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PubMed1994

Characterization of the main egg envelope proteins of the sea bass Dicentrarchus labrax L. (Teleostea, Serranidae).

Fish eggs are surrounded by a resistant acellular coat commonly called the chorion or zona radiata. This study characterizes the eggshell proteinaceous content of unfertilized eggs of the sea bass Dicentrarchus labrax, with a view to the preparation of immunogens. Solubilization of the purified eggshells was achieved in 8 M urea followed by one- and two-dimensional gel electrophoresis. Glycoproteins were detected using concanavalin-A in one and two-dimensional gels, and the principal glycoproteins had a molecular weight of 47 kDa and 170 kDa. Partial purification of a few polypeptides in the 45 kDa to 55 kDa range was achieved by gel filtration chromatography. Although whole eggshells were relatively insoluble even in 8 M urea, partial purification of these polypeptides enable them to dissolve completely in solutions at low ionic strength.

Animals↗

PubMed1990

Geographical distribution of contaminants and productivity measures of herring gulls in the Great Lakes: Lake Erie and connecting channels 1978/79.

The distribution and size of colonies, residue levels of DDE, DDT, HCB, dieldrin, mirex and PCBs in eggs, productivity and eggshell thickness were determined for herring gulls at 14 sites in Lake Erie and connecting channels. The centre of distribution for breeding herring gulls was the Western Basin where approximately 90% of the 6200 nests in the study area were located. Seven of 22 colonies showed an average annual population increase of 48.3%. Most of the increase in breeding herring gulls on Lake Erie is directly associated with sites that have undergone habitat modification by man. Levels of PCBs and DDE ranged from 35 to 140 ppm (wet weight) and from 2.8 to 9.4 ppm, respectively; all other residues were less than 0.49 ppm. Most organochlorine residue levels were highest in eggs from colonies in or near the Niagara or Detroit Rivers. Mirex residues were greatest in the Niagara River and decreased significantly to the west. PCB residues were greatest in the Detroit River and decreased significantly to the east. The lowest levels generally came from colonies in the Sandusky Basin and near Pelee Island in western Lake Erie. Discriminant function analysis of six organochlorine contaminants correctly classified 90% or more of the eggs from up to four colonies in one or more years. Levels of PCBs and HCB appeared to have the greatest discriminating power. Herring gull productivity at all colonies (1-1.7 young gulls/pair) was normal and showed no significant geographical variation. Eggshell thickness was greatest in colonies in the Sandusky Basin and least in colonies in the Detroit River and extreme west end of the lake; mean eggshell thickness was 0.350 +/- 0.02 mm (6.7% thinning), which was weakly, but significantly correlated to DDE concentration. The variation in contaminants in herring gull eggs on a Basin basis (i.e., Western, Eastern, Sandusky, etc.) paralleled those known for sediments, water and fish. Thus, we suggest that in addition to its role as an indicator of lake-wide contamination of the Great Lakes, the herring gull, under some circumstances, may function as an indicator of "regional" contamination. This is an important distinction as it improves the geographical specificity of the herring gull as an indicator species on the Great Lakes, where it is a non-migratory species.

Animals↗

PubMed1993

Chitinolytic activities in Heligmosomoides polygyrus and their role in egg hatching.

The occurrence of chitin in the eggshell of Heligmosomoides polygyrus has been determined by histochemical and biochemical techniques. Approximately 5% of the egg dry weight was chitin. Staining with Calcofluor white showed the chitin in the eggshell to be more accessible to the stain after hatching or rupturing of the eggshell. Chitinolytic activity has been detected using fluorescent substrates in extracts of adult males (at low levels), females and eggs. Enzyme activity in situ, within the developing larvae, was visualised with the same substrates. It was localized in discrete granules about 1 micron in diameter which occurred as groups in areas of about 5 microns in diameter, in the posterior third of the larvae. The chitinolytic activity in the eggs increased with the age of the egg and was released into the medium when the eggs hatched. The chitinase activities were very sensitive to inhibition by allosamidin, a specific chitinase inhibitor, with an IC50 for the crude egg extract of 2.2 nM. However, treatment of eggs with 250 microM allosamidin resulted in a slowing but not cessation of egg hatching.

