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Is there lipid peroxidation induced malondialdehyde production during egg shell formation?

The marked increase observed in malondialdehyde concentration of the blood plasma and liver of laying hens during egg shell formation in a previous experiment (Mézes and Lencsés, 1985) were explained as a possible consequence of physiologically controlled lipid peroxidation. In this experiment, 54-week-old laying hens were treated intrauterinally with indomethacin (1.0 and 2.0 mg/kg body mass) after oviposition. Two other groups of hens were treated per os with excess amounts of vitamin E (100 and 200 mg/bird). The higher dose of indomethacin significantly decreased the PGF2 alpha production and malondialdehyde content of the sell gland, as well as the malondialdehyde concentration of the plasma. The excess amount of vitamin E had the same effect. The results suggest that during egg shell formation malondialdehyde is derived from prostanoid biosynthesis rather than from a free-radical initiated lipid peroxidative process. On the other hand, the excess amount of vitamin E inhibited prostaglandin biosynthesis as well as malondialdehyde production.

Animals↗

Calcium reserve assembly: a basic structural unit of the calcium reserve system of the hen egg shell.

A well-defined proper subregion of the mammillary knob is described, consisting of baseplate (BP), calcium reserve body (CRB), and an integral but distinct CRB cover. The BP extends 14 to 20 microns into the outer shell membrane and rises a few microns above its surface. The organic matrix of the BP is a composite of modified membrane fibers and adjacent components. The distinct CRB matrix is attached to the BP and extends outward in attenuated from, much like the fractal aggregates associated with filter "cakes." The three regions comprise a unit designated as the CRA (calcium reserve assembly, or assemblies). The CRA are fully developed and mineralized by 10.5 to 11 h postoviposition of the previous egg. By 13 h postoviposition a new structure, the crown, is evident. The classical mammillary knob consists of the CRA/crown complex. By hatch time the CRA are partially decalcified, except in the air cell region. Consequently, a zone of separation develops in the affected CRA about midway through the CRB. Estimates of the calcium recovered from the egg shell and of the calcium found in the CRA, excluding the CRA in the air cell region, indicate that more than enough calcium is present in the CRA to meet the drawdown of calcium from the egg shell by the developing embryo. It is concluded that the CRA that are available to the action of the chorioallantoic membrane represent the calcium reserve of the hen egg shell.

Animals↗

The magnetic field effect affecting on the metallic chemical components of egg shell as demonstrated for breeding chicken.

The chickens were raised under fixation of the permanent magnet (80 mT) and the effect of extra magnetic field, which affects on the content of the several inorganic chemical elements of egg shell, was investigated by X-ray fluorescence spectroscopy. From 3 days after fixing the magnet, the relative concentration of Ca begun to increase. After 10 days, the relative contents of Ca were decreased, while conversely the contents of K were increased. Though there was a linear relationship between Ca content and P content in inner layer of chicken egg shell, the relationship was greatly hindered by applying the static magnetic field. The experimental result suggests that both Ca and K ion transport can be perturbed by addition of extra magnetic field in the incubation processes of chicken egg, and that Ca, P and K ions play the important role in egg shell formation.

Journal Article↗

Regulation of net intestinal calcium uptake in hens laying obligatory soft-shelled eggs.

1. Individually caged laying hens had a loop of thread inserted into the shell gland. This resulted in the laying of soft shelled eggs. 2. A balance study was performed for a one week period before and after the operation. After the operation birds with threads consumed less calcium than before. Their requirements for calcium for eggshells decreased, resulting in increases in both calcium excreted and calcium retained. 3. Net calcium extraction in the digestive tract was measured in groups of birds with threads and intact controls, when shelling or not, by examining ratios of Ca to TiO2 in different gut segments. Observations were made during the period following premature oviposition in birds with threads, but within the normal shelling period of control birds. The period of study was at least two weeks after the operation. 4. Birds with threads absorbed less calcium than control birds up to the upper jejunum. 5. Control birds secreted calcium between the upper jejunum and colon, but birds with threads showed little change in absorption in this part of the digestive tract. 6. The increase in calcium absorption in intact birds was a response to the stimulus of shelling an egg or replacing calcium in medullary bone during a pause day, rather than of ovulation.

Animals↗

Isolation of Salmonella typhimurium Tn5 mutants defective for survival on egg shell surface using transposon footprinting.

