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.     

Photodynamic antimicrobial activity of avian eggshell pigments

Pigmentation in avian eggshells appears to be associated with shell strength, temperature regulation, and camouflage. The pigments found in eggshells are mainly porphyrins, which have been utilized therapeutically as photosensitizers. Here, we examined the photoinactivation of gram-positive (Staphylococcus aureus, Bacillus cereus) and gram-negative bacteria (Escherichia coli, Salmonella enteritidis) by hen eggshells and their pigments. The results indicated that eggshells have a light-dependent antimicrobial activity against gram-positive, but not gram-negative, bacteria. Our results indicate the possibility that the natural pigments used therapeutically have evolved in nature as a defence system.     

Molecular preservation in Late Cretaceous sauropod dinosaur eggshells

Exceptionally preserved sauropod eggshells discovered in Upper Cretaceous (Campanian) deposits in Patagonia, Argentina, contain skeletal remains and soft tissues of embryonic Titanosaurid dinosaurs. To preserve these labile embryonic remains, the rate of mineral precipitation must have superseded post-mortem degradative processes, resulting in virtually instantaneous mineralization of soft tissues. If so, mineralization may also have been rapid enough to retain fragments of original biomolecules in these specimens. To investigate preservation of biomolecular compounds in these well-preserved sauropod dinosaur eggshells, we applied multiple analytical techniques. Results demonstrate organic compounds and antigenic structures similar to those found in extant eggshells.     

A re-evaluation of aff. Megaloolithidae eggshell fragments from the uppermost Cretaceous of the Pyrenees and implications for crocodylomorph eggshell structure

The Upper Cretaceous outcrops of the Pyrenees yield one of the most extensive and continuous records of paleoological remains anywhere in the world. Most of eggs and eggshells have been referred to the oofamily Megaloolithidae. In this study, we present a revision of eggshell fragments from the Blasi 2 locality, lattermost Maastrichtian in age, previously assigned to aff. Megaloolithidae. The presence of a blocky extinction pattern and basal knobs supports a crocodilian affinity of these materials. We classify them as Krokolithidae indet. Three structural layers can be recognised in the Blasi 2 eggshells, a feature that is shared with other recent eggshells (e.g. Crocodylus porosus and Crocodylus niloticus) and fossil crocodylomorph eggshells (Krokolitheswilsoni), which were previously described as single layered. The new proposed affinity of the Blasi 2 eggshells reduces the Megaloolithidae oodiversity of the last few million years of the Cretaceous in the Pyrenees to only two valid ootaxa, Megaloolithusmamillare and Megaloolithusbaghensis. The lack of more complete material precludes the erection of new ootaxa based on the Blasi 2 material.     

一种从雉类孵化卵壳中提取DNA的方法

从鸟类已孵化的卵壳中提取DNA属于一种非损伤性取样技术,在鸟类分子生态学研究中具有广阔的应用前景。本实验以河南董寨自然保护区白冠长尾雉(Syrmaticus reevesii)和环颈雉(Phasianuscolchicus)已孵化的卵壳为材料,利用红细胞破碎液、蛋白酶K及RNA酶等试剂,对卵壳膜内的总DNA进行了提取,建立了一种提取高质量DNA的新方法。琼脂糖凝胶电泳检测显示,采用新改进的方法提取出的DNA条带清晰。同时,利用聚合酶链式反应(PCR)技术成功地从总DNA中扩增出两种雉类的线粒体DNA控制区(CR)片段,测序后与GenBank中同一物种的CR序列进行比对,结果证实了PCR产物的真实性。文中利用卵壳膜提取出的DNA,对一窝环颈雉的雏鸟进行了性别鉴定,其结果与根据形态特征进行鉴定的结果完全一致,均为3雄4雌,从而证实了从卵壳膜中提取DNA的真实性。该种DNA提取方法在雉类研究中将具有广泛的应用。     

BIOMECHANICAL PROPERTIES OF DINOSAUR EGGSHELLS (Ⅲ)──THE MECHANIC ANALYSIS OF THE BABY DINOSAURS' EMERGING FROM THEIR EGGS

