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PubMed2004

Crystal structure of ovocleidin-17, a major protein of the calcified Gallus gallus eggshell: implications in the calcite mineral growth pattern.

Ovocleidin-17 (OC17) from Gallus gallus is one of the best candidates to control and regulate the deposition of calcium carbonate in the calcified eggshell layer. Here, the crystal structure of monomeric OC17, determined at a resolution of 1.5 A, was refined to a crystallographic R-factor of 20.1%. This is the first protein directly involved in a non-pathological biomineralization process resolved by x-ray diffraction to date. The protein has a mixed alpha/beta structure containing a single C-type lectin-like domain. However, although OC17 shares the conserved scaffold of the C-type lectins, it does not bind carbohydrates. Nevertheless, in vitro OC17 modifies the crystalline habit of calcium carbonate (CaCO3) and the pattern of crystal growth at intervals of 5-200 microg/ml. Determining the three-dimensional structure of OC17 contributes to a better understanding of the biological behavior of structurally related biomolecules and of the mechanisms involved in eggshell and other mineralization processes.

Amino Acid Sequence↗
PubMed1995

Calcium solubilization and retention in the gastrointestinal tract in chicks (Gallus domesticus) as a function of gastric acid secretion inhibition and of calcium carbonate particle size.

In chicks, immature pullets and laying hens, the inhibition of gastric acid secretion by omeprazole, an H+,K(+)-transporting ATPase (EC 3.6.1.36) inhibitor, greatly increased proventricular and gizzard pH values. Consequently, gizzard soluble Ca concentration deceased and the insoluble Ca fraction increased. Inhibition of acid secretion increased duodenal pH values in immature pullets and laying hens but not in chicks. Duodenal soluble and ionic Ca concentrations were lowered by gastric acid inhibition in chicks and to a larger extent in immature pullets and laying hens. The use of Ca of coarse particle size increased the gizzard insoluble Ca fraction in chicks and pullets. However, it did not influence its soluble Ca fraction in chicks but tended to reinforce the negative effect of omeprazole on soluble Ca in the gizzard and duodenum of chicks and laying hens. Coarse particles of Ca led to an increase in gizzard and duodenal soluble Ca at the end of eggshell calcification in laying hens. An enhancement in the level of Ca in the diet from 10 to 36 g/kg increased gizzard soluble Ca and duodenal soluble and ionic Ca concentrations in immature and adult hens. Intestinal Ca retention and bone mineralization was unaffected by gastric acid inhibition in chicks but were largely diminished by the use of coarse particles of Ca. Gastric acid inhibition was associated in laying hens with decreased Ca retention to a small extent and with reduced eggshell quality. These observations confirm that gastric acid secretion is of importance for CaCO3 solubilization but question its role as a prerequisite for intestinal Ca retention in chicks and even in hens fed on a high Ca diet.

Animals↗
PubMed2002

Positive effects of a chicken eggshell powder-enriched vitamin-mineral supplement on femoral neck bone mineral density in healthy late post-menopausal Dutch women.

Although bone metabolism is largely under genetic control, the role of nutrition is considerable. The present study evaluates the effects of chicken eggshell powder, a new source of dietary Ca, and purified CaCO3 on bone mineral density (BMD) of the lumbar spine and hip. Besides BMD we also looked at biochemical markers of bone and Ca metabolism. Both Ca sources were provided in combination with minerals and vitamins including Mg, cholecalciferol and phylloquinone. We designed a randomised, double-blind, placebo-controlled study to take place over 12 months. Healthy Caucasian women (n 85), selected by age (> or =50 and <70 years), from the databases of general practitioners were recruited by telephone calls. They had to be at least 5 years post-menopausal, with lumbar spine T-score being > - 2.5. At baseline, their mean habitual daily Ca intake was adequate. The women were randomly allocated to: eggshell powder-enriched (group A; n 24), purified CaCO3-enriched (group B; n 22), or a placebo product (group C; n 27). BMD was measured at baseline and then after 6 and 12 months of supplementation as were the biochemical markers bone-specific alkaline phosphatase, amino-terminal propeptide extension of type I collagen, deoxypyridinoline, calcitonin, intact parathyroid hormone, calcidiol, and urinary Ca. After 12 months of supplementation, only mean BMD of the femoral neck in group A was significantly increased (P=0.014) by 1.75% (95% CI 0.18, 3.32) compared with a decrease of -0.60% (95% CI -1.92, 0.72) in group C. This increase coincided with significant decreases in markers of bone resorption and formation. No significant changes were seen in BMD at other sites, including lumbar spine, nor in groups B and C. No differences were found between groups A and B, or B and C. The present study indicates that healthy late post-menopausal women with an adequate Ca intake at baseline may increase BMD of the hip within 12 months following supplementation with the chicken eggshell powder-enriched supplement.

