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The effect of avian uterine fluid on the growth behavior of calcite crystals.

Eggshell formation takes place on the eggshell membrane in an acellular medium, the uterine fluid that contains the inorganic minerals and precursors of the organic matrix. The high degree of eggshell structure could be due to an interaction between calcium carbonate and the organic matrix. The aim of this study was to demonstrate such an interaction by measuring the effect in vitro of uterine fluid collected at various phases of shell formation on precipitation kinetics, size, and morphology of calcite crystals. The SDS-PAGE profiles of the organic constituents differed between the different phases of eggshell formation. The predominant constituents were ovalbumin and ovotransferrin at the initial phase and lysozyme, ovocleidin-17, ovocalyxin-32, 36- and 21-kDa bands, and ovocleidin-116 at the growth phase. These proteins were numerous in the terminal phase and showed an increased staining of the 32- and 66-kDa bands and appearance of very low molecular weight bands. The precipitation lag time was shortened in proportion to the protein concentration at the initial stage. The effect was observed with a lower magnitude in the presence of constituents of growth and terminal phases. Crystal size was smaller in the presence of constituents from the three stages compared with the control. Components from the initial phase induced the formation of twinned crystals and of rounded corners in the rhombohedric crystals. The presence of components from the growth and terminal phases strongly modified the morphology of the calcite crystals. The majority of the corners became rough and developed curved faces. These observations confirm the interaction of the uterine fluid with calcite and its contribution to eggshell structure.

Animals↗

Effects of S6-strain Mycoplasma gallisepticum inoculation at ten, twenty-two, or forty-five weeks of age on the performance characteristics of commercial egg laying hens.

Experimental inoculation of commercial laying hens, maintained under controlled conditions, with the S6-strain of Mycoplasma gallisepticum (S6MG) at 10 wk of age has previously been shown to affect the lengths and weights of various portions of the reproductive tract without affecting subsequent performance. Two trials were conducted to compare the effects of S6MG inoculation at 10 wk of age (prior to lay), 22 wk of age (onset of lay), and 45 wk of age (during lay) on performance characteristics in commercial layers housed and maintained under controlled conditions, as in previous studies. In each trial, BW, mortality, egg production, egg weight, eggshell weight per unit of surface area, percentage eggshell weight, percentage albumen weight, percentage yolk weight, and yolk weight per albumen weight ratio were examined at various ages throughout an entire laying cycle. Across wk 47 and 58 (age periods after the last 45 wk inoculation), eggshell weight per unit of surface area and percentage eggshell weight were significantly reduced in birds that had received an S6MG inoculation at 45 wk of age when compared with birds that had not received an S6MG inoculation or had been inoculated with S6MG at either 10 or 22 wk of age. Alterations in eggshell quality in response to S6MG may become evident only in older birds that are experiencing declines in production when housed under controlled conditions.

Aging↗

Fluorosis in the laying hen.

One hundred fifty Single Comb White Leghorn hens of two strains previously selected for either high eggshell strength (HES strain) or low eggshell strength (LES strain) were treated for 8 weeks with sodium fluoride added to a practical-type corn-soybean meal basal diet at 0, 300, 600, 900, or 1200 ppm. The HES strain was found to be more sensitive to fluorosis than the LES strain. High levels of dietary fluoride produced greater decreases in feed consumption, egg production, eggshell breaking strength, eggshell weight, and eggshell thickness in the HES strain than in the LES strain, despite the fact that the absolute intake of fluoride was greatest in the LES strain. Fluoride treatment significantly increased bone magnesium content in both strains and plasma phosphorus in the HES strain but did not significantly affect bone breaking strength or the concentrations of total calcium, ionized calcium, or magnesium in plasma of either strain. High fluoride treatment (1200 ppm) reduced the growth of HES progeny after 8 weeks of treatment of the hens, but LES progeny were not affected. These results suggest that the decreased sensitivity of the LES strain to fluoride may relate to its relatively greater (ca. 14%) volume of bone in which most absorbed fluoride is deposited.

