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Helicoidal architecture of fish eggshell.

Previous publications show arced patterns in electron micrographs of either microfibrils or canals in sectioned fish eggshells, but these have been misinterpreted. We show here that such patterns in the inner layer of cod (Gadus morrhua), plaice (Pleuronectes platessa) and trout (Salmo gairdneri) eggs arise from a helicoidal structure. This consists of a laminate of protein microfibrils, with the direction of ply processing like the steps of a spiral staircase and with the same sense as a left-handed corkscrew. Mechanically, this is an ideal way to strengthen a spherical shell, to resist deforming forces equally from any direction. Radial canals which traverse this layer are forced into flattened and twisted ribbons. Both the helicoidal microfibrillar structure and the canal shape in fish eggshells show remarkable convergent evolution with similar structures in insect cuticles. Trout eggs were resistant to deforming forces as high as 380,000 N/m2. Fish eggshells, like those of many other organisms, are mechanically well designed.

Animals

Changes in whole blood oxygen affinity and eggshell permeability in high altitude chickens translocated to sea level.

High altitude (HA; n = 5) chickens (Gallus gallus) with a high oxygen hemoglobin (Hb) affinity were transported from their birthplace (Puno, Perú 4,000 m) down sea level (Lima, Perú). The in vivo whole blood oxygen affinity (P50) and the eggshell permeability (P) were studied after several months living at sea level and in the first (F1) and second (F2) generations born at sea level. Our approach was to analyze changes in Hb affinity and eggshell permeability, considered as indicators of HA adaptation in birds. Our results show an increase of the P50 values (a decrease in Hb affinity) towards sea-level values. The results in P indicate that this variable increases towards sea level values in the F2 generation. We conclude that in the Andean chicken, a relative "newcomer" to high altitude (no more than 500 years), neither the Hb affinity for oxygen nor the eggshell permeability are invariable indicators of HA adaptation, in contrast with other native high altitude mammals and birds.

Acclimatization

Eggshells are shaped by a precise spatio-temporal arrangement of sequentially deposited macromolecules.

The avian eggshell is a composite bioceramic which is formed by a controlled interaction of an organic and an inorganic phase. The organic phase contains, among other constituents, type X collagen and proteoglycans, mainly keratan and dermatan sulfate. Understanding the principles governing the synthesis and temporo-spatial distribution of such macromolecules, and their influence on the organization of the crystalline phase, is an essential aspect of establishing the biological basis of the quality of eggshell, both as an embryonic chamber and as a natural food package. In the present study, we have examined the process of eggshell formation by immunohistochemistry, scanning electron microscopy and energy dispersive X-ray microanalysis. Precise sites and timing of secretion were established for the deposition of particular macromolecules. Type X collagen is detected at the very first moment of shell membrane formation. The appearance of keratan sulfate coincides with the appearance of mammillae, while dermatan sulfate is deposited later, coincident with shell matrix deposition. We propose that keratan sulfate, due to its precise localization, temporal appearance and calcium-binding affinity, relates to the maintenance of calcium reserve bodies, the primary source of calcium for the embryo. On the other hand, dermatan sulfate may control crystal growth, resulting in a preferential orientation of calcite crystals within the palisade layer.

Animals

Heavy metals in avian eggshells: another excretion method.

Birds can rid their bodies of heavy metals through both excretion and deposition in feathers, and females can also eliminate heavy metals in the contents of their eggs. In this paper the levels of heavy metals (lead, cadmium, mercury, selenium, manganese, chromium) in the contents and shells of eggs of roseate terns (Sterna dougallii) and herring gulls (Larus argentatus) nesting at Cedar Beach, Long Island, are reported. For both species, metal concentrations were significantly higher in the contents compared to the shells for lead, mercury, selenium, and chromium. For herring gulls, metal levels were higher in the shells for cadmium and manganese. Levels of cadmium, mercury, and selenium were significantly higher in roseate tern egg contents than for herring gulls. In eggshells, lead, cadmium, mercury, and selenium were significantly higher in roseate terns compared to herring gulls. For both species, eggshells account for about 7-8% of the egg by weight, but less than 1% of the egg burden for mercury, 1-5% for lead, selenium, and chromium, and 7-11% for manganese. For cadmium, shells account for only 5% of the egg burden for roseate terns, but 29% for herring gulls. These data suggest that, except for mercury, eggshells provide another method of excretion of metals in these two species of birds.