Acetylglucosamine↗

PubMed2002

Early reproductive success of western bluebirds and ash-throated flycatchers: a landscape-contaminant perspective.

Eggshell quality, clutch size, sex ratio, and hatching success of western bluebirds (Sialia mexicana) and ash-throated flycatchers (Myiarchus cinerascens) were studied on a landscape-soil contaminant gradient at Los Alamos National Laboratory (LANL) in New Mexico from 1997 to 1999. A variety of contaminants (heavy metals, chemicals, insecticides, polychlorinated biphenyls (PCBs), organochlorines, and radioactive isotopes) range across different spatial scales and concentrations on LANL land. This study is an example of a monitoring program over a large area with varying degree of contamination that is used to highlight locations of concern for future research. There were two locations where the flycatcher had a lower hatching success. The bluebirds at Sandia wetland, a location of concern for PCBs. had a thinner eggshell thickness index (RATCLIFFE) and the eggs were smaller than at other locations. The flycatcher had thinner eggshells than bluebirds, which could add to sensitivity to exposure to contaminants. There was no variation in clutch size or sex ratio between locations or areas closer to contaminant release sites for both species. Percent females in the clutch ranged from 0 to 100% in the WEBL and from 33 to 67% for ATFL.

Animals↗

PubMed2002

Mineral status of embryos of domestic fowl following exposure in vivo to the carbonic anhydrase inhibitor acetazolamide.

Eggs of domestic fowl were given daily injections of vehicle (DMSO) or vehicle plus acetazolamide, a potent inhibitor of the enzyme carbonic anhydrase, beginning on day 12 of incubation. Embryos were removed from eggs on days 16 and 18, and carcasses and yolks were analyzed for calcium, magnesium, and phosphorus. Treatment with acetazolamide did not affect the quantity of calcium or phosphorus in carcasses and the effect, if any, on magnesium in carcasses was small. However, calcium content of yolk was reduced substantially by acetazolamide both on day 16 and day 18. The reduction in calcium content of yolk led, in turn, to a reduction in the total quantity of calcium in eggs on days 16 and 18. Embryos exposed to acetazolamide seemingly mobilized less calcium from the eggshell than did control embryos. When faced with a shortfall in the availability of calcium from the eggshell, embryos defended carcass calcium, and the shortfall was reflected in a reduction in the quantity of calcium deposited in yolk. The results of this study support the concept that the enzyme carbonic anhydrase plays a role in solubilization of the eggshell and provision of calcium to embryos.

Acetazolamide↗

PubMed1999

Evaluation of a protocol for studying the chick chorioallantoic membrane in vitro.

Explants of eggshell with and without the chorioallantoic membrane were taken from fertile chicken eggs on day 16 of incubation and exposed in vitro to inhibitors (acetazolamide and benzolamide) of carbonic anhydrase to determine if enzyme inhibition affected release of calcium from the shell. A separate experiment examined the effect of the metabolic poison dinitrophenol (DNP) on release of calcium from explants. Explants with the chorioallantois in situ released more calcium than those lacking the epithelium, but neither the enzyme inhibitors nor DNP affected release of calcium. The lack of effect of the enzyme inhibitors could indicate that activity of carbonic anhydrase is not as important to the release of calcium from the eggshell as has been assumed. However, the absence of an effect of DNP instead indicates that release of calcium mediated by the chorioallantois in vitro simply lacks physiological relevance. Thus, results of this investigation raise doubts that the mechanism underlying release of calcium from the eggshell can be assessed in vitro.

Allantoin↗

PubMed2001

Cathepsin L is essential for embryogenesis and development of Caenorhabditis elegans.

