Effect of Refrigeration on In Vitro Penetration of Salmonella Enteritidis through the Egg Yolk Membrane

2006 ◽  
Vol 69 (6) ◽  
pp. 1426-1429 ◽  
Author(s):  
RICHARD K. GAST ◽  
PETER S. HOLT ◽  
RUPA GURAYA
Keyword(s):  
Treatment Group ◽  
Salmonella Enteritidis ◽  
Egg Yolk ◽  
Vitelline 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°C, 15% of samples refrigerated after 6 h of storage at 30°C, and 40% of samples stored at 30°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.

Multiplication and Motility of Salmonella enterica Serovars Enteritidis, Infantis, and Montevideo in In Vitro Contamination Models of Eggs

2006 ◽  
Vol 69 (5) ◽  
pp. 1012-1016 ◽  
Author(s):  
TOSHIYUKI MURASE ◽  
KAZUHIKO FUJIMOTO ◽  
RUI NAKAYAMA ◽  
KOICHI OTSUKI
Keyword(s):  
Salmonella Enterica ◽  
Salmonella Enteritidis ◽  
Egg Yolk ◽  
Vitelline Membrane ◽  
Human Patient ◽  
Polycarbonate Membrane ◽  
Specific Pathogen ◽  
Egg Albumen ◽  
Petri Plate

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°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 106 CFU/ml was observed in 48 of the 125 albumen samples. An inoculum of 1,000 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-μm pores, was filled with egg yolk and placed into the albumen at the center of the dish, which was maintained at 25°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 (>108 CFU/ml). The present study suggests that Salmonella organisms in egg albumen are unlikely to actively move toward the yolk, although deposition on or near the vitelline membrane can be advantageous for proliferation.

Multiplication of Salmonella Enteritidis in Egg Yolks after Inoculation outside, on, and inside Vitelline Membranes and Storage at Different Temperatures

2010 ◽  
Vol 73 (10) ◽  
pp. 1902-1906 ◽  
Author(s):  
RICHARD K. GAST ◽  
RUPA GURAYA ◽  
JEAN GUARD ◽  
PETER S. HOLT
Keyword(s):  
Bacterial Growth ◽  
Salmonella Enteritidis ◽  
Egg Yolk ◽  
Vitelline Membrane ◽  
Salmonella Enterica Serovar Enteritidis ◽  
Exterior Surface ◽  
Different Temperatures ◽  
Subsequent Storage ◽  
Storage 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.

Effect of Temperature on the Growth Response of Salmonella Enteritidis Inoculated onto the Vitelline Membranes of Fresh Eggs

2003 ◽  
Vol 66 (8) ◽  
pp. 1368-1373 ◽  
Author(s):  
G. J. FLEISCHMAN ◽  
C. L. NAPIER ◽  
D. STEWART ◽  
S. A. PALUMBO
Keyword(s):  
Salmonella Enteritidis ◽  
Growth Response ◽  
Effect Of Temperature ◽  
Vitelline Membrane ◽  
Growth Responses ◽  
In Ovo ◽  
Maximum Increase ◽  
Comparison Of The Results ◽  
As If

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°C. Comparison of the results obtained for in vitro growth showed that at 4, 8, and 15°C, SE behaved as if it were in the albumen, with its numbers decreasing over time. At 27 and 37°C, SE grew as if it were in yolk, with a maximum increase of 4.5 log CFU after 2 days at 37°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.

Multiplication of Salmonella Enteritidis on the Yolk Membrane and Penetration to the Yolk Contents at 30°C in an In Vitro Egg Contamination Model

2008 ◽  
Vol 71 (9) ◽  
pp. 1905-1909 ◽  
Author(s):  
RICHARD K. GAST ◽  
RUPA GURAYA ◽  
JEAN GUARD-BOULDIN ◽  
PETER S. HOLT
Keyword(s):  
Salmonella Enterica ◽  
Salmonella Enteritidis ◽  
Vitelline Membrane ◽  
Critical Aspect ◽  
Salmonella Enterica Serovar Enteritidis ◽  
Exterior Surface ◽  
Increased Risk ◽  
The Mean ◽  
Uncommon Location

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°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.

Evaluation of Snake Venom’s PhospholipaseA2 Enzyme Inhibition Activity of Cyphostemma adenocoule

2020 ◽  
Vol 14 (3) ◽  
pp. 196-202
Author(s):  
Atul Kaushik ◽  
Teamrat S. Tesfai ◽  
Daniel K. Barkh ◽  
Furtuna K. Ghebremeskel ◽  
Habtom G. Zerihun ◽  
...  
Keyword(s):  
Enzyme Inhibition ◽  
Snake Venom ◽  
Ethanol Extract ◽  
Egg Yolk ◽  
Chloroform Extract ◽  
Inhibition Activity ◽  
Traditional Use ◽  
Snake Bites ◽  
Ethanol Extracts

