The relationship of pores and mammillae on the inner surface of the eggshell of the alligator (Alligator mississippiensis)

The morphology of the eggshell of the alligator, Alligator mississippiensis, is similar to that of birds. In many avian species there is a positive linear correlation between the numbers of pores and mammillae on the inner surfaces of eggshells, indicating that the distribution and density of mammillae may determine the porosity of the shell. It is not known, however, if a relationship exists between pores and mammillae on the shell of the alligator. Using a scanning electron microscope, we counted pores and mammillae on the inner surfaces of pieces of shell from the middle of fertile and infertile eggs from wild and captive, pen-reared alligators. Data were analyzed by ANOVA, Duncan's multiple range tests and linear regression equations. Results demonstrate a positive linear correlation between the numbers of pores and mammillae on the shells of unincubated fertile and infertile eggs from wild and captive alligators; however, there is no correlation between pores and mammillae on shells of eggs that were incubated for 55 days. It is suggested that initially the porosity of the eggshell of the alligator is related to the density of mammillae on the inner surface of the shell and that erosion of the shell during incubation destroys the original relationship between pores and mammillae.     

Determinants of avian eggshell porosity

The manner in which the porosity of avian eggshells varies with egg weight is discussed in relation to the number of pores, their cross-sectional area and length. Shell porosity and ways in which this can change to satisfy different environmental demands are discussed on the basis of a simple model which envisages the shell as being composed of columns of calcite.     

ULTRASTRUCTURAL AND FUNCTIONAL MORPHOLOGY OF EGGSHELLS SUPPORTS THE IDEA THAT DINOSAUR EGGS WERE INCUBATED BURIED IN A SUBSTRATE

Abstract:  The reproductive biology of dinosaurs is of great interest, particularly in light of the many fossil eggs assigned to this group. The ultrastructural characteristics of dinosaur eggshells are examined in order to calculate water vapour conductance, which indicates the nesting environment. Data were mainly derived from the literature but new values are also presented. Allometric analyses were carried out on a variety of shell parameters against predicted egg mass, and comparison was made with allometric equations for bird eggs. Shell thickness was generally larger than seen for extant birds. Total pore number and pores per unit area were similar to values predicted from bird eggs. Total pore area showed an isometric increase with egg mass, parallel to the relationship for birds, but the constant value was an order to magnitude higher than the bird values. Pore radius was unaffected by egg mass. Water vapour conductance showed an allometric increase with egg mass, parallel to the bird values, but for any given egg mass values for dinosaurs were an order of magnitude higher. Mass-specific water vapour conductance was unaffected by egg mass but was an order of magnitude higher than the bird values. Water vapour conductance per pore showed an allometric decrease with egg mass but again the predicted values were an order of magnitude higher than for bird eggs. The ultrastructural characteristics of dinosaur eggshells indicate that the nesting environment had to be saturated with water vapour and that dinosaur eggs had to be fully buried in a substrate. In this sense, therefore, dinosaur eggs resemble more those of modern reptiles than those of birds. As a consequence, maintenance of incubation conditions would have depended on the prevailing environment.     

内蒙古巴音满都呼晚白垩世棱齿龙蛋化石的发现

本文记述的恐龙蛋化石标本,采自内蒙古乌拉特后旗巴音满都呼上白垩统牙道黑达组中。蛋化石在蛋窝中排列的方式和蛋壳的显微结构特征与北美发现的含有可鉴定为棱齿龙胚胎骨骼的蛋化石基本相似,但还有一些差别,如蛋壳外表面不具纵向细纹,柱状层中鱼骨型纹饰不明显等。因此,应为棱齿龙科中另一新的属种代表。     

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.     

Magellanic Penguin eggshell pores: does number matter?

Most penguin species lay two eggs 3–4 days apart that hatch 1–2 days apart. Hypotheses to explain the shorter incubation period for the second egg include differences in porosity of the eggshells. Eggs with more or larger pores and/or thinner shells have higher gas exchange rates and faster embryonic development. We used eggshells from hatchling Magellanic Penguins Spheniscus magellanicus to test whether eggshell thickness and pore density affect incubation period. We expected: (i) second eggs to have thinner shells and/or more pores than first eggs; (ii) eggs to have thinner shells and/or more pores if they were laid by older females, later in the season, or in burrow nests (more humid than open nests under bushes). We found no support for these hypotheses. Egg pores and shell thickness were not related to incubation period. Eggs from the same clutch were similar in eggshell thickness and pore density. Female age, laying date and nest type were poor predictors of eggshell thickness and pore density. Egg pores, we conclude, have little explanatory power for differences in incubation time of eggs in Magellanic Penguins, and suggest that synchronous onset of incubation matters.     

