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.     

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.     

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.     

Populations of Salmonella enteritidis in artificially inoculated chicken eggs as influenced by the temperatures under which eggs might be held from the day of lay until the day of processing

This study was undertaken to determine the levels of Salmonella Enteritidis in artificially inoculated eggs as affected by the temperatures under which eggs might be held from the day of lay until the day of processing. Unprocessed chicken eggs of different sizes (n=1920, with 480 being laid in each season) were inoculated in the albumen with a five-strain mixture of Salmonella at 102 CFU per egg. The eggs were stored at 4, 10, and 22 degrees C for 3 weeks and sampled twice a week to determine the populations of Salmonella and total aerobic bacteria. The season in which eggs were laid did not significantly impact the growth of the pathogen (P > 0.05). The mean populations of the inoculated Salmonella were not significantly different in eggs stored at 4 versus 10 degrees C (P > 0.05). Eggs stored at 22 degrees C had a mean Salmonella population that was 3.71 or 3.37 log higher than the Salmonella population of eggs stored at 4 or 10 degrees C (P > 0.05). The mean Salmonella population at 22 degrees C increased from the initial 2.12 log CFU/ml to 3.36 log CFU/ml after 2 weeks of storage and to 7.84 log CFU/ml after 3 weeks of storage. A sharp increase in the population of Salmonella occurred after 2 to 2.5 weeks of storage at 22 degree C. This study provided a scientific basis for the current egg handling and transporting temperature requirements and reinforced the importance of maintaining low temperatures in controlling and preventing the growth of Salmonella Enteritidis in eggs from the day of lay until the day of processing.     

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.     

贮藏温度变化对壳蛋新鲜度的影响

该文以新鲜罗曼粉壳蛋为研究对象,研究4个贮藏温度变化处理组壳蛋新鲜度指标的变化,分别于0、10、20、30 d检测所有实验组壳蛋失重率、气室高度、哈夫单位、蛋黄指数、浓稀蛋白比和蛋清pH。结果表明,贮藏10～30 d,处理组2、处理组3、处理组4的失重率显著升高(P<0.05),而气室高度和浓稀蛋白比显著降低(P<0.05),除对照组外,所有处理组壳蛋pH呈现先升高后下降的趋势,处理组3和处理组4的蛋黄指数下降趋势更为明显。对照组和处理组1壳蛋的哈夫单位、蛋黄指数和浓稀蛋白比显著高于(P<0.05)处理组3和处理组4。在贮藏10～20 d时,处理组2的壳蛋pH显著高于(P<0.05)对照组和处理组1。因此,如果前10 d常温贮藏温度≥20℃时,引起壳蛋失重率的增加和哈夫单位的下降,与低温(4℃)贮藏相比,采用贮藏温度变化(常温-低温)贮藏壳蛋更容易引起壳蛋新鲜度指标的下降。     

贮藏温度-时间变化对壳蛋新鲜度影响研究

该文以新鲜罗曼粉壳蛋为研究对象,研究5个贮藏温度-时间变化组壳蛋新鲜度指标变化,分别于0、10、20、30 d检测所有实验组壳蛋失重率、气室高度、哈夫单位、蛋黄指数、浓稀蛋白比、蛋清pH。结果表明,贮藏期间壳蛋失重率、气室高度和蛋清pH显著升高（P<0.05）,哈夫单位、蛋黄指数和浓稀蛋白比显著下降（P<0.05）。贮藏20 d时,处理组5壳蛋哈夫单位和蛋黄指数显著高于（P<0.05）对照组1、处理组1、处理组2和处理组3,而失重率、气室高度、蛋清pH较低（P<0.05）。贮藏30 d时,处理组5壳蛋失重率和气室高度显著低于对照组1、处理组1和处理组3（P<0.05）,蛋黄指数显著高于对照组1、处理组2、处理组3、处理组4（P<0.05）。除失重率、哈夫单位外,对照组2与处理组5壳蛋新鲜度指标整体差别不大。因此,除了壳蛋自身品质外,需要尽可能延长低温（4 ℃）贮藏时间,延缓壳蛋新鲜度指标劣变。     

The effect of short term storage on the albumen quality of shell eggs

Certain physical changes occur naturally in the albumen of shell eggs when stored under cool conditions which cause the thick white to become thin. The whipping characteristics of the albumen are changed during this process but the functional performance of the albumen as measured by the angel food cake method is not affected to any marked degree. Three 10-day storage trials were undertaken using day-old shell eggs which demonstrated that a significant loss of albumen performance did not occur until the storage temperature reached 18 ºC. This conclusion clearly indicates the importance of maintaining shell eggs under cool conditions between collection at the farm and commercial distribution by the egg floors, particularly during the summer season.     

