Incubation of supplemented egg contents pools to support rapid detection of Salmonella enterica serovar Enteritidis

Detecting internal contamination of eggs with Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) is an important aspect of efforts to identify infected laying flocks. When egg contents pools are tested for Salmonella Enteritidis, a preliminary incubation step is often employed to allow small initial populations of contaminants to multiply to more easily detectable numbers. Consistent detection of Salmonella Enteritidis in egg pools by direct plating requires the presence of at least 10(5) CFU/ml, whereas some very rapid methods can require as many as 10(7) CFU/ml. The present study determined the rates at which initial inocula of approximately 10 Salmonella Enteritidis cells multiplied in 10-egg pools, some of which were supplemented with concentrated nonselective enrichment broth or with a source of iron. At 37 degrees C, Salmonella Enteritidis concentrations in supplemented egg pools usually reached 10(5) CFU/ml within 12 h and 10(7) CFU/ml by 12 to 15 h of incubation. At 25 degrees C, Salmonella Enteritidis concentrations in supplemented egg pools typically attained 10(5) CFU/ml by 18 to 27 h and 10(7) CFU/ml by 27 to 36 h of incubation. At both temperatures, Salmonella Enteritidis multiplication was significantly slower in unsupplemented pools. Accordingly, the length of incubation time necessary for consistent detection of small numbers of Salmonella Enteritidis in egg contents pools depends on the incubation temperature used, on whether the egg pools are supplemented to increase the rate of bacterial multiplication, and on the sensitivity of subsequent tests applied to the incubated pools.     

Comparison of homogenization methods for recovering Salmonella Enteritidis from eggs

For Salmonella Enteritidis (SE) detection, shell eggs have been homogenized with stomachers, with electric blenders, and by hand massaging. However, to date, there have been no published reports addressing whether the method of homogenization affects the recovery of SE from raw eggs. Three inoculum levels (10, 126, and 256 SE cells per pool of 10 eggs) were used to conduct three experiments. The 10-egg pools were homogenized by one of four homogenization methods--mechanical stomaching, electric blending, hand massaging, and hand stirring-for 30 s. The homogenized eggs were then incubated at 37 degrees C, and SE colonies were enumerated after 24 and 48 h of incubation. After 24 h of incubation, no SE was recovered from egg samples from stomached or electrically blended pools inoculated with <10 cells, while levels of 106 CFU/ml were found for samples from whipped or hand-massaged pools inoculated with <10 cells. Similarly, after 24 h of incubation, the numbers of SE cells recovered from hand-massaged or hand-stirred egg pools inoculated with 126 cells were significantly larger than the numbers recovered from stomached or electrically blended egg pools inoculated with 126 cells. The number of SE cells recovered from samples homogenized with a blender was still significantly smaller than the numbers recovered from samples homogenized by the other three methods when the inoculum level was increased to 256 CFU per pool. However, the SE count for all samples approached 9 log10 CFU/ml after 48 h of incubation. It is concluded that the detection of small SE populations in shell egg samples could be improved with the use hand massaging and hand stirring for homogenization.     

Multiplication of Salmonella enteritidis on the yolk membrane and penetration to the yolk contents at 30 degrees C in an in vitro egg contamination model

Refrigeration to limit bacterial multiplication is a critical aspect of efforts to control the transmission of Salmonella enterica serovar Enteritidis (SE) to consumers of contaminated eggs. Although the nutrient-rich yolk interior is an uncommon location for SE contamination in freshly laid, naturally contaminated eggs, migration across the vitelline membrane could lead to rapid bacterial multiplication even when the initial site of deposition is outside the yolk. Multiplication on the yolk membrane (before, or in addition to, multiplication within the yolk contents) could be another source of increased risk to consumers. The present study used an in vitro egg contamination model to compare the abilities of four strains of SE to either multiply in association with the yolk membrane or migrate through that membrane to reach the yolk contents during 36 h of incubation at 30 degrees C. After inoculation onto the exterior surface of intact, whole yolks, all four SE strains penetrated the vitelline membrane to reach the yolk contents (at an overall frequency of 11.5%) after 12 h of incubation. The mean log concentration of SE was significantly higher in whole yolks (including yolk membranes) than in yolk contents at both 12 h (0.818 versus 0.167 CFU/ ml) and 36 h (2.767 versus 1.402 CFU/ml) of incubation. These results demonstrate that SE multiplication on the vitelline membrane may both precede and exceed multiplication resulting from penetration into the yolk contents during the first 36 h of unrefrigerated storage, reinforcing the importance of rapid refrigeration for protecting consumers from egg-transmitted illness.     