The objective of this study was to screen the genome of Salmonella typhimurium for genes potentially required for survival on egg shell surfaces using a novel transposon footprinting method based on a previously developed Tn5 mutagenesis technique. Since the amplified DNA fragment length should usually be unique for each mutant, the polymerase chain reaction products separated on an agarose gel generate a transposon footprint with each band in the footprint representing the corresponding Tn5 mutant. By comparing the footprints from the pools of S. typhimurium Tn5 mutants before and after inoculation on egg shell surfaces, Tn5 mutants not recovered after selection were rapidly identified for potential isolation and genetic analysis.

Animals↗

Introduction of a fluorescence-microscopic technique for the detection of eggs, egg shells, and mites in scabies.

A technique using the fluorescence microscope can prove helpful in the laboratory diagnosis of scabies. Specimens from fifteen patients with scabies were used in this study. All of the specimens were embedded with glycerine instead of potassium hydroxide (KOH) solution. The specimens were examined at 0 min, 30 min, 1 hr, 6 hrs, 24 hrs, and one week after mounting under light and fluorescence microscopes. Specimens embedded with non-fluorescent glycerine were not clear immediately after mounting but became so after about 1 hr. Eggs and egg shells were easily counted in the specimens under the fluorescence microscope but were very hard to identify under the light microscope. Mites were absent in half of the specimens; only eggs and egg shells were present in those specimens found by the fluorescence microscope. The above findings suggest that the detection of egg shells by the use of fluorescence microscope may be helpful for the diagnosis of scabies, in particular with mite negative specimens. Slides prepared with non-fluorescent glycerine were more stable and could be preserved for a long time. However, this method is time-consuming and requires expensive equipment.

Animals↗

Lysyl oxidase coupled with catalase in egg shell membrane.

The activity of lysyl oxidase was found in egg shell membrane (ESM) of hens. The activity was determined by measuring the enzymatic conversion of n-butylamine and Nalpha-acetyl-L-lysine to n-butyraldehyde and Nalpha-acetyl-L-allysine, respectively. ESM lysyl oxidase was significantly inhibited by beta-aminopropionitrile, chelating agents, and deoxygenation, consistent with the known properties of lysyl oxidase. Nevertheless, ESM lysyl oxidase was insoluble in urea solution, suggesting that it complexes with ESM. These findings support previous reports indicating the presence of lysine-derived cross-links in ESM and the necessity of lysyl oxidase located in the isthmus of the hen oviduct for the biosynthesis of ESM. Lysyl oxidase secreted around the egg white from the isthmus may initiate the cross-linking reaction of ESM protein, and remain as the constituent of ESM. Moreover, the H(2)O(2) released by lysyl oxidase in ESM was completely decomposed by coexisting catalase activity. ESM lysyl oxidase activity was greatly elevated in the presence of H(2)O(2), probably due to the O(2) produced by catalase. These findings indicate that lysyl oxidase is coupled with catalase in ESM. This coupling enzyme system was considered to be involved in the biosynthesis of ESM and to protect the embryo against H(2)O(2).

Animals↗

Microbial barrier properties of hen egg shells.

Scanning electron micrographs of shell surfaces revealed a highly fissured outer layer with few open pores on brown eggs but many on white eggs. Total removal of the cuticle with solvents was difficult but partial removal with surface etching was possible using concentrated nitric acid. Staining methods to estimate the number of pores were unsatisfactory but it was possible to detect and count pores on micrographs, as pore mouths were usually associated with depressions and cuticle disruptions. Additionally, porosity could be estimated by measuring the distance between pores along fractured edges of samples. Egg shell contents were replaced with nutrient agar and incubated in Escherichia coli broth. Colonies were subsequently isolated from the agar, indicating that the barrier properties of shells and membranes had been compromised. Experiments with isolated inner shell membranes showed that these posed no significant barrier to E. coli. The exposure of whole intact eggs to E. coli broth followed by seep filtration and microbiological analysis of egg contents, indicated that bacteria had entered the eggs. The degree of infection was correlated with the pole of the egg in contact with the E. coli broth and was attributed to the increased porosity of the blunt pole of the egg compared with that of the apex. Similar experiments immersing eggs into broth inoculated with a Salmonella strain resulted in contamination of the egg contents with this organism.

Animals↗

The development of the oxygen permeability of the avian egg shell and its membranes during incubation.

