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Complex and at times extreme environments have pushed many bird species to develop unique eggshell surface properties to protect the embryo from external threats. Because microbes are usually transmitted into eggs by moisture, some species have evolved hydrophobic shell surfaces that resist water absorption, while also regulating heat loss and the exchange of gases. Here, we investigate the relationship between the wettability of eggshells from 441 bird species and their life-history traits. We measured the initial contact angle between sessile water droplets and the shell surface, and how far the droplet spread. Using phylogenetic comparative methods, we show that body mass, annual temperature and eggshell maculation primarily explained variance in water contact angle across eggshells. Species nesting in warm climates were more likely to exhibit highly hydrophobic eggshells than those nesting in cold climates, potentially to reduce microbial colonization. In non-passerines, immaculate eggs were found to have more hydrophobic surfaces than maculate eggshells. Droplets spread more quickly on eggshells incubated in open nests compared to domed nests, likely to decrease heat transfer from the egg. Here, we identify clear adaptations of eggshell wettability across a diverse range of nesting environments, driven by the need to retain heat and prevent microbial adhesion.

Eggshells are a potential tool for nonlethally sampling contaminant concentrations in bird eggs, yet few studies have examined their utility to represent mercury exposure. We assessed mercury concentrations in eggshell components for 23 bird species and determined whether they correlated with total mercury (THg) in egg contents. We designed a multi-experiment analysis to examine how THg is partitioned into eggshell components, specifically hardened eggshells, material adhered to the eggshells, and inner eggshell membranes. The THg concentrations in eggshells were much lower than in egg contents, and almost all of the THg within the eggshell was contained within material adhered to eggshells and inner eggshell membranes, and specifically not within calcium-rich hardened eggshells. Despite very little mercury in hardened eggshells, THg concentrations in hardened eggshells had the strongest correlation with egg contents among all eggshell components. However, species with the same THg concentrations in eggshells had different THg concentrations in egg contents, indicating that there is no global predictive equation among species for the relationship between eggshell and egg content THg concentrations. Furthermore, for all species, THg concentrations in eggshells decreased with relative embryo age. Although the majority of mercury in eggshells was contained within other eggshell components and not within hardened eggshells, THg in hardened eggshells can be used to estimate THg concentrations in egg contents, if embryo age and species are addressed. Environ Toxicol Chem 2017;36:2417-2427. Published 2017 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America., (© 2017 SETAC.)

In this study, wild hatched eggshells were collected from the nests of threatened Wattled Crane and South Ground-Hornbill in an attempt to determine their total Hg concentrations. A total of fourteen eggshell samples from both bird species were collected from different study areas in the Mpumlanga and KwaZulu-Natal Provinces of South Africa. The eggshells were acid digested under reflux and their total Hg concentrations were determined using cold-vapour atomic absorption spectrophotometry (CV-AAS). The observed total Hg levels for the South Ground-Hornbill samples ranged from 1.31 to 8.88 µg g(-1) dry weight (dw), except for one outlier which had an elevated 75.0 µg g(-1) dw. The levels obtained for the Wattled Crane samples were relatively high and these ranged from 14.84 to 36.37 µg g(-1) dw. Generally, all the measured total Hg concentrations for the Wattled Crane samples exceeded the estimated total Hg levels derived for eggshell which were known to cause adverse reproductive effects in avian species from previous studies. Based on these findings, it is, therefore, possible that the exposure of these birds to elevated Hg may have contributed to their present population decline., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Birds have evolved an astonishing diversity of egg pigmentation and this variation is driven by a complex interplay of proximate and ultimate factors. In ecological research, egg pigmentation is often measured at randomly selected points or regions of an eggshell, but it remains poorly explored whether and how spottiness and colouration are spatially consistent within a single egg. Here, we aimed to quantify consistency (repeatability) of spottiness patterns and background/spot colouration traits among the opposite lateral regions of an avian eggshell. For this purpose, we collected digital images of eggshells from three non-passerine birds showing different patterns of egg spottiness and colouration, i.e. Eurasian Coot Fulica atra, Black-headed Gull Chroicocephalus ridibundus, and Common Tern Sterna hirundo. We found an excellent within-egg repeatability (mean R > 0.90) of both background and spot colouration in all three species. In contrast, within-egg repeatability of spottiness traits was significantly lower, but the patterns of spatial variability were trait- and species-specific. While some spottiness traits showed relatively high within-egg repeatability (e.g. the total spot area), others were either poorly or moderately repeatable, depending on the species. Our results may have important practical implications for future research on eggshell colouration and spottiness, since measurements of reflectance spectra within a single region (side) are likely to be representative for the entire eggshell, while reliable characterisation of maculation may require replicate measurements across different eggshell regions. We recommend that the adaptive significance of spatial (within-egg) heterogeneity in avian eggshell pigmentation should be subject to further investigation under broader comparative framework. [ABSTRACT FROM AUTHOR]

Avian females can alter nest‐site selection and breeding behaviour in response to changes in ambient temperature. However, whether females alter eggshell characteristics in response to high ambient temperature is underexplored. To fill this gap, wild‐derived and domesticated Zebra Finches were bred at temperatures of 18 °C and 30 °C or 35 °C. Here we show that eggs from finches bred at 18 °C and 30 °C had similar characteristics, but females exposed to 35 °C laid eggs that were significantly thinner compared with the 18 °C group. These data suggest that a temperature that is ecologically relevant to the species in the arid zone of Australia can influence eggshell thickness, which is one of the important factors for hatchability in birds. [ABSTRACT FROM AUTHOR]

People have always been fascinated by birds. In the Roman period, birds (particularly chicken) were still largely employed in several rituals. Although Roman writers, such as Cicero, Livy, Pliny the Elder, and Aelian, inform us about the use of birds in a number of rituals, the archaeological evidence rarely allows us to associate the collected zooarchaeological remains with a single deity. In 2017, a well‐preserved archaeological context located in the vicinity of the temple of Isis in Pompeii and related to a single ritual event (dated to the mid‐1st century CE), revealed a number of faunal remains as well as ceramic and archaeobotanical finds. Chicken dominates the assemblage, followed by goose, pigeon, and turtle dove. A few remains of pig and a marine bivalve, the truncate donax, were also recovered. Skeletally young chickens were largely employed in the ritual, whereas the other bird taxa are only represented by mature bones. Chicken eggshell, taxonomically identified using ancient protein analysis, was also found. Taphonomic evidence indicates that bird bones were still covered with meat when exposed to the ritual fire. The results obtained have been linked to relevant written and iconographic sources dated to the Roman period. This work confirms that bird sacrifice was an important part of the Isis rituals. Among the birds involved, goose could be considered a possible marker of that goddess. The evidence from Pompeii adds new insights into our knowledge of the animals in the Isis cult during the 1st century CE. [ABSTRACT FROM AUTHOR]

The domestic chicken is a species of bird that has been extensively studied in regard to its biology and as a model organism for science. The reproduction of the species is by the laying of fertilized eggs, which in a period of 21 days will develop a chick inside. Several methods have been described to develop embryos ex-ovo, allowing the observation and manipulation of the organism. This work has the propose to standardize a method that allows the development of the embryos inside the artificial incubation system, which has a low cost and is easy to make. In this work, 100 chicken eggs were used to study the effects of humidity, mineral supplementation, and the preincubation time of the egg on the incubation ex-ovo of the embryos. Embryo development was documented through the different days. Pulverized eggshell was selected as an optimal source to provide calcium, magnesium, phosphorus, and other minerals to the developing embryo. By providing 900-1200 mg of pulverized eggshell, 40 mL of the 0.001 % solution of benzalkonium chloride, and a preincubation time of approximately 56 h, the embryos were able to develop until 19 days, and even though they did not reach hatching, the incubation conditions that allowed the survival and development of embryos until late stages were achieved. Thus, due to the conditions established for calcium, humidity and preincubation time, in the present work, the chicks reached 19 days of development. [ABSTRACT FROM AUTHOR]