Cysteine proteases play critical biological roles in both intracellular and extracellular processes. We characterized Ce-cpl-1, a Caenorhabditis elegans cathepsin L-like cysteine protease. RNA interference with Ce-cpl-1 activity resulted in embryonic lethality and a transient delayed growth of larvae to egg producing adults, suggesting an essential role for cpl-1 during embryogenesis, and most likely during post-embryonic development. Cpl-1 gene (Ce-cpl-1:lacZ) is widely expressed in the intestine and hypodermal cells of transgenic worms, while the fusion protein (Ce-CPL-1::GFP) was expressed in the hypodermis, pharynx, and gonad. The CPL-1 native protein accumulates in early to late stage embryos and becomes highly concentrated in gut cells during late embryonic development. CPL-1 is also present near the periphery of the eggshell as well as in the cuticle of larval stages suggesting that it may function not only in embryogenesis but also in further development of the worm. Although the precise role of Ce-CPL-1 during embryogenesis is not yet clear it could be involved in the processing of nutrients responsible for synthesis and/or in the degradation of eggshell. Moreover, an increase in the cpl-1 mRNA is seen in the intermolt period approximately 4 h prior to each molt. During this process Ce-CPL-1 may act as a proteolytic enzyme in the processing/degradation of cuticular or other proteins. Similar localization of a related cathepsin L in the filarial nematode Onchocerca volvulus, eggshell and cuticle, suggests that some of the Ce-CPL-1 function during development may be conserved in other parasitic nematodes.

Amino Acid Sequence↗

PubMed1999

Gordon Memorial Lecture. An egg ist ein ei, es un huevo, est un oeuf.

1. Many of the mechanical tests devised to measure shell quality are inadequate because they fail to recognise the complex interaction between the organic and inorganic aspects of the eggshell. 2. Twelve structural modifications have been observed at the level of the mammillary layer and their presence correlated with a variety of environmental stress events. Occurring as they do in the basal layers of the shell, these morphological variants influence its mechanical properties. 3. The organic matrix proteins which complex with the calcium carbonate derive from a variety of sites within the oviduct and vary in their location within the fully formed shell. In vitro mineralisation reveals the significance of these proteins in the crystal growth mechanism. 4. The isolation and identification of the protein moiety in well-structured eggshells is an essential prerequisite to understanding the abnormalities in crystal growth observed in the shells of older birds challenged by disease and other undesirable 'on farm' events. 5. The eggshell is the daily indicator of the bird's harmony with its environment and as such provides a readily accessible and non invasive measure of welfare. The integration of these data with those derived from behavioural and biochemical testing should provide industry with a reliable numerical welfare index.

Animal Feed↗

PubMed1989

A female sterile screen of the Drosophila melanogaster X chromosome using hybrid dysgenesis: identification and characterization of egg morphology mutants.

We have conducted a hybrid dysgenic screen of the X chromosome for mutations affecting female fertility, with particular attention to those causing abnormal egg and eggshell morphology. In a screen of 4017 dysgenic strains, 398 mutants derived from 168 different germ lines were isolated and assigned to eight classes according to their diverse phenotypes. One interesting class consists of mutants that block oogenesis at specific stages. Our analysis has focused on mutations affecting eggshell formation, including mutants that lay morphologically abnormal sterile eggs as well as those that lay no eggs but exhibit blocks in the late stages of oogenesis. A subset of 48 mutants was assorted into 30 allelic groups by inter se complementation and genetically localized by interval mapping. Two multiallele complementation groups, de1 (7 alleles) and ne1 (8 alleles), were identified as well as five two-allele complementation groups. A search for alleles among mutants generated in other female sterile screens was unsuccessful, pointing to the distinctive nature of the dysgenic mutant collection. The single case of allelism determined in this study was one with a lethal allele of the Broad-Complex, l(1)npr, suggesting a possible involvement of ecdysone in choriogenesis. A subset of 18 dysgenic strains was analyzed for P element hybridization and 16 of these were found to have hybridization signals in the appropriate cytogenetic interval. By examining these signals in two or more alleles of the same complementation group, we have been able to tentatively localize two mutations. Light and electron microscopy of the eggshell in 43 different strains has revealed a variety of effects. The respiratory appendages were defective in 27 of these mutants. Effects on the ultrastructure of the main body of the endochorion were not strongly correlated with the appendage defects, and could be classified as minor (14 mutants) or major (16 mutants). Although 13 mutants showed no ultrastructural chorion defects, six of these had defective respiratory appendages.

