Sources and timing of calcium mobilization during embryonic development of the corn snake, Pantherophis guttatus

Embryos of oviparous Reptilia (=turtles, lepidosaurs, crocodilians and birds) extract calcium for growth and development from reserves in the yolk and eggshell. Yolk provides most of the calcium to embryos of lizards and snakes. In contrast, the eggshell supplies most of the calcium for embryonic development of turtles, crocodilians and birds. The yolk sac and chorioallantoic membrane of birds recover and transport calcium from the yolk and eggshell and homologous membranes of squamates (lizards and snakes) probably transport calcium from these two sources as well. We studied calcium mobilization by embryos of the snake Pantherophis guttatus during the interval of greatest embryonic growth and found that the pattern of calcium transfer was similar to other snakes. Calcium recovery from the yolk is relatively low until the penultimate embryonic stage. Calcium removal from the eggshell begins during the same embryonic stage and total eggshell calcium drops in each of the final 2 weeks prior to hatching. The eggshell supplies 28% of the calcium of hatchlings. The timing of calcium transport from the yolk and eggshell is coincident with the timing of growth of the yolk sac and chorioallantoic membrane and expression of the calcium binding protein, calbindin-D28K, in these tissues as reported in previous studies. In the context of earlier work, our findings suggest that the timing and mechanism of calcium transport from the yolk sac of P. guttatus is similar to birds, but that both the timing and mechanism of calcium transport by the chorioallantoic membrane differs. Based on the coincident timing of eggshell calcium loss and embryonic calcium accumulation, we also conclude that recovery of eggshell calcium in P. guttatus is regulated by the embryo.     

Accumulation of calcium and phosphorus in pigeon (Columba livia) embryos

Summary Calcium and phosphorus were measured in the yolk and albumen of fertile pigeon (Columba livia) eggs incubated for 0–17 days, and in embryos and hatchlings. Shell provided most of the calcium for skeletal mineralization of the embryos, whereas phosphorus was derived from the yolk and albumen. Mobilization of calcium from the shell to the embryo commenced at approximately day 11 of incubation, accumulating both in the embryo and the yolk sac. There was 1.4 times more calcium in squab yolk sacs than that contained in newly laid egg yolks. The results suggest that whereas general patterns of calcium and phosphorus accumulation during embryogenesis in altricial birds closely resemble those of precocial birds, calcium mobilization from the shell begins later, proceeds at a slower rate and results in a less mineralized hatchling.CIDA/NSERC Visiting Research Associate Permanent address: Department of Animal Science, University of Peradeniya, Peradeniya, Sri Lanka     

Environmental modulation of calcium and phosphorus metabolism in embryonic snapping turtles (Chelydra serpentina)

Summary Eggs of common snapping turtles (Chelydra serpentina) were incubated on wet (–150 kPa water potential) and dry (–950 kPa) substrates in a laboratory study assessing the effects of the hydric environment on patterns of mobilization of calcium and phosphorus by developing embryos. We found that embryos developing in wet environments withdrew nutrients from their yolk faster, grew more rapidly, and incubated longer than embryos exposed to dry environments. Turtles developing in both environments absorbed calcium from the yolk at similar rates and depleted the yolk of almost its entire reserve of calcium prior to hatching. Calcium withdrawn from the yolk was supplemented with calcium mobilized from the eggshell, but embryos in wet environments obtained substantially more calcium from the eggshell than did those in dry settings. Embryos obtained all of the phosphorus used in skeletogenesis from the yolk, but those incubating in wet environments mobilized phosphorus from this compartment more rapidly than did those in dry settings. Exposing embryonic snapping turtles to wet environments apparently allows them to make more efficient use of the transitory source of calcium in the eggshell than is possible in dry environments. However, the residual yolk in hatchlings from both wet and dry environments contains too little calcium to support the growth of hard and soft tissues in neonates at rates similar to those characterizing the growth phase of development in embryogenesis.     