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The avian eggshell is a remarkable biomineral, which is essential for avian reproduction; its properties permit embryonic development in the desiccating terrestrial environment, and moreover, are critically important to preserve unfertilized egg quality for human consumption. This calcium carbonate (CaCO3) bioceramic is made of 95% calcite and 3.5% organic matrix; it protects the egg contents against microbial penetration and mechanical damage, allows gaseous exchange, and provides calcium for development of the embryonic skeleton. In vertebrates, eggshell occurs in the Sauropsida and in a lesser extent in Mammalia taxa; avian eggshell calcification is one of the fastest known CaCO3 biomineralization processes, and results in a material with excellent mechanical properties. Thus, its study has triggered a strong interest from the researcher community. The investigation of eggshell biomineralization in birds over the past decades has led to detailed characterization of its protein and mineral constituents. Recently, our understanding of this process has been significantly improved using high-throughput technologies (i.e., proteomics, transcriptomics, genomics, and bioinformatics). Presently, more or less complete eggshell proteomes are available for nine birds, and therefore, key proteins that comprise the eggshell biomineralization toolkit are beginning to be identified. In this article, we review current knowledge on organic matrix components from calcified eggshell. We use these data to analyze the evolution of selected matrix proteins and underline their role in the biological toolkit required for eggshell calcification in avian species. Amongst the panel of eggshell-associated proteins, key functional domains are present such as calcium-binding, vesicle-binding and protein-binding. These technical advances, combined with progress in mineral ultrastructure analyses, have opened the way for new hypotheses of mineral nucleation and crystal growth in formation of the avian eggshell, including transfer of amorphous CaCO3 in vesicles from uterine cells to the eggshell mineralization site. The enrichment of multi-omics datasets for bird species is critical to understand the evolutionary context for development of CaCO3 biomineralization in metazoans, leading to the acquisition of the robust eggshell in birds (and formerly dinosaurs). [ABSTRACT FROM AUTHOR]

Runs of homozygosity (ROH) are chromosomal stretches that in a diploid genome appear in a homozygous state and display identical alleles at multiple contiguous loci. This study aimed to systematically compare the genomic distribution of the ROH islands among five populations of wild vs. commercial chickens of both layer and broiler type. To this end, we analyzed whole genome sequences of 115 birds including white layer (WL, n = 25), brown layer (BL, n = 25), broiler line A (BRA, n = 20), broiler line B (BRB, n = 20) and Red Junglefowl (RJF, n = 25). The ROH segments varied in size markedly among populations, ranging from 0.3 to 21.83 Mb reflecting their past genealogy. White layers contained the largest portion of the genome in homozygous state with an average ROH length of 432.1 Mb (618.7) per bird, despite carrying it in short segments (0.3-1 Mb). Population-wise inbreeding measures based on Wright's (Fis) and genomic (FROH) metrics revealed highly inbred genome of layer lines relative to the broilers and Red Junglefowl. We further revealed the ROH islands, among commercial lines overlapped with QTL related to limb development (GREM1, MEOX2), body weight (Meis2a.1, uc_338), eggshell color (GLCCI1, ICA1, UMAD1), antibody response to Newcastle virus (ROBO2), and feather pecking. Comparison of ROH landscape in sequencing resolution demonstrated that a sizable portion of genome of commercial lines segregates in homozygote state, reflecting many generations of assortative mating and intensive selection in their recent history. In contrary, wild birds carry shorter ROH segments, likely suggestive of older evolutionary events. [ABSTRACT FROM AUTHOR]

Living birds are unique among vertebrates in the formation of a female-specific bone tissue called medullary bone (MB) that is strictly associated with reproductive activity. MB is a rapidly mobilized source of calcium and phosphorus for the production of eggshell. Among living taxa, its skeletal distribution can be highly extensive such that it even exists in the ribs of some species. Due to its ephemeral nature, MB is rarely fossilized and so little is understood with regard to the origin of MB and its skeletal distribution in early taxa. Here we describe a new Early Cretaceous enantiornithine bird, Mirusavis parvus , gen. et. sp. nov. indicating that skeleton-wide distribution of MB appeared early in avian evolution. We suggest that this represents the plesiomorphic condition for the Aves and that the distribution of MB observed among extant neornithines is a product of increased pneumatization in this lineage and natural selection for more efficient distribution of MB. [ABSTRACT FROM AUTHOR]

The exceptional eggshell assemblage from Çatalhöyük was studied using an integrated approach combining morphology (by optical and scanning electron microscopy) and palaeoproteomics (by mass spectrometry). We provide taxonomic classification for 90 fragments, of which only 11 remain undetermined. The striking predominance of Anseriformes (probably including greylag geese, as well as ducks and swans) in all types of deposits examined, particularly middens and burial fills, suggests that these eggs were exploited as food and, at the same time, had a special significance for the inhabitants of the site. We detected the presence of crane eggshell in contexts associated with both the world of the living and the world of the dead (consistent with the well-known importance of this bird at Çatalhöyük), as well as the possible but infrequent occurrence of bustards and herons. Overall, we suggest that eggshell analysis, together with osteological data and the archaeological context, can provide the basis for a nuanced understanding of the relationship between humans and birds in the past. [ABSTRACT FROM AUTHOR]

In this study, we compared the antibacterial effectivity of the eggs of six precocial and four altricial bird species using Escherichia coli, based on their eggshell traits. The ultrastructure of eggshell was observed using a scanning electron microscope (SEM). According to SEM results, eggs from precocial birds (chicken, turkey, quail, duck, ostrich, and goose) had cuticle on the eggshells, while eggs from altricial birds (pigeon, budgerigar, munia, and canary) did not. The environment/selection pressure may induce the divergent evolution process in eggs of precocial and altricial birds. The E. coli experiment results showed that chicken, turkey, quail, duck, and goose eggs, with a high cuticle opacity, exhibited a much lower E. coli penetration rate. In contrast, the eggs with poor (ostrich) or without (pigeon, budgerigar, munia, and canary) cuticle exhibited a higher penetration rate. It is suggested that cuticle is a main barrier against bacterial penetration in precocial birds’ eggs. Turkey and quail eggs showed the lowest E. coli contamination rate (3.33% and 2.22%, respectively), probably because of the tightly connected nanosphere structure on their cuticle. As for altricial birds’ eggs, the eggs of budgerigar, munia, and canary with small pore diameter (0.57 to 1.22 μm) had a lower E. coli penetration rate than pigeon eggs (45.56%, 66.67%, 50%, and 97.78%, respectively, P < 0.05), indicating that pore diameter played a significant role in defending against bacterial trans-shell invasion. We found that eggshell thickness and pore area decreased with egg size. The cuticle quality had no relationship with egg size, but was closely related to the bird species. The E. coli penetration rate of altricial birds’ eggs was significantly higher than that of precocial birds’ eggs, mainly because the pores are exposed on the eggshell surface and cuticle protection is absent. This study provides detailed information on the eggshell cuticle, which gives insight into the cuticle evolution process that occurred in precocial and altricial bird species. Moreover, the results of E. coli penetration may help understanding the antibacterial behavior in birds. [ABSTRACT FROM AUTHOR]

The present study assessed heavy metals content in the eggs of the (house sparrows (Passer domesticus) and laughing dove (Spilopelia senegalensis) to give baseline data about metals concentration in two species from urban environments in the Basrah city(south of Iraq). The house sparrow is strappingly connected with human surroundings, and be able to live in urban or rural locations. Laughing dove live successfully in the urban environment . Although, it is one of the great inhabitants of the avian world. However, using common species with broad allocations prevents disagreeable effects on an endangered species population and may permit comparing concentrations of metals gathering between different environments. Concentrations of pb and cr in eggs of the birds were determined using Atomic Absorption spectrometry. Result established that eggshell in two birds species were collected high concentrations of lead metal arrived at (25.6,18.6ug/g) respectively, while, egg content was accumulate lowest level of lead ranged between (3.5 ug/g )and (2.5ug/g), respectively . Conversely, level of chromium was low in eggshell and egg content of two birds species. Positive relationships were observed between pb and cr level in eggshell and egg content in house sparrow. [ABSTRACT FROM AUTHOR]

