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 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.     

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.     

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.     

Outgrowth of Salmonellae and the physical property of albumen and vitelline membrane as influenced by egg storage conditions

This study was undertaken to determine the influence of storage time and temperature on the volume, weight, and pH of egg albumen, the physical strength of vitelline membrane, and the fate of Salmonella Enteritidis artificially inoculated into egg albumen. A fiber-optic probe was used for inoculation with Salmonella Enteritidis at 10(2), 10(4), or 10(6) cells per egg. Both fresh and inoculated eggs were stored at 4, 10, and 22 degrees C for 6 weeks. Five fresh uninoculated eggs from each storage group were collected each week, and the weight, volume, and pH of the egg albumen were measured. The forces, energies, and degrees of membrane deformation required to rupture the vitelline membranes also were determined from either albumen-free yolks or yolks surrounded by albumen. In separate experiments, five inoculated eggs were evaluated each week for populations of Salmonella Enteritidis. When the eggs were stored at 4 degrees C, the albumen retained significantly more volume and weight and had a relatively lower pH. The vitelline membranes from eggs stored at 4 and 10 degrees C required more force and energy for rupture. Salmonellae flourished at 22 degrees C, even in the albumen with the lowest initial population, 10(2) cells per egg. Storage at 4 and 10 degrees C inhibited the growth of salmonellae in the albumen of eggs with initial populations of 10(2), 10(4), or 10(6) cells per egg. In eggs with initial Salmonella populations of 10(6) cells per egg that were stored at 22 degrees C, the populations of reached as high as 10(10) cells per egg after 4 weeks of storage. Storage at 4 and perhaps 10 degrees C postponed the aging process of chicken eggs, preserved the antimicrobial agents of the albumen, and maintained the integrity of vitelline membrane. Low-temperature storage therefore had a significant impact on the safety and overall quality of the eggs.     

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.     

Multiplication and motility of Salmonella enterica serovars enteritidis, infantis, and montevideo in in vitro contamination models of eggs

The invasive ability of Salmonella enterica serovars Enteritidis, Infantis, and Montevideo in eggs was examined. Strains of these serovars originating from egg contents, laying chicken houses, and human patients were experimentally inoculated (0.1-ml dose containing 78 to 178 cells) onto the vitelline membrane of eggs collected from specific-pathogen-free chickens and incubated at 25 degrees C. The test strains were detected in 25 of 138 yolk contents by day 6, indicating the penetration of Salmonella organisms through the vitelline membrane. There were no significant differences in overall rates of penetration between serovars. The organisms were also detected in the albumen from 125 of 138 eggs tested by day 6. Growth to more than 10(6) CFU/ml was observed in 48 of the 125 albumen samples. An inoculum of 1000 Salmonella cells was added to 15 ml of albumen at the edge of a petri plate. A 10-mm-diameter cylindrical well, the bottom of which was sealed with a polycarbonate membrane with 3.0-microm pores, was filled with egg yolk and placed into the albumen at the center of the dish, which was maintained at 25 degrees C. Experiments were performed in triplicate with each strain. Salmonella organisms in all the albumen samples were detected by day 11. However, motility of the organisms toward the yolk was observed in only two dishes inoculated with the Salmonella Enteritidis strain from a human patient and in one dish inoculated with the Salmonella Infantis strain from liquid egg. The albumen samples obtained from the dishes inoculated with the Salmonella Enteritidis strain had high numbers of bacteria (>10(8) CFU/ml). The present study suggests that Salmonella organisms in egg albumen are unlikely to actively move toward the yolk, although depositionon or near the vitelline membrane can be advantageous for proliferation.     

In vitro Survival and Growth of Salmonella Typhimurium inoculated on Yolk Membrane after long Term refrigerated Storage of Shell Eggs