Animals↗

Influence of diet-mediated maternal thyroid alterations on functional properties of turkey eggs.

Maternal thyroid status was altered by means of diet to determine its effect on functional properties of turkey eggshells. Hens were fed a control diet (CON), the CON diet containing .5 ppm triiodothyronine (T3), the CON diet containing 2.1 ppm iodine as potassium iodide (KI) or the CON diet containing .1% thiouracil (THIO). Feeding T3 decreased plasma thyroxine but elevated plasma T3 concentrations compared to CON. The KI diet had no effect on plasma thyroid hormone concentrations, but feeding THIO depressed plasma thyroxine with no effect on T3, resulting in an elevated ratio of the two hormones compared to the CON ratio. Feeding KI decreased egg volume and T3 increased egg density compared to CON, but no effects on egg weight, surface area, width, or length were noted. Dietary T3 depressed eggshell water vapor conductance compared to CON. Dietary iodine resulted in thinner eggshells with fewer pores than the CON, whereas THIO caused significantly more pores in eggshells than CON but had no effects on shell thickness. Dietary KI had no effects on maternal plasma thyroid hormone concentrations, suggesting that the effects were due to iodine availability rather than to thyroid hormones. It is concluded that the availability of iodine to turkey breeder hens may influence eggshell characteristics.

Animals↗

Research note: responses of laying hens on saline drinking water to dietary supplementation with various zinc compounds.

Production variables, eggshell defects, eggshell quality, the concentration of calcium-binding protein (CaBP), and the activity of carbonic anhydrase (CA) in the shell gland mucosa were determined in hens receiving town water (10 mg Na/L), or town water supplemented with 2 g NaCl/L (796 mg Na/L). Five treatments were examined. Control hens received town water and a proprietary layer mash containing .17% Na. The remaining four treatments received the water containing NaCl (2 g/L). Hens of Treatment 2 were fed the proprietary layer mash and those of Treatments 3, 4, and 5 received, respectively, the layer mash containing supplements of Zn-methionine (Zinpro-200; .5 g/kg), Zn sulfate (ZnSO4.7H2O; .46 g/kg), or chelated Zn-EDTA (.54 g/kg) to supply the same concentration of Zn (.1 g/kg). The treatments were applied for 6 wk. Hens receiving the saline drinking water without any dietary Zn supplement produced significantly (P < .05) more eggs with shell defects than hens receiving the town water. This increase in the incidence of eggshell defects was associated with significant reductions in eggshell breaking strength, the concentration of CaBP, and the activity of CA. Supplementing the saline drinking water with any of the three Zn compounds significantly reduced the incidence of eggshell defects and in some cases improved shell breaking strength, the concentration of CaBP, and the activity of CA.

Animal Feed↗

Salmonella contamination in commercial eggs and an egg production facility.

Egg samples were collected from various stages of an egg processing operation and from the attached production facility. Salmonella was isolated from 72.0% of all samples collected from the laying house environment. Recovery of Salmonella from flush water, ventilation fan, egg belt, and egg collector samples were (positive samples/total samples collected): 2/2, 4/4, 16/22, and 14/22, respectively. Salmonella was found on 7 of the 90 eggshells sampled before processing and 1 of 90 eggshells sampled after processing, but Salmonella was not found in the 180 eggs analyzed for internal contamination following processing. The one eggshell found positive for Salmonella following processing was detected when the pH of wash water samples was lowest (10.19). The 60 isolates from production facilities included the following Salmonella serotypes: S. agona, S. typhimurium, S. infantis, S. derby, S. heidelberg, S. california, S. montevideo, S. mbandaka, and untypable. The 22 isolates obtained from eggshells prior to processing were serotyped as S. heidelberg and S. montevideo. All five isolates obtained from eggshells after processing were serotyped as S. heidelberg. These data suggest that although the shells of about 1% of commercial eggs are contaminated with Salmonella, contamination of the internal contents of eggs with Salmonella is a rare event.

