Thickness Of Mammillary Layer Research Articles

Comparative Analysis of the Ultrastructure, Bubble Pores, and Composition of Eggshells of Dwarf Layer-White and Guinea Fowl.

The decrease in eggshell quality seriously affects production efficiency. Guinea fowl (GF) eggs possess strong eggshells because of their unique crystal structure, and few systematic studies have compared laying hen and GF eggs. Sixty eggs were collected from both 40-week-old Dwarf Layer-White (DWL-White) laying hens and GF, and the eggshell quality, ultrastructure, bubble pores, and composition were measured. The results showed that the DWL-White eggs had a higher egg weight and a lower eggshell strength, strength per unit weight, thickness, and ratio than the GF eggs (p < 0.01). There were differences in the mammillary layer thickness ratio, the effective layer thickness ratio, the quantity of bubble pores (QBPs), the ratio of the sum of the area of bubble pores to the area of the eggshell in each image (ARBE), and the average area of bubble pores (AABPs) between the DWL-White and GF eggs (p < 0.01). The composition analysis demonstrated that there were differences in the organic matter, inorganic matter, calcium, and phosphorus between the DWL-White and GF eggs (p < 0.01). There were positive associations between the mammillary knob number in the image and the QBPs and ARBE and a negative correlation with the AABPs in the DWL-White eggs (p < 0.01). This study observed distinctions that offer new insights into enhancing eggshell quality.

Effects of manganese glycine on eggshell quality, eggshell ultrastructure, and elemental deposition in aged laying hens

Poor eggshell quality of eggs laid by aged laying hens is the major problem affecting the length of the rearing period in the laying hen industry. Trace elements are required and play vital roles in the eggshell quality of laying hens. Appropriate dose of organic microelements is environmentally friendly and sufficient to satisfy the needs of hens because of their greater bioavailability and lower excretion than inorganic forms. The aim of this experiment was to investigate the effects of manganese (Mn) glycine (MG) on eggshell quality, elemental deposition, and eggshell ultrastructure in aged laying hens. A total of 720 Hy-Line Brown hens 70 weeks old were assigned equally to four groups with six replicates of 30 birds each. The hens were fed basal diets (without Mn supplementation) supplemented with 120 mg/kg of Mn from manganese sulfate monohydrate (MSM), or 40, 80, or 120 mg/kg Mn from MG for 12 weeks. Dietary supplementation with 80 mg/kg Mn from MG resulted in the greatest eggshell strength after 6 weeks of treatment (P = 0.047), and in greater eggshell strength than observed in the MSM control after 12 weeks of treatment (P = 0.025). After 12 weeks of treatment, the eggs of hens in the MG groups showed lower mammillary layer thickness in the blunt end, equator, and acute end than observed in the MSM control group (P < 0.001). With the exception of the blunt ends of eggs from hens in the 120 mg/kg MG group, the eggs of hens in the MG groups, compared with the MSM control group, exhibited a lower mammillary layer ratio, and greater palisade layer ratio and effective layer ratio in the blunt end, equator, and acute end (P < 0.001). Dietary supplementation with 80 mg/kg Mn from MG, compared with the MSM control and 40 and 120 mg/kg MG, resulted in the greatest palisade layer thickness and effective layer thickness, and the lowest mammillary layer thickness in the equator (P < 0.001, P = 0.001, P < 0.001, respectively). Furthermore, supplementation with 80 mg/kg Mn from MG exhibited the greatest ratio of the palisade layer and effective layer, and the lowest mammillary layer ratio in the blunt end and equator (all P < 0.001). The Mn content of eggshells in hens-fed diets supplemented with 80 and 120 mg/kg Mn from MG was greater than that in the MSM control and 40 mg/kg MG groups (P = 0.035). Dietary supplementation with 80 or 120 mg/kg Mn from MG resulted in greater tibia Mn content than observed in the 40 mg/kg MG group (P = 0.019), and greater yolk Mn content than observed in the 40 mg/kg MG and MSM control groups (P = 0.018). In conclusion, dietary supplementation with 80 mg/kg Mn from MG, compared with the MSM control (120 mg/kg Mn), may increase the deposition efficiency of Mn, alter eggshell elemental composition, improve eggshell ultrastructure, and enhance eggshell strength in aged laying hens.

산란후기 사료 내 가수분해 효모의 첨가 급여가 생산성과 계란 품질에 미치는 영향

An experiment was conducted to investigate the effects of varying levels of hydrolyzed yeast on egg production and egg quality in aged laying randomly allotted to three dietary treatments such