Calcium mobilization, water balance, and growth in embryos of the agamid lizard Amphibolurus barbatus

Embryos of the agamid lizard Amphibolurus barbatus are at developmental stage 29 of Dufaure and Hubert at the time of oviposition. Mobilization of calcium and other nutrients from the yolk proceeds slowly for the first half of incubation, during which time embryonic growth also proceeds slowly. During the second half of incubation, however, embryos withdraw calcium and other nutrients from the yolk very rapidly, and growth rates are correspondingly high. Approximately 60% of the calcium used by developing embryos is obtained from the yolk, but fully 40% of their requirement is met by calcium mobilized from the eggshell. Very little calcium remains in residual yolk of hatchlings, so this yolk must be used in maintenance metabolism rather than in growth of neonates. No dichotomy exists among oviparous, amniotic vertebrates with respect to sources of calcium used by developing embryos, but one does exist with respect to patterns of mobilization of this element. Whereas calcium is extracted from yolk of embryonic reptiles throughout incubation, it actually is deposited in yolk of embryonic birds after the midpoint in development.     

Selenium distribution in eggs of avian species

We studied the effect of egg mass of eight different avian species on Se distribution between egg components and the effect of incubation on Se accumulation by chicken eggshell and shell membrane. Eight groups of birds received a diet without Se supplementation. Unfertile eggs were collected after 35 days of feeding; yolk, albumen, shell and shell membrane were assayed separately for Se. All avian species studied showed identical Se concentration in yolk–albumen complex equal to 38.7 μg Se/100 g, reflecting a linear correlation between yolk–albumen mass and Se content. Shells and shell membrane Se accumulation showed quadratic correlation with the appropriate mass thus explaining unusually high Se concentration in ostrich shell and shell membrane, that reached values 1785 and 1904 μg Se/kg respectively. Incubation of fertile eggs decreased eggshell Se content, the effect being more expressed in eggs from hens fed sodium selenite compared to organic Se utilization (Sel-Plex). It was concluded that shell might be an additional Se source for an embryo.     

THE PHYSIOLOGICAL ECOLOGY OF REPTILIAN EGGS AND EMBRYOS. AND THE EVOLUTION OF VIVIPARITY WITHIN THE CLASS REPTILIA

1. Eggs of Crocodilia and Chelonia, like those of birds, have a pair of egg membranes separating a thick layer of albumen from the calcareous shell. In contrast, eggs of oviparous Lepidosauria have only a single shell membrane, upon which relatively small amounts of calcium carbonate are deposited; and the volume of albumen in eggs is extraordinarily small at the time of oviposition. 2. With the possible exception of certain geckos and some chelonians, eggs of oviparous reptiles seem always to absorb water from the substrate during the course of normal incubation. In so far as the rate of water absorption exceeds the rate of water loss by transpiration from exposed surfaces, the eggs swell during incubation. The term ‘cleidoic’ cannot be used to describe eggs of this type. 3. Embryos of lizards and snakes influence the water potential of extra-embryonic fluids contained within their eggs, thereby maintaining or increasing the gradient in water potential that drives water absorption. 4. Embryos of Crocodilia and Chelonia obtain a substantial portion of the calcium used in ossification of skeletal elements from the inner surfaces of the eggshell. In contrast, embryonic lizards and snakes draw upon extensive reserves of calcium present in the yolk, and obtain little (if any) calcium from the eggshell. 5. All reptilian embryos seem to produce substantial quantities of urea as a detoxification product of protein catabolism. Contrary to expectation, uricotelism may not be common among reptilian embryos, even in those few instances where development takes place within a hard, calcareous egg. 6. In eggs of Crocodilia and Chelonia, respiratory gases seem to pass by diffusion through pores in the calcareous eggshell and through spaces between the fibres of the pair of egg membranes. No pores have been observed in the shell of lepidosaurian eggs, and so gases presumably diffuse between the fibres of the single (multilayered) shell membrane. 7. Metabolism of reptilian embryos is temperature-dependent, as is true for most ectothermic organisms. For each species, there appears to be a particular temperature at which embryonic development proceeds optimally, and departures from this optimum elicit increases in developmental anomalies and/or embryonic mortality. 8. Viviparity has evolved on numerous occasions among species of the Squamata, but seemingly never among Crocodilia or Chelonia. Since the evolution of viviparity entails a progressive reduction in the eggshell, only those organisms whose embryos do not depend upon the eggshell as a source of calcium may have the evolutionary potential to become viviparous. 9. Evolutionary transitions from oviparity to viviparity could have been driven by selection related to (i) thermal benefits to embryos consequent upon retention of eggs within the body of a parent capable of behavioural thermoregulation; (ii) protection of the eggs from nest predators and/or soil microbes; and (iii) more effective exploitation of a seasonal food resource by early emerging young.     

