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Cleidoic eggs possess very efficient and orchestrated systems to protect the embryo from external microbes until hatch. The cuticle is a proteinaceous layer on the shell surface in many bird and some reptile species. An intact cuticle forms a pore plug to occlude respiratory pores and is an effective physical and chemical barrier against microbial penetration. The interior of the egg is assumed to be normally sterile, while the outer eggshell cuticle hosts microbes. The diversity of the eggshell microbiome is derived from both maternal microbiota and those of the nesting environment. The surface characteristics of the egg, outer moisture layer and the presence of antimicrobial molecules composing the cuticle dictate constituents of the microbial communities on the eggshell surface. The avian cuticle affects eggshell wettability, water vapor conductance and regulates ultraviolet reflectance in various ground-nesting species; moreover, its composition, thickness and degree of coverage are dependent on species, hen age, and physiological stressors. Studies in domestic avian species have demonstrated that changes in the cuticle affect the food safety of eggs with respect to the risk of contamination by bacterial pathogens such as Salmonella and Escherichia coli . Moreover, preventing contamination of internal egg components is crucial to optimize hatching success in bird species. In chickens there is moderate heritability (38%) of cuticle deposition with a potential for genetic improvement. However, much less is known about other bird or reptile cuticles. This review synthesizes current knowledge of eggshell cuticle and provides insight into its evolution in the clade reptilia. The origin, composition and regulation of the eggshell microbiome and the potential function of the cuticle as the first barrier of egg defense are discussed in detail. We evaluate how changes in the cuticle affect the food safety of table eggs and vertical transmission of pathogens in the production chain with respect to the risk of contamination. Thus, this review provides insight into the physiological and microbiological characteristics of eggshell cuticle in relation to its protective function (innate immunity) in egg-laying birds and reptiles.

Calcified eggshells protect developing embryos against environmental stress and contribute to reproductive success 1 . As modern crocodilians and birds lay hard-shelled eggs, this eggshell type has been inferred for non-avian dinosaurs. Known dinosaur eggshells are characterized by an innermost membrane, an overlying protein matrix containing calcite, and an outermost waxy cuticle 2 – 7 . The calcitic eggshell consists of one or more ultrastructural layers that differ markedly among the three major dinosaur clades, as do the configurations of respiratory pores. So far, only hadrosaurid, a few sauropodomorph and tetanuran eggshells have been discovered; the paucity of the fossil record and the lack of intermediate eggshell types challenge efforts to homologize eggshell structures across all dinosaurs 8 – 18 . Here we present mineralogical, organochemical and ultrastructural evidence for an originally non-biomineralized, soft-shelled nature of exceptionally preserved ornithischian Protoceratops and basal sauropodomorph Mussaurus eggs. Statistical evaluation of in situ Raman spectra obtained for a representative set of hard- and soft-shelled, fossil and extant diapsid eggshells clusters the originally organic but secondarily phosphatized Protoceratops and the organic Mussaurus eggshells with soft, non-biomineralized eggshells. Histology corroborates the organic composition of these soft-shelled dinosaur eggs, revealing a stratified arrangement resembling turtle soft eggshell. Through an ancestral-state reconstruction of composition and ultrastructure, we compare eggshells from Protoceratops and Mussaurus with those from other diapsids, revealing that the first dinosaur egg was soft-shelled. The calcified, hard-shelled dinosaur egg evolved independently at least three times throughout the Mesozoic era, explaining the bias towards eggshells of derived dinosaurs in the fossil record. Molecular analyses of newly discovered, embryo-bearing ornithischian and sauropod dinosaur eggs suggest that the ancestral dinosaur egg was soft-shelled, and that hard-shelled eggs evolved independently at least three times in the major dinosaur lineages.

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.

The higher availability of zinc (Zn) from organic than inorganic sources is already established, but more assertive and cost-friendly protocols on the total replacement of inorganic with organic Zn sources for laying hens still need to be developed. Because some discrepancy in the effects of this replacement in laying hen diets is noticeable in the literature, the objective of this meta-analysis was to properly quantify the effect size of total replacing inorganic Zn with organic Zn in the diet of laying hens on their laying performance, egg quality, and Zn excretion. A total of 2340 results were retrieved from Pubmed, Scielo, Scopus, WOS, and Science Direct databases. Of these, 18 primary studies met all the eligibility criteria and were included in this meta-analysis. Overall, the replacement of inorganic Zn with organic Zn, regardless of other factors, improved (p < 0.01) egg production by 1.46%, eggshell thickness by 0.01 mm, and eggshell resistance by 0.11 kgf/cm2. Positive results of the same nutritional strategy on egg weight and Zn excretion were only observed at specific conditions, especially when organic Zn was supplemented alone in the feed, not combined with other organic minerals. Therefore, there is evidence in the literature that the total replacement of inorganic Zn with organic Zn improves egg production, eggshell thickness, and eggshell resistance. Factors such as hen age and genetics, organic Zn source, concentration of Zn in the feed, and the strategy of its supplementation have to be more carefully considered in protocols designed to address egg weight and Zn excretion by the hen.