The eggshell blueness is an interesting object for chicken genetic studies and blue-shelled chicken industry, especially after the discovery of the causative mutation of chicken blue eggshell. In the present study, genome wide association study (GWAS) was conducted in Chinese Dongxiang blue-shelled chicken underlying four traits of blue eggshell pigments: quantity of biliverdin (QB), quantity of protoporphyrin (QP), quantity of total pigment (QT), and color density trait (CD). A total of 139 individuals were randomly collected for GWAS. We detected two SNPs in genome-wise significance and 35 in suggestive significance, 24 out of the 37 SNP were located either within intron/exon or near 15 genes in a range of ~1.17 Mb on GGA21. For further confirmation of the identified SNP loci by GWAS, the follow-up replication studies were performed in two populations. A total of 146 individuals of the second generation derived from the former GWAS population, as well as 280 individuals from an alternative independent population were employed for genotyping by MALDI-TOF MS in a genotype-phenotype association study. Eighteen SNPs evenly distributed on the GGA21 significant region were successfully genotyped in the two populations, of which 4 and 6 SNP loci were shown significantly associated with QB, QT and QP in the two repeat populations, respectively. Further, the SNPs were narrowed down to a region of ~ 653.819 Kb on GGA21 that harbors five candidate genes: AJAP1, TNFRSF9, C1ORF174, CAMTA1, and CEP104. Shell gland of chickens laying dark and light blue eggshell was chosen for detection of mRNA expression of the five candidate genes. The results showed differential expression levels of these genes in the two groups. The specific function of these genes has not yet been defined clearly in chickens and further in-depth studies are needed to explore the new functional role in chicken eggshell blueness. [ABSTRACT FROM AUTHOR]

Avian eggshells are composed of several layers made of organic compounds and a mineral phase (calcite), and the general structure is basically the same in all species. A comparison of the structure, crystallography, and chemical composition shows that despite an overall similarity, each species has its own structure, crystallinity, and composition. Eggshells are a perfect example of the crystallographic versus biological concept of the formation and growth mechanisms of calcareous biominerals: the spherulitic—columnar structure is described as “a typical case of competitive crystal growth”, but it is also said that the eggshell matrix components regulate eggshell mineralization. Electron back scattered diffraction (EBSD) analyses show that the crystallinity differs between different species. Nevertheless, the three layers are composed of rounded granules, and neither facets nor angles are visible. In-situ analyses show the heterogeneous distribution of chemical elements throughout the thickness of single eggshell. The presence of organic matrices other than the outer and inner membranes in eggshells is confirmed by thermograms and infrared spectrometry, and the differences in quality and quantity depend on the species. Thus, as in other biocrystals, crystal growth competition is not enough to explain these differences, and there is a strong biological control of the eggshell secretion. [ABSTRACT FROM AUTHOR]

Birds are arguably the most evolutionarily successful extant vertebrate taxon, in part because of their ability to reproduce in virtually all terrestrial habitats. Common guillemots, Uria aalge, incubate their single egg in an unusual and harsh environment; on exposed cliff ledges, without a nest, and in close proximity to conspecifics. As a consequence, the surface of guillemot eggshells is frequently contaminated with faeces, dirt, water and other detritus, which may impede gas exchange or facilitate microbial infection of the developing embryo. Despite this, guillemot chicks survive incubation and hatch from eggs heavily covered with debris. To establish how guillemot eggs cope with external debris, we tested three hypotheses: (1) contamination by debris does not reduce gas exchange efficacy of the eggshell to a degree that may impede normal embryo development; (2) the guillemot eggshell surface is self-cleaning; (3) shell accessory material (SAM) prevents debris from blocking pores, allowing relatively unrestricted gas diffusion across the eggshell. We showed that natural debris reduces the conductance of gases across the guillemot eggshell by blocking gas exchange pores. Despite this problem, we found no evidence that guillemot eggshells are selfcleaning, but instead showed that the presence of SAM on the eggshell surface largely prevents pore blockages from occurring. Our results demonstrate that SAM is a crucial feature of the eggshell surface in a species with eggs that are frequently in contact with debris, acting to minimise pore blockages and thus ensure a sufficient rate of gas diffusion for embryo development. [ABSTRACT FROM AUTHOR]

Organochlorine compounds (OCs) pose a serious threat towards the wildlife due to their well-known adverse effects. India is the second largest producer of pesticides in Asia, with DDT production still ongoing, and is ranked amongst the leading countries of pesticide consumption. However, a significant data gap in avian biomonitoring studies has been identified in Asia. The objective of this review is to compile and discuss the available literature on concentrations of organochlorine pesticides and PCBs in Indian birds. The review of 18 articles showed that DDTs were the OCs most frequently analysed, followed by HCHs and PCBs (highest hepatic mean values: 11.6, 1.8 and 1.03 µg/g ww, respectively). The most frequently analysed matrix was whole body homogenates, followed by internal tissues. Plasma, eggs, feathers and guano were poorly sampled. The range of sampling years was 1980-2007. In general, hepatic OC concentrations were below the level known to cause adverse effects, although p,p’-DDE in eggs was found in concentrations associated with eggshell thinning. Most of the studies were carried out in Southern India (Tamil Nadu). Out of 106 species studied, house crow (Corvus splendens) was the most frequently monitored. However, the number of individuals sampled per species is generally low and different sample types are used, thus, huge limitations to compare OC exposure exist. This review shows that there is a clear deficit of data on OC concentrations and sublethal effects that needs to be addressed to understand the status of OC exposure, spatio-temporal trends and potential impacts in Indian avifauna. [ABSTRACT FROM AUTHOR]

The Gekkota is an important clade in the evolution of calcified eggshells in that some of its families lay rigid eggshells like archosaurs. However, the fundamental differences and similarities between the mechanism of rigid eggshell formation of the Gekkota and Archosauria have not been investigated thoroughly due to the lack of knowledge of gekkotan eggshells. Here, we report for the first time a comprehensive analysis of morphological, chemical compositional, and crystallographic features of rigid and soft gekkotan eggshells. Exhaustive morphological description provided common characters for gekkotan eggshells, as well as unique features of each species. We found that elemental distribution of rigid gekkotan eggshells is different from that of avian eggshells, especially in the case of Mg and P. In addition, the crystallographic features (size, shape, and alignment of calcite grains) of gekkotan eggshells are completely different from those of archosaur eggshells. The result of this study suggests that soft gekkotan eggshells are morphologically more similar to tuatara eggshells rather than soft eggshells of derived squamates. The chemical compositional analysis suggests that the eggshell may act as a mineral reservoir for P and F as well as Ca. More importantly, all chemical compositions and crystallographic features imply that the gekkotan eggshell formation may begin at the outer surface and growing down to the inner surface, which is opposite to the direction of the archosaur eggshell formation. This character would be crucial for identifying fossil gekkotan eggs, which are poorly known in paleontology. All these lines of evidence support that soft gekkotan and tuatara eggshells share the primitive characters of all lepidosaurid eggshells. Finally, gekkotan and archosaur rigid eggshells represent a typical example of convergent evolution in the lineage of the Sauropsida. [ABSTRACT FROM AUTHOR]