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.     

Optimization of iron supplementation for enhanced detection of Salmonella Enteritidis in eggs

Mixed raw egg contents were inoculated with approximately 10 CFU of Salmonella Enteritidis and supplemented with 0 to 7 mg of FeSO4 per g of egg contents. Egg contents were then incubated at 37 degrees C, and Salmonella Enteritidis colonies were enumerated for up to 106 h. Iron supplementation significantly enhanced the growth of Salmonella Enteritidis. Within the first 24 h of incubation, the optimum iron level for Salmonella Enteritidis growth in egg contents was between 0.2 and 2 mg of FeSO4 per g of egg contents. After 24 h of incubation at 37 degrees C. Salmonella Enteritidis counts in eggs supplemented with 0.5 mg of FeSO4 per g of egg contents consistently reached approximately 1 x 10(9) CFU/ml, whereas Salmonella Enteritidis counts in eggs without iron supplementation varied from less than 5 CFU/ml to 8.4 x 10(6) CFU/ml. A 3 by 3 factorial design was used to study the effect of type of preenrichment and level of iron supplementation on the growth of Salmonella Enteritidis in egg contents. No significant differences in Salmonella Enteritidis counts between preenrichment and nonpreenrichment treatments were observed when egg contents were supplemented with 0.5 mg of FeSO4 per g of egg contents. It was concluded that preenrichment was not necessary for isolation of Salmonella Enteritidis from eggs. The effect of iron supplementation on the sensitivity of detection by the direct plating method was investigated. The direct plating method detected a significantly higher percentage of Salmonella Enteritidis in raw egg contents supplemented with 0.5 mg of FeSO4 per g of egg contents (90%) than in raw egg contents without iron supplementation (63.3%).     

Effect of refrigeration on in vitro penetration of Salmonella enteritidis through the egg yolk membrane

Internally contaminated eggs have been implicated as leading sources of transmission of Salmonella Enteritidis (SE) to humans. Although SE is not often deposited inside the nutrient-rich yolks of naturally contaminated eggs, penetration through the vitelline membrane to reach the yolk contents could result in rapid bacterial multiplication. In previous studies, such penetration has been observed occasionally at warm temperatures during experiments with in vitro egg contamination models. The present study was conducted to determine whether refrigeration affects the frequency of in vitro SE penetration of the egg yolk membrane. After inoculation of small numbers of SE onto the outside of the vitelline membranes of intact yolks, immediate refrigeration of contaminated samples prevented the penetration of SE into the egg yolk contents during 24 h of storage. However, SE penetrated inside the yolk contents in 4% of contaminated egg samples refrigerated after 2 h of storage at 30 degrees C, 15% of samples refrigerated after 6 h of storage at 30 degrees C, and 40% of samples stored at 30 degrees C for 24 h (48 samples per treatment group). These results highlight the value of prompt refrigeration for restricting the opportunities for SE to multiply to high numbers inside the yolks of contaminated eggs.     

Growth of Salmonella enterica serovar Enteritidis in albumen and yolk contents of eggs inoculated with this organism onto the vitelline membrane