Age Factors↗
PubMed2001

Utilisation of different phosphorus sources in relation to their fluorine content for broilers and layers.

1. The relative utilisation of different phosphorus sources in relation to their fluorine (F) content was studied in commercial broilers (5 to 40 d) and White Leghorn layers (252 to 364 d). The phosphorus (P) sources tested were bonemeal (BM), low fluorine (LFRP) and high fluorine (HFRP) rock phosphates and a commercial mineral mixture (CMM). The P sources were incorporated in broiler and layer diets by replacing dicalcium phosphate (DCP) on a P basis. 2. The F contents of diets based on BM, LFRP, CMM and HFRP were 53, 365, 622 and 1383 mg/kg in the broiler experiment and 34, 242, 437 and 967 mg/kg in the layer experiment, respectively. F was not detected in DCP based diets. 3. In broilers, body weight gain, food intake, gain: food, P retention and serum inorganic P content on P sources (BM and LFRP) containing F up to 365 mg/kg diet were similar to those on DCP. Body weight gain, food intake, serum calcium and inorganic P contents and retention of P were depressed in groups fed on CMM and HFRP, which may have been due to the toxic effects of F (622 and 1383 mg/kg) present in diets based on these P sources. 4. Bone ash and its P content were not affected by feeding diets containing F up to 1383 mg/kg from various P sources. The amount of F deposited in tibia increased significantly with increases in dietary F concentration. 5. In layers, egg production and food intake were not affected by F up to 437 mg/kg in diets containing BM, LFRP or CMM as the sole source of supplemental P. Egg production and food intake were depressed significantly in layers given the diet containing 967 mg F/kg from HFRP. 6. Egg mass: food, egg weight, shell quality (shell thickness and shell weight) and serum calcium and inorganic P levels were not affected by F up to 967 mg/kg in diets containing different P sources. 7. It may be concluded that the performance of broilers and layers was not affected by feeding various P supplements with dietary levels of F up to 365 and 437 mg/kg, respectively. The reduced performance in broilers and layers observed with some of the P sources may have been due to poor availability of P and/or toxic effects of F (622 and 967 mg/kg, respectively).

Animals↗
PubMed2002

Eggshell conductance, embryonic growth, hatchability and embryonic mortality of broiler breeder eggs dipped into ascorbic acid solution.

1. Eggs from a broiler breeder flock between 25 and 29 weeks of age were dipped into solutions of various concentrations of ascorbic acid (AA) for up to 2 min, in order to evaluate its effects on eggshell conductance (EC), amniotic fluid pH, albumen height and pH, embryonic weight and hatchability. 2. Three experiments were conducted and the following treatments were used: control, non-dipped (CND); control, water-dipped (CWD); 10 g AA/l (AA1); 20 g AA/l (AA2) and 30 g AA/l (AA3). Measurements of EC before and after dipping were made in experiments 1 and 2 in eggs from hens at 25 and 27 weeks of age, respectively. Albumen height and pH were measured after dipping the eggs in experiment 2. In experiment 3, egg weight loss, embryonic weight and amniotic fluid pH at 14 d of age, hatchability percentage (HP), and embryonic mortality were measured in eggs from hens at 29 weeks of age. 3. AA treatment of eggs and dipping period of time (1, 1.5 and 2 min) increased EC of eggs from hens at 25 and 27 weeks of age. 4. AA3 treatment for 2 min reduced albumen height and increased albumen pH of eggs when compared with the CND treatment in eggs from hens at 27 weeks of age. 5. HP of AA1-treated eggs from hens at 29 weeks of age was higher than that of the CND treatment. AA2 and AA3 treatments of eggs reduced HP and increased the percentage of non-pips when compared with any of the other treatments. 6. AA2 and AA3 treatments of eggs increased egg weight loss when compared with the control treatments (CND and CWD). Dipping treatment did not influence amniotic fluid pH or embryo weight expressed as a percentage of initial egg weight. 7. It is concluded that dipping hatching eggs into AA solution increased EC of eggs. Dipping eggs into 10 g AA/l for a period of 2 min increased HP in eggs from hens at 29 weeks of age, although this was not associated with a significant increase in egg weight loss.