Regulation of Membrane Transport Sites for Amino Acids in Myogenic Cells

Abstract. Myoblasts from 12-day chick embryos in cell culture transport the nonmetabolizable amino acid α-aminoisobutyric acid (AIB) two to three-fold more rapidly than multinucleated myotubes which form from them. This decrease in transport is due to a relative decrease in the number of transport sites per unit area of cell surface suggesting a compositional change in the plasma membrane during myogenesis. In studies reported here, AIB transport was monitored throughout myogenesis and correlated with other aspects of differentiation. During myogenesis the number of amino acid transport sites remains constant per myotube nucleus. As myogenesis proceeds, there is a marked increase in cellular protein and cell surface without a commensurate increase in amino acid transport sites. The net consequence of the surface area change is fewer amino acid transport sites per unit area of myotube membrane surface. The decrease in membrane transport sites for AIB per unit area of membrane is not a result of length of time in culture per se, medium depletion, or cell density, but is a result of differentiation, since blocking myoblast fusion by deprivation of calcium delays the decrease in AIB transport sites per unit cell surface area while reversal of the calcium deprivation block is accompanied by a rapid decrease in the number of AIB transport sites per unit cell surface area. Thus, the decrease in AIB transport sites is an aspect of differentiation which accompanies the marked elaboration of surface membrane during myogenesis.     

Regulation of membrane transport sites for amino acids in myogenic cells. A differentiation dependent phenomenon

Myoblasts from 12-day chick embryos in cell culture transport the nonmetabolizable amino acid alpha-aminoisobutyric acid (AIB) two to three-fold more rapidly than multinucleated myotubes which form from them. This decrease in transport is due to a relative decrease in the number of transport sites per unit area of cell surface suggesting a compositional change in the plasma membrane during myogenesis. In studies reported here, AIB transport was monitored throughout myogenesis and correlated with other aspects of differentiation. During myogenesis the number of amino acid transport sites remains constant per myotube nucleus. As myogenesis proceeds, there is a marked increase in cellular protein and cell surface without a commensurate increase in amino acid transport sites. The net consequence of the surface area change is fewer amino acid transport sites per unit area of myotube membrane surface. The decrease in membrane transport sites for AIB per unit area of membrane is not a result of length of time in culture per se, medium depletion, or cell density, but is a result of differentiation, since blocking myoblast fusion by deprivation of calcium delays the decrease in AIB transport sites per unit cell surface area while reversal of the calcium deprivation block is accompanied by a rapid decrease in the number of AIB transport sites per unit cell surface area. Thus, the decrease in AIB transport sites is an aspect of differentiation which accompanies the marked elaboration of surface membrane during myogenesis.     

Electroporation in dense cell suspension--theoretical and experimental analysis of ion diffusion and cell permeabilization

Electroporation is a process where increased permeability of cells exposed to an electric field is observed. It is used in many biomedical applications including electrogene transfection and electrochemotherapy. Although the increased permeability of the membrane is believed to be the result of pores due to an induced transmembrane voltage U(m), the exact molecular mechanisms are not fully explained. In this study we analyze transient conductivity changes during the electric pulses and increased membrane permeability for ions and molecules after the pulses in order to determine which parameters affect stabilization of pores, and to analyze the relation between transient pores and long-lived transport pores. By quantifying ion diffusion, fraction of transport pores f(per) was obtained. A simple model, which assumes a quadratic dependence of f(per) on E in the area where U(m)>U(c) very accurately describes experimental values, suggesting that f(per) increases with higher electric field due to larger permeabilized area and due to higher energy available for pore formation. The fraction of transport pores increases also with the number of pulses N, which suggest that each pulse contributes to formation of more and/or larger stable transport pores, whereas the number of transient pores does not depend on N.     

Structure of the shell and tertiary membranes of eggs of softshell turtles (Trionyx spiniferus)

Eggs of the turtle Trionyx spiniferus are rigid, calcareous spheres averaging 2.5 cm in diameter. The eggshell is morphologically very similar to avian eggshells. The outer crystalline layer is composed of roughly columnar aggregates, or shell units, of calcium carbonate in the aragonite form. Each shell unit tapers to a somewhat conical tip at its base. Interior to the crystalline layer are two tertiary egg membranes: the outer shell membrane and the inner shell membrane. The outer shell membrane is firmly attached to the inner surface of the shell, and the two membranes are in contact except at the air cell, where the inner shell membrane separates from the outer shell membrane. Both membranes are multi-layered, with the inner shell membrane exhibiting a more fibrous structure than the outer shell membrane. Numerous pores are found in the eggshell, and these generally occur at the intersection of four or more shell units.     