Vitamin D and chick embryonic yolk calcium mobilization: identification and regulation of expression of vitamin D-dependent Ca2(+)-binding protein, calbindin-D28K, in the yolk sac

The developing chick embryo acquires calcium from two sources. Until about Day 10 of incubation, the yolk is the only source; thereafter, calcium is also mobilized from the eggshell. We have previously shown that during normal chick embryonic development, vitamin D is involved in regulating yolk calcium mobilization, whereas vitamin K is required for eggshell calcium translocation by the chorioallantoic membrane. We have studied here the biochemical action of 1,25-dihydroxy vitamin D3 in the yolk sac by examining the expression and regulation of the cytosolic vitamin D-dependent calcium-binding protein, calbindin-D28K. Two types of embryos are used for this study, normal embryos developing in ovo and embryos maintained in long-term shell-less culture ex ovo, the latter being dependent solely on the yolk as their calcium source. Our findings are (1) calbindin-D28K is expressed in the embryonic yolk sac, detectable at incubation Days 9 and 14; (2) the embryonic yolk sac calbindin-D28K resembles that of the adult duodenum in both molecular weight (Mr 28,000) and isoelectric point, as well as the presence of E-F hand Ca2(+)-binding structural domains; (3) systemic calcium deficiency caused by shell-less culture of chick embryos results in enhanced expression of calbindin-D28K in the yolk sac during late development; (4) yolk sac calbindin-D28K expression is inducible by 1,25-dihydroxy vitamin D3 treatment in vivo and in vitro; and (5) immunohistochemistry revealed that yolk sac calbindin-D28K is localized exclusively to the cytoplasm of the yolk sac endoderm. These findings indicate that the chick embryonic yolk sac is a genuine target tissue of 1,25-dihydroxy vitamin D3.     

Ultrastructure of avian eggshell during resorption following egg fertilization

For skeletal mineralization, the avian embryo mobilizes calcium from its calcitic eggshell. This occurs through dissolution of specific interior regions of the shell in a process that also weakens the shell to allow hatching. Here, we have examined eggshell ultrastructure during dissolution occurring between laying of a fertilized egg (with incubation) and hatching of the chick (Gallus gallus). We have focused on changes in shell mammillae where the majority of dissolution takes place. Using scanning electron microscopy, we describe differences in matrix–mineral structure and relationships not observed in unfertilized eggs (unresorbed eggshell). We document changes in the calcium reserve body – an essential sub-compartment of mammillae – consistent with it being an early, primary source of calcium essential for embryonic skeletal growth. Dissolution events occurring in the calcium reserve sac and in the base plate of the calcium reserve body, and similar changes in surrounding bulk mammillae structure, all correlate with advancing skeletal embryonic calcification. The changes in mammillae sub-structures can generally be characterized as mineral dissolutions revealing fine surface topographies on remaining mineral surfaces and the exposure of an extensive, intracrystalline (occluded) organic matrix network. We propose that this mineral-occluded network regulates how shell mineral is dissolved by providing dissolution channels facilitating calcium release for the embryonic skeleton.     