Animals↗

Regulation of extraembryonic calcium mobilization by the developing chick embryo.

During development, the chick embryo mobilizes the calcium it needs from two extraembryonic sources, first the yolk and then the eggshell. Since previous studies have strongly suggested that vitamins D and K may regulate chick embryonic calcium metabolism, we have examined here how these vitamins might be involved in regulating the calcium mobilization processes. We used as our experimental system chick embryos which were maintained in long-term in vitro culture in the absence of the eggshell. Our results showed that exogenous vitamin D3, in the form of the active 1,25-dihydroxylated metabolite, was hypercalcaemic in both control embryos and the calcium-deficient, shell-less embryos. Since the eggshell was absent in the latter, the vitamin D-induced hypercalcaemia must involve mobilization of calcium from the yolk and, or, the embryonic skeleton. The latter was unlikely since concomitant hyperphosphataemia was not observed. By radiolabelling the yolk with 45Ca2+ and subsequently monitoring its distribution, we showed that vitamin D3 stimulated yolk calcium mobilization. However, exogenous vitamin D3 did not appear to influence the calcium uptake activity of the chorioallantoic membrane (CAM), the tissue responsible for translocating eggshell calcium. On the other hand, when embryos were rendered vitamin K deficient by the administration of its antagonist, Warfarin, CAM calcium activity was significantly depressed, an effect which was remedied by vitamin K supplementation. We conclude that, during normal chick embryonic development, vitamin D is primarily involved in regulating yolk calcium mobilization whereas vitamin K is required for eggshell calcium translocation by the CAM.

Allantois↗

Effects of PCBs, DDT, and mercury compounds in chickens and Japanese quail.

In well-controlled experiments using white leghorn chickens and Japanese quail, dietary polychlorinated biphenyls (PCBs), DDT and related compounds produced no detrimental effects on eggshell quality. A drastic reduction in hatchability of chicks occurred with 10-20 ppm PCBs, but no detrimental effects on eggshell quality, egg production or hatchability were found with 0.5 and 1.0 ppm PCBs, or DDT up to 100 ppm. Dietary PCBs potentiated a vitamin E-selenium deficiency in the chick, increased exudative diathesis, and decreased plasma glutathione peroxidase levels. Dietary PCBs induced hepatic microsomal benzopyrine hydroxylase. Dietary levles of 100 or 200 ppm inorganic mercury as HgSO4 or HgCl2 had little effect on egg production, hatchability, shell quality, morbidity and mortality. Methylmercury chloride, however, at levels providing 10 or 20 mg Hg/kg of diet, severely affected all of these parameters. Even though the present experiments demonstrate that neither DDT nor PCBs has any effect on eggshell quality in chickens and Japanese quail, they may cause thinning of eggshells in other species. Controlled experiments are lacking. Eagles, ospreys and pelicans all consume fish which in many areas of the world are known to contain methyl mercury. The thinning of eggshells in the species in the wild may have been due, at least in part, to environmental contamination with methylmercury rather than DDT, DDE or PCBs, as has been claimed.

Animals↗

Effects of DDE, TDE, and PCBs on shell thickness of western grebe eggs, Bear River Migratory Bird Refuge, Utah--1973-74.

DDE, TDE, and polychlorinated biphenyls (PCBs) as Aroclors 1260 and 1254 were detected in low concentrations in eggs of western grebes (Aechmophorus occidentalis) from Bear River Migratory Bird Refuge, Utah. DDE was the only contaminant which was both negatively correlated with eggshell thickness and a significant variable in a multiple regression model for predicting eggshell thickness. The eggshell thickness index for western grebe decreased 2.3 percent from pre- to post-DDT-use periods. Incubation stage appeared to have no measurable correlation with eggshell thickness. The small amount of eggshell thinning seen in western grebe eggs at Bear River Migratory Bird Refuge appeared to have no detectable effect on reproduction.

Animals↗

Integration of epithelial patterning and morphogenesis in Drosophila ovarian follicle cells.