Effects of polyploidy on photosynthesis

In polyploid plants the photosynthetic rate per cell is correlated with the amount of DNA per cell. The photosynthetic rate per unit leaf area is the product of the rate per cell times the number of photosynthetic cells per unit area. Therefore, the photosynthetic rate per unit leaf area will increase if there is a less than proportional increase in cell volume at higher ploidal levels, or if cell packing is altered to allow more cells per unit leaf area. In autopolyploids (Medicago sativa, C₃ species, and Pennisetum americanum, C₄ species) there is a doubling of photosynthesis per cell and of cell volume in the tetraploid compared to the diploid. However, there is a proportional decrease in number of cells per unit leaf area with this increase in ploidy such that the rate of photosynthesis per leaf area does not change. There is more diversity in the relationship between ploidal level (gene dosage) and photosynthetic rates per unit leaf area in allopolyploids. This is likely to reflect the effects of natural selection on leaf anatomy, and novel genetic interactions from contributed genomes which can occur with allopolyploidy. In allopolyploid wheat (C₃ species) a higher cell volume per unit DNA at the higher ploidal level is negatively correlated with photosynthesis rate per unit leaf area. Although photosynthesis per cell increases with ploidy, photosynthesis per leaf area decreases, being lowest in the allohexaploid, cultivated bread wheat (Triticum aestivum). Alternatively, doubling of photosynthetic rate per cell with doubling of DNA, with apparent natural selection for decreased cell volume per unit DNA, results in higher rates of photosynthesis per leaf area in octaploid compared to tetraploid Panicum virgatum (C₄) which may be a case of allopolyploidy. Similar responses probably occur in Festuca arundinacea. Therefore, in some systems anatomical factors affecting photosynthesis are also affected by ploidal level. It is important to evaluate that component as well as determining the effect on biochemical processes. Current information on polyploidy and photosynthesis in several species is discussed with respect to anatomy, biochemistry and bases for expressing photosynthetic rates.Abbreviations Chl chlorophyll - RuBPC ribulose-1,5-bisphosphate carboxylase     

PROPERTIES OF AVIAN EGG SHELLS AND THEIR ADAPTIVE VALUE

I. An arbitrary classification of avian eggshells is proposed.
2. The role of the eggshell in conserving the water in eggs at oviposition is discussed. There is as yet no correlation between this property and the pore systems in avian eggshells.
3. The pore systems may act as diffusion pathways and the hypothesis has been advanced that in many eggs the shells are adapted so that restriction of gaseous diffusion by mud, preening oils and nest debris is prevented.
4. The mechanical properties of the shell are considered in the novel context of defence against (a) attrition that could lead to the pores being blocked with dust, and (b) cracking that would destroy the diffusion pathways noted in 3.
5. The overall objective of the review was to discuss the concept that avian eggshell are adapted to fit an egg to the nest environment.     

Structure of the Photosynthetic Apparatus in Leaves of Freshwater Hydrophytes: 1. General Characteristics of the Leaf Mesophyll and a Comparison with Terrestrial Plants

The structure of photosynthetic elements was investigated in leaves of 42 boreal plant species featuring different degrees of submergence (helophytes, neustophytes, and hydatophytes). The mesophyll structure types were identified for all these species. Chlorenchyma tissues and phototrophic cells were quantitatively described by such characteristics as the sizes of cells and chloroplasts in the mesophyll and epidermis, the abundance of cells and chloroplasts in these tissues, the total surface area of cells and chloroplasts per unit leaf area, the number of plastids per cell, etc. The hydrophytes typically had thick leaves (200–350 m) with a well-developed aerenchyma; their specific density per unit area (100–200 mg/dm²) was lower than in terrestrial plants. Mesophyll cells in aquatic plants occupied a larger volume (5–20 × 10³m³) than epidermal cells (1–15 × 10³m³). The number of mesophyll cells per unit leaf area was nearly 1.5 times higher than that of epidermal cells. Chloroplasts were present in the epidermis of almost all species, including emergent leaves, but the ratio of the chloroplast total number to the number of all plastids varied depending on the degree of leaf submergence. The total number of plastids per unit leaf area (2–6 × 10⁶/cm²) and the surface of chloroplasts per unit leaf area (2–6 cm²/cm²) were lower in hydrophytes than in terrestrial plants from climatically similar habitats. The functional relations between mesophyll parameters were similar for hydrophytes and terrestrial plants (a positive correlation between the leaf weight per unit area, leaf thickness, and the number of mesophyll cells per unit leaf area), although no correlation was found in hydrophytes between the volume of mesophyll cells and the leaf thickness. Phototrophic tissues in aquatic plants contributed a larger fraction to the leaf weight than in terrestrial plants, because the mechanical tissues were less developed in hydrophytes. The CO₂assimilation rates by leaves were lower in hydrophytes than in terrestrial plants, because the total surface area of chloroplasts per unit leaf area is comparatively small in hydrophytes, which reduces the conductivity for carbon dioxide diffusion towards the carboxylation sites.     