Fetal nutrition in lecithotrophic squamate reptiles: Toward a comprehensive model for evolution of viviparity and placentation

The primary pattern of embryonic nutrition for squamate reptiles is lecithotrophy; with few exceptions, all squamate embryos mobilize nutrients from yolk. The evolution of viviparity presents an opportunity for an additional source of embryonic nutrition through delivery of uterine secretions, or placentotrophy. This pattern of embryonic nutrition is thought to evolve through placental supplementation of lecithotrophy, followed by increasing dependence on placentotrophy. This review analyzes the relationship between reproductive mode and pattern of embryonic nutrition in three lecithotrophic viviparous species, and oviparous counterparts, for concordance with a current model for the evolution of viviparity and placentation. The assumptions of the model, that nutrients for oviparous embryos are mobilized from yolk, and that this source is not disrupted in the transition to viviparity, are supported for most nutrients. In contrast, calcium, an essential nutrient for embryonic development, is mobilized from both yolk and eggshell by oviparous embryos and reduction of eggshell calcium is correlated with viviparity. If embryonic fitness is compromised by disruption of a primary source of calcium, selection may not favor evolution of viviparity, yet viviparity has arisen independently in numerous squamate lineages. Studies of fetal nutrition in reproductively bimodal species suggest a resolution to this paradox. If uterine calcium secretion occurs during prolonged intrauterine egg retention, calcium placentotrophy evolves prior to viviparity as a replacement for eggshell calcium and embryonic nutrition will not be compromised. This hypothesis is integrated into the current model for evolution of viviparity and placentation to address the unique attributes of calcium nutrition. The sequence of events requires a shift in timing of uterine calcium secretion and the embryonic mechanism of calcium retrieval to be responsive to calcium availability. Regulation of uterine calcium secretion and the mechanism of embryonic uptake of calcium are important elements to understanding evolution of viviparity and placentation. J. Morphol., 2013. © 2013 Wiley Periodicals, Inc.     

Expression of calbindin-D28K by yolk sac and chorioallantoic membranes of the corn snake, Elaphe guttata

The yolk splanchnopleure and chorioallantoic membrane of oviparous reptiles transport calcium from the yolk and eggshell to the developing embryo. Among oviparous amniotes, the mechanism of calcium mobilization to embryos has been studied only in domestic fowl, in which the mechanism of calcium transport of the yolk splanchnopleure differs from the chorioallantoic membrane. Transport of calcium is facilitated by calbindin-D(28K) in endodermal cells of the yolk splanchnopleure of chickens but the chorioallantoic membrane does not express calbindin-D(28K). We used immunoblotting to assay for calbindin-D(28K) expression in yolk splanchnopleure and chorioallantoic membrane of the corn snake, Elaphe guttata, to test the hypothesis that the mechanism of calcium transport by extraembryonic membranes of snakes is similar to birds. High calbindin-D(28K) expression was detected in samples of yolk splanchnopleure and chorioallantoic membrane during late embryonic stages. We conclude that calbindin-D(28K) is expressed in these extraembryonic membranes to facilitate transport of calcium and that the mechanism of calcium transport of the chorioallantoic membrane of the corn snake differs from that of the chicken. Further, we conclude that calbindin-D(28K) expression is developmentally regulated and increases during later embryonic stages in the corn snake.     

Seasonal patterns of nutrient deposition in a Quercus douglasii woodland in central California