Effect of various refrigeration temperatures on quality of shell eggs

BACKGROUND: The objective of this study was to evaluate the effects of low storage temperatures on shell egg quality. RESULTS: Approximately 2100 shell eggs were collected and stored at ? 1.1, 0.6, 2.2, 3.9, 5.6 and 7.2 °C for up to 4 weeks. Eighteen eggs at each storage temperature were evaluated after 0, 2, 7, 14, 21 and 28 days of storage. Haugh units (HU), yolk index (YI), albumen pH (pHA), yolk pH (pHY) and angel food cake density (CD) were measured. Shell egg quality tended to be preserved better at below 2.2 °C, as high HU and YI values relative to eggs stored at 7.2 °C were determined on day 28. However, storage at ? 1.1 °C tended to cause the opposite effect, especially highly declined HU values over time. Significantly different HU values of shell eggs were measured after 14 days of storage, with eggs stored at 0.6 and 2.2 °C having the highest HU values, 80.42 and 77.97 respectively. CONCLUSION: A lower temperature limit for shell egg storage could be established between 0.6 and 2.2 °C, as both temperatures showed the highest HU values, 77.88 and 77.60 respectively, after 28 days of storage. Copyright © 2011 Society of Chemical Industry     

Changes in Heat-Induced Rheological Properties During Cold Storage of Egg Albumen

Two groups of eggs were stored at 15°C for 0, 5 and 10 days. One group was stored, with the shell removed, in sterile Whirl-Pak bags. A second group was stored as intact whole eggs. Viscosity index, apparent elasticity, and initial penetration force were determined for heat-induced (80°C, 80 min) gels of the thick, thin and mixed albumens from each group. The rheological parameters increased with storage time, with the greater increase exhibited by the gels of the albumens that were stored as shell eggs. Solution pH was a major factor controlling these rheological properties of heated egg albumen.     

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.     

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.     

EFFECT OF RAPID COOLING ON THE GROWTH AND PENETRATION OF SALMONELLA ENTERITIDIS INTO EGG CONTENTS

Shell eggs were inoculated internally with approximately 10 cells of Salmonella enterica serovar enteritidis (S. enteritidis) and subjected to three different cooling treatments. Eggs were cooled from an initial temperature of 27C to approximately 7.2C. After cooling, eggs were stored at approximately 7.2C for 36 days, or stored at 5.7–9.5C for 30 days plus 6 days at 37C to simulate temperature abuse. Rapid cooling and subsequent storage of eggs at approximately 7.2C inhibited the growth of S. enteritidis in eggs. Slow cooling, and/or temperature abuse promoted growth of S. enteritidis in eggs. The penetration study indicated that rapid cooling and subsequent storage at 7.2C for 30 days did not affect the penetration of S. enteritidis into egg contents. The S. enteritidis isolated from the eggshell with shell membranes was significantly higher (P < 0.05) than from the internal egg contents, indicating that most of the S. enteritidis cells were trapped within the shell pores and/or shell membranes.     

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.     

不同气体配比气调包装对鸡蛋保鲜效果的影响

为延长鸡蛋保鲜期,研究在室温(25℃)条件下,不同配比CO2、O2、N2三元混合气体的气调包装对鸡蛋保鲜效果影响。结果表明:鸡蛋采用体积分数50%CO2、7%～11%O2、39%～43%N2的气调包装,在室温贮藏,质量损失率、蛋白pH值保持相对较低水平,哈夫单位、蛋白凝胶强度保持相对较高水平,贮藏30d鸡蛋仍保持AA级,而不包装的对照组降到了B级。     

Incubation of egg contents pools at an elevated temperature (42 degrees C) does not improve the rapid detection of Salmonella enteritidis phage type 14b

Detecting internal Salmonella Enteritidis (SE) contamination in eggs is essential for protecting public health. Pooling together > or = 10 eggs for sampling allows many eggs to be screened for contamination, but such pools must be incubated (usually at 25 to 37 degrees C) to permit small numbers of SE to multiply before further testing. The present study determined whether incubating egg contents pools at an elevated temperature (42 degrees C) could increase the rate of multiplication of a phage type 14b strain of SE sufficiently to support the detection of contamination by a rapid lateral flow immunodiffusion method within a single day. Pools of 10 eggs were contaminated with approximately 10 CFU of SE, supplemented with concentrated broth enrichment medium, and incubated at either 37 or 42 degrees C. Incubation of contaminated egg pools at 42 degrees C resulted in significantly higher SE levels after 6, 8, 10, and 12 h. However, incubation at 42 degrees C could only generate a mean log SE concentration of 4.21 CFU/ml within a single working day (8 h), inadequate to support efficient detection by most rapid assays. Detection of SE contamination in egg pools by a rapid lateral flow immunodiffusion test was not achieved at a high frequency until 12 h of incubation at 42 degrees C.     