Determining the functional significance of ornamental traits and whether they are shaped by sexual selection requires understanding how they vary among individuals, and their relationship with other measures of quality. Conspicuous eggshell coloration is a trait that may have evolved through sexual selection as a signal of female quality. This hypothesis is based on the possibility that the pigment used to colour blue-green eggs is limited and potentially costly to allocate to eggshells. We investigated the signalling potential of blue-green eggshell colour by quantifying patterns of within- and among-clutch colour variation in Mountain Bluebirds (Sialia currucoides) to determine if pigment was limited and if egg colour traits are intrinsic to individual females. We also explored relationships over two breeding seasons between eggshell colour and other female traits and investment in eggs. Eggshell colour was highly repeatable within clutches for most measures, and relatively consistent between breeding events in different years, indicating that eggshell colour is an intrinsic trait. Eggs in second clutches within a season had less saturated blue-green colour than in first clutches, and within clutches, later-laid eggs were brighter but generally less blue-green in colour than earlier laid eggs, providing evidence of pigment limitation. However, associations between eggshell colour and other female traits and investment in eggs were less clear, and inconsistent among study years. Our findings suggest that there is potential for blue-green egg colour to function as a signal of female quality, but further study is needed to determine if blue-green eggshell colour is a truly informative signal. [ABSTRACT FROM AUTHOR]

Ornithobacterium rhinotrachealeis a bacterium that causes respiratory disease in birds and it has been isolated in countries with a large poultry production, including Mexico. The pathogenicity mechanisms of this bacterium have not been completely elucidated yet. The capacity of the bacterium to adhere to epithelial cells of chickenin vitrohas been evidenced, and since this bacterium has been isolated from the lungs and air sacs of several avian species, the aim of this study was to determine if this bacterium can adhere to chicken lung cells. We used fiveO. rhinotrachealereference serovars (A–E) that were in contact with primary lung cells cultured from a 19-day-old chicken embryo.O. rhinotrachealeadherence was evaluated through optical and transmission electron microscopies. The results revealed thatO. rhinotrachealeis capable of adhering to chicken embryo lung cells within 3 h of incubation with a diffuse adherence pattern. The adherence percentages of the chicken embryo lung cells were 51–96% according to the serovar of the bacterium. Relative adherence was from 4 to 8 bacteria per cell. Transmission electron microscope data revealed intracellular bacteria inside a vacuole in less than 3 h of incubation. [ABSTRACT FROM AUTHOR]

The green eggshell color produced by ducks is a threshold trait that can be influenced by various factors, such as hereditary, environment and nutrition. The aim of this study was to investigate the genetic regulation of the formation of eggs with green shells in Youxian ducks. We performed integrative analysis of mRNAs and miRNAs expression profiling in the shell gland samples from ducks by RNA-Seq. We found 124 differentially expressed genes that were associated with various pathways, such as the ATP-binding cassette (ABC) transporter and solute carrier supper family pathways. A total of 31 differentially expressed miRNAs were found between ducks laying green eggs and white eggs. KEGG pathway analysis of the predicted miRNA target genes also indicated the functional characteristics of these miRNAs; they were involved in the ABC transporter pathway and the solute carrier (SLC) supper family. Analysis with qRT-PCR was applied to validate the results of global gene expression, which showed a correlation between results obtained by RNA-seq and RT-qPCR. Moreover, a miRNA-mRNA interaction network was established using correlation analysis of differentially expressed mRNA and miRNA. Compared to ducks that lay white eggs, ducks that lay green eggs include six up-regulated miRNAs that had regulatory effects on 35 down-regulated genes, and seven down-regulated miRNAs which influenced 46 up-regulated genes. For example, the ABC transporter pathway could be regulated by expressing gga-miR-144-3p (up-regulated) with ABCG2 (up-regulated) and other miRNAs and genes. This study provides valuable information about mRNA and miRNA regulation in duck shell gland tissues, and provides foundational information for further study on the eggshell color formation and marker-assisted selection for Youxian duck breeding. [ABSTRACT FROM AUTHOR]

The common cuckoo (Cuculus canorus) is an avian brood parasite, laying its eggs in the nests of other bird species, where these hosts incubate the parasitic eggs, feed and rear the nestlings. The appearance of a cuckoo egg in a host nest may change the bacterial community in the nest. This may have consequences on the hatchability of host eggs, even when hosts reject the parasitic egg, typically within six days after parasitism. The present study revealed the bacterial community of cuckoo eggshells and those of the great reed warbler (Acrocephalus arundinaceus), one of the main hosts of cuckoos. We compared host eggs from non-parasitized clutches, as well as host and cuckoo eggs from parasitized clutches. As incubation may change bacterial assemblages on eggshells, we compared these egg types in two stages: the egg-laying stage, when incubation has not been started, and the mid-incubation stage (ca. on days 5–7 in incubation), where heat from the incubating female dries eggshells. Our results obtained by the 16S rRNA gene sequencing technique showed that fresh host and cuckoo eggs had partially different bacterial communities, but they became more similar during incubation in parasitized nests. Cluster analysis revealed that fresh cuckoo eggs and incubated host eggs in unparasitized nests (where no cuckoo effect could have happened) were the most dissimilar from the other groups of eggs. Cuckoo eggs did not reduce the hatchability of great reed warbler eggs. Our results on the cuckoo-great reed warbler relationship supported the idea that brood parasites may change bacterial microbiota in the host nest. Further studies should reveal how bacterial communities of cuckoo eggshells may vary by host-specific races (gentes) of cuckoos. [ABSTRACT FROM AUTHOR]

Ten reference genes were investigated for normalization of gene expression data in the shell gland of laying hens. Analyses performed with geNorm revealed that hypoxanthine phosphoribosyltransferase 1 (HPRT1) and hydroxymethylbilane synthase (HMBS) were the two most stable reference genes in response to post-oviposition time alone (POT) or with nicarbazin treatment (POT+N) of laying hens. NormFinder analyses showed that the two most stable reference genes in response to POT and POT+N were 18S ribosomal RNA (18S rRNA), ribosomal protein L4 (RPL4) and HMBS, RPL4, respectively. BestKeeper analyses showed that 18S rRNA, RPL4 and HPRT1, HMBS were the two most stable reference genes for POT, and POT+N, respectively. Of the ten reference genes, all except B2M showed geNorm M <0.5, suggesting that they were stably expressed in the shell gland tissue. Consensus from these three programs suggested HPRT1 and HMBS could be used as the two most stable reference genes in the present study. Expression analyses of four candidate target genes with the two most and the two least stable genes showed that a combination of stable reference genes leads to more discriminable quantification of expression levels of target genes, while the least stable genes failed to do so. Therefore, HMBS and HPRT1 are recommended as the two most stable reference genes for the normalization of gene expression data at different stages of eggshell formation in brown-egg laying hens. Available statistical programs for reference gene ranking should include more robust analysis capability to analyse the gene expression data generated from factorial design experiments. [ABSTRACT FROM AUTHOR]

Avian egg shape is generally explained as an adaptation to life history, yet we currently lack a global synthesis of how egg-shape differences arise and evolve. Here, we apply morphometric, mechanistic, and macroevolutionary analyses to the egg shapes of 1400 bird species. We characterize egg-shape diversity in terms of two biologically relevant variables, asymmetry and ellipticity, allowing us to quantify the observed morphologies in a two-dimensional morphospace. We then propose a simple mechanical model that explains the observed egg-shape diversity based on geometric and material properties of the egg membrane. Finally, using phylogenetic models, we show that egg shape correlates with flight ability on broad taxonomic scales, suggesting that adaptations for flight may have been critical drivers of egg-shape variation in birds. [ABSTRACT FROM AUTHOR]