Drosophila oogenesis involves the coordinated development of germ cells and an overlying follicular epithelium. The follicle cells provide a genetically tractable system to investigate the cell biology of patterning and morphogenesis. Follicle cells initially form a cuboidal epithelium surrounding a syncytium of nurse cells and oocyte. Epithelial structure is maintained as these cells reorganize to create the three dimensional architecture of the eggshell. Both long-range and short-range cell-cell communications pattern the domains of follicle cells that will create specific eggshell structures. After terminal differentiation to deposit the eggshell proteins, the follicle cells die. This review summarizes recent progress in understanding the cell-cell communication that orchestrates follicle cell patterning and migrations. DE-cadherin-mediated adhesion is important at several steps in egg chamber formation and follicle cell migration. Notch signaling is critical during each successive round of patterning and migration. Integration of bone morphogenetic protein (BMP) and epidermal growth factor (EGF) signals patterns the elaborate structures of the dorsal-anterior eggshell.

Animals↗

Molecular analysis and rescue of a vitelline membrane mutant in Drosophila.

The eggshell in Drosophila is produced by ovarian follicle cells during the later stages of oogenesis. Eggshell formation involves the ordered synthesis and assembly of several protein components. Genes encoding the most abundant eggshell proteins have been identified by molecular cloning studies. Morphological examination of eggs produced by females carrying female sterile mutations on the X and third chromosomes have revealed additional loci involved in chorion formation. In this study we screened a collection of female sterile mutants carrying EMS-induced mutations on the second chromosome for eggshell mutants. A class of six mutants with potential vitelline membrane defects was identified on the basis of the response of mutant eggs to hypochlorite solutions. Biochemical analysis showed that one mutant, fs(2)QJ42, failed to produce a major vitelline membrane protein, sV23. The mutation was mapped cytogenetically to 26A, a region previously implicated in vitelline membrane formation by molecular cloning studies. Northern blot analysis using a cloned copy of the sV23 gene as probe showed a 10- to 15-fold reduction of sV23 RNA levels in the mutant. sV23 synthesis and fertility were restored when a normal copy of the sV23 gene was introduced into the mutant via germ line transformation. Transposons carrying the sV23 gene with as little as 147 bp of 5' flanking DNA were capable of restoring fertility and sV23 protein to wild type levels.

Animals↗

Prevalence and antimicrobial resistance of Salmonella enteritidis and other salmonellas in the eggs and egg-storing trays from retail markets of Coimbatore, South India.

A study was conducted to determine the incidence of Salmonella enterica serovar Enteritidis and other Salmonella serovars on eggshell, egg contents and on egg-storing trays. A total of 492 eggs and 82 egg-storing trays were examined over a period of 1 year from different retail outlets of a residential area of Coimbatore city, South India. Salmonella contamination was recorded in 38 of 492 (7.7%) eggs out of which 29 was in eggshell (5.9%) and 9 in egg contents (1.8%). Around 7.5% of the egg-storing trays were also found to be contaminated with Salmonella. Serotyping of the Salmonella strains showed that 89.7% of the strains from eggshell, 100% of the strains from egg contents and 71.4% of the strains from egg-storing trays were Salmonella Enteritidis. Other serovarvars encountered were S. Cerro, S. Molade and S. Mbandaka from eggshell and S. Cerro from egg-storing trays. Seasonal variations in the prevalence pattern were identified with, a higher prevalence during monsoon months followed by post-monsoon and premonsoon. Further examination of the Salmonella strains was carried out by testing their antimicrobial sensitivity against 10 commonly used antimicrobials. Results revealed high prevalence of multiple antimicrobial resistance among these strains suggesting possible prior selection by use of antimicrobials in egg production.

Animals↗

Ultrastructure of eggs of Paracapillaria (Crossicapillaria) philippinensis and evidence related to its life cycle.