Thepurposeofthispaperistoanalyzethestressofdinosaureggshellloadedbythehatch1ingswhentheyhatch,andfourtypesofdinosaureggshellsareavaila-ble(seeTablelofZhaoeta1.,l994;TablelofMaandZhao,1994).Mostofthelivingreptileshaveflexible,parchment-likeeggshellscomposedoffibresandrelativelysma1lamountsofcalciumcarbonate.Thchatchlings,eachequ-ippedwithasmalI,sharp"eggtooth",cracktheireggshel1stoemerge'However,theyoungbirdspokeatthebluntendoftheeggwiththeirbeak,thenstruggleoutfromtheireggs.Thehatchingproce…     

Prevalence of potentially pathogenic culturable bacteria on eggshells and in cloacae of female Pied Flycatchers in a temperate habitat in central Spain

ABSTRACT Bacteria grow on avian eggshells and thus can potentially cause diseases in developing embryos. Little is known about culturable bacteria colonizing avian eggshells in free‐living birds, with most studies restricted to poultry. Our objective was to examine the culturable bacterial array growing on eggshells during incubation that could negatively affect hatching success of Pied Flycatchers (Ficedula hypoleuca) in a temperate montane habitat in central Spain. Cloacal culturable bacteria of females were also analyzed because bacteria can be vertically transmitted from females to eggs. We used fecal samples as surrogates of cloacal samples due to the small size of sampled birds. We found that eggshells and female cloacae of Pied Flycatchers harbored 24 and 40 bacterial families and species, respectively, but only a few in each clutch and each cloaca. Rod‐shaped gram‐negative bacteria and bacteria in the family Pseudomonadaceae were the most common bacteria on eggshells during early and late incubation and in female cloacae. Although based on small sample sizes, we found that females with rod‐shaped gram‐negative bacteria in their cloacae laid eggs that also had these bacteria, providing possible evidence for vertical transmission. We found no evidence for vertical transmission of Pseudomonadaceae, suggesting a possible environmental source for these bacteria. The prevalence of bacterial morphological types and major taxonomical categories on eggshells did not vary from early to late stages of incubation, providing support for the hypothesis that incubation may have bacteriostatic effects on bacterial proliferation on eggshells. Despite being primary egg invaders in poultry, we detected no effects of culturable Pseudomonadaceae or Pseudomonas luteola on hatching success. Our study represents the first to examine the culturable bacteria growing on the eggshells of a wild bird in a temperate habitat and additional studies based on culture‐independent techniques are required to confirm our results.     

Fossile Reptil-Eischalen (Chelonia,Crocodilia, Dinosauria) aus dem unteren Barremium von Galve (Provinz Teruel,SE-Spanien)

The Lower Cretaceous vertebrate bearing sediments of Galve (province of Teruel, SE Spain) have yielded some hundred fossil eggshell fragments, all slightly diagenetically altered. Concerning their microstructure, an assignment of the material to different taxa of reptiles was possible. The eggshell assemblage of Galve consists of a-c) three types of chelonian eggshells, one of them probably representing the subfamily Batagurinae, which would extend this taxon back into the Lower Cretaceous, d) the oldest known crocodilian eggshells, with lacunae systems on the outer surface, comparable to Recent taxa, and e-f) two different types of dinosaurian eggshells. In correspondence to the suggested fluviatile paleoenvironment of Galve, the fossil eggshells described herein are fragmented and represent mostly turtles and crocodiles.     

Synthesis of three-dimensional calcium carbonate nanofibrous structure from eggshell using femtosecond laser ablation

Background  

Natural biomaterials from bone-like minerals derived from avian eggshells have been considered as promising bone substitutes owing to their biodegradability, abundance, and lower price in comparison with synthetic biomaterials. However, cell adhesion to bulk biomaterials is poor and surface modifications are required to improve biomaterial-cell interaction. Three-dimensional (3D) nanostructures are preferred to act as growth support platforms for bone and stem cells. Although there have been several studies on generating nanoparticles from eggshells, no research has been reported on synthesizing 3D nanofibrous structures.     