Animals

Ultrastructure and characteristics of eggshells of the olive ridley turtle (Lepidochelys olivacea) from Gahirmatha, India.

The structure of a chelonian eggshell is of prime importance for the developing embryo. It acts as a protective covering as well as mediator in the exchange of heat and water. The fresh eggshell of the olive ridley (Lepidochelys olivacea) turtle displays a variety of structural forms in their aragonite framework. Its netted substrate, loose texture and poorly organised crystallites favour the easy exchange of air and water during its development. The shell consists mostly of calcium in carbonate form and K, Mg, Fe, Ni, Pb, Zn, Co, Cr, Cu, Mn and Cd in traces. Ba and Sr in traces are also a feature of the thermal analysis graph. Phosphorous was not detected in the shell and its absence eliminates an earlier suggestion of it being a key factor in the development of aragonite crystals in the inorganic structure of marine turtle eggshells. Its combustion characteristics, which have not been previously reported, are recorded.

Animals

Eggshell calcification of lymph nodes: an update.

Peripheral calcification of lymph nodes, "eggshell calcification," commonly occurs in patients with silicosis and coal-worker's pneumoconiosis. Sarcoidosis, postirradiation Hodgkin disease, blastomycosis, and scleroderma are other reported causes. Two not previously described causes, amyloidosis and histoplasmosis, are presented, and disorders that simulate eggshell calcification are listed. The explanation for the eggshell pattern of calcification is not known.

Amyloidosis

The presence of desmosine and isodesmosine in eggshell membrane protein.

Investigation of eggshell membrane by amino acid analysis and radioimmunoassay established the presence of the crosslinking amino acids desmosine and isodesmosine at levels of 28 microgram/100 mg dry weight. Specific enzymatic and chemical hydrolysis indicates that these crosslinks are present as constituents of eggshell membrane protein and do not reflect the presence of small amounts of elastin. The eggshell membrane protein has approximately 10 times the level of lysine-derived aldehyde found in elastin.

Amino Acids

Role of type X collagen on experimental mineralization of eggshell membranes.

Type X collagen is a transient and developmentally regulated collagen that has been postulated to be involved in controlling the later stages of endochondral bone formation. However, the role of this collagen in these events is not yet known. In order to understand the function of type X collagen, if any, in the process of biomineralization, the properties of type X collagen in eggshell membranes were further investigated. Specifically, calvaria-derived osteogenic cells were tested for their ability to mineralize eggshell membranes in vitro. Immunohistochemistry with specific monoclonal antibodies was used to correlate the presence or absence of type X collagen or its propeptide domains with the ability of shell membranes to be mineralized. The extent of mineralization was assessed by Von Kossa staining, scanning electron microscopy and energy-dispersive spectroscopy. The results indicate that the non-helical domains of type X collagen must be removed to facilitate the cell-mediated mineralization of eggshell membranes. In this tissue, intact type X collagen does not appear to stimulate or support cell-mediated mineralization. We postulate that the non-helical domains of type X collagen function in vivo to inhibit mineralization and thereby establish boundaries which are protected from mineral deposition.

Amino Acid Sequence

Optimal dietary level of 1 alpha,25-dihydroxycholecalciferol for eggshell quality in laying hens.