Animals↗

PubMedDate not supplied

Trait association of a genetic marker near the IGF-I gene in egg-laying chickens.

The insulin-like growth factor I (IGF-I) gene was screened for genetic variants associated with trait means and trait correlations. Analysis of an unselected randomly mated White Leghorn population revealed a PstI restriction fragment length polymorphism (RFLP) in the 5' region of the gene which segregated at a frequency of 0.83 for the PstI(+) allele (presence of a PstI restriction site). A comparison of the three genotypic classes revealed that the PstI(-/-) genotype was associated with a significantly lower egg weight measured in three different time periods, while the PstI(+/-) genotype was significantly associated with a higher eggshell weight estimated from the egg weight and egg specific gravity. For eggshell weight, the effect was age dependent and significant only for the last two periods of egg laying. No genotype associations were found for body weight, feed consumption, and egg laying rates. Significant dominance effects of the IGF-I genotype were observed for two of the egg weight measurements and three of the eggshell weight estimates. Partial correlation analyses in the two most frequent genotypic classes, PstI(+/+) and PstI(+/-), revealed the presence of a regulatory loop between feed consumption, body weight, egg weight, and the rate of egg laying. Several aspects of this regulatory loop were different between the two genotypic classes. In particular, for the PstI(+/+) genotype, feed consumption was positively associated with egg weight, while there was no significant association for the PstI(+/-) genotype. Further, the degree of association of body weight with egg weight decreased with age in the genotypic class PstI(+/-), while it was constant for the PstI(+/+) genotype. The results indicated that the marker in the IGF-I gene was not only associated with changes in some trait means, but also with changes in the stability of the coordination between feed intake, body weight, and egg production traits.

Animals↗

PubMed1999

Packaging effects on shell egg breakage rates during simulated transportation.

Shell eggs were packaged in either expanded polystyrene (EPS) foam or molded paper pulp (MPP) one dozen cartons, then were bulk packaged in either polypropylene crates or corrugated boxes. The packages were then subjected to a well-defined computer-simulated vibration test on an electrohydraulic test machine. The percentage and the location on the egg (side, top, bottom) of breakage was determined in the secondary (corrugated box or polypropylene crate) and primary (EPS or MPP carton) package after 15, 75, and 180 min. For each of three trials, 60 dozen Grade A large eggs were randomly assigned to each primary package and cross-stacked in a secondary container that contained three cartons in a row and a total of five layers. When cartons were packed in 15-dozen corrugated boxes, no significant difference was found in total eggshell damage rates between the MPP carton and the EPS carton. However, when eggs were packed in 15-dozen plastic crates, the MPP cartons caused significantly less eggshell damage than the EPS cartons. The EPS cartons packed in corrugated boxes had the lowest breakage (4.63%), whereas the EPS foam cartons packed in plastic crates had the highest breakage (12.59%). When the effect of secondary packaging and vibration time were not considered, no significant difference was found between MPP and EPS cartons. In addition, when the effect of primary packaging was not taken into account, the corrugated boxes had significantly lower breakage rates than the plastic crates. Nearly 55% of the breakage occurred in the bottom section of the eggshell as compared to the side and top. When the test periods were compared, the EPS cartons packed in plastic crates had the highest breakage (16.28%) at 180 min.

Animals↗

PubMed2000

Effects of breeder age and dietary fat source and level on broiler hatching egg characteristics.