卵壳的超微结构特征

本文运用扫描电镜研究了美洲鸵鸟,鹂鹋,非洲鸵鸟,普通家鸡,环颈雉,绿头鸭,王企鹅等七种现生鸟蛋壳和更新世安氏鸵鸟蛋壳以及六种白垩纪恐龙蛋壳（长形长形蛋Ｅｌｏｎｇａｔｏｏｌｉｔｈｕｓｅｌｏｎｇａｔｕｓ,安氏长形蛋Ｅｌｏｎｇａｔｏｏｌｉｔｈｕｓａｎｄｒｅｗｓｉ,瑶屯巨形蛋Ｍａｃｒｏｏｌｉｔｈｕｓｙａｏｔｕｎｅｎｓｉｓ粗皮巨形蛋Ｍａｃｒｏｏｌｉｔｈｕｓｒｕｇｕｓｔｕｓ,将军顶圆形蛋Ｓｐｈｅｒｏｏｌｉｔｈ     

Two new theropod egg sites from the Late Jurassic Lourinhã Formation,Portugal

Two new Late Jurassic (uppermost Late Kimmeridgian) dinosaur eggshell sites are described, Casal da Rola and Porto das Barcas, both near Lourinhã, central-west Portugal. Casal da Rola yields eggshells with an obliquiprismatic morphotype comparable to those from a nest with the associated fossil embryos from Paimogo, tentatively assigned to the theropod Lourinhanosaurus antunesi. The Porto das Barcas eggshells have a dendrospherulitic morphotype with a prolatocanaliculate pore system. This morphotype was also recognised in eggshells from a clutch with associated Torvosaurus embryos at the Porto das Barcas locality. A preliminary cladistic analysis of eggshell morphology suggests theropod affinities for the Casal da Rola eggs, but is unable to resolve the phylogenetic position of the Porto das Barcas eggs. The eggshells at both sites are preserved in distal flood plain mudstones and siltstones. Carbonate concretions within the deposits indicate paleosol development.     

三种海鸟卵壳的超微结构和无机成分的研究

应用扫描电镜和ＴＮ－５５００能谱仪对三种海鸟卵壳的超微结构和基本无机元素进行研究分析,结果表明,基本结构相似,但表层突起和裂纹,乳锥和锥本形态,以及气孔在单位面积内的数量,壳膜与卵壳的锚连方式,壳膜元素组成等都存在着不同,这既反映其遗传,繁殖生理功能上的差异,也显示了不同种鸟卵壳之间的相似和相异性,因而对研究严分类和地理分布具有一定差异价值。     

颍上八里河自然保护区鹭卵3种重金属残留分析

2005年5月,采集了淮河地区颍上八里河集群繁殖的夜鹭(Nycticorax nycticorax)和小白鹭(Egrettagarzetta)鸟卵共18枚,用原子吸收法测定了卵壳、内容物中重金属Cd、Pb、Cr的残留量。结果表明,所有卵壳样品中均检出一定水平的Cd、Pb和Cr残留量,夜鹭卵壳中重金属残留量为Pb>Cr>Cd,小白鹭卵壳中重金属残留量为Cr>Pb>Cd,卵壳中Pb和Cr残留量的种间差异都不显著,但Cd的种间差异显著;在2种鹭卵内容物中,Cr都被检出,在夜鹭卵内容物样品中,Pb只被部分检出,Cd在2种鹭卵内容物样品中都没有被检出,3种重金属在内容物中残留量的种间差异都不显著。2种鹭卵壳中Pb和Cd残留量极其显著地高于卵内容物,但Cr的这种残留分布上的差异不显著。本研究进一步证明卵壳可用作重金属污染物的指示物,监测和评价湿地生态系统中重金属的污染状况。     

Palaeoenvironmental reconstruction of dinosaur nesting sites based on a geochemical approach to eggshells and associated palaeosols (Maastrichtian, Provence Basin, France)