Salmonella enterica serovar Typhimurium has been isolated from commercial egg production facilities in the United States. Given its importance as a causative organism for food-borne salmonellosis, identifying approximate timelines for bacterial invasion of the egg is needed. The objective of this study was to examine net growth of S. Typhimurium in egg components over time. In trial 1 eggs were collected over a 24 hour period from a flock of single comb white leghorn hens while in trial 2 eggs were picked up from a commercial laying source once a week over the course of eight weeks and stored. Eggs were held at refrigeration temperature and each week, subsets of eggs were cracked, separated into yolk and albumen components, and inoculated with 10⁸ CFU/ml of novobiocin and nalidixic acid (NO/NA) resistant S. Typhimurium onto the vitelline membrane of the egg. Yolks were then covered with albumen. Eggs were incubated for twenty-four hours at 25°C. After incubation eggs were again separated into albumen, yolk, and vitelline membrane samples. In trial 1, S. Typhimurium net growth occurred in albumen by the second week and continued from 4 to 8 weeks while in trial 2 net growth only occurred at week 5 and 7. S. Typhimurium net growth on vitelline membranes occurred by 2 weeks and continued from 4 to 8 weeks in trial 1 while no net growth occurred in trial 2 over the 8 week period. Yolk samples showed no net increases in S. Typhimurium populations over the 8 week period. An erratum to this article is available at .     

In vitro Survival and Growth of Salmonella Typhimurium inoculated on Yolk Membrane after long Term refrigerated Storage of Shell Eggs

Salmonella enterica serovar Typhimurium has been isolated from commercial egg production facilities in the United States. Given its importance as a causative organism for food-borne salmonellosis, identifying approximate timelines for bacterial invasion of the egg is needed. The objective of this study was to examine net growth of S. Typhimurium in egg components over time. In trial 1 eggs were collected over a 24 hour period from a flock of single comb white leghorn hens while in trial 2 eggs were picked up from a commercial laying source once a week over the course of eight weeks and stored. Eggs were held at refrigeration temperature and each week, subsets of eggs were cracked, separated into yolk and albumen components, and inoculated with 10⁸ CFU/ml of novobiocin and nalidixic acid (NO/NA) resistant S. Typhimurium onto the vitelline membrane of the egg. Yolks were then covered with albumen. Eggs were incubated for twenty-four hours at 25°C. After incubation eggs were again separated into albumen, yolk, and vitelline membrane samples. In trial 1, S. Typhimurium net growth occurred in albumen by the second week and continued from 4 to 8 weeks while in trial 2 net growth only occurred at week 5 and 7. S. Typhimurium net growth on vitelline membranes occurred by 2 weeks and continued from 4 to 8 weeks in trial 1 while no net growth occurred in trial 2 over the 8 week period. Yolk samples showed no net increases in S. Typhimurium populations over the 8 week period.     

Growth of Salmonella enteritidis in artificially contaminated eggs: the effects of inoculum size and suspending media.

Growth profiles of two isolates of Salmonella enteritidis phage type (PT) 4 inoculated into either the albumen of whole shell eggs or into separated albumen were found to be markedly affected by the size of the inoculum and the composition of the medium used to suspend the cells prior to inoculation. Using our model with an inoculum of two cells, multiplication of the Salmonella was not seen in 93% of eggs held at 20 degrees C for 8 days. In approximately 7% of eggs, however, growth occurred during the 8 days of storage. If the inoculum equaled or exceeded 25 cells per egg when eggs were subsequently stored at 20 degrees C, or 250 cells per egg when eggs were stored at 30 degrees C, high levels of growth of Salmonella in the egg occurred significantly more frequently than when the inoculum was two cells. High levels of growth were also seen more frequently if the inoculum was suspended in buffered peptone water or maximal recovery diluent rather than in phosphate buffered saline. Growth of Salmonella in separated albumen occurred very infrequently (1.1% of samples) at low inoculum levels and did not become significant until the inoculum was 250 cells or greater. Growth in the albumen was unaffected by the composition of the suspending medium. Provided that the inoculum was approximately 2 cells per egg and the bacteria were suspended in PBS, observed growth profiles of S. enteritidis inoculated into the albumen of whole eggs resembled those in naturally contaminated eggs.     

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.     