The varied ultrastructure of the eggshell of Paracapillaria (Crossicapillaria) philippinensis, collected from a human sample, is reported from a scanning electron microscopy study. Two distinct egg shapes were identified: typical peanut-shaped and swollen peanut-shaped. Both thick and thin eggshells were detected. Thick eggshells are either fairly smooth or bear a beam-like network in relation to the pillars in their surface ultrastructure. Thin eggshells are transparent allowing visibility of the coiled larva within. Presence of the thin shell provides supportive evidence of autoinfection involved in the life cycle of this medically important parasite.

Animals↗

Mosquito embryos and eggs: polarity and terminology of chorionic layers.

The development of genetically modified vectors refractory to parasites is seen as a promising strategy in the future control of endemic diseases such as malaria. Nevertheless, knowledge of mosquito embryogenesis, a pre-requisite to the establishment of transgenic individuals, has been presently neglected. We have here studied the eggs from two neotropical malaria vectors. Eggs from Anopheles (Nyssorhynchus) albitarsis and Anopheles (Nyssorhynchus) aquasalis were analyzed by laser scanning microscopy and scanning electron microscopy and compared to those of Drosophila melanogaster. We verified basic conflicting data such as mosquito egg polarity and ultrastructure of eggshell layers. A 180 degrees rotation movement of the mosquito embryo along its longitudinal axis, a phenomenon not conserved among all Diptera, was confirmed. This early event is not taken into account by several present groups, leading to a non-consensual assignment of eggshell dorsal and ventral poles. Since embryo and egg polarities, defined during oogenesis, are the same, we propose to consider the flattened egg side as the dorsal one. The structure of Anopheles eggshell was also examined. Embryos are covered by a smooth endochorion or inner chorion layer. Outside this coat lies the compound exochorion or outer chorion layer, assembled by a thin basal lamellar layer and external tubercles. The terminology related to eggshell layers is discussed.

Journal Article↗

The effects of nest environment on calcium mobilization by leatherback turtle embryos (Dermochelys coriacea) during development.

We investigated the effect of sand moisture content and sand temperature on developmental success and the mobilization of calcium during development using laboratory incubated eggs (n=251) collected from leatherbacks nesting at Parque Nacional Marino Las Baulas, Costa Rica. Calcium concentrations of egg components [eggshell, yolk plus albumen (Y+A) and embryo] changed significantly through incubation for both viable and undeveloped eggs. In developed eggs, eggshell calcium content decreased 42.9% by day 60 of incubation. The Y+A calcium decreased by 20.8% until the last quarter of incubation, and then increased to 0.99% above initial Y+A calcium concentrations just prior to hatching. In undeveloped eggs, eggshell calcium content decreased by 25.7%, with the rate of decrease slowing significantly beyond day 30 of incubation. In contrast, Y+A calcium increased steadily through the 60-day incubation period. Embryos incorporated a higher proportion of calcium when incubated at a lower sand moisture content (5% H(2)O>12% H(2)O) and at lower sand temperatures (28.5 degrees C, 29.5 degrees C>31.0 degrees C). The total wet mass of freshly oviposited eggs was negatively correlated with calcium concentration per gram of eggshell (r=-0.569; P<0.001). Thus, each yolked egg, regardless of initial wet mass, had an average of 1.23 g (+/-0.43 g) of calcium per egg (Mean egg mass: 76.24+/-1.21 g). Both developmental success (24.1%) and hatching success (7.4%) of laboratory-incubated eggs were dependent to a greater extent on temperature than on moisture, with an increase in mortality as sand temperature increased. For natural nests on Playa Grande, developmental success (37.4%) and hatching success (19.8%) were similar in magnitude to the results obtained from the laboratory. The recent ENSO (El Niño Southern Oscillation) event and increased tidal activity may be responsible for the high embryonic mortality measured during the 1997-1998 nesting season.

Animals↗

Genetic variation for egg production, egg quality and bone strength in selected and traditional breeds of laying fowl.