The monthly deposition of total nitrogen, phosphorus, potassium, calcium and magnesium via canopy throughfall, and various components of the litterfall was measured for 31 months under mature Quercus douglasii and in the bulk precipitation in the surrounding open grassland. Seasonal patterns of nutrient concentration in leaf litter, throughfall, and precipitation were also measured. Total annual subcanopy deposition exceeded open precipitation deposition by approximately 45–60x for nitrogen, 5–15x for phosphorus, 30–35x for potassium, 25–35x for calcium, and 5–10x for magnesium. Total annual subcanopy deposition was low in comparison to other oak woodland sites reported in the literature. Throughfall and leaf litter were the primary sources of nutrients and thus determined the seasonal peaks of nutrient deposition. The first autumn rains and leaf fall were associated with one peak in nutrient deposition, and throughfall during early spring leaf emergence was associated with a second peak in potassium, magnesium and phosphorus. Non-leaf plant litter (excluding acorns) provided approximately 15–35% of most nutrients, with twigs and bark depositing over 12% of the annual calcium flux in 1987–1988, and flower litter depositing over 8% of the annual nitrogen flux in 1986–1987. Acorns had high concentrations of phosphorus and nitrogen and during the mast season of 1987–1988 they contained a large proportion of the total subcanopy annual flux of these elements. With acorns excluded, total annual nutrient deposition was similar between years, but timing of nutrient deposition differed. Late summer leaf fall associated with drought, variation in precipitation, and variation in deposition of non-leaf parts were associated with seasonal differences in nutrient deposition between years.     

Yolk utilization inScyliorhinus canicula,an oviparous dogfish

Synopsis Using plastic embedding techniques and semithin sections, in order to overcome the difficult sectioning of yolky eggs, we have been able to carry out histological study of the external yolksac from fertilization until birth in the oviparous dogfishScyliorhinus canicula. The endoderm and its contacting giant yolk nuclei remained very flat, seemingly inactive, during the larger part of development. They became activated only when the external yolksac (EYS) began to shrink. This activation increased along a vegetal-animal gradient in the EYS, but it was essentially restricted to the parts located near the yolk stalk. A statistical study of oocyte, yolk, embryo and newborn fresh and dry weights confirmed that the mass of dry tissue in the embryo (30mg) and in EYS wall (<1 mg) at mid-development were still very low compared to 0.8–1.5 g mass of yolk available for development. This explains why yolk weight remained practically the same during the first half of development. The end of this first period was marked by entry into the pre-hatching state at 85–115 days under laboratory conditions (14–16°C). At this time, yolk began to enter the spiral gut, where it was digested during the second half of development and during one week period after eclosion. Eclosion occurred 170–220 days after egg laying or extraction from oviduct. Two internal storage organs were studied biometrically in the newborn: the internal yolksac (IYS), and the liver, which was fully developed at birth. Both IYS and liver dry weights corresponded to about 10% of the original yolk, while the gut was only 2%, and the rest of the newborn body 58%. Thus, about 20% of yolk dry mass was consumed during development, a figure that is low for oviparous animals.     

Some analyses of artificially incubated eggs and hatchlings of Green and Loggerhead sea turtles

Eggs of the Green sea turtle ( Chelonia mydas ) and the Loggerhead sea turtle ( Caretta caretta ) were incubated in coral sand or silica sand moistened with distilled water. The eggs and hatchlings were analysed for calcium, magnesium and phosphorus. There was less magnesium but more calcium in the hatchlings than in the original egg contents but this was not influenced by the type of sand used during incubation. The results are interpreted as indicating that the eggshell is the major source of calcium for the developing embryos. The Logger-head sea turtle shows a greater efficiency in the incorporation of yolk magnesium and phosphorus into the embryo than does the Green sea turtle.     

The effect of replacing soya bean oil with glycerol in diets on performance,egg quality and egg fatty acid composition in laying hens

The objective of this experiment was to replace soya bean oil with glycerol in laying hen diets and assess the change’s effect on performance, parameters of egg quality and the egg fatty acid profile. A total of 60 44-week-old Hy-Line W36 laying hens were distributed according to a completely randomised experimental design into four treatments consisting of glycerol substitutions for soya bean oil dietary at varying inclusion levels (0%, 25%, 50% and 75%), with five replicates of three birds each. Dietary treatments had no significant effect on BW change, egg production, feed intake, feed conversion ratio, egg weight and egg mass of laying hens. The inclusion of glycerol in the diet of laying hens had no significant effect on egg specific gravity, eggshell breaking strength, eggshell weight, eggshell thickness, egg shape index, albumen index, yolk index, haugh unit, albumen pH, yolk pH and egg yolk colour values. The inclusion of glycerol in the diet of laying hens had no significant effect on palmitic, palmitoleic, stearic, oleic and linolenic acid contents of the egg yolk. The linoleic acid and polyunsaturated fatty acid contents of the egg yolk significantly decreased with the higher levels of dietary glycerol supplementation (P<0.05). The results of this study show that it is possible to replace 75% of soya bean oil (4.5% in diet) with glycerol.     