The optimal dietary level of 1 alpha,25-dihydroxycholecalciferol [1,25-(OH)2D3] for eggshell quality was established. White Leghorn hens, 59 wk of age, were fed one of eight diets that contained the same basal ingredients, including 3.1% calcium, but different levels (microgram/kg) or forms of calciferol supplements: no calciferol supplement of any form (56 hens); 27.5 (control) or 55.0 micrograms of cholecalciferol (56 hens each); 3, 5, or 7 micrograms of 1,25-(OH)2D3 (28 hens each); 5 micrograms of 24,25-dihydroxycholecalciferol [24,25-(OH)2D3] with 28 hens; 5 micrograms each of 1,25-(OH)2D3 and 24,25-(OH)2D3 (28 hens). All groups were fed the control diet prior to the 21-wk treatment. The group fed 5 micrograms 1,25-(OH)2D3/kg diet ranked first in specific gravity (SG), e.g., 1.081 versus 1.077 for the control group at Week 21 (P less than .05). The group fed 7 micrograms 1,25-(OH)2D3/kg consumed 30% less feed and laid 20% fewer eggs than the control, but shell quality was not affected. The groups receiving no calciferol supplement or receiving only 24,25-(OH)2D3 laid eggs with significantly lower SG than the control after 2 wk of treatment (1.072 or less versus 1.082 at Week 2). The rest of the treatment groups mentioned were comparable to the control in eggshell quality and egg production. Groups fed the combination of 1,25-(OH)2D3 and 24,25-(OH)2D3 per kilogram of feed, or 1,25-(OH)2D3 alone at 5 micrograms/kg, had significantly higher tibial weights relative to the control group. All groups receiving the diets without cholecalciferol supplementation had markedly reduced hatchability. It was concluded that the optimal dietary level of 1,25-(OH)2D3 for improving eggshell quality without affecting egg production was approximately 5 micrograms/kg and the toxic level was 7 micrograms/kg.

Animals

Effect of cimetidine on eggshell quality and plasma 25-hydroxycholecalciferol in laying hens.

Experiments were conducted to investigate the effect of feeding cimetidine (CIMET), ranging from 0 to 750 mg/kg, on vitamin D3 metabolism and eggshell calcification in laying hens fed two levels of vitamin D3 (500 and 2,000 ICU/kg). Final BW and feed intake were not significantly affected by either CIMET or vitamin D3 level. Feeding 500 and 750 mg of CIMET significantly decreased total egg production in hens fed either level of vitamin D3, but no differences were observed at lower CIMET levels. Tibia ash decreased significantly in hens fed 150 to 750 mg of CIMET, regardless of the vitamin D3 level. Plasma Ca and inorganic P concentrations were decreased in hens fed high CIMET levels (500 and 750 mg/kg) at Week 2, but no differences were observed at Week 4. Feeding CIMET (500 and 750 mg/kg) significantly decreased plasma 25-hydroxycholecalciferol (25-OHD3) levels at Week 2 in hens fed both vitamin D3 diets but not at Week 4. Eggshell breaking force, shell thickness, and percentage shell weight were decreased significantly by CIMET in all experiments; however, in one experiment, shell quality recovered by Week 8. These results suggest that the CIMET-induced reduction in bone mineralization, eggshell quality, and plasma 25-OHD3 levels could be due to interference of CIMET with vitamin D3 metabolism in vitamin D3-replete laying hens. Shell quality decreased in CIMET-treated hens fed the higher vitamin D diet even though 250-HD3 plasma levels were three times higher than in hens fed the lower vitamin D diet, suggesting that CIMET affected shell quality through some mechanism other than inhibition of 250-HD3 synthesis.

Animal Feed

Influence of vitamin D3, 1 alpha-hydroxyvitamin D3, and 1,25-dihydroxyvitamin D3 on eggshell quality, tibia strength, and various production parameters in commercial laying hens.