By using an in vitro model simulating the potential opportunities for Salmonella enterica serovar Enteritidis (SE) to proliferate within eggs contaminated with this organism following oviposition, we investigated growth of SE in eggs. Seventy to 140 CFU of one of three SE strains originating either from egg contents, chicken meat, or a human infection were experimentally inoculated onto the vitelline membrane of eggs collected from specific-pathogen-free flocks of chickens and incubated at 25 degrees C. SE organisms were detected in 6 of 71 yolk contents of the eggs inoculated with any of the test strains attaining levels ranging from 2.0 x 10(2) to 4.2 x 10(8) CFU/ml by day 6. The organisms were also detected in the albumen from 38 of 55 eggs tested, growing to levels ranging from 1.0 x 10(2) to 4.3 x 10(8) CFU/ml by day 6 after inoculation. An additional three yolk contents and 15 albumen samples were culture positive for SE following enrichment. There was no correlation between the number of the organisms in the yolk contents and that in the albumen from each of the eggs. When 73 to 91 CFU of the egg strain were inoculated into samples of separated albumen obtained from eggs that were stored at 4 degrees C for 1 to 4 weeks or at 25 degrees C for 1 week, slight growth (3.0 x 10(2) to 7.4 x 10(3) CFU/ml) was found in only 3 of the 60 albumen samples by day 6 after inoculation, but the organisms were recovered from 52 samples following enrichment. The results suggest that the environment on or near the vitelline membrane can be conducive to SE proliferation over time.     

Effect of temperature on the growth response of Salmonella enteritidis inoculated onto the vitelline membranes of fresh eggs

The growth response of Salmonella Enteritidis (SE) on the vitelline membrane in vitro was studied with the use of a special tube devised specifically for the inoculation of SE onto the vitelline membrane and for the sampling of the yolk near the inoculation site. This latter ability allowed the detection of the movement of SE into the yolk. The growth of SE on the membrane was compared with that of SE inoculated into yolk and albumen in vitro and in ovo in fresh in-shell eggs. The incubation time was 2 days, and the incubation temperatures were 4, 8, 15, 27, and 37 degrees C. Comparison of the results obtained for in vitro growth showed that at 4, 8, and 15 degrees C, SE behaved as if it were in the albumen, with its numbers decreasing over time. At 27 and 37 degrees C, SE grew as if it were in yolk, with a maximum increase of 4.5 log CFU after 2 days at 37 degrees C. In no experiments involving growth on the vitelline membrane did SE appear in the yolk. Comparisons between in vitro and in ovo growth responses of SE in yolk and albumen indicate that SE growth on the membrane parallels that in the in-shell egg.     

Influence of eggshell condensation on eggshell penetration and whole egg contamination with Salmonella enterica serovar enteritidis

Shells of agar-filled and whole eggs were inoculated with 10(3) to 10(4) CFU of Salmonella enterica serovar Enteritidis per eggshell. The agar-filled eggs were used to study bacterial eggshell penetration, and the whole egg results were used to characterize contamination of the egg contents. In each group, half of the eggs were stored for 21 days at 20 degrees C and 60% relative humidity (RH), and the other half was stored for 24 h at 6 degrees C and then for 20 days at 20 degrees C. The latter conditions resulted in condensation on the eggshell for 30 min from the moment the eggs were placed in the 20 degrees C chamber. Taking into account the ages at which hens were studied (39, 53, and 67 weeks), an average of 62% of the eggshells with condensate were penetrated compared with 43% for the control group; this difference was significant (P < 0.01). No significant difference in whole egg contamination was found; 18% of the control eggs were contaminated compared with 22% of the condensate eggs. Whole egg contamination was significantly higher for eggs from the hens at an older age (67 weeks). This difference probably was not due to a higher penetration potential because differences were not observed for the corresponding agar-filled eggs. Condensation on the eggshell seemed to encourage bacterial penetration of the eggshell but had a smaller impact on whole egg contamination.     

A robotic DNA purification protocol and real-time PCR for the detection of Campylobacter jejuni in foods