The effects of breeder age and added dietary fat source and level on broiler hatching egg characteristics were evaluated. Diets included no added fat (NAF) or 3.0% added poultry fat (PF) for peak energy intakes of 430 and 467 kcal/hen-day (PCD) or 1.5% PF or 3.0% corn oil (CO) at 449 PCD. As added dietary fat was changed from CO to PF, the percentage of unsaturated dietary fatty acids, including linoleic acid, decreased. Feeding of experimental diets was initiated when breeders were 22 wk old. Total fresh egg weight; eggshell weight; percentages of yolk (PYK), albumen (PAB), and eggshell (PSHL) weights; and yolk:albumen ratio were measured at various weeks between 26 and 47 wk of age. Egg weight increased progressively with hen age. Significant increases in yolk:albumen ratio occurred between Weeks 26 and 31 and between Weeks 31 and 35. Low (430 PCD) dietary energy levels significantly reduced PYK at 35 wk and increased PAB across breeder age. Eggshell weight was lower in birds fed moderate (449 PCD) compared to low energy levels at Week 26, moderate compared to high (467 PCD) energy levels at Week 41, and PF compared to CO across fat level at Week 31. At Weeks 31 and 41, PSHL was increased by the use of 3.0% PF compared to 1.5%, and PSHL was increased at Weeks 26 and 41 by using added PF compared to CO across fat level. Increased dietary energy decreased PAB and the use of added dietary CO rather than PF decreased PSHL in broiler breeders between 26 and 47 wk of age.

Age Factors↗

PubMed1992

Alligators provide evidence for the evolution of an archosaurian mode of oviparity.

The female reproductive tract of birds is different from that of other oviparous amniotes in that the eggshell membranes and calcareous layer are formed in separate regions of the uterus; the isthmus and shell gland, respectively. Phylogenetically, birds are included among the archosaurs, along with crocodilians and dinosaurs. Many dinosaurs were oviparous, producing hard-shelled eggs, yet the reproductive system of dinosaurs has proven difficult to investigate, due to poor preservation of soft anatomy. In this study, we examined functional morphology and eggshell formation in a reptilian archosaur, the American alligator, and demonstrated that the crocodilian reproductive tract has separate uterine regions for formation of the eggshell membranes and calcareous layer. These uterine regions are ultrastructurally comparable to the isthmus and shell gland of birds, and may be homologous. This similarity of reproductive functional morphology between crocodilians and birds may implicate the evolution of an archosaurian mode of oviparity that may shed light on dinosaur reproduction.

Alligators and Crocodiles↗

PubMed1994

Preventing heel pressure sores: a comparison of heel pressure relieving devices.

It has been widely recognized that elderly patients with an orthopaedic problem are predisposed to developing heel pressure sores. In this study four pressure-reducing devices, commonly used in the prevention of heel ulcers, were objectively compared for their ability to decrease or remove pressure on the heels of patients with fractured necks of femurs and fractured femurs. Forty-one patients were randomly allocated a pressure relieving device. The efficacy of the device was evaluated by continuously assessing the skin integrity of both heels on a daily basis over a period of 12 days. Data were collected over a 30 month period. The four devices were foam splints, eggshell foam, duoderm and heel protector boots. Foam splints and eggshell foam proved to be more effective devices in relieving pressure exerted on the heel. This study recognizes that meticulous nursing care remains the critical element in prevention of heel ulcers; however, the use of eggshell foam and foam splints in conjuction with this enhances the maintenance of skin integrity.

Aged↗

PubMed1988

Indexes of vitamin D deficiency in Japanese quail embryos.