Quality of shell eggs pasteurized with heat or heat-ozone combination during extended storage

The physical quality and functionality of shell eggs, pasteurized with heat or a combination of heat and ozone, were assessed during eight weeks of storage at 4 or 25 °C. Shell eggs were treated as follows: (1) immersion heating that mimics commercial pasteurization processes (egg internal temperature of 56 ± 0.1 °C for 32 min), or (2) a newly developed combination process comprised of heating (56 ± 0.1 °C, internal, for 10 min) followed by gaseous ozone treatment. Eggs were tested for yolk index, Haugh units, albumen pH, albumen turbidity, and percent overrun. Additionally, albumen samples were assayed for lysozyme activity and free sulfhydryl group content, and were analyzed using differential scanning calorimetry and Fourier transform infrared (FTIR) spectroscopy. Both processed and unprocessed eggs maintained superior quality when stored at 4 °C, as opposed to 25 °C. Pasteurization, regardless of method, led to superior maintenance of Haugh units during storage but also increased albumen opacity and decreased albumen overrun. Detrimental effects on quality markers were more severe in heat-pasteurized eggs than those treated with the ozone-based process. Pasteurization of shell eggs by either process did not affect lysozyme activity or sulfhydryl group content. Changes in protein secondary structure, as indicated by FTIR analysis, suggest that the ozone-based process is less damaging to albumen proteins than is the heat-alone process. In conclusion, heat-ozone pasteurization, by virtue of its less severe heat treatment, yields a safe final product that more closely resembles untreated shell eggs.     

Variations in external and internal microbial populations in shell eggs during extended storage

The current project was conducted to determine the microbial quality of commercially processed shell eggs during extended storage. Unwashed eggs were collected at the accumulator before entering the processing line. Washed eggs were retrieved after placement in flats. All eggs were stored on pulp flats at 4 degrees C for 10 weeks. Twelve eggs from each treatment were rinsed on the day of collection and during each week of storage. After rinsing, eggs were sanitized in ethanol, and contents were aseptically collected. Total aerobes, yeasts and molds, Enterobacteriaceae, and pseudomonads were enumerated from shell rinses and pooled egg contents. During storage, no differences were found between unwashed and washed eggs for Enterobacteriaceae and pseudomonads in either shell rinses or contents. No differences were found between treatments for population levels of total aerobes or yeasts and molds in the egg contents throughout the storage period. Significant differences between treatments were found at each week of storage for external shell contamination by total aerobes. The highest unwashed egg contamination occurred at week 8 of storage and the lowest was at weeks 0 and 1 of storage. The highest shell contamination with aerobic bacteria on the washed eggs was found at week 0 of storage and the lowest was at week 7. Yeast and mold contamination determined by shell rinses was also significantly different between treatments at each week of storage. Commercially washed eggs were significantly less contaminated than were unwashed eggs for the populations monitored.     

Storage effects on dielectric properties of eggs from 10 to 1800 MHz

ABSTRACT:  The dielectric constant and loss factor of egg albumen and egg yolk over the frequency range from 10 to 1800 MHz were measured at 24 °C at weekly intervals during 5-wk storage at 15 °C. Moisture and ash contents of albumen and yolk, as well as Haugh unit and yolk index, were also measured. The dielectric constant and loss factor of albumen were higher than those of yolk. Linear relationships were evident between the log of frequency, below about 1000 MHz, and the log of loss factor of albumen as well as that of yolk. The dielectric constants of albumen and yolk at 10 MHz were lower than those of fresh albumen and yolk when eggs were stored at 15 °C for 1 wk. However, after 2 wk in storage these dielectric constants rose and remained at higher levels for the rest of the 5-wk period. At frequencies of 100 MHz and higher, the dielectric constant was essentially constant during the entire storage period. Storage had much less influence on the loss factor of either albumen or yolk. In general, the moisture content and ash content of albumen and yolk decreased slightly as eggs aged. The moisture content of yolk increased somewhat with storage, and there was a corresponding decrease in albumen moisture content. The freshness qualities, Haugh unit and yolk index, also decreased as eggs aged. No obvious correlation between dielectric properties and moisture content, ash content, Haugh unit, or yolk index was observed.