Medullary bone (MB) is a special endosteal tissue forming in the bones of female birds during egg laying to serve as a labile calcium reservoir for building the hard eggshell. Therefore, the presence of MB reported in multiple nonavian dinosaurs is currently considered as evidence that those specimens were sexually mature females in their reproductive period. This interpretation has led to further inferences on species-specific growth strategies and related life-history aspects of these extinct vertebrates. However, a few studies questioned the reproductive significance of fossil MB by either regarding the tissue pathological or attributing alternative functions to it. This study reviews the general inferences on extinct vertebrates and discusses the primary role, distribution, regulation and adaptive significance of avian MB to point out important but largely overlooked uncertainties and inconsistencies in this matter. Emerging discordancy is demonstrated when the presence of MB vs. trade-off between growth and reproduction is used for interpreting dinosaurian growth curves. Synthesis of these data suggests that fossil MB was related to high calcium turnover rates but not exclusively to egg laying. Furthermore, revised application of Allosaurus growth data by modelling individual-based growth curves implies a much higher intraspecific variability in growth strategies, including timing of sexual maturation, than usually acknowledged. New hypotheses raised here to resolve these incongruences also propose new directions of research on the origin and functional evolution of this curious bone tissue. [ABSTRACT FROM AUTHOR]

Avian eggshell color seems to fulfill multiple functions, some of them being structural and others signaling. In this study, we tested whether or not eggshell coloration may play a role in sexual selection of Tree Sparrows ( Passer montanus). According to the 'Sexually selected eggshell coloration' hypothesis, eggshell coloration signals female, egg or chick quality and males adjust parental investment according to this signal. Eggs of this species are covered with brown spots and patches, and variation between clutches is high. We found that eggshell coloration correlates with both protoporphyrin and biliverdin, but protoporphyrin concentrations are ten times higher. Eggshell coloration reflects egg and offspring quality, but not female quality. Thus, eggshell coloration may signal female postmating investment in offspring rather than female quality. Furthermore, differential allocation in terms of maternal investment is supported by the fact that females lay more pigmented clutches when mated to males with bigger melanin-based ornaments relative to their own. Moreover, males invested proportionally more to chicks that hatched from more pigmented clutches. Our correlative results thus seem to support a role of sexual selection in the evolution of eggshell coloration in birds laying brown eggs, pigmented mainly by protoporphyrin. [ABSTRACT FROM AUTHOR]

Archaeologists and ecologists have increasingly developed new tools to help us understand the character of past ecosystems as well as human influences upon them. This study advances a refined method for the identification of avian eggshell fragments recovered through archaeological, ecological, and paleoecological fieldwork. The method is based upon high-resolution scanning electron microscopic (SEM) imaging of the inner surface of the eggshell and a system to describe the diagnostic features. These include the shape and texture of mammillae (the protruding ends of the calcareous columns that make up most of the eggshell) and the microrelief of intermammillary spaces and contact points. We also propose a method to measure the spatial density of mammillae. Finally, we provide a visual reference collection of eggshell from 25 bird species, most of which are widespread in the North Atlantic region, where our research is focused. By focusing upon one taxonomic group in particular (ducks, 10 species) we affirm that eggshell microstructure is species specific. This methodology is easily transportable to other regions and species assemblages and we advocate cataloging eggshell micromorphology on a larger scale to facilitate the use of eggshell fragments in archaeological and ecological studies. • This work presents a method for the identification of avian eggshell using scanning electron microscopy. • Qualitative and quantitative characteristics are catalogued along with micrographs of eggshell of 25 avian species. • The method will contribute to the growth and accessibility of archaeological and ecological research on avian eggshell. [ABSTRACT FROM AUTHOR]

Brown eggs are popular in many countries and consumers regard eggshell brownness as an important indicator of egg quality. However, the potential regulatory proteins and detailed molecular mechanisms regulating eggshell brownness have yet to be clearly defined. In the present study, we performed quantitative proteomics analysis with iTRAQ technology in the shell gland epithelium of hens laying dark and light brown eggs to investigate the candidate proteins and molecular mechanisms underlying variation in chicken eggshell brownness. The results indicated 147 differentially expressed proteins between these two groups, among which 65 and 82 proteins were significantly up-regulated in the light and dark groups, respectively. Functional analysis indicated that in the light group, the down-regulated iron-sulfur cluster assembly protein (Iba57) would decrease the synthesis of protoporphyrin IX; furthermore, the up-regulated protein solute carrier family 25 (mitochondrial carrier; adenine nucleotide translocator), member 5 (SLC25A5) and down-regulated translocator protein (TSPO) would lead to increased amounts of protoporphyrin IX transported into the mitochondria matrix to form heme with iron, which is supplied by ovotransferrin protein (TF). In other words, chickens from the light group produce less protoporphyrin IX, which is mainly used for heme synthesis. Therefore, the exported protoporphyrin IX available for eggshell deposition and brownness is reduced in the light group. The current study provides valuable information to elucidate variation of chicken eggshell brownness, and demonstrates the feasibility and sensitivity of iTRAQ-based quantitative proteomics analysis in providing useful insights into the molecular mechanisms underlying brown eggshell pigmentation. [ABSTRACT FROM AUTHOR]

In Australia and other parts of the world, table eggs with uniform brown eggshell color are well regarded by consumers. Brown eggshell color has been positively correlated with certain egg characteristics such as shell strength and egg specific gravity, along with specific antibacterial functions. In the current study, the effect of hen oviposition time, flock age, and egg position in-clutch on the intensity of brown eggshell color was studied in commercial laying hens. The collected eggs were processed to measure egg weight, shell reflectivity, shell color (L*a*b*), quantification of protoporphyrin IX (PP IX), and shell thickness. Hen oviposition time had a statistically significant effect (P < 0.05) on egg weight, L*a*b*, amount of PP IX, and shell thickness. L* increased from 59.72 in the first half hour after lights on to 61.67 6 hours later, and PP IX per gram of eggshell decreased from 1.32?10-7 mM to 1.26?10-7 mM. Flock age had a significant effect on egg weight, L*a*b*, shell reflectivity, PP IX, and shell thickness. The mean egg weight increased from 55.4 g at 25 wk of flock age to 63.3 g at 75 wk of flock age. PP IX per gram of eggshell was 1.45?10-7 mM at 25 wk and declined to 1.31?10-7 mM at 75 wk of flock age. Individual hen clutch length was highly variable, ranging from 22 to 123 eggs in a single clutch. Egg position in a clutch had a significant effect on all egg quality variables measured; however, the R² values for each variable measured were low. The eggshell color declined to a greater extent with increasing position in a clutch for long clutches compared with short and medium clutches. In conclusion, hen oviposition time affected brown eggshell color with darker brown eggs laid early in the d and lighter colored brown eggs laid later in the morning. The intensity of brown color decreased with flock age, and egg position in-clutch had relatively little effect on brown eggshell color. [ABSTRACT FROM AUTHOR]

How microbial symbionts are established and maintain on their hosts is a leading question with important consequences for the understanding of the evolution and functioning of mutualistic relationships. The acquisition by hosts of mutualistic microbial symbionts can be considered as colonization processes of environments (i.e., host) by symbionts. Colonization processes can be explored by characterizing the nestedness of communities, but this approach has rarely been applied to communities of microbial symbionts. We used this approach here, and estimated the nestedness of bacterial communities of hoopoes ( Upupa epops), a species with symbiotic bacteria in their uropygial gland that are expected to colonize eggshells where they protect embryos from pathogens. Bacterial communities were characterized by ARISA (Automated rRNA Intergenetic Spacer Region) and studied the nestedness characteristics of bacterial communities living in the uropygial secretion, bill, belly and eggshells of each sampled female hoopoes. We detected a consistent nested pattern of bacterial communities of hoopoes; from the uropygial gland to the eggshell. We also found evidence of study year and reproductive events influencing the level of nestedness of bacterial communities of hoopoes. These results indicate that bacterial communities of eggshells and body parts of female hoopoes are at least partially conditioned by the symbiotic community in the uropygial gland. We discuss the importance of these results for understanding this host-microbial mutualism functioning and evolution. [ABSTRACT FROM AUTHOR]