Composition, accumulation and utilization of yolk lipids in teleost fish

Lipid reserves in teleost eggs are stored in lipoprotein yolk and, in some species, a discrete oil globule. Lipoprotein yolk lipids are primarily polar lipids, especially phosphatidylcholine and phosphatidylethanolamine (PE), and are rich in (n–3) polyunsaturated fatty acids, especially 22:6(n–3) (docosahexaenoic acid, DHA). Oil consists of neutral lipids and is rich in monounsaturated fatty acids (MUFA). Egg lipids are derived from dietary fatty acid, fatty acid mobilized from reserves and possibly fatty acid synthesized de novo. There is selective incorporation of essential fatty acids, particularly DHA, into yolk lipids and discrimination against incorporation of 22:1(n–11). Lipid is delivered to the oocyte by vitellogenin, which is rich in polar lipids, and likely also by other lipoproteins, especially very low density lipoprotein, which is rich in triacylglycerol (TAG). All classes of lipid may be used as fuel during embryonic and larval development and MUFA are preferred fatty acids for catabolism by embryos. Catabolism of oil globules is frequently delayed until latter stages of development. In some species, DHA derived from hydrolysis of phospholipid may be conserved by transfer to the neutral lipid. Recent work has expanded knowledge of the role of DHA in membrane structure, especially in neural tissue, and molecular species analysis has indicated that PE containing sn-1 oleic acid is a prime contributor to membrane fluidity. The results of this type of study provide an explanation for the selection pressures that influence yolk lipid composition. Future work ought to expand knowledge of specific roles of individual fatty acids in embryos along with knowledge of the ecological physiology of ovarian recrudescence, environmental influences on vitellogenin and yolk lipid composition, and the control of yolk lipid accumulation and utilization.     

Sources and timing of calcium intake during reproduction in flycatchers

Calcium availability may limit the reproductive output of birds and snail shells are considered to be the main source of calcium in many passerine species. This study of collared (Ficedula albicollis) and pied (F. hypoleuca) flycatchers evaluates calcium intake of a natural diet in Central Europe, and sex differences in the utilization of experimentally supplemented sources of calcium during the entire breeding period in aviary birds. The study provides the first evidence that successful reproduction of these species depends on the availability of woodlice (Isopoda) and millipedes (Diplopoda). Each of these two components provided about 3 times more calcium than the snail shells contained in a natural nestling diet. The breeding performance of aviary birds was poor when only snail shells and the fragments of eggshells were provided in food, i.e., irregular laying, smaller clutches, eggshell defects (25 of 53 eggs), and eggs dried-up during incubation. In contrast, no defective eggshell or dried-up eggs were found and the overall breeding performance increased 2–3 times when woodlice were added to the food. Females increased their intake of woodlice during both the pre-laying and laying periods, and both sexes did so during the nestling period. Both sexes took more woodlice in the evening than in the morning, independent of the nesting stage. Intake by females was low until 4 days before laying the first egg, then increased to the highest level, dropping immediately after laying the last egg. Intake of woodlice by both sexes increased steadily from hatching until the nestling age of about 10–12 days decreasing thereafter, which corresponds with the period of rapid skeletal growth. In contrast, the intake of mealworms increased until the nestling age of 13–14 days leveling off thereafter which corresponds with the growth curve of nestling body mass.Due to an error in the citation line, this revised PDF (published in December 2003) deviates from the printed version, and is the correct and authoritative version of the paper.     