Variability of the stable isotopic (δ¹³C and δ¹⁸O) and trace element (Mg, Sr, Fe, and Mn) content from dinosaur eggshells from the Provence Basin has been addressed by considering the stratigraphic distribution of the eggshells, the palaeoenvironments, and the chemical content of associated sediments. Preferentially, in situ eggshells were analysed. Samples were collected from two sections, one belonging to the floodplain (Roques Hautes) and the other to the channel belt (Rousset-Bréguières), as determined from sedimentological criteria. On a regional scale, this combined analysis provided new insights into palaeohydrology, vegetation cover and dinosaur behaviour. Different sources of drinking water were identified: (1) the main stream waters with significant salinity levels, (2) lateral feeders with water composition close to that of precipitation. Water composition is reflected in the eggshell trace element contents: a high Sr content for the eggshells from the channel belt (mean 1200 ppm) contrasting with the very low values for the floodplain (mean 250 ppm). During the first part of the Maastrichtian, evaporation rates were quite significant, as seen from the 1.5‰ difference between the eggshell δ¹⁸O (drinking water) and the palaeosol carbonate nodules (precipitation). This contrast diminished and became nearly insignificant in the second half of the Maastrichtian. Vegetation was composed of C₃ plants and two ecosystems are reconstructed: a riparian forest extending along the channel belt and open vegetation in the floodplain. The contrasted isotopic and trace element content of the eggshells was a crucial factor in identifying animal migration from one site to another. Migration was quite limited, suggesting that the animals settled for some time at the places where they laid their eggs. Intraspecific variations among modern birds are well correlated with the environment and may be greater than interspecies variations, which limits the potential of geochemical analysis to characterise dinosaur species.     

Changes in femoral morphology during egg-laying in Alligator mississippiensis

Birds and many reptiles are egg-layers. Birds provide calcium for the formation of eggshells by resorbing medullary bone, which is laid down before ovulation. Turtles do not possess this mechanism and resorb structural bone to form eggshells. Femora from three groups of alligators (egg-laying females; quiescent, immature, or barren females; and males) were examined to determine if alligators, which are closely related to birds in evolution, resorb structural bone during the formation of eggshells as do turtles. Microradiographs of cross sections from femoral mid-shafts were analyzed for porosity, and the robusticity index of each femur was determined. Scanning electron micrographs of anorganic endosteal and periosteal femoral surfaces were analyzed to determine numbers of entrances of vascular canals, numbers of lacunae of osteoblasts, and types of femoral surfaces. Femora from egg-laying females were significantly less robust than those of other females or males, and sections of bone from the egg-layers were significantly more porous than those of the other groups. Scanning electron microscopy of anorganic femoral endosteal surfaces from egg-laying females revealed significantly more resorption areas when compared with males or non egg-laying females. Periosteal surfaces from egg-layers had significantly more resting and less bone-forming surface than those from the other groups. Results indicated that apposition of periosteal bone may have been reduced in egg-layers and that egg-laying alligators, like turtles, resorb endosteal structural bone, which may be used as a source of calcium for the formation of eggshells.     

Titanosaur eggshells from the Tremp Formation (Upper Cretaceous,Southern Pyrenees,Spain)

Hundreds of megaloolithid eggshells have been found at the Costa de la Coma site (Tremp Fm, South-Central Pyrenees, Spain), the microstructural features (spherulitic eggshell units, thickness, ornamentation and pore system) of which agree with the Megaloolithus oogenus diagnosis. Comparisons with other megaloolithids from the Tremp Basin, southern France and South America, however, showed that these eggshells share many features with the South American oospecies Megaloolithus patagonicus. A cladistic analysis of the qualitative eggshell features of the main dinosaur taxa was performed. The results grouped the Costa de la Coma eggshells with Megaloolithus patagonicus, forming a clade separated from other megaloolithids, and revealed a polytomy of megaloolithids (associated with sauropods) and spheroolithids (associated with hadrosaurs). The finding of Megaloolithus cf. patagonicus in the Tremp Fm agrees with skeletal evidence provided by four indeterminate titanosaur forms from the Formation's late Cretaceous deposits. Other Megaloolithus oospecies of the Tremp Fm may belong to hadrosaurs, the skeletal remains of which are also abundant in the Tremp Basin.     