Eggshells are significant part of hatchery waste which consist of calcium carbonate crust, membranes, and proteins and peptides of embryonic origins along with other entrapped contaminants including microbes. We hypothesized that using this product as a nutritional additive in poultry diet may confer better immunity to the chickens in the paradigm of mammalian milk that enhances immunity. Therefore, we investigated the effect of hatchery eggshell membranes (HESM) as a short term feed supplement on growth performance and immunity of chickens under bacterial lipopolysaccharide (LPS) challenged condition. Three studies were conducted to find the effect of HESM supplement on post hatch chickens. In the first study, the chickens were fed either a control diet or diets containing 0.5% whey protein or HESM as supplement and evaluated at 5 weeks of age using growth, hematology, clinical chemistry, plasma immunoglobulins, and corticosterone as variables. The second and third studies were done to compare the effects of LPS on control and HESM fed birds at 5 weeks of age following at 4 and 24 h of treatment where the HESM was also sterilized with ethanol to deplete bacterial factors. HESM supplement caused weight gain in 2 experiments and decreased blood corticosterone concentrations. While LPS caused a significant loss in body weight at 24 h following its administration, the HESM supplemented birds showed significantly less body weight loss compared with the control fed birds. The WBC, heterophil/lymphocyte ratio, and the levels of IgG were low in chickens fed diets with HESM supplement compared with control diet group. LPS challenge increased the expression of pro-inflammatory cytokine gene IL-6 but the HESM fed birds showed its effect curtailed, also, which also, favored the up-regulation of anti-inflammatory genes compared with control diet fed chickens. Post hatch supplementation of HESM appears to improve performance, modulate immunity, and increase resistance of chickens to endotoxin. [ABSTRACT FROM AUTHOR]

Skin and feather characteristics, which play a critical role in body temperature maintenance, can be affected by incubation circumstances, such as incubation temperature. However, no study to date has assessed the influence of incubation temperature during the fetal stage on morphometric characteristics and vascular development of the skin, feather characteristics, and their relationship to hormone levels and preferred temperature in later life in chickens. Broiler breeder eggs were exposed to low (36°C), control (37.5°C), or high (39°C) temperatures (treatments LT, CK, and HT, respectively) from day 13 of incubation onward, because it is known that the endocrine axes are already established at this time. During this period, eggshell temperature of HT eggs (38.8±0.33°C) was higher than of LT (37.4±0.08°C) and CK eggs (37.8 ±0.15°C). The difference between eggshell and incubator air temperature diminished with the increasing incubation temperature, and was approximately zero for HT. HT hatchlings had higher surface temperature on the head, neck, and back, and thinner and more vascularized skin than did CK and LT hatchlings. No differences were found among treatments for body weight, total feather weight, number and length of barbs, barbule length, and plasma T4 concentration. LT hatchlings showed lower plasma T3 and GH, as well as lower T3/T4 ratio and decreased vascularity in the neck, back, and thigh skin compared to CK hatchlings. On the other hand, HT hatchlings had decreased skin thickness and increased vascularity, and preferred a higher ambient temperature compared to CK and HT hatchlings. In addition, for all treatments, surface temperature on the head was higher than of the other body regions. We conclude that changes in skin thickness and vascularity, as well as changes in thyroid and growth hormone levels, are the result of embryonic strategies to cope with higher or lower than normal incubation temperatures. Additionally exposure to increased temperature during incubation is an environmental factor that can exert early-life influence on ambient temperature preference of broiler hatchlings in later life. [ABSTRACT FROM AUTHOR]

The hypothesis has been tested that evolution has resulted in lower thermal emissivity of eggs of birds breeding openly in cold climates than of eggs of birds that nest under protective covering or in warmer climates. Directional thermal emissivity has been estimated from directional–hemispherical reflectance spectra. Due to several methodological difficulties the absolute emissivity is not accurately determined, but differences between species are obvious. Most notably, small waders of the genus Calidris , breeding in cold climates on the tundra, and in most cases with uniparental nest attendance, have low directional emissivity of their eggshells, about 0.92 when integration is carried out for wavelengths up to 16 μm. Species belonging to Galloanserinae have the highest directional emissivity, about 0.96, of their eggs. No differences due to climate or breeding conditions were found within this group. Eggs of most other birds tested possess intermediate emissivity, but the values for Pica pica and Corvus corone cornix are as low as for Calidris . Large species-dependent differences in spectral reflectance were found at specific wavelengths. For instance, at 4.259 μm the directional–hemispherical reflectance for galliforms range from 0.05 to 0.09, while for Fratercula arctica and Fulmarus glacialis it is about 0.3. The reflection peaks at 6.5 and 11.3 μm due to calcite are differentially attenuated in different species. In conclusion, the hypothesis that evolution has resulted in lower thermal emissivity of bird eggs being exposed in cold climates is not supported by our results. The emissivity is not clearly related to nesting habits or climate, and it is unlikely that the small differences observed are ecologically important. The spectral differences between eggs that nevertheless exist should be taken into account when using infrared thermometers for estimating the surface temperature of avian eggs. [ABSTRACT FROM AUTHOR]

The striking variation in colour and maculation of bird eggs has fascinated biologists since centuries, and many hypotheses based on mechanical, physiological or signalling functions have been proposed for its evolution by natural and sexual selection. Protoporphyrin is the main eggshell pigment found in brown maculated eggs, and is assumed to function as a mediator of these selection processes. It is a precursor of heme with pro-oxidant properties, and hence a link between brown maculation and female condition has been proposed and tested in a number of studies, albeit with contrasting results. A variety of different visual methods have been used to quantify outer eggshell pigmentation, which has been assumed to correspond to overall quantity of protoporphyrin in the shell. Yet, this relationship has rarely been tested. The aim of this study was to apply four commonly used methods to assess pigmentation in great tit eggs with protoporphyrin as the predominant eggshell pigment, and to compare the results of these methods. Specifically, we 1) ranked eggshell pigmentation by human naked eye, 2) applied a granularity approach and 3) measured spectrophotometric reflectance of eggshell pigments. Second, we estimated the relationship between outer eggshell pigmentation (i.e. estimated by three different methods above) and true protoporphyrin concentration deposited in the entire shell measured by HPLC. Among-method estimates were significantly correlated for the traits describing pigment ' darkness ' only. While the model including scores based on human naked eyes explained 16% of the variance of pigment concentration in the entire shell, spectrometry explained 27%, and the granularity approach explained 40%. Thus, the estimation of true pigment concentration in the entire shell from the visible outer side of the shell is most reliable with the granularity approach. It is relevant for studies where the maintenance of the integrity of the eggs is essential. [ABSTRACT FROM AUTHOR]

Obligate brood parasitic birds exploit their hosts to provide care for unrelated young in the nest. Potential hosts can reduce the cost of parasitism by rejecting foreign eggs from the nest. Observational, comparative, and experimental studies have concluded that most hosts use the coloration and patterning of eggshells to discriminate between own and foreign eggs in the nest. However, an alternative hypothesis is that birds use the colour contrasts between eggshells and the nest lining to identify parasitic eggs (egg-nest contrast hypothesis). In support of this hypothesis, we found that the avian perceivable chromatic contrasts between dyed eggs and unmanipulated nest linings significantly and negatively covaried with the rejection rates of different dyed eggs of the great reed warbler Acrocephalus arundinaceus, a frequently parasitized host of the common cuckoo Cuculus canorus. To experimentally test whether egg-nest contrasts influence rejection, we reciprocally dyed both eggs and the nest lining of this host species with one of two colours: orange and green. Contrary to the egg-nest contrast hypothesis, host rejection patterns in response to dyed eggs were not altered by dyeing nests, relative to unmanipulated control eggs and nests. In turn, experimental egg colour was the only significant predictor of egg rejection rate. Our results demonstrate that egg-nest contrast is a collateral, not a causal factor in egg rejection, and confirm the conclusions of previous studies that hosts can rely on the parasitic egg's appearance itself to recognize the foreign egg in the nest. [ABSTRACT FROM AUTHOR]