Albumins↗
PubMed2005

Effect of intensity of eggshell pigment and illuminated incubation on hatchability of brown eggs.

The effects of intensity of brown eggshell pigment (light (LBP), medium (MBP) and dark (DBP)) and light intensity during incubation (low and high, 900 to 1380 and 1430 to 2080 lux, respectively) on eggshell characteristics, embryonic growth, hatchability traits, chick hatching weight and hatching time were investigated using eggs from a meat-type breeder (Hybro) flock at 32, 36 and 41 weeks of age in three trials. With eggs of similar weights the intensity of brown pigment was not associated with eggshell weight and thickness, and did not influence embryo weight and egg weight loss during incubation. The shade of brown pigment of eggs laid by young hens influenced the percentage hatchability (HP) of eggs incubated under light. Illuminated incubation improved HP of LBP eggs (compared with MBP and DBP eggs) from 32- and 36-week-old hens, but had no significant effect on HP of eggs from 41-week-old hens. Light intensity during incubation did not influence egg weight loss. High intensity of light during incubation reduced HP and increased early death percentage (EDP) in the LBP and MBP groups, and did not influence HP and EDP in the DBP group. Brown eggshell pigment and intensity of light during incubation did not influence hatching time. It is concluded that the shade of brown pigment, intensity of light during incubation and age of the breeder hens influenced the hatchability performance of embryos from brown eggs. Light during incubation improved the hatchability of embryos in light brown eggs laid by young hens and the shade of brown pigment of eggs laid by older hens did not influence hatchability under illuminated incubation. High intensity of light during incubation reduced hatchability of light and medium brown eggs, but not the dark brown eggs.

Animals↗
PubMed1986

Excessive cholecalciferol in a layers diet: decline in some aspects of reproductive performance and increased bone mineralisation of progeny.

Feeding hens a diet containing 5,000 micrograms (200,000 ICU)/kg of cholecalciferol for four 28-d periods had no adverse effect on hen-day egg production or hatchability. Egg weight, shell quality, food consumption and fertility were significantly decreased in hens fed 5,000 micrograms/kg. of cholecalciferol compared with those fed 24 micrograms (960 ICU) cholecalciferol/kg diet. Plasma calcium increased significantly as the concentration of cholecalciferol was increased in the diet. However, no histologically detectable changes in the viscera or changes in the proportion of bone ash were observed with any concentration of the vitamin. Chicks hatched from dams receiving excessive doses of cholecalciferol (5,000 micrograms/kg) and maintained on a rachitogenic diet for 4 weeks had a significantly higher proportion of tibial ash but there was no effect on either body weight or tibial calcium.

Animals↗
PubMed1996

Soluble matrix of hen's eggshell extracts changes in vitro the rate of calcium carbonate precipitation and crystal morphology.