Four hundred 53-wk-old Hyline W36 laying hens were randomly allocated to 10 treatments. The effects of feeding two vitamin D3 metabolites, 1 alpha-hydroxyvitamin D3 [1 alpha-(OH) D3] and 1,25-dihydroxyvitamin D3 [1,25-(OH)2 D3], each at five dietary levels (0, .75, 1.50, 3.00, and 4.50 micrograms/kg of feed) were determined on eggshell quality and tibia strength in commercial laying hens (Experiment 1). In Experiment 2, 1,440 Hyline W36 65-wk-old laying hens were used to determine the effects of four levels of vitamin D3 (0, 500, 1,000, and 1,500 ICU vitamin D3/kg) and three levels of dietary 1,25-(OH)2 D3 (0, .5, and 1.0 microgram/kg of feed) on eggshell quality, tibia strength, and egg production. In Experiment 1, neither 1,25-(OH)2 D3 nor 1 alpha-(OH) D3 affected eggshell quality or production criteria. Tibia weight was increased by adding either 1,25-(OH)2 D3 or 1 alpha-(OH) D3. In Experiment 2, 1,25-(OH)2 D3 increased percentage of shell, shell weight, and egg breaking strength when 0 ICU D3/kg was fed but had no effect at higher levels of vitamin D3. Egg production, feed consumption, and egg weight were also increased with supplemental 1,25-(OH)2 D3 when 0 ICU D3/kg was fed. Tibia weight and tibia breaking strength were also increased by adding 1,25-(OH)2 D3 to the diet. The commercial laying hen metabolizes sufficient 1,25-(OH)2 D3 from dietary vitamin D3 to maintain shell quality but not enough to maintain tibia strength.

Animals

Effects of selection for growth and selection diet on eggshell quality and embryonic development in Japanese quail.

Decreased hatchability in Japanese quail following selection for growth and relative improvements in hatchability following selection during goitrogen treatment were investigated. An unselected quail line (Line C) and lines selected for high 4-wk body weight while being fed diets containing either 20% CP and .2% thiouracil (TU) (Line T), or 28% CP and no TU (Lines P and H-CD) were used. Egg weight loss between 0 and 4, and 0 and 14 days of incubation, hatchability, and developmental stage of embryonic mortalities were determined in Experiment 1. Length of the incubation period, and percentages of body water and dry body weight were determined for 10- and 14-day embryos and 18-day-old chicks in Experiment 2. Eggshell quality was determined in Experiment 3. Hatchability was lower in selected than in unselected quail due to increased early and late embryonic mortality. Egg weight loss during incubation was greater in Line P than in Line C eggs. Weight loss, measured across lines, was lower in eggs that hatched than in those in which embryos died early. Percentage body water was higher in Line C than in Lines P and T at 14 and 18 days of incubation; however, percentage body water in Line T was lower than Line P at 14 days. Percentage of dry body weight was greater in Lines P and T than in Line C at 14 and 18 days. Eggshell thickness was greater in Line H-CD when compared with Line C. Increases in eggshell permeability occurring during selection for growth were associated with increased embryonic mortality and decreased hatchability.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Inheritance of tinted eggshell colors in white-shell stocks.

The present study was conducted to study the genetics of tinted eggshell colors in two breeds of chickens laying white-shell eggs. Reciprocal crosses were made between an inbred White Leghorn line (ES) and an inbred Ancona line (ANC). The F1 birds were intercrossed and F1 females were backcrossed to each of the original lines. Eggshell color from each resultant group was measured using a Minolta chromometer, and a value (e) representing the color intensity adjusted by hue and saturation was used as the measurement criterion. Age, hatch group, and crosses each contributed significantly to the variation seen in eggshell color. Distribution comparisons indicated that two major autosomal loci affected the trait in these lines: one gene having incomplete dominance controls the amount of pigment deposition; the second completely inhibits pigment deposition when homozygous recessive. A test of goodness of fit supported this hypothesis. Genetic components were estimated by linear models. Epistasis, dominance, and additive effects contributed significantly to this trait. No sex-linked effects were noted.

Analysis of Variance

Collection and separation of Aedes taeniorhynchus eggshells from mangrove soil.