Background: Eggshell is subject to quality loss with aging process of laying hens, and damaged eggshells result in economic losses of eggs. However, the genetic architecture underlying the dynamic eggshell quality remains elusive. Here, we measured eggshell quality traits, including eggshell weight (ESW), eggshell thickness (EST) and eggshell strength (ESS) at 11 time points from onset of laying to 72 weeks of age and conducted comprehensive genome-wide association studies (GWAS) in 1534 F2 hens derived from reciprocal crosses between White Leghorn (WL) and Dongxiang chickens (DX). Results: ESWs at all ages exhibited moderate SNP-based heritability estimates (0.30 ~ 0.46), while the estimates for EST (0.21 ~ 0.31) and ESS (0.20 ~ 0.27) were relatively low. Eleven independent univariate genome-wide screens for each trait totally identified 1059, 1026 and 1356 significant associations with ESW, EST and ESS, respectively. Most significant loci were in a region spanning from 57.3 to 71.4 Mb of chromosome 1 (GGA1), which together account for 8.4 ~ 16.5% of the phenotypic variance for ESW from 32 to 72 weeks of age, 4.1 ~ 6.9% and 2.95 ~ 16.1% for EST and ESS from 40 to 72 weeks of age. According to linkage disequilibrium (LD) and conditional analysis, the significant SNPs in this region were in extremely strong linkage disequilibrium status. Ultimately, two missense SNPs in GGA1 and one in GGA4 were considered as promising loci on three independent genes including ITPR2, PIK3C2G, and NCAPG. The homozygotes of advantageously effective alleles on PIK3C2G and ITPR2 possessed the best eggshell quality and could partly counteract the negative effect of aging process. NCAPG had certain effect on eggshell quality for young hens. Conclusions: Identification of the promising region as well as potential candidate genes will greatly advance our understanding of the genetic basis underlying dynamic eggshell quality and has the practical significance in breeding program for the improvement of eggshell quality, especially at the later part of laying cycle. [ABSTRACT FROM AUTHOR]

In birds, the duration of egg incubation (the time from incubation onset to hatching) can affect multiple components of nest success, but what affects incubation duration? Previous studies suggest that incubation duration is affected by both parental behavior and components of the egg, which have yet to be determined. One egg component that may be related to incubation behavior and the time until hatching is eggshell porosity, which affects the exchange of metabolic gasses and water vapor across the shell and, thus, the speed of embryonic development and incubation duration. We tested whether eggshell porosity was associated with the timing of incubation onset by female House Wrens ( Troglodytes aedon Vieillot, 1809), and whether porosity varied within clutches in a manner that might be associated with incubation periods and hatching patterns (i.e., synchronous vs. asynchronous hatching). Eggshell porosity was unrelated to the onset of maternal incubation and did not differ between early- and later-laid eggs within clutches, but differed significantly among females and covaried with egg size. We conclude that producing all eggshells of similar porosity within clutches, while adjusting incubation onset once most or all eggs are laid, provide facultative maternal control over variation in hatching patterns. [ABSTRACT FROM AUTHOR]

Eggshell mechanical property traits such as eggshell breaking strength (ESS), eggshell thickness (EST) and eggshell weight (ESW) are most common and important indexes to evaluate eggshell quality in poultry industry. Uterine ion transporters involve in eggshell formation and might be associated with eggshell mechanical property traits. In this study, 99 SNPs in 15 ion transport genes were selected to genotype 976 pedigreed hens of Rhode Island Red. ESS, EST and ESW were measured for each bird at 55 weeks of age. The association study showed that 14 SNPs in 8 genes were significantly related (p < 0.05) with at least one trait, and their contributions to phenotypic variance ranged from 0.23% to 4.14%. Both ATP2A3 and SLC4A5 had a significant effect on all the three traits. Strong linkage disequilibrium (LD) was detected among SNPs in four genes: ATP2A3, ITPR1, SLC8A3, SCNN1a. The significant effects of those diplotypes on eggshell mechanical property traits were found, and their contributions to phenotypic variance ranged from 0.50% to 0.70%. It was concluded that the identified SNPs and diplotypes in this study were potential markers influencing the eggshell mechanical properties, which could contribute to the genetic improvement of eggshell quality. [ABSTRACT FROM AUTHOR]

Animals live in a bacterial world, and detecting and exploring adaptations favouring mutualistic relationships with antibiotic-producing bacteria as a strategy to fight pathogens are of prime importance for evolutionary ecologists., Uropygial secretion of European hoopoes ( Upupa epops, Linnaeus) contains antimicrobials from mutualistic bacteria that may be used to prevent embryo infection. Here, we investigated the microscopic structure of hoopoe eggshells looking for special features favouring the adhesion of antimicrobial uropygial secretions., We impeded female access to the uropygial gland and compared microscopic characteristics of eggshells, bacterial loads of eggs and of uropygial secretion, and hatching success of experimental and control females. Then, we explored the link between microbiological characteristics of uropygial secretion and these of eggs of hoopoes, as well as possible fitness benefits., The microscopic study revealed special structures in hoopoes' eggshells (crypts). The experimental prevention of females' gland access demonstrated that crypts are filled with uropygial secretion and that symbiotic enterococci bacteria on the eggshells come, at least partially, from those in the female's uropygial gland. Moreover, the experiment resulted in a higher permeability of eggshells by several groups of bacteria and in elimination of the positive relationships detected for control nests between hatching success and density of symbiotic bacteria, either in the uropygial secretion of females or on the eggshell., The findings of specialized crypts on the eggshells of hoopoes, and of video-recorded females smearing secretion containing symbiotic bacteria at a high density onto the eggshells strongly support a link between secretion and bacteria on eggs. Moreover, the detected associations between bacteria and hatching success suggest that crypts enhancing the adhesion of symbiont-carrying uropygial secretion likely protect embryos against infections. [ABSTRACT FROM AUTHOR]

SUPPLEMENTAL DATA—Supplemental materials are available for this article for free atwww.tandfonline.com/UJVP [ABSTRACT FROM AUTHOR]

Microorganisms are closely associated with eggs and may play a determinant role in embryo survival. Yet, the majority of studies focusing on this association relied on culture-based methodology, eventually leading to a skewed assessment of microbial communities. By targeting the 16 S r RNA gene and internal transcribed spacer ( ITS) region, we, respectively, described bacterial and fungal communities on eggshells of the homing pigeon Columba livia. We explored their structure, abundance, and composition. Firstly, we showed that sampling technique affected the outcome of the results. While broadly used, the egg swabbing procedure led to a lower DNA extraction efficiency and provided different profiles of bacterial communities than those based on crushed eggshell pieces. Secondly, we observed shifts in bacterial and fungal communities during incubation. At late incubation, bacterial communities showed a reduction in diversity, while their abundance increased, possibly due to the competitive advantage of some species. When compared to their bacterial counterparts, fungal communities also decreased in diversity at late incubation. In that case, however, the decline was associated with a diminution of their overall abundance. Conclusively, our results showed that although incubation might inhibit microbial growth when compared to unincubated eggs, we observed the selective growth of specific bacterial species during incubation. Moreover, we showed that fungi are a substantial component of the microbial communities associated with eggshells and require further investigations in avian ecology. Identifying the functional roles of these microorganisms is likely to provide news insights into the evolutionary strategies that control embryo survival. [ABSTRACT FROM AUTHOR]