1. Extra and intramineral eggshell matrix proteins were solubilised before and after demineralisation by sequential extractions using guanidine hydrochloride and EDTA. 2. The intramineral electrophoretic profile of SDS-PAGE showed the presence of 80, 66, 43, 36 and 15 kDa bands with a predominance of a 17 kDa band. In the extramineral part, the major protein was the 15 kDa band. 3. The introduction of intramineral extract to a metastable solution of calcium carbonate delayed the rate of crystal growth. The delay in the rate of precipitation was elicited by a single fraction (MW 50-80 kDa), isolated by gel filtration chromatography, of eggshell extracts. Extramineral extracts had no effect. 4. Addition in vitro of intramineral eggshell extracts modified the morphology of calcite; the crystals aggregated and showed irregular surfaces. 5. These observations suggest that constituents of the eggshell matrix are involved in the control of calcite growth and crytallographic structure of the hen's eggshell.

Animals↗
PubMed2006

Productivity of tree swallows (Tachycineta bicolor) exposed to PCBs at the Kalamazoo River superfund site.

A 123-km stretch of the Kalamazoo River in Michigan, was designated a Superfund site in 1990 due to historical releases of effluent containing polychlorinated biphenyl (PCB)-contaminated paper waste. Risk to bird species in the river ecosystem was evaluated using the tree swallow (Tachycineta bicolor) as a monitor for possible effects due to PCB exposure at two nesting locations, one in the Superfund site and one in an upstream reference location that is less contaminated with PCBs. In 2 of the 3 years of the study, clutch size at the contaminated location was 3.7 +/- 1.4 and 4.8 +/- 0.73 eggs per nest (mean +/- SD), which was significantly less than the clutch size at the reference location (5.0 +/- 1.1 and 5.3 +/- 1.1 eggs per nest). However, there were no statistically significant differences in fledging success, predicted brood size, predicted number of fledglings, or growth of nestlings between the Kalamazoo River Superfund site and an upstream reference location with lesser concentrations of PCBs in the sediments and riparian soils. Productivity and hatching success comparisons between these same sites were also not significantly different; however, the power of these conclusions was less (p < .10). The reduction in clutch size at the co-contaminated location could not be attributed to PCBs due to a number of confounding factors, including Co-cocontaminants, habitat structure, and food availability. Other reproductive parameters were not significantly impaired, and the size of the newly established colony at the Kalamazoo River Superfund site continued to grow over the period of the study. These site-specific observations, combined with multiple lines of evidence approach that considered results reported for the effects of both total PCBs and 2,3,7,8 tetrachlorodibenzo-p-dioxin equivalents (TEQ) on tree swallows at other locations, suggest that there were no significant population-level effects of PCBs on tree swallows at the Kalamazoo River Superfund site.

Animals↗
PubMed1977

Effects of DDT on eggshell quality and calcium adenosine triphosphatase.

Adult mallard ducks were fed a diet containing 50 ppm DDT for 6 months. Eggs laid during this period were collected and eggshell weight, thickness, and calcium were determined. Chronic ingestion of DDT resulted in production of eggshells that were significantly thinner and lighter than those of controls. Total calcium of thinned eggshells was also reduced; however, calcium per gram of eggshell was not altered, indicating that other eggshell constituents were not incorporated as well. Calcium adenosine triphosphatase activity in the microsomal fraction of eggshell gland epithelium was assayed in control and DDT-fed ducks. Enzyme activity in DDT-fed ducks was reduced to 65% of control values. Since Ca-ATPase has been shown to be associated with calcium transport, enzyme inhibition may be responsible for decreased eggshell weight and thickness. Electron microscopic evaluation of microsomal fractions showed elements of the plasma membrane, including cilia and microvilli, as well as rough and smooth endoplasmic reticulum. Inhibition of calcium transport at the plasma membrane of mucosal epithelium is proposed as a possible mechanism of DDT-induced eggshell thinning.

Adenosine Triphosphatases↗
PubMed1991

Great Lakes embryo mortality, edema, and deformities syndrome (GLEMEDS) in colonial fish-eating birds: similarity to chick-edema disease.