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.     

Cues for communal egg‐laying in lizards (Bassiana duperreyi,Scincidae)

Animals may aggregate either because the presence of conspecifics provides information about habitat suitability, or because the presence of conspecifics directly enhances individual viability. For a female lizard, the advantage of laying her eggs in a communal nest may entail either information transfer (hatched eggshells show that the site has been successful in previous seasons) or direct physiological benefits (recently‐laid eggs can enhance water availability to other eggs). We tested the relative importance of these two mechanisms in the three‐lined alpine skink (Bassiana duperreyi Gray, 1838) by offering gravid females a choice between sites with hatched eggshells versus freshly‐laid eggs. Females selectively oviposited beside fresh eggs. In this species, early‐nesting females use information transfer (i.e. the presence of old eggshells) as a nest‐site criterion, whereas later nesters switch to a reliance on direct benefits of conspecific presence (i.e. the presence of freshly‐laid eggs). © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110 , 839–842.     

Evidence for a calcium-binding site on the eggshell of Globodera rostochiensis with a role in hatching

Two inhibitors of hatching in Globodera rostochiensis, ruthenium red and lanthanum, have been shown to bind to the eggshell using the techniques of microdensitometry for ruthenium red and X-ray microanalysis for lanthanum. Neither inhibitor penetrated or adhered to unhatched or hatched viable juveniles. Scatchard analysis for binding of lanthanum and ruthenium red to eggshells gave dissociation constants (K) of K_La 32.5 ± 14.0 μM and K_Rured 33.5 ± 5.0 μM respectively. Both values are within the 95% fiducial limits of those shown to cause 50% inhibition of hatch in previous work. Pretreatment with sodium hypochlorite separated an outer part of the eggshell from an inner region which exclusively bound ruthenium red. It is the inner lipoprotein layer that is believed to include the membranes controlling the permeability of the tylenchid eggshell. The rate of binding of ruthenium red was similar for intact and isolated eggshells with 50% binding occurring after 6.11 ± 0.91 min and 4.95 ± 2.38 min but the latter gave a significantly higher maximum binding suggesting that rupture of the eggshells made available additional binding sites on their inner surface. The binding of ruthenium red to the eggshells was pH dependant over most of the range pH 2.8–8.5 with 50% binding, given with its standard deviation, occurring at pH 5.75 ± 0.85. Competitive binding of lanthanum influenced the binding of ruthenium red to the eggshells from which Scatchard analysis gave K_la of 176 ± 79 μM. Similarly, calcium influenced the binding but this caused a biphasic plot with high and low affinity binding sites of K“_ca of 0.423 ± 1.16 μm and K‘_ca of 1078 ± 462 μM. The existence of a high affinity site for calcium that also binds ruthenium red, suggests that the eggshell membrane includes a calcium binding glycoprotein as found in some other receptor mechanisms.     

Morphometric analysis of dinosaur eggshells: constraints of size on shape

Studies on fossil eggs and eggshells have been traditionally focused on analysis of microstructure and systematic classification, but few attempts have been made to examine the variability of eggshell characters. This work examines the variability of one diagnostic character, shape of the calcitic units, of some dinosaur eggshells by using outline shape analysis (elliptic Fourier analysis). The studied material belongs to five different oospecies of the parafamily Megaloolithidae Zhao, 1979. To test shape variation within and between oospecies, an analysis on 129 outlines was carried out and their variability was summarised by principal component analysis. We then tested if the shape of megaloolithid units changes according to size (i.e. allometry). A regression analysis between eggshell unit shape and two different size measurements (centroid size and eggshell thickness) showed that the thinnest eggshells (thickness ≤ 1 mm) have different shapes than thicker ones ( ≥ 2.5 mm). Our results stress that the variability in the shape of eggshell units in Megaloolithus oospecies is correlated to size (i.e. eggshell thickness), a phenomenon that should be then considered carefully when determining oospecies, since a single oospecies can show both a wide range of eggshell thickness and combined with different eggshell unit shapes.