Primers designed to amplify a Campylobacter jejuni cadF gene sequence were used in an SYBR Green I real-time PCR assay as an alternative to conventional bacteriological methods for the rapid detection of C. jejuni in foods. Twenty-five grams of chicken skin (breast and thigh) was contaminated by adding approximately 1, 10, or 50 CFU of C. jejuni ATCC 35560. Twenty-five grams of pork and 25-ml aliquots of milk were also inoculated with 1 and 10 CFU of the pathogen. The samples were incubated in Bolton broth for different periods at 37 and 42 degrees C under microaerophilic conditions. Using a commercial robotic DNA purification system, DNA was extracted and purified from 1-ml aliquots of the enrichment cultures before and after centrifugation of the 250-ml enrichment broth at 15,900 x g for 10 min at 4 degrees C. The DNA was used as the template in a real-time PCR assay. C. jejuni was detected after 12 h of enrichment from samples inoculated with about 50 CFU/25-g sample. After centrifugation, an enrichment step of 8 h was sufficient to allow detection of pathogen in samples inoculated with 10 CFU/25 g. However, 24 h of enrichment was necessary to detect pathogen in samples inoculated with approximately 1 CFU/25 g. The real-time PCR protocol developed in this study significantly reduced the detection time of C. jejuni in foods.     

Laying season and egg shell cracks on the growth of Salmonella enteritidis in the egg albumen during storage

We studied the effects of laying seasons and egg shell cracks on the ability of egg albumen to support the growth of Salmonella Enteritidis (SE) in eggs. Hens eggs used were those laid in February, June, and October in a farm in Japan and stored at 10, 20, and 30 degrees C, and at 30 degrees C after storage at 10 degrees C, immediately after receipt or after cracking the shell. At several-day intervals during storage, the egg contents were poured into a dish, SE was inoculated into albumen, and then the growth of SE during 3 days incubation at 18 degrees C was measured. The results demonstrated that storage temperature and laying season affected the growth of SE in the egg albumen. The proportion of eggs upon which albumen allowed the growth of SE was higher in the eggs stored at 30 degrees C than those stored at 10 degrees C. The growth of SE in eggs was lowest in the following order of laying: February, October, and June. SE grew preferably in albumen of cracked eggs than intact eggs.     

Identification of Enterobacteriaceae from washed and unwashed commercial shell eggs

To evaluate the effect of processing on the safety and quality of retail shell eggs, a storage study was conducted with unwashed and commercially washed eggs. This work demonstrated that commercial processing decreased microbial contamination of eggshells. To know which species persisted during storage on washed or unwashed eggs, Enterobacteriaceae isolates were selected and identified biochemically. For each of three replications, shell eggs were purchased from a commercial processing plant, transported back to the laboratory, and stored at 4 degrees C. Once a week for 6 weeks, 12 eggs for each treatment (washed and unwashed control) were rinsed in sterile phosphate-buffered saline. A 1-ml aliquot of each sample was plated onto violet red bile glucose agar with overlay and incubated at 37 degrees C for 24 h. Following incubation, plates were observed for colonies characteristic of the family Enterobacteriaceae. A maximum of 10 isolates per positive sample were streaked for isolation before being identified to the genus or species level using commercially available biochemical strips. Although most of the isolates from the unwashed control eggs belonged to the genera Escherichia or Enterobacter, many other genera and species were identified. These included Citrobacter, Klebsiella, Kluyvera, Pantoea, Providencia, Rahnella, Salmonella, Serratia, and Yersinia. Non-Enterobacteriaceae also recovered from the unwashed egg samples included Xanthomonas and Flavimonas. Very few washed egg samples were contaminated with any of these bacteria. These data provide useful information on the effectiveness of processing in removing microorganisms from commercial shell eggs.     

Automated rapid screening of foods for the presence of salmonellae

We previously reported an accelerated method to detect salmonellae in foods. It consisted of a preenrichment step (6 h) followed by immunomagnetic separation (IMS) and automated detection, based on positive biochemical characteristics, during overnight incubation. We describe here a simplified procedure with a detection limit of 2 CFU/25 g of food, a procedure that does not require the IMS step. Liquid eggs, milk, ice cream, or poultry were inoculated with Salmonella or non-Salmonella Enterobacteriaceae organisms (2 to 100 CFU/25 g). Following preenrichment for 6 h at 35 degrees C, samples (3 ml) were added to selective liquid media (6 ml). Automated light transmittance was recorded during incubation at 42 degrees C in the BioSys instrument. Salmonella-positive samples were identified by a sharp drop in transmittance (caused by black discoloration of the media) within 18 h of incubation following the formation of hydrogen sulfide and its reaction with iron ions. Detection of 2 to 10 CFU/25 g of the tested foods was completed in 24 h. A total of 24 Salmonella spp., including heat-injured cells, and 37 non-Salmonella Enterobacteriaceae were tested using the procedure.     

