prevalence survey, the approach presented here provides insight to the relative contribution of various pathways leading to contaminated eggs. A Monte Carlo model with four branches is developed. The first branch predicts the proportion of all US flocks that are SE-affected. The second branch apportions SE-affected flocks into three categories (high, moderate, and low level affected flocks) based on population-adjusted epidemiologic data. The third branch predicts the proportion of affected flocks that are molted and producing eggs during a high risk period subsequent to molt. The fourth branch predicts the fraction of contaminated eggs produced by flocks of the type described by the pathway (e.g. high level affected flocks that are not molted) based on egg sampling evidence from naturally infected flocks. The model is simulated to account for uncertainty in the data used to estimate the branch probabilities. Correlation analysis is used to estimate the sensitivity of model output to various model inputs. The output of this model is an uncertainty distribution for the fraction of all eggs that are SE-contaminated during 1 year of production in the US. The expected value of this distribution is approximately one SE-affected egg in every 20,000 eggs annually produced, and the 90% certainty interval is between one SE-contaminated egg in 30,000 eggs, and one SE-contaminated egg in 12,000 eggs. The model estimates that an average of 14% of all eggs (i.e. contaminated and not contaminated) from affected flocks are produced by high level, non-molted affected flocks, but these flocks are estimated to account for more than two-thirds of the total fraction of contaminated eggs produced annually. Sensitivity analysis also suggests that the proportion of affected flocks that are high level flocks - and the egg contamination frequency for these types of flocks - are the most sensitive model inputs. The model's pathways provide a framework for evaluating interventions to reduce the number of contaminated eggs produced in the US. Furthermore, sensitivity analysis of the model identifies those inputs whose uncertainty is most influential on the model's output. Future farm-level research priorities can be established on the basis of this analysis, but public policy decisions require a fuller exposure assessment and dose-response analysis to account for microbial growth dynamics, meal preparation, and consumption demographics among US egg consumers.     

Salmonella Enteritidis in shell eggs: Current issues and prospects for control

Foodborne illness caused by Salmonella spp. is a worldwide problem. In the United States Salmonella Enteritidis is the second most commonly isolated serotype from human illness, and is known to be strongly associated with shell eggs and egg containing products. Eggs can become contaminated internally either by penetration through the shell or directly during formation in the reproductive tract. This review begins with a brief account of the physiology of egg production and the various physical and chemical barriers the egg possesses to prevent bacterial contamination. Factors involved in vertical and horizontal transmission of S. Enteritidis are examined, as well as the role of forced molt in colonization of the hen. Pre- and post-harvest mitigation strategies are also discussed.     

Prevalence of Salmonella spp. in poultry broilers and shell eggs in Korea

This study was conducted to determine the presence of Salmonella spp. in raw broilers and shell eggs in Korea. In total, 135 dozen shell eggs and 27 raw broilers were tested. None of the egg yolks were found to contain Salmonella organisms but Escherichia coli, Escherichia hermanii, and Citrobacter freundii were isolated from egg shells. Salmonella spp. were detected in 25.9% of raw broilers, and Salmonella serotypes isolated from raw broilers were Salmonella Enteritidis, Salmonella Virchow, and Salmonella Virginia. D-values and antibiotic resistance of Salmonella isolates were also investigated. D-values of Salmonella enteritidis, Salmonella Virginia, and Salmonella Virchow in tryptic soy broth at 55 degrees C were 2.36, 2.13, and 0.70 min and 0.53, 0.37, and 0.20 min at 60 degrees C, respectively. All Salmonella isolates showed multiple antibiotic resistance patterns and were resistant to penicillin and vancomycin. One strain of Salmonella Enteritidis showed resistance to 12 antibiotics used in this study.     

Inactivation of Salmonella enteritidis and Salmonella senftenberg in liquid whole egg using generally recognized as safe additives, ionizing radiation, and heat

The effect of combining irradiation and heat (i.e., irradiation followed by heat [IR-H]) on Salmonella Enteritidis and Salmonella Senftenberg inoculated into liquid whole egg (LWE) with added nisin, EDTA, sorbic acid, carvacrol, or combinations of these GRAS (generally recognized as safe) additives was investigated. Synergistic reductions of Salmonella populations were observed when LWE samples containing GRAS additives were treated by gamma radiation (0.3 and 1.0 kGy), heat (57 and 60 degrees C), or IR-H. The presence of additives reduced the initial radiation Dgamma -values (radiation doses required to eliminate 90% of the viable cells) by 1.2- to 1.5-fold, the thermal decimal reduction times (D,-values) by up to 3.5- and 1.8-fold at 57 and 60 degrees C, respectively, and the thermal D,-values after irradiation treatments by up to 3.4- and 1.5-fold at 57 and 60 degrees C, respectively, for both Salmonella serovars. Of all the additives investigated, nisin at a concentration of 100 IU/ml was the most effective at reducing the heat treatment times needed to obtain a 5-log reduction of Salmonella. Thus, while treatments of 21.6 min at 57 degrees C or of 5 min at 60 degrees C should be applied to achieve a 5-log reduction for Salmonella in LWE, only 5.5 min at 57 degrees C or 2.3 min at 60 degrees C after a 0.3-kGy radiation pretreatment was required when nisin at a concentration of 100 IU/ml was used. The synergistic reduction of Salmonella viability by IR-H treatments in the presence of GRAS additives could enable LWE producers to reduce the temperature or processing time of thermal treatments (current standards are 60'C for 3.5 min in the United States) or to increase the level of Salmonella inactivation.