Reproduction is a critical period for birds as they have to cope with many stressful events. One consequence of an acute exposure to stress is the release of corticosterone, the avian stress hormone. Prolonged stress can have negative impacts on the immune system, resulting in, for example, increased oxidative stress. Through maternal effects, females are known to modulate their investment in eggs content according to their own physiological condition. Less is known about maternal investment in eggshells, especially in pigments. The two main eggshell pigments may possess opposite antioxidant properties: protoporphyrin (brown) is a pro-oxidant, whereas biliverdin (blue-green) is an antioxidant. In Japanese quail, we know that the deposition of both pigments is related to female body condition. Thus, a chronic stress response may be reflected in eggshell coloration. Using female Japanese quails that lay brown-spotted eggs, we explored whether physiological exposure to corticosterone induces a change in female basal stress and antioxidant factors, and eggshell pigment concentration, spectrophotometric reflectance, and maculation coverage. We supplemented adult females over a 2 week period with either peanut oil (control) or corticosterone (treatment). We collected pre- and post-supplementation eggs and analysed the effect of corticosterone treatment on female physiology and eggshell appearance parameters. Except for corticosterone-fed birds which laid eggs with brighter spots, supplementation had no significant effect on female physiology or eggshell pigment concentration, reflectance and maculation. The change in eggshell spot brightness was not detected by a photoreceptor noise-limited color opponent model of avian visual perception. Our data confirms that eggshell reflectance in spotted eggs varies over the laying sequence, and spot reflectance may be a key factor that is affected by females CORT exposure, even if the changes are not detected by an avian visual model. [ABSTRACT FROM AUTHOR]

Defence mechanisms against parasites and pathogens are some of the most elaborate biological systems in animals. The oily secretion of the avian uropygial gland has been suggested to serve as a chemical defence against feather and eggshell bacteria. Yet, the traits associated with uropygial gland oil production are not well understood. We conducted a phylogenetic analysis comprising 132 European bird species aiming to test: (1) whether life-history and ecological traits drive gland size evolution by potentially promoting microbial infestation and (2) how these traits affects change in the gland size throughout the annual cycle. We show that the size of the uropygial gland is dynamic (i.e. increasing from the nonbreeding to the breeding season, independent of sex). Furthermore, we found that the year-round size of the gland was similar between sexes and was correlated with different ecological and life-history traits promoting microbial infection throughout the annual cycle. During the breeding season, the total eggshell surface area in a clutch correlated significantly and positively with the gland size, suggesting the importance of oil in protecting eggs from microbes. Social species exhibited a larger gland size increase during the breeding season compared to nonsocials; a change that was also predicted by the total eggshell surface area. Aquatic, riparian and non-migratory species had larger glands than terrestrials and migrants, respectively. The findings of the present study suggest that aquatic environments may promote the production of gland oil, through either the need of waterproofing the plumage and/or defending it against the intensified feather degradation in these moist conditions. Finally, we found a negative effect of the incubation period on uropygial gland size, which may suggest an energetic constraint imposed by other development-connected costly activities. Our results show that the role of the uropygial gland dynamically varies during the annual cycle, potentially in response to seasonal variation in parasitic infection risk. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110, 543-563. [ABSTRACT FROM AUTHOR]

Understanding how resources are allocated to form eggs is crucial to our better understanding of avian reproductive strategies. However, little is currently known about how egg synthesis in wild birds might be constrained by the availability of specific micronutrients. Here, we investigated the potential role of calcium (Ca) in constraining egg synthesis in the Audouin's Gull ( Ichthyaetus audouinii (Payraudeau, 1826)). In particular, we evaluated the relationship between plasma Ca levels (mg/dL) in incubating females (as an indicator of the physiological response of females to increased Ca demand associated with clutch production) and several fitness-related egg traits such as egg size (i.e., egg volume), egg shape, and eggshell thickness from three-egg clutches. Egg size was positively related with incubating female plasma Ca levels, with the slope of this relationship being significantly higher for later-laid eggs. The observational nature of this study and reversed timing precludes causal inferences, but observed relationships supported the constraining role of Ca in egg synthesis and suggested that Ca may also have a role in modulating the intraclutch pattern of egg-size variation typical of this gull species. [ABSTRACT FROM AUTHOR]

Abstract: Avian eggshell is a common component of many archaeological deposits, but its archaeological potential remains largely unexplored. The most obvious reasons are two-fold. Firstly, despite its abundance on many sites, eggshell is often overlooked during excavation. Even when it is recovered, small fragmented remains are difficult to identify taxonomically. Here we introduce a minimally destructive qualitative analytical technique for taxonomic identification of eggshell fragments based on highly sensitive mass spectrometry and peptide mass fingerprinting (ZooMS), and illustrate its application to eggshell recovered from the Viking Age urban site at Hungate, York. We adopt a more extreme version of the method of bleach treating used to prepare ancient eggshell for DNA analysis, followed by conventional peptide mass fingerprinting using MALDI-ToF mass spectrometry. The development of this technique will allow future research to make better use of eggshell fragments recovered from archaeological sites. [Copyright &y& Elsevier]

1. Performance traits were measured on 1908 Japanese quail and egg quality traits assessed on 1800 eggs at 10 wk of age. 2. Genetic and phenotypic correlations were estimated using a bivariate animal model with restricted maximum likelihood using ASREML software. 3. Body weight at different ages showed positive genetic correlations with egg weight and most of the internal egg quality traits, whereas their genetic correlations with eggshell thickness, eggshell strength and eggshell percentage were negative. 4. Genetic correlations of age at sexual maturity and egg number with most of external and internal traits were negative. 5. It was concluded that selection for higher body weight will result in heavier and better quality eggs. Because of the negative genetic correlation between BW and egg shell quality, a selection index including BW and eggshell strength would be the best breeding strategy for genetic improvement of egg quality in Japanese quail. [ABSTRACT FROM PUBLISHER]

Selection pressures due to parasitism play an important role in driving the evolution of life history traits of birds in general and of behaviour at the nest in particular. Eggshell bacterial load has been shown to predict hatching failure (i.e. the probability of embryo infection) but the relationships between the bacterial environment of the nest and life history characteristics of birds remain poorly investigated. We explored interspecific variation in eggshell bacterial load of mesophilic bacteria, Enterococcus spp., Staphylococcus spp. and Enterobacteriaceae groups across 24 bird species and assessed whether bacterial load is associated with breeding traits. Interspecific variation was much higher than intraspecific variation for all measures of bacterial load even after controlling for annual variation. Thus, we were able to assess the correlation between bacterial community characteristics and life history traits. After correcting for phylogenetic effects, we found that nest type, the use of feathers or plants as lining material, and incubation behaviour explained a significant proportion of the variance in bacterial communities on eggshells. The strength of these associations depended on study year, suggesting an important role of environmental conditions for eggshell bacterial load or community. Overall, these results suggest that bacteria on eggshells are associated with bird species traits, probably because birds are mediating the deleterious effect of eggshell microbes through behavioural traits that modify bacterial load. [ABSTRACT FROM AUTHOR]

Brood parasitism imposes several fitness costs on the host species. To reduce these costs, hosts of avian brood parasites have evolved various defenses, of which egg rejection is the most prevalent. In the face of variable host-parasite mimicry and the costs of egg discrimination itself, many hosts reject only some foreign eggs. Here, we experimentally varied the recognition cues to study the underlying cognitive mechanisms used by the Chalk-browed Mockingbird ( Mimus saturninus) to reject the white immaculate eggs laid by the parasitic Shiny Cowbird ( Molothrus bonariensis). Immaculate eggs are the only parasite eggs rejected by this host, as it accepts all polymorphic, spotted eggs laid by cowbirds. Using a within-breeding pair experimental design, we tested for the salience of spotting, UV reflectance, and brightness in eliciting rejection. We found that the presence of spotting significantly decreased the probability of rejection while increments in brightness significantly increased rejection frequencies. The cognitive rules underlying mockingbird rejection behavior can be explained by a decision-making model which predicts changes in the levels of rejection in direct relation to the number of relevant attributes shared between host and parasite eggs. [ABSTRACT FROM AUTHOR]