The 02-permeability of the avian egg shell and shell membranes is initially low (approximately equal to 0.1-10(-6) ml O2 STP-sec-1cm-2-mmHg-1) during incubation but increases about 10-fold after the first week. This increase correlates with a decline in water content of the membranes. A major increase in colloid osmotic pressure (up to 50 cm Hg) occurs and is the suggested cause for the onset of rapid removal of water from the membranes. A high membrane water content correlates with low O2-permeability and vice versa. The final degree of membrane hydration reached during incubation is independent of ambient humidity; that is, the O2-permeability of the shell and its membranes is controlled by conditions inside the egg. An hypothesis is presented to explain the profound increase in O2-permeability while the rate of water loss from the egg remains stable during incubation. The removal of water from the shell membranes by the increased colloid osmotic pressure will increase the number of gas filled channels in the membranes which in turn will increase the O2-permeability. In spite of the change in water content, the water vapor pressure in the shell membranes will always be very close to that of a free water surface and water loss will hence be determined by the ambient humidity and the porosity of the mineral shell.

Animals↗

[Structure of the hen's egg shell and its changes during incubation. A scanning electron microscope study].

The structure of the hen's egg shell was studied during the incubation. The local differences in the structure of the shell were estimated during the incubation in dependence to the development of the allantochorial vascular system. On the external surface of the shell there is a thin layer, homogenous in the SEM. It remains changeless during the whole period of incubation. The main part of shell is formed by long prisms with their long axes oriented perpendicularly to the external surface of the shell. These prisms are partially twisted round each other, similarly as in the case of the teeth enamel. The inner layer of the shell is formed by a system of fibers forming the three-dimensional network, similar to an air filter. In the middle of incubation, when the embryo's skeleton is still predominantly cartilaginous, is the structure of the shell identical with that one at the beginning of incubation. At the end of incubation, when the embryo's skeleton is already predominantly ossificated, the inner layer of the shell disappears without any trace. By resorption of the shell are the peaks of the prisms of the middle layer of the shell destroyed as well. The above described changes are the best visible in the equator region of the shell. Less developed are they in the region of the sharp pole of the egg. They are not detectable in the region of the blunt pole of the egg. In this region the egg's shell has the same structure during the whole period of incubation. The changes of structure of the shell--the resorption of the inner layer together with partial destruction of the prisms of the middle layer - exactly correspond with extension of the allantochorial vascular system.

Animals↗

Nutritional and environmental factors involved in egg shell quality of laying hens.

1. Two experiments were conducted to assess the effects of lighting regime, dietary calcium source and sodium bicarbonate on production variables and egg shell quality of White Leghorn hens. In both experiments, hens were assigned to one of three lighting programmes that provided evening, morning or intermittent (midnight) lighting supplements in addition to natural daylight. Experimental diets used in the first study were formulated to contain (1) ground oyster shell flour, (2) limestone flour, or (3) and (4) the same +2/3 of the calcium source as hen-size oyster shell grit. The same 4 diets plus those containing hen-size limestone or hen-size limestone and oyster shells were used in experiment II. Additionally, diets in the first experiment contained either 0 or 10 g/kg sodium bicarbonate. 2. Hen-day egg production and food consumption were not affected by any of the experimental treatments. Hens fed on oyster shell diets or exposed to intermittent lighting regimes laid eggs of the highest specific gravity. Shell quality, as measured by specific gravity, was not affected by the addition of dietary sodium bicarbonate. 3. As expected, elevated temperatures (greater than 32 degrees C) significantly reduced egg shell quality. However, this effect was variable particularly in experiment II which used younger hens. 4. The shell quality of eggs from hens exposed to intermittent lighting in experiment II was significantly higher in each of the 4 sampling periods: morning (08.00-12.00), afternoon (12.00-16.00), evening (16.00-20.00) and night (20.00-08.00). 5. It is suggested that midnight lighting programmes provide a means of supporting egg shell quality of older laying hens during the summer months without a significant reduction in egg production.

Animal Nutritional Physiological Phenomena↗

The isolation and characterisation of hyaluronic acid in egg shell.

Analysis showed that the organic part of the chicken's egg shell consisted of a series of proteins and polysaccharides, probably present as glycoproteins and glycosaminoglycans. A purified preparation of a glycosaminoglycan (minimum mol. wt. 25 000), homogeneous by sedimentation velocity analysis and sedimentation to equilibrium in a density gradient, contained equimolar amounts of N-acetylglucosamine (36.3% s/w) and glucuronic acid 35.6% w/w). Digestion with testicular and streptomyces hyaluronidases and identification of the degradation products showed the glycosaminoglycan to be hyaluronidases and identification of the degradation products showed the glycosaminoglycan to be hyaluronic acid.