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 serovars in hens’ egg albumen as affected by storage prior to inoculation

Different Salmonella enterica serovars, including Enteritidis, were tested for growth at 20°C in separated albumen upon inoculation with 39 cfu ml⁻¹. The albumen was fresh or stored for up to 3 weeks prior to inoculation (p.i.) either in the shell egg or separated from the yolk. The serovar Enteritidis did not behave differently than the other serovars indicating that the association between human S. Enteritidis infections and eggs is not due to its growth behaviour in albumen. A pronounced growth occurred more frequently and up to a one-log unit higher level in fresh albumen than in albumen stored p.i. This was at least partly explained by a pH effect. Since growth in the separated albumen was similar when the albumen had been stored p.i. in the absence or presence of yolk, we have no indication that nutrients or factors negating the inhibitory properties of the albumen leak out from the yolk during storage. Growth of Salmonella inoculated at a level of 8 cfu in the albumen of fresh and stored whole shell eggs was studied to simulate a more natural situation. In this case, growth also occurred more frequently when inoculated in the albumen of fresh eggs compared to eggs stored p.i. It can be concluded from our study that cooling practices are recommended shortly after lay to prevent Salmonella from growing in eggs.     

Thermal death time measurement using thin flexible sleeves: a new experimental approach for determining microbial destruction kinetics in fluids of arbitrary viscosity

The thermal death time kinetics of Salmonella Enteritidis (SE) was measured in buffer, egg yolk, and albumen using thin layer plastic sleeves. The sleeves allowed for the loading and sampling of liquids of high or unusual viscosity, as in the case of yolk and albumen, and accepted relatively large volumes (2 to 3 ml) of fluid. The sleeves maintained the volume of the fluid in a thin layer and could be easily handled for heat exposure. The thin layer maintained one-dimensional heat transfer and minimized temperature gradients, thus preventing parts of the fluid from experiencing different heating rates. A representative strain of SE associated with an egg-based salmonellosis outbreak was used in this study. The D- and z-values of the chosen strain, H7037, were measured in buffer, yolk, and albumen. In buffer, SE had the following mean (±standard deviation) D-values: D(55°C) = 3.51 ± 0.30 min, D(57°C) = 1.75 ± 0.13 min, and D(60°C) = 0.25 ± 0.06 min. In yolk, D(58°C) = 0.90 ± 0.05, D(60°C) = 0.26 ± 0.03, and D(62°C) = 0.20 ± 0.02. In albumen, D(55°C) = 1.26 ± 0.31, D(56°C) = 0.68 ± 0.10, and D(57°C) = 0.44 ± 0.04. The z-values for SE calculated from these D-values were 4.29 ± 0.39°C in buffer, 6.12 ± 0.26°C in yolk, and 4.63 ± 1.14°C in albumen. The sleeves allowed one consistent approach to determining thermal death time kinetics regardless of viscosity.     

储存温度对鸡蛋微生物及品质的影响

为了解温度对鸡蛋中各组分微生物生长的影响,对鸡蛋在8、10、20、25℃储存30d内的蛋壳、蛋清和蛋黄的细菌总数变化规律做初步研究,并分析储存终点时鸡蛋的气室深度、蛋黄指数、浓蛋白含量、挥发性盐基氮含量品质指标。结果表明：储存温度对鸡蛋各部分细菌总数影响较大。8℃储存30d时蛋壳、蛋清和蛋黄细菌总数分别为3.5×103、2.4×103、1.7×103CFU/g,而25℃储存时则分别达4.6×106、7.5×104、5.3×105CFU/g。8℃和10℃储存10d内,蛋清和蛋黄中均未检测到细菌,25℃储存时则分别在第5天和第10天从蛋清和蛋黄中检测到细菌。鸡蛋各项品质指标的测定结果也表明储存温度越高鸡蛋品质越差。     

Multiplication in egg yolk and survival in egg albumen of Salmonella enterica serotype Enteritidis strains of phage types 4, 8, 13a, and 14b

Refrigeration of eggs is vital for restricting the multiplication of Salmonella enterica serotype Enteritidis contaminants, but differences between Salmonella Enteritidis strains or phage types in their survival and multiplication patterns in egg contents might influence the effectiveness of refrigeration standards. The present study compared the abilities of 12 Salmonella Enteritidis isolates of four phage types (4, 8, 13a, and 14b) to multiply rapidly in egg yolk and to survive for several days in egg albumen. The multiplication of very small numbers of Salmonella Enteritidis inoculated into yolk (approximately 10(1) CFU/ml) was monitored during 24 h of incubation at 25 degrees C, and the survival of much larger numbers of Salmonella Enteritidis inoculated into albumen (approximately 10(5) CFU/ml) was similarly evaluated during the first 3 days of incubation at the same temperature. In yolk, the inoculated Salmonella Enteritidis strains multiplied to mean levels of approximately 10(3) CFU/ml after 6 h of incubation and 10(8) CFU/ml after 24 h. In albumen, mean levels of approximately 10(4) CFU/ml or more of Salmonella Enteritidis were maintained through 72 h. Although a few differences in multiplication and survival were observed between individual isolates, the overall range of values was relatively narrow, and no significant differences (P < 0.05) were evident among phage types.     