1. A multi-breed experiment was conducted with 25 commercial and traditional lines of laying fowl to determine the extent of between-breed genetic variation for adult body weight, sexual maturity, rate of lay, egg weight and egg composition to 55 weeks of age. The genetic variability for bone strength and eggshell strength was determined at 55 weeks of age and a comparison of commercially selected and traditional breeds was performed. 2. The proportion of the total variation that was associated with breed or line of origin was high (> 0.8) for body weight, sexual maturity and shell colour; moderately high (0.4 to 0.7) for rate of lay, early and late egg weight, weights of egg yolk, albumen and shell at 55 weeks; and low (< 0.4) for egg weight at 42 to 45 weeks, albumen quality and the number of egg inclusions. 3. There were no detectable differences between breed within category (traditional and commercial lines) for rate of lay, and estimates of breed variation for egg weight and egg components were substantially decreased within category compared with the overall analysis. 4. Commercial lines displayed earlier sexual maturity, greater rates and persistency of lay, and higher egg weights at earlier (32 to 35 weeks) and later (52 to 55 weeks) ages. At 55 weeks, the larger eggs from commercial birds contained more albumen of higher quality and paler yolks of similar weight to those from traditional breeds. The relative weight of the eggshell was similar in both categories. 5. There was considerable genetic variation between commercial lines for bone density and a moderate proportion of genetic variability for bone strength. Commercial lines had very weak bones compared with traditional lines but there was relatively little genetic variation for eggshell strength. The results suggest that eggshell quality is maintained in genetically selected lines at the expense of bone strength and bone radiographic density.

Animals↗

Frustrated nesting behaviour: relation to extra-cuticular shell calcium and bone strength in White Leghorn hens.

1. The inability of hens to express normal nesting behaviour in battery cages results in frustration, manifesting itself as stereotyped pacing. Frustration can cause hens to retain their eggs beyond the expected time of lay, resulting in an extra-cuticular layer of calcium on eggshells. 2. This study investigated whether frustrated nesting behaviour caused extra-cuticular calcification of eggshells and whether this loss of non-structural calcium to eggshells may have subsequently contributed to weak bones in spent hens. 3. One hundred and twenty hens were assigned to one of three treatments: NB (hens given nest boxes in their cages), NN (no nest box) and R (access to nest box periodically restricted). 4. NB hens spent the least amount of time pacing in the hour before oviposition. NN and R hens spent similar amounts of time pacing. 5. Contrary to expectation, eggshells from hens under different treatments yielded the same amount of extra-cuticular calcium. Similarly tibia strength did not significantly differ among treatments.

Animals↗

Effect of lighted incubation on embryonic growth and hatchability performance of two strains of layer breeder eggs.

1. Eggs from two layer-type breeder flocks (ISA-W vs Leghorn) between 30 and 45 weeks of age were used in 4 trials to study the effects of lighted incubation on embryonic growth from 5 to 18 d of age and hatchability per cent (HP). The physical dimensions, eggshell characteristics and conductance (EC) of eggs of the two strains were compared. 2. Eggs were set in an incubator on trays either in the dark-control or under two tubes of 20-watt white fluorescent light during the first 18 d of incubation. The light intensity ranged from 1230 to 1790 lux at the surface of the eggs. Eggs were transferred to dark hatching compartments at d 19 of incubation. 3. The genetic make-up of birds influenced the physical dimensions and eggshell characteristics of eggs. ISA-W eggs had higher weight, surface area (ESA), volume (EV), width, EC, shell volume and HP, and lower ESA:EV ratio, per cent shell, shell density and dead embryos than those of Leghorn eggs. 4. Lighted incubation increased daily embryonic growth (mg/d) and HP by 3.9 and 5.9%, respectively, when compared with the dark-control incubation. 5. The physical dimensions and eggshell characteristics of eggs influenced the effects of lighted incubation on embryonic growth and HP. Lighted incubation increased embryonic growth and HP of ISA-W eggs. The increase in embryonic growth and HP of the Leghorn eggs was not significant. 6. The genetic make-up of birds influenced the physical dimensions and eggshell characteristics of their eggs and these differences in the characteristics of hatching eggs influenced embryonic growth and HP when incubated under light.

Animals↗