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 Ni?o Southern Oscillation) event and increased tidal activity may be responsible for the high embryonic mortality measured during the 1997-1998 nesting season.     

Eggshell biliverdin concentration does not sufficiently predict eggshell coloration

Avian eggshell coloration may have arisen due to selection on the biological, chemical, or physical properties of the pigments embedded within the eggshell, or due to selection on the coloration that emerges due to pigment deposition. Within both hypothetical frameworks, pigment‐based eggshell coloration would be related to metrics of egg quality; however, no one has evaluated the relative strength of coloration and pigment concentration in predicting egg quality. Here, we examined 66 European starling Sturnus vulgaris eggs and quantified eggshell biliverdin concentration (an antioxidant that produces the eggshell's blue coloration) and used 28 different coloration metrics derived from both photographic and spectrophotometric data. We also measured egg size, eggshell thickness, concentration of carotenoids in the yolk, and concentration of lysozyme in the albumen to capture variation in egg quality. We found that throughout the laying sequence, biliverdin concentration increased while eggshell thickness, yolk carotenoid concentration, and lysozyme concentration in the albumen all decreased, but this variation was not captured by any eggshell coloration metric. Both eggshell coloration and biliverdin concentration were negatively associated with yolk carotenoid concentration, but only eggshell biliverdin concentration was negatively associated with yolk mass. Lastly, biliverdin concentration explained, at most, only 46% of the variation for all eggshell coloration metrics. Our results suggest biliverdin concentration is a better predictor of egg quality than egg coloration in European starlings, supporting the hypothesis that eggshell pigment concentration per se may be the target of selection.     

Effects of egg size on Double-crested Cormorant (Phalacrocorax auritus) egg composition and hatchling phenotype

Maternal investment of yolk and albumen in avian eggs varies with egg mass and contributes to variation in hatchling mass. Here we use the natural variation in mass and composition of Double-crested Cormorant (Phalacrocorax auritus) eggs to examine consequences of variation in yolk and albumen mass on hatchling phenotype. The Double-crested Cormorant, a large bird with altricial young, lays eggs ranging in mass from 40 to 60 g and containing an average of 82% albumen and 18% yolk. Variation in Cormorant egg mass arises primarily from variation in the amount of albumen and water in the eggs; yolk mass remains relatively constant, contributing only 10% to egg mass variation. Likewise, variation in hatchling mass correlates positively with albumen mass and albumen solids contribute to hatchling dry mass. Thus, variation in Cormorant egg mass is primarily the result of variation in the amount of egg albumen, which contributes most to variation in hatchling mass. Similarities in egg composition of altricial birds, along with data presented here, suggest that variation in hatchling mass of all altricial birds may depend most on the amount of egg albumen, unlike species with precocial young that hatch from eggs with substantially more yolk.     

Egg Turning During Incubation has no Effect Upon the Growth of Embryos of Alligator mississippiensis

Alligator eggs are not turned during incubation, instead the embryo adheres to the top inside of the shell. Turning is alleged to shear off the embryo and kill it. Avian egg turning allegedly facilitates embryonic development by stimulating growth of the area vasculosa and minimizing the effects of unstirred yolk and albumen layers. From day 10 to day 45 of incubation, alligator eggs were experimentally turned, gently, through ± 60° in an hourly cycle. This turning regime killed only 6 out of 25 embryos. Compared with unturned controls, no significant effects were observed on the growth, production of extraembryonic fluids or utilization of albumen and yolk for those embryos that survived turning. The protein concentration of amniotic fluid at various stages of alligator development was examined in eggs incubated at 30 and 33°C. The fluid contained very little protein (max <8 mg) at any time: the protein concentration did not change consistently as development progressed. Differences in response to egg turning in birds and reptiles may be associated with the length of the incubation period, the protein content of the albumen and the mechanism of albumen utilization.