Metallic lithium anodes are highly promising for revolutionizing current rechargeable batteries because of their ultrahigh energy density. However, the application of lithium metal batteries is considerably impeded by lithium dendrite growth. Here, a biomacromolecule matrix obtained from the natural membrane of eggshell is introduced to control lithium growth and the mechanism is motivated by how living organisms regulate the orientation of inorganic crystals in biomineralization. Specifically, cryo-electron microscopy is utilized to probe the structure of lithium at the atomic level. The dendrites growing along the preferred < 111 > crystallographic orientation are greatly suppressed in the presence of the biomacromolecule. Furthermore, the naturally soluble chemical species in the biomacromolecules can participate in the formation of solid electrolyte interphase upon cycling, thus effectively homogenizing the lithium deposition. The lithium anodes employing bioinspired design exhibit enhanced cycling capability. This work sheds light on identifying substantial challenges in lithium anodes for developing advanced batteries. Inspired by the role of proteins in regulating eggshell mineralization, here Tao, Liu and colleagues apply trifluoroethanol modified eggshell membrane to combat lithium dendrite. Cryo-electron microscopy reveals that the growth along the most favored crystallographic direction is suppressed.

Signs of aging in facial skin correlate with lifespan and chronic disease; however, the health of aging skin has not been extensively studied. In healthy young skin, the dermis forms a type III collagen-rich dermal papilla, where capillary vessels supply oxygen and nutrients to basal epidermal cells. Chicken eggshell membranes (ESMs) have been used as traditional medicines to promote skin wound healing in Asian countries for many years. Previously, we designed an experimental system in which human dermal fibroblasts (HDFs) were cultured on a dish with a solubilized ESM (S-ESM) bound to an artificial phosphorylcholine polymer; we found that genes that promoted the health of the papillary dermis, such as those encoding type III collagen, were induced in the S-ESM environment. The present study found that a gel with a ratio of 20% type III/80% type I collagen, similar to that of the baby skin, resulted in a higher elasticity than 100% type I collagen (p < 0.05) and that HDFs in the gel showed high mitochondrial activity. Thus, we decided to perform further evaluations to identify the effects of S-ESM on gene expression in the skin of hairless mice and found a significant increase of type III collagen in S-ESM. Picrosirius Red staining showed that type III collagen significantly increased in the papillary dermis after S-ESM treatment. Moreover, S-ESM application significantly improved human arm elasticity and reduced facial wrinkles. ESMs may have applications in extending lifespan by reducing the loss of tissue elasticity through the increase of type III collagen.

The avian eggshell is a natural protective envelope that relies on the phenomenon of biomineralization for its formation. The shell is made of calcium carbonate in the form of calcite, which contains hundreds of proteins that interact with the mineral phase controlling its formation and structural organization, and thus determine the mechanical properties of the mature biomaterial. We describe its mineralogy, structure and the regulatory interactions that integrate the mineral and organic constituents. We underline recent evidence for vesicular transfer of amorphous calcium carbonate (ACC), as a new pathway to ensure the active and continuous supply of the ions necessary for shell mineralization. Currently more than 900 proteins and thousands of upregulated transcripts have been identified during chicken eggshell formation. Bioinformatic predictions address their functionality during the biomineralization process. In addition, we describe matrix protein quantification to understand their role during the key spatially- and temporally- regulated events of shell mineralization. Finally, we propose an updated scheme with a global scenario encompassing the mechanisms of avian eggshell mineralization. With this large dataset at hand, it should now be possible to determine specific motifs, domains or proteins and peptide sequences that perform a critical function during avian eggshell biomineralization. The integration of this insight with genomic data (non-synonymous single nucleotide polymorphisms) and precise phenotyping (shell biomechanical parameters) on pure selected lines will lead to consistently better-quality eggshell characteristics for improved food safety. This information will also address the question of how the evolutionary-optimized chicken eggshell matrix proteins affect and regulate calcium carbonate mineralization as a good example of biomimetic and bio-inspired material design.