The vitelline membrane: Dynamics of cholesterol metabolism in hens' eggs

Changes in cholesterol levels in egg contents were studied to indicate trends and dynamics of processes of penetration through the vitelline membrane from yolk to egg white. Investigations were carried out on eggs stored for a period of 35 days at room temperature (22°C). Levels of free and esterified cholesterol were determined by the spectrophotometric method at 7-day intervals. The selective character of the vitelline membrane depends on conditions and period of storage. The study demonstrated that the vitelline membrane loses its selective nature between the twentieth and thirtieth days under particular conditions of storage. Following this period, a spontaneous penetration of cholesterol into the egg white is observed—i.e. the barrier equilibrium of the membrane becomes unstable. The presence of free cholesterol in egg white may be indicative of loss of egg freshness.     

大蒜、葱白和花椒复配物涂膜对鸡蛋在高温下贮藏保鲜效果的影响

以新鲜鸡蛋为原料,大蒜、葱白和花椒按质量比20:10:1复配制备保鲜原液,原液加水等质量比稀释至原液的50%和25%,分别用上三个浓度复配物涂膜鸡蛋,并设置对照组,在温度为37℃的恒温培养箱中贮藏,检测复配物对鸡蛋感官品质、失重率、哈夫单位、蛋黄指数和蛋清pH的影响。结果表明:对照组蛋品质量在第10 d已严重下降,第20 d全部散黄,而复配物涂膜组均有不同程度保鲜效果。其中大蒜、葱白浸泡花椒原汁(即100%)复配物37℃环境中贮藏20 d后感官性状良好,失重率4.61%,哈夫单位63.77,蛋黄指数0.38,蛋清pH为9.67,保鲜效果最好。     

Survival and growth of salmonella in high-moisture pecan nutmeats, in-shell pecans, inedible nut components, and orchard soil

Outbreaks of salmonellosis associated with almonds have raised interest in better understanding the behavior of Salmonella on other tree nuts. We undertook a study to determine the survival and growth characteristics of Salmonella on high-moisture (water activity of 0.96 to 0.99) pecan nutmeats, in-shell pecans, and inedible components (shuck, shell, and middle septum tissue) of in-shell pecans. Salmonella did not grow on high-moisture nutmeat halves, pieces, or granules stored at 4°C for up to 48 h. Growth did occur, however, at 21, 30, and 37°C. Increases of 1.77 to 5.87 log CFU/g of nutmeats occurred within 48 h at 37°C; the order in which nutmeats supported growth was granules > pieces > halves. Populations of Salmonella on and in high-moisture in-shell pecans (kernel water activity of 0.94) stored at 4, 21, 30, and 37°C for 8 days decreased by 0.52 to 1.19 log CFU/g. The pathogen grew on the surface of high-moisture (water activity of 0.99) pecan shucks and shells but died on middle septum tissue stored at 21, 30, and 37°C for up to 6 days. Salmonella died in water extracts of shucks and in pecan orchard soil saturated with water or shuck extract, but survived well for at least 18 weeks in dry soil. The ability of the pathogen to grow on high-moisture nutmeats and some of the inedible components of pecans emphasizes the importance of controlling or limiting the time pecans are exposed to water in preharvest and postharvest environments.     

Adaptation of Listeria in liquid egg containing sucrose resulting in survival and outgrowth

Survival of Listeria species was tested in liquid egg products such as albumen, yolk, whole egg and whole egg containing 25% sucrose. At an incubation temperature of 4 degrees C Listeria survived in all products, and even increased slightly in yolk, whole egg and whole egg containing 25% sucrose. However, at 20-22 degrees C, in whole egg containing sucrose, a rapid an dramatic decrease in numbers of Listeria was observed. Following this initial decrease, an increase in the numbers of Listeria was observed. The ability to grow in liquid egg containing sucrose was maintained upon reinoculation into a freshly prepared product. However, this property disappeared after subculturing in brain heart infusion broth. The significance of these findings is discussed.