Two methods to separate eggshells of Aedes taeniorhynchus from mangrove soil were compared. Selective sieving, using nested sieves with 0.185 and 0.170-mm screen openings, and water flotation both removed over 99% of the soil. However, water flotation recovered a significantly greater percentage of eggshells (62% vs. 34%). There was no significant difference in the recovery rate of viable eggs and new and old eggshells using water flotation.

Aedes

F strain Mycoplasma gallisepticum vaccination of post-production-peak commercial Leghorns and its effect on egg and eggshell quality.

Forty-five-week-old commercial leghorns negative for antibodies to Mycoplasma gallisepticum (MG) and M. synoviae were vaccinated with high-passage F strain MG (FMG). Hens were confined in modified Horsfall-Bauer isolation units through 60 weeks of age. Egg production (% hen day) and parameters of egg and eggshell quality were monitored, including egg weight, eggshell strength, Haugh unit score, pimpling, and blood/meat spot incidence. Egg production was significantly lower (P less than 0.05) for FMG vaccinates than controls (down 5.76% and 5.80% in Trials 1 and 2, respectively). However, vaccinates and controls did not differ significantly in eggshell strength, shell thickness, pimpling, or blood/meat spot incidence. Haugh unit scores were significantly (P less than 0.05) greater for FMG vaccinates. At necropsy, all reproductive tracts appeared grossly normal. These studies suggest that high-passage FMG vaccination of post-production-peak hens does not adversely affect oviduct function.

Animals

Supplemented eggshell restores calcium transport in chorioallantoic membrane of cultured shell-less chick embryos.

It was previously reported (Tuan, 1980a) that the development-specific expression of calcium transport and related functions in the chick embryonic chorioallantoic membrane (CAM) requires the continuous presence of the eggshell, the calcium source of the embryo. To further understand the mechanism of action of the eggshell on the CAM functions, this study reports the effects of eggshell supplementation on chick embryos maintained in shell-less cultures. The cultured embryos were able to accumulate and utilize the exogenous shell calcium, applied directly onto the CAM, for skeletal formation. In the region of the CAM directly adhering to the added shell, calcium transport activity, calcium-binding protein (CaBP) activity, and vitamin K-dependent gamma-glutamyl carboxylase activity were significantly restored. These results strongly suggest that the proximity of shell calcium may regulate expression of calcium transport and related functions in the chick embryonic CAM.

Allantois

Oocyte surface proteins EGG-1 and EGG-2 are required for eggshell integrity in Caenorhabditis elegans.

Metazoan eggs are surrounded by a specialized coat of extracellular matrix that mediates sperm-egg interactions. This coat is rapidly remodeled after fertilization to form a barrier that prevents polyspermy, protects against environmental insults, and provides structural support to the developing embryo. In C. elegans several oocyte surface proteins have been identified that mediate these events. However, whether two of these proteins, EGG-1 and EGG-2, are required for fertilization or downstream events has been unclear. Here, we address this question using more recent advances in genome editing tools through the creation of egg-1 egg-2 deletions of the endogenous loci. We found that egg-1 egg-2 oocytes are fertilization competent and form rudimentary eggshells. While the integrity of the egg-1 egg-2 eggshells are compromised and often rupture within the uterus, surprisingly, some embryos are capable of undergoing several rounds of cell division. Overall, our findings demonstrate that EGG-1 and EGG-2 are not required for fertilization but are involved in post-fertilization processes.

Caenorhabditis elegans

Eggshell aspiration in infants.

Two infants inhaled eggshell fragments. The first aspirated a small fragment into the upper part of the trachea and resuscitation attempts resulted in descent of the foreign body to the right lower lobe bronchus. Two attempts at bronchoscopy were unsuccessful and spontaneous disintegration occurred. The second infant aspirated an eggshell fragment to the entrance of the larynx where it was removed by laryngoscopy. Laryngeal or tracheal foreign body in infants under 1 year of age are rare, and the diagnosis and removal are difficult.

Animals