Acetylglucosamine↗

Selection for egg shell strength in laying hens using shell membrane characteristics.

1. Divergent selection for attachment strength between the shell membrane and the calcium shell was performed in a White Leghorn strain. Multivariate analysis was used to estimate genetic parameters for shell membrane measurements and shell thickness. The aim was to investigate the possibility of improving shell strength in laying hens by selecting for increased attachment strength. 2. A significant direct selection response for attachment strength was achieved. There was a favourable correlated selection differential in the frequency of cracked eggs which resulted in a nonsignificant but favourable, correlated response. Selection for a strong attachment resulted in a thinner egg shell, and vice versa. Heritabilities were all relatively high (0.30 to 0.70). Most of the genetic correlations were in agreement with the achieved correlated responses. 3. Two factors that probably contributed to the responses in egg shell thickness were an unfavourable genetic correlation between attachment strength and shell thickness, and natural selection against changes in attachment strength, in the form of increased mortality during incubation and hatching. Problems connected with breeding for increased shell strength are discussed.

Animals↗

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

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

Animals↗

A nutrient re-evaluation of shell eggs.

Most data on the nutrient composition of shell eggs were obtained before 1950. Since then, management systems for egg production and analytical methods for many components have changed. Freeze-dried whole egg, yolk and white samples were prepared from eggs obtained from a single source of 15 month old White Leghorns. The sample were packaged under nitrogen and held at -20 degrees C. until analyzed. Conversion factors are included for expressing the amount of each component on a per egg as well as a per 100 g. edible liquid basis. This re-evaluation of nutrient data included total solids, lipid, protein, cholesterol, ash, calories, amino acids, fatty acids, vitamins and minerals.

Amino Acids↗

Modulation of quail intestinal and egg shell gland calbindin (Mr 28,000) gene expression by vitamin D3, 1,25-dihydroxyvitamin D3 and egg laying.

The effects of vitamin D3 sources, egg production and egg cycle on the genomic expression of calbindin (Mr 28,000) in the intestine and egg shell gland (ESG) of quail were characterized by Northern blot and solution hybridization, using synthetic oligonucleotide probe. In vitamin D3- or 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3)-fed quail, onset of egg production induced duodenal and ESG calbindin mRNA and calbindin synthesis. Duodenal calbindin mRNA was slightly higher during the period of shell calcification as compared with the period during which shells were not formed (ESG inactivity). ESG calbindin mRNA was markedly higher during the period of shell calcification than of ESG inactivity. Increasing dietary intake of [3H]1 alpha-hydroxyvitamin D3 increased the duodenal, but not ESG, content of 1,25-(OH)2D3 and calbindin. Duodenal calbindin and its mRNA were absent in vitamin D-deficient quail and were not affected by egg laying. ESG calbindin in the vitamin D-deficient quail was not affected by egg laying, but calbindin mRNA increased in the vitamin D-deficient birds during shell calcification. The results suggest that: (a) intestinal calbindin mRNA and calbindin are induced and/or regulated, either directly or indirectly, by 1,25-(OH)2D3; (b) intestinal calbindin and its mRNA are further induced at the onset of egg laying by an additional stimulator besides 1,25-(OH)2D3; (c) 1,25-(OH)2D3 is required for the expression of the latter stimulator; (d) ESG calbindin mRNA and calbindin are induced in egg-laying birds by a stimulator associated with the egg cycle; and (e) the induction of ESG calbindin mRNA does not need vitamin D metabolites, but 1,25-(OH)2D3 is required for the translation of the mRNA.

Animals↗

Embryonated eggs compared with fragments of chorioallantois attached to egg shell for isolation of Newcastle disease virus.

The feasibility of using fragments of chorioallantois attached to egg shell (C-S) as an alternative to embryonated eggs for isolating Newcastle disease virus (NDV) was assessed. In paired titrations of 6 strains of virus, embryonated eggs detected an average of 6-to-7-fold greater amounts of virus than did C-S fragments. The efficiency of isolation of viscerotropic velogenic NDV from 240 swab samples was 97% in embryonated eggs and 84% in C-S fragments. Sensitivity differences between the 2 host systems were revealed only in tests of samples expected to contain small amounts of virus. These results suggest that C-S fragments can be reliably used in some but not all instances, thereby relieving logistic constraints sometimes associated with the use of embryonated eggs.

Allantois↗