In this research work, the conventional single slope still (CSS) with egg shells of breed Gallus gallus domesticus cascara as sensible heat storage (SHS) material are studied experimentally to enhance the yield. In this experimental investigation, the proposed single slope still (PSS) with SHS material was made in comparison with the CSS to evaluate the productivity of fresh water under the same ambient conditions. Comparatively, this PSS has higher thermal conductivity than the CSS. The yield obtained from the PSS is 2.46 L/m 2 , while the yield from the CSS is 2.07 L/m 2 . The average rate at which the rise of output fresh water obtained from the PSS is 18% more than the fresh water output obtained from the CSS. The daily energy efficiency of the PSS is 26.07%, and for the CSS, it is only 22.25%. The daily exergy efficiency of the PSS is 2.36%, and for the CSS, it is only 1.67%. Since using the egg shell will employ as organic waste management and modification in this still is economical, less initial, and maintenance cost.

Chicken eggshell (ES) waste is a rich source of calcium carbonate (CaCO 3 ); however, the potential of ES as dietary calcium (Ca) in old laying hens has not been explored. This study compared the effects of feeding limestone, cockle shell, oyster shell, fine ES, and coarse ES as the sole Ca source on production performance, egg quality, blood biochemical constituents, and tibia characteristics in old laying hens. A total of 450 ISA-Brown laying hens at 73 wk of age with similar egg production rate (EPR) were randomly assigned to 5 treatment groups (90 hens/group, 9 hens/replicate) for 7 wk. Dietary treatment groups comprised a corn-soybean meal based diet containing different Ca sources: (i) limestone (LS; < 2 mm and 2–4 mm mixed in the ratio of 3:7) as control, (ii) cockle shell (CS; 1–4 mm), (iii) oyster shell (OS; 3–16 mm), (iv) ES fine particles (ESF; < 1 mm), and (v) ES coarse particles (ESC; 3–5 mm). Results indicated that dietary inclusion of coarse ES particles significantly increased average egg weight ( P  < 0.001) and daily egg mass ( P  < 0.05), and decreased feed conversion ratio ( P  < 0.001) as compared with the other treatments. However, no significant differences in EPR, feed intake, cracked egg proportion, and mortality were observed among the dietary treatments ( P  > 0.05). Notably, the use of ESF led to a lower proportion of cracked eggs than ESC ( P  < 0.05). ESC fed hens produced the heaviest eggs whereas CS fed hens produced the lightest ( P  < 0.001); the particle size of ES also affected the egg weight ( P  < 0.05). The eggs from OS and ESC fed hens showed a greater albumen height in comparison to eggs from CS group ( P  < 0.05); but no significant difference was observed among the LS, OS, ESF, and ESC groups (P > 0.05). The yolk color was darker in the eggs of group ESF as compared with other dietary groups ( P  < 0.01). However, no significant effects on Haugh units and shell properties were observed among the treatments ( P  > 0.05). The blood biochemistry results were not affected by the dietary Ca ( P  > 0.05) except for lower levels of high-density lipoprotein percentage (HDL %) in OS and ESC fed hens ( P  < 0.05). The tibia characteristics including weight, length, width, and breaking strength did not differ among the dietary groups ( P  > 0.05). However, the ESC and OS fed hens showed higher tibia bone mineral density (BMD) than the other groups ( P  < 0.001). In conclusion, coarse ES as a sole Ca source had beneficial effects on the production performance, egg quality, and tibia BMD in old laying hens.

Today’s awareness of environmental pollution and the idea of creating a reuse area for waste is one of the trend topics. The base opinion, regarding the reuse of any material is that all resources are inherently limited in the world. In the ethical approach, it is a moral issue that people take responsibility for future generations to protect nature. Eggshells are one of the most used natural biomasses. And natural harmless eggshells deserve to be researched to reveal their potential. The aim of this study, blending the characterization processes with the research published until now, finding criteria for the unique structures and application capacities of eggshell species, ensures choosing the right type as biomass in the industry, and directing the eggshell usage to the appropriate applications and industries. In many industries, finding benchmarks of eggshell types in their unique structures and application capacity gives a clue to selecting the right type and directing the eggshells to a suitable place. In this study, different species of eggshell ( Coturnix Coturnix Japonica , Anser Anser , Denizli Hen , Alectoris Chukar , and Struthio Camelus ) were characterized by XRD, FTIR, AFM, Stereo Microscope, SEM, XRF, and TGA analysis. Calcined forms of eggshell samples were characterized by XRD, FTIR, and XRF analysis. TGA analysis results are used as a precursor to determine the temperature of calcination (800–900°). XRD results show that the CaCO 3 peak is 2Ɵ=29.58° for all eggshells. The reason why this peak is not observed after the calcination process is that the entire CaCO 3 structure is converted to CaO. In FTIR results, the C-O stretching band which is observed at 1424 cm − 1 is the main characteristic band of selected eggshell species. When AFM images are examined, it is seen that the surface of small eggshells is rougher, while the surface of the eggshell becomes smoother as the egg size increases. SEM and stereo microscope images show that the shell thickness increases as the egg size increases.