Dynamic predictive model for growth of Salmonella enteritidis in egg yolk

ABSTRACT:  Salmonella Enteritidis (SE) contamination of poultry eggs is a major human health concern worldwide. The risk of SE from shell eggs can be significantly reduced through rapid cooling of eggs after they are laid and their storage under safe temperature conditions. Predictive models for the growth of SE in egg yolk under varying ambient temperature conditions (dynamic) were developed. The growth of SE in egg yolk under several isothermal conditions (10, 15, 20, 25, 30, 35, 37, 39, 41, and 43 °C) was determined. The Baranyi model, a primary model, was fitted with growth data for each temperature and corresponding maximum specific growth rates were estimated. Root mean squared error (RMSE) values were less than 0.44 log₁₀ CFU/g and pseudo- R ² values were greater than 0.98 for the primary model fitting. For developing the secondary model, the estimated maximum specific growth rates were then modeled as a function of temperature using the modified Ratkowsky's equation. The RMSE and pseudo- R ² were 0.05/h and 0.99, respectively. A dynamic model was developed by integrating the primary and secondary models and solving it numerically using the 4th-order Runge–Kutta method to predict the growth of SE in egg yolk under varying temperature conditions. The integrated dynamic model was then validated with 4 temperature profiles (varying) such as linear heating, exponential heating, exponential cooling, and sinusoidal temperatures. The predicted values agreed well with the observed growth data with RMSE values less than 0.29 log₁₀ CFU/g. The developed dynamic model can predict the growth SE in egg yolk under varying temperature profiles.     

Influence of the conditions of storage and cooking on growth,invasion and survival of Salmonella enteritidis in eggs

Basal studies for the confirmation of sanitary rules in the kitchen were performed, focusing on preventing an outbreak of food poisoning due to eggs contaminated with Salmonella Enteritidis (SE), using hen and quail eggs. SE did not grow at 5 degrees C but grew markedly at 25 degrees C in eggs. The invasion and growth of SE were marked under very humid conditions regardless of whether the eggshell was damaged. The invasion of SE into egg also occurred when eggs were taken in and out of the refrigerator. Moreover, SE was spread immediately to all non-contaminated eggs when SE-contaminated eggs were cracked into a bowl with non-contaminated eggs. In homemade mayonnaise containing 15% vinegar, sterilization took several hours to occur. On a stainless-steel bowl, SE survived for 2 weeks or more. These findings suggest that it is necessary to pay attention to secondary contamination.     

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.     

Multiplication of Salmonella enteritidis in egg yolks after inoculation outside, on, and inside vitelline membranes and storage at different temperatures

Prompt refrigeration to restrict bacterial growth is important for reducing eggborne transmission of Salmonella enterica serovar Enteritidis (SE). The nutrient-rich yolk interior is a relatively infrequent location for initial SE deposition in eggs, but migration across the vitelline membrane can result in rapid bacterial multiplication during storage at warm temperatures. The objective of the present study was to measure the multiplication of SE in yolks after introduction at three different locations and subsequent storage at a range of temperatures. Using an in vitro egg contamination model, approximately 100 CFU of SE was inoculated either inside yolks, onto the exterior surface of vitelline membranes, or into the adjacent albumen. After storage of samples from each inoculation group at 10, 15, 20, and 25°C for 24 h, SE was enumerated in yolks. For all three inoculation locations, the final SE levels in yolks increased significantly with increasing storage temperatures. At all storage temperatures, significant differences in SE multiplication were observed between inoculation sites (yolk inoculation>vitelline membrane inoculation>albumen inoculation). At 25°C, final log concentrations of 7.759 CFU of SE per ml (yolk inoculation), 2.014 CFU/ml (vitelline membrane inoculation), and 0.757 CFU/ml (albumen inoculation) were attained in yolks after storage. These results demonstrate that, even when the initial site of SE deposition is outside the egg yolk, substantial multiplication supported by yolk nutrients can occur during the first day of storage and the risk of bacterial growth increases at higher ambient storage temperatures.     