Background: A snake bite is fundamentally an injury often resulting in puncture wounds meted out by the animal's fangs and occasionally resulting in envenomation. Rate of snake bites around 5,400,000 bites per year leads to over 2,500,000 envenomings and around 125,000 fatal cases annually. Snake venom enzymes are rich in metalloproteinases, phospholipaseA2, proteinases, acetylcholinesterases and hyaluronidases. Objective: Cyphostemma adenocoule is traditionally being used for the treatment of snake bites in Eritrea. The present research was aimed at evaluating the snake venom enzyme inhibition activity of C. adenocoule against puff adder venom and developing a base for the traditional use of the plant against snakebites in Eritrea. Methods: The anti-venom activity of C. adenocoule was assessed in-vitro through phospholipaseA2 enzyme inhibition assay using egg yolk as a cell. The ethanol and chloroform extracts of C. adenocoule showed in vitro anti phospholipase A2 activity, whereas the water extracts of the plant showed no activity. Results: Among the extracts of C. adenocoule, the highest percentage of inhibition was obtained from chloroform extract (95.55% at 100mg/ml). The extract showed prominent activity at different concentrations (34.7% at10mg/ml, 48.8% at 20mg/ml, 54.8% at 40mg/ml, 60.9% at 60mg/ml, 80.5% at 80mg /ml). The ethanol extract also showed certain activity at various concentrations (25.22% at10mg/ml, 14.78% at 20mg/ml, 2.6% at40mg/ml). The activity of the chloroform extracts increases as concentration increases, whereas the activity of the ethanol extracts decreases as concentration increases. The aqueous extract of C. adenocoule did not show any activity at all concentrations. Conclusion: In this study, the chloroform and ethanol extracts of the plant inhibited the enzyme of interest and thus proved the efficacy of anti-snake venom activity of the plant.

Combined application of phenolic acids and essential oil components against Salmonella Enteritidis and Listeria monocytogenes in vitro and in ready-to-eat cooked ham

LWT ◽  
2021 ◽  
pp. 111881
Author(s):  
Jessica Audrey Feijó Corrêa ◽  
João Vitor Garcia dos Santos ◽  
Alberto Gonçalves Evangelista ◽  
Anne Caroline Schoch Marques Pinto ◽  
Renata Ernlund Freitas de Macedo ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Essential Oil ◽  
Phenolic Acids ◽  
Salmonella Enteritidis ◽  
Combined Application ◽  
Cooked Ham ◽  
Oil Components

Mechanism of the adverse effect of hyaluronidase used for oocyte denudation on early development of bovine embryos

Zygote ◽  
2021 ◽  
pp. 1-5
Author(s):  
Shiori Ashibe ◽  
Kanade Irisawa ◽  
Ken Yokawa ◽  
Yoshikazu Nagao
Keyword(s):  
Treatment Group ◽  
Assisted Reproductive Technologies ◽  
Cumulus Cells ◽  
Reproductive Technologies ◽  
Control Group ◽  
Free Calcium ◽  
Parthenogenetic Development ◽  
Early Embryos ◽  
Bovine Oocytes

Summary Hyaluronidase is widely used in animal and human assisted reproductive technologies (ARTs) to remove cumulus cells around oocytes. However, adverse effects of hyaluronidase treatment, such as increased rates of degeneration and parthenogenesis, have been found after treatment of human and mouse oocytes. Currently, the mechanism(s) of the detrimental effects are unclear. The present study was initiated to identify the mechanism of adverse responses to hyaluronidase treatment in bovine oocytes and early embryos. Cumulus cells were removed from cumulus–oocyte complexes (COCs) with or without hyaluronidase and the oocytes were subjected to intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF). Significantly lower rates of blastocyst formation were obtained in the hyaluronidase treatment group after ICSI (22.4%) and IVF (21.2%) compared with the non-hyaluronidase control groups: 36.1% after ICSI and 30.4% after IVF. Next, we examined the effect of hyaluronidase on parthenogenetic development rates and on the cytoplasmic levels of free calcium ions (Ca2+), reactive oxygen species (ROS) and reduced glutathione (GSH). No differences in parthenogenesis rates were found between treated and untreated groups. Ca2+ levels in oocytes from the hyaluronidase treatment group indicated using mean fluorescence intensity were significantly higher (68.8 ± 5.3) compared with in the control group (45.0 ± 2.5). No differences were found in the levels of ROS or GSH between the treated and untreated groups. We conclude that hyaluronidase might trigger an increase in Ca2+ levels in oocytes, resulting in a decreased potential for normal embryonic development.

Outgrowth of Salmonellae and the Physical Property of Albumen and Vitelline Membrane as Influenced by Egg Storage Conditions

2005 ◽  
Vol 68 (12) ◽  
pp. 2553-2558 ◽  
Author(s):  
JINRU CHEN ◽  
HILARY SHALLO THESMAR ◽  
WILLIAM L. KERR
Keyword(s):  
Salmonella Enteritidis ◽  
Antimicrobial Agents ◽  
Physical Property ◽  
Storage Conditions ◽  
Vitelline Membrane ◽  
Initial Population ◽  
Fiber Optic Probe ◽  
Physical Strength ◽  
Egg Storage ◽  
Egg Albumen

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 102, 104, or 106 cells per egg. Both fresh and inoculated eggs were stored at 4, 10, and 22°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°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°C required more force and energy for rupture. Salmonellae flourished at 22°C, even in the albumen with the lowest initial population, 102 cells per egg. Storage at 4 and 10°C inhibited the growth of salmonellae in the albumen of eggs with initial populations of 102, 104, or 106 cells per egg. In eggs with initial Salmonella populations of 106 cells per egg that were stored at 22°C, the populations of reached as high as 1010 cells per egg after 4 weeks of storage. Storage at 4 and perhaps 10°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.

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