Relationship between age and colour of hatched eggshells of Ochlerotatus vigilax (Skuse) (Diptera: Culicidae) on an Australian saltmarsh

Abstract The presence of aedine mosquito eggshells in soil has recently been used to delineate oviposition patterns in coastal wetlands. However, the colour of eggshells may reveal information about the oviposition activity of mosquitoes that is not evident from the density and distribution of eggshells alone. This study describes the rate of colour change of newly hatched eggshells of Ochlerotatus (Ochlerotatus) vigilax , the common saltmarsh mosquito. In March 1996, eggshells were buried in soil of five mosquito-breeding sites on Kooragang Island, NSW. Eggshells were graded by colour using Munsell^® soil-colour charts. Seventy-five percent of newly hatched eggshells were black and 25% were dark brown in colour. Black eggshells faded quickly to lighter shades of brown, so that none were recovered 20 months after commencement of the study. Eggshells faded to a median of dark brown after 13 months and medium brown after 2 years. In another study, significant differences in colour frequency between existing saltmarsh and two recently created oviposition sites were found; the latter site being characterised by numerous black and dark-brown eggshells and very few light-brown eggshells. The results suggest that active and inactive oviposition sites may be identified using the frequency distribution of eggshell colours based on large numbers of eggshells, whereas the colour of individual eggshells only coarsely reflects age since eclosion.     

Structure of shells from eggs of kinosternid turtles

Shells from eggs of five species of kinosternid turtle (Sternotherus minor, Kinosternon flavescens, K. baurii, K. Hirtipes, and K. alamosae) were examined with light and scanning electron microscopy. Except for possible differences among species in thickness of eggshells, structure of shells from all eggs was similiar. In general, kinosternid turtles lay eggs having a rigid calcareous layer composed of calcium carbonate in the form of aragonite. The calcareous layer is organized into individual shell units with needlelike crystallites radiating from a common center. Most of the thickness of the eggshell is attributable to the calcareous layer, with the fibrous shell membrane comprising only a small fraction of shell thickness. Pores are found in the calcareous layer, but they are not numereous. The outer surface of the eggshells is sculptured and may have a thick, organic layer in places. The outer surface of the shell membrane of decalcified eggshells is studded with spherical cores which presumably nucleate growth of shell units during shell formation. The shell membrane detaches from eggs incubated to hatching, carrying with it remnants of the calcareous layer. Such changes in shell structure presumably reflect withdrawal of calcium from the eggshell by developing embryos.     

Ultrastructural morphology of the shell and shell membrane of eggs of common snapping turtles (Chelydra serpentina)

Common snapping turtles (Chelydra serpentina) lay nearly spherical, flexible-shelled eggs having an outer mineral layer composed of calcium carbonate in the aragonite form. The mineral layer is arranged into loosely organized groups of nodular shell units, with numerous spaces (or pores) between adjacent shell units. Shell units are structurally complex, consisting of an inner tip that is morphologically distinct from the main body of the shell unit. Contained within an intact shell unit at the interface of the tip and the main part of the shell unit is the central plaque, an apparent modification of the shell membrane that may serve to nucleate calcification of shell units during shell formation. The tips of shell units are firmly attached to a single, multilayered shell membrane throughout much of incubation. The calcareous layer begins to detach from the shell membrane about half-way through incubation, and changes in shell morphology attending this detachment indicate that snapping turtles may use the shell as a source of calcium during embryogenesis. The arrangement of the mineral layer into groups of shell units, the large number of spaces between shell units, and little or no interlocking of crystallites of adjacent shell units apparently are factors contributing to the ability of these eggs to swell as they absorb water.     