Growth ofSalmonella enteritidis(SE) in egg contents from hens vaccinated with an SE bacterin

Vaccination of hens withSalmonella enteritidis(SE) bacterin has become an important industry management tool to reduce both the incidence of SE in flocks and the production of SE-contaminated eggs. After vaccination, antibodies to SE can be found in both the serum and egg yolks. The current study was undertaken to examine whether the antibodies deposited in eggs after vaccination would have any effect on thein vitrogrowth of 10 SE seeded into pooled egg contents and incubated at 37°C for 24 h. The SE inoculum grew well in egg contents from control-vaccinated hens, and a high percentage of these samples were culture positive for SE. Conversely, significantly fewer egg contents from vaccinated hens were positive for SE at weeks 2, 3, 4, and 5 post-primary vaccination and weeks 1, 2, and 4 post-secondary vaccination. After 24 h incubation, the SE inoculum grew to 10⁸–10⁹organisms ml⁻¹in egg contents from control hens, while growing to less than 10⁴organisms ml⁻¹in egg contents from vaccinated hens. No inhibition of growth was observed for a second organism,Proteus mirabilis, in egg contents from bacterin-vaccinated hens, indicating antigen-specificity of the activity. Inhibition of SE growth was observed in egg contents from vaccinated hens diluted 1:5 in eggs from control hens. Mixed results were observed for egg contents receiving a 10-fold higher SE inoculum with significant inhibition of growth observed in one trial and minimal inhibition observed in a second trial. Supplementing the egg samples with an iron source abrogated the inhibition, suggesting that the iron- restricted environment found in eggs may play a role in the observed inhibition.     

Growth and survival of Escherichia coli O157:H7 and Listeria monocytogenes in egg products held at different temperatures

Growth and survival of Escherichia coli O157:H7 and Listeria monocytogenes in steamed eggs and scrambled eggs held at different temperatures (5, 18, 22, 37, 55, and 60 degrees C) were investigated in the present study. Among the holding temperatures tested, both pathogens multiplied best at 37 degrees C followed by 22, 18, and 5 degrees C. In general, E. coli O157:H7 grew better in the egg products than L. monocytogenes did at all the storage temperatures tested except at 5 degrees C. E. coli O157:H7 did not grow in steamed eggs and scrambled eggs held at 5 degrees C. L. monocytogenes showed a slight population increase of approximately 0.6 to 0.9 log CFU/g in these egg products at the end of the 36-h storage period at 5 degrees C. The population of both pathogens detected in the egg products was affected by the initial population, holding temperature, and length of the holding period. It was also noted that L. monocytogenes was more susceptible than E. coli O157:H7 in steamed eggs held at 60 degrees C. After holding at 60 degrees C for 1 h, no detectable viable cells of L. monocytogenes with a population reduction of 5.4 log CFU/g was observed in steamed eggs, whereas a lower population reduction of only approximately 0.5 log CFU/ml was noted for E. coli O157:H7.     

Behavior of Listeria monocytogenes in pasteurized milk during fermentation with lactic acid bacteria

The behavior of Listeria monocytogenes in pasteurized milk during fermentation with starter and nonstarter lactic acid bacteria was investigated. Pasteurized milk was co-inoculated with approximately 10(4) CFU/ml of L. monocytogenes and 10(6) CFU/ml of Lactococcus lactis, Lactococcus cremoris, Lactobacillus plantarum, Lactobacillus bulgaricus, or Streptococcus thermophilus. Inoculated milks were incubated at 30 degrees C or 37 degrees C for 24 to 72 h. Listeria monocytogenes survived and also grew to some extent during incubation in the presence of all starter cultures; however, inhibition ranged from 83 to 100% based on maximum cell populations. During incubation with L. bulgaricus and L. plantarum, L. monocytogenes was completely inactivated after 20 h and 64 h of incubation at 37 degrees C and 30 degrees C, respectively. The pH of the fermenting milks declined steadily throughout the fermentation periods and was approximately 4.2 at the conclusion of the experimental period regardless both of the starter culture and pathogen combination or the temperature of incubation.