Effects of vitamin D deficiency (-D) on mineral homeostasis were investigated in Japanese quail embryos. The -D embryos from 1,25(OH)2D3-fed hens became progressively calcium deficient, as documented by hypocalcemia and reduced calcium accumulation by the skeleton, yolk sac, and allantoic fluid. Plasma phosphate was progressively elevated between days 11 and 15. Increased calcium accumulation by the skeleton, yolk sac, and allantoic fluid occurred between days 12 and 15 in +D embryos. Phosphate and adenosine 3',5'-cyclic monophosphate (cAMP) concentrations of allantoic fluid increased in +D embryos during the period of shell calcium mobilization. Further increases in phosphate and cAMP excretion into allantoic fluid occurred in -D embryos, although no calcium was absorbed from the shell. Renal 25(OH)D-1-hydroxylase activity increased between days 11 and 13, whereas the adenylate cyclase response to parathyroid hormone was lost in -D embryos by day 14. These changes in renal function are indicative of secondary hyperparathyroidism in the -D embryos. Differentiation of villus cavity and capillary covering cells occurred in the chorionic epithelium of -D embryos, but eggshell calcium was apparently not absorbed. In contrast, 75% of the total body calcium of newly hatched (+D) chicks was obtained from the eggshell. Thus the dissolution and/or transport of eggshell calcium is dependent on vitamin D in quail embryos.

Adenylyl Cyclases↗

PubMed2002

Modeling and computational analysis of EGF receptor-mediated cell communication in Drosophila oogenesis.

Autocrine signaling through the Epidermal Growth Factor Receptor (EGFR) operates at various stages of development across species. A recent hypothesis suggested that a distributed network of EGFR autocrine loops was capable of spatially modulating a simple single-peaked input into a more complex two-peaked signaling pattern, specifying the formation of a pair organ in Drosophila oogenesis (two respiratory appendages on the eggshell). To test this hypothesis, we have integrated genetic and biochemical information about the EGFR network into a mechanistic model of transport and signaling. The model allows us to estimate the relative spatial ranges and time scales of the relevant feedback loops, to interpret the phenotypic transitions in eggshell morphology and to predict the effects of new genetic manipulations. We have found that the proposed mechanism with a single diffusing inhibitor is sufficient to convert a single-peaked extracellular input into a two-peaked pattern of intracellular signaling. Based on extensive computational analysis, we predict that the same mechanism is capable of generating more complex patterns. At least indirectly, this can be used to account for more complex eggshell morphologies observed in related fly species. We propose that versatility in signaling mediated by autocrine loops can be systematically explored using experiment-based mechanistic models and their analysis.

Animals↗

PubMed1993

Transepithelial calcium transport in the chick chorioallantoic membrane. I. Isolation and characterization of chorionic ectoderm cells.

The chicken eggshell supplies approximately 80% of the calcium found in the hatchling chick. The mobilization of eggshell calcium into the developing embryo involves the transepithelial transport of large amounts of calcium in a development-specific manner. The cells responsible for the transport of eggshell calcium into the embryonic circulation are the ectodermal cells of the chorioallantoic membrane. In this report, we present a method for the isolation and culture of chorioallantoic membrane ectodermal cells, which are amenable to direct experimental manipulation. Cell preparations are characterized with respect to the expression of an ectoderm-specific cell surface marker (transcalcin, a calcium-binding protein), and a specific enzymatic activity (elevated Ca(2+)-activated ATPase). Functional assessment of in vitro cellular calcium uptake by 45Ca2+ tracer kinetics indicates the persistence of a temperature-sensitive, rapid-influx pathway similar to that observed in vivo. The preparations of primary ectodermal cells present an in vitro system applicable to the experimental analysis of calcium metabolism and transport by the chick chorioallantoic membrane.

Allantois↗

PubMed1984

Calcium metabolism in embryos of the oviparous snake Coluber constrictor.

Total calcium in embryos of an oviparous, colubrid snake (Coluber constrictor L.) rises rapidly during the last half of incubation as the embryos increase in size. Although most of this calcium is drawn from stores in the yolk, hatchlings contain more calcium than was present in yolk of eggs at oviposition. Because shells from eggs incubated to hatching contain less calcium than do shells from freshly-laid eggs, the extra calcium appears to be drawn from the eggshell. Indeed, approximately 20% of the calcium required for development in this snake is obtained from the eggshell, with the remainder coming from the yolk. Thus, embryos of oviparous snakes, like embryonic chelonians, crocodilians and birds, withdraw calcium from their eggshells and do not rely exclusively on calcium supplied in their yolk for support of growth and development.

Animals↗