Histochemical studies on eggshell formation in Paramphistomum cervi (Digenea: Paramphistomatidae)

The formation of the egg mass in the ootype, containing the oocyte, vitelline cells, numerous spermatozoa and Mehlis' gland secretion marks the beginning of eggshell formation. Mehlis' gland cells pour their lipoprotein secretion into the ootype; this secretion forms a thin template around the egg mass and upon this template the shell globules are deposited. Eggshell formation begins in the proximal uterus and is completed in the middle uterus. The eggshells are devoid of phenols and phenolase indicating that hardening is not by quinone tanning. The eggshells contain keratin and are stabilized by disulphide linkages.     

A Mechanobiology-based Algorithm to Optimize the Microstructure Geometry of Bone Tissue Scaffolds

Complexity of scaffold geometries and biological mechanisms involved in the bone generation process make the design of scaffolds a quite challenging task. The most common approaches utilized in bone tissue engineering require costly protocols and time-consuming experiments. In this study we present an algorithm that, combining parametric finite element models of scaffolds with numerical optimization methods and a computational mechano-regulation model, is able to predict the optimal scaffold microstructure. The scaffold geometrical parameters are perturbed until the best geometry that allows the largest amounts of bone to be generated, is reached. We study the effects of the following factors: (1) the shape of the pores; (2) their spatial distribution; (3) the number of pores per unit area. The optimal dimensions of the pores have been determined for different values of scaffold Young''s modulus and compression loading acting on the scaffold upper surface.Pores with rectangular section were predicted to lead to the formation of larger amounts of bone compared to square section pores; similarly, elliptic pores were predicted to allow the generation of greater amounts of bone compared to circular pores. The number of pores per unit area appears to have rather negligible effects on the bone regeneration process. Finally, the algorithm predicts that for increasing loads, increasing values of the scaffold Young''s modulus are preferable.The results shown in the article represent a proof-of-principle demonstration of the possibility to optimize the scaffold microstructure geometry based on mechanobiological criteria.     

Incubation reduces microbial growth on eggshells and the opportunity for trans-shell infection

Avian eggshells harbour microbes shortly after laying, and under appropriate ambient conditions they can multiply rapidly, penetrate through shell pores, infect egg contents and cause embryo mortality. We experimentally examined how incubation affects bacterial processes on the eggshells of pearl-eyed thrashers Margarops fuscatus nesting in tropical montane and lowland forests in Puerto Rico. Bacteria and fungi grew rapidly on shells of newly laid, unincubated eggs exposed to ambient conditions, but declined to low levels on shells of eggs incubated by thrashers. Divergence in bacterial growth between incubated and exposed eggs was more marked at the montane forest than at the lowland site. Pathogenic microorganisms became increasingly dominant on shells of exposed eggs, but these groups were relatively rare on incubated eggs, where more benign, less invasive groups prevailed. Some incubation during laying may be necessary to decrease the probability of trans-shell infection by reducing the growth of harmful bacteria and fungi on eggshells, although it may increase hatching asynchrony and the likelihood of brood reduction.     

Evolutionarily stable resource allocation for production of wind-dispersed seeds

We developed a game-theoretic model for wind-dispersed seed production to examine the seed mass–dispersal ability relationship and the evolutionarily stable distance of seed dispersal in terms of exploitation of safe sites. We assumed trade-offs between masses of the embryo (including albumen) and the wind-dispersal structures per seed, and also between seed mass and number of seeds per parent. We showed that ESS wing-loading is independent of embryo mass; that is, heavy seeds are not poor dispersers if the cost of producing wind-dispersal structures per unit area is constant. The ESS embryo mass per seed depends only on the factors which determine the probability of a seedling being established from a seed. However, wing-loading was found to increase with embryo mass when the change in length was isometric and there was a negative correlation between seed mass and dispersal ability. Thus, the area–mass relationship in wind-dispersal structures may have large effects on the ESS production of wind-dispersed seeds. On the other hand, given that only a limited number of adults can be established at a safe site, the ESS seed dispersal distance depends on the relative degree of sib to non-sib competition. A parent disperses its seeds over a wide area to exploit many safe sites if sib competition is strong. However, it disperses its seeds within a narrow area if the mean number of parents per unit area is large, or if non-sib competition is strong. Thus, in addition to an upper limit on the number of adults per safe site, the degree of sib and non-sib competition may be important for the ESS dispersal distance in wind-dispersed seeds. This revised version was published online in July 2006 with corrections to the Cover Date.