Several species of colonial fish-eating birds nesting in the Great Lakes basin, including herring gulls, common terns and double-crested cormorants, have exhibited chronic impairment of reproduction. In addition to eggshell thinning caused by high levels of DDT and metabolites, the reproductive impairment is characterized by high embryonic and chick mortality, edema, growth retardation, and deformities, hence the name Great Lakes embryo mortality, edema, and deformities syndrome (GLEMEDS). The hypothesis has been advanced that GLEMEDS in colonial fish-eating birds resembles chick-edema disease of poultry and has been caused by exposure to chick-edema active compounds that have a common mode of action through the cytochrome P-448 system. Detailed evidence has been collected from the following three groups of studies on herring gulls in the lower Great Lakes during the early 1970s; Forster's terns in Green Bay, Wisconsin in 1983; and double-crested cormorants and Caspian terns in various locations in the upper Great Lakes from 1986 onwards. It has proved difficult to establish not only the onset of the disease in the various species at various locations but also the period in which chick-edema active compounds were released. Anecdotal evidence suggested that serious egg mortality in Lake Ontario herring gulls first occurred in 1966, through the signs of chick-edema disease were not looked for until 1974. Only indirect evidence is available on the date of the release of one of the presumed causal agents, 2,3,7,8-tetrachlorodibenzo-p-dioxin, but highest levels may have occurred in the early to mid 1960s. More reliable data show that the onset of the improvement of reproduction of Lake Ontario herring gulls coincided with the declines in organochlorine compounds and particularly 2,3,7,8-TCDD and PCB. Similarly, information on the onset of the disease and exposures in the Forster's tern and double-crested cormorants in Green Bay is uncertain but bird banders did not observe deformities until the 1970s, which corresponds with the onset of high levels of PCB. If the disappearance of the Caspian tern from Saginaw Bay in the mid 1960s corresponds with the onset of GLEMEDS at that location, then there is a close temporal relationship to the onset of high PCB levels. Chick-edema disease is difficult to diagnose because there is no specific lesion, but rather there is a suite of lesions.(ABSTRACT TRUNCATED AT 400 WORDS)

Abnormalities, Drug-Induced↗
PubMed2000

Ultrastructural analysis of the eggshell: contribution of the individual calcified layers and the cuticle to hatchability and egg viability in broiler breeders.

1. Scanning electron microscopy was used to assess the relationship between the layers of the eggshell and egg viability. The relative thickness (absolute and %) of the mammillary, palisade, vertical crystal and cuticle layers relative to the total eggshell were measured over a 30-week laying period in 2 broiler breeder lines having different hatchability rates. 2. A significant difference in hatchability of 5.5% between the breeder lines was observed. However, there was no significant difference in either egg viability or fertility between lines. 3. The relative thickness of the individual calcified layers and the cuticle did not differ between lines. Therefore, variations in the thickness of the individual eggshell layers are unlikely to explain the 5.5% difference in hatchability rate between lines. 4. The thickness of the calcified region did not alter as the broiler hen aged. However, the percent contribution of each calcified layer significantly differed over the egg production period. 5. The cuticle was significantly thinner at the beginning and at the end of the laying period compared to 38 weeks of age. 6. No significant correlation was observed between egg viability and the relative thicknesses of the mamillary, palisade or cuticle layers. In contrast, there was a positive relationship between egg viability and the thickness of the vertical crystal layer. 7. The percent contribution of the mammillary and palisade layers was similar in the thinnest and thickest eggshells, suggesting conservation of the proportions of these layers independent of eggshell thickness.

Animals↗
PubMed2001

Stability of dietary ascorbic acid and the effect of supplementation on reproductive performance of broiler breeder chickens.

1. The purpose of the study was to determine the stability of dietary ascorbic acid and the reproductive responses of broiler breeder chickens to supplemental 75 mg ascorbic acid/kg diet. 2. Six breeder flocks of 13,000 birds each were studied. Egg production, eggshell porosity, fertility, hatchability and plasma ascorbic acid were measured. 3. Storage of the diets under dry heat resulted in a linear decrease in ascorbic acid content and the rate of decline was 5-fold higher in the supplemented diet. 4. Differences were not detected between treatments in egg production, egg weight, eggshell porosity, fertility, hatchability or plasma ascorbic acid. 5. The results did not provide evidence of a beneficial reproductive response to the inclusion of ascorbic acid in commercial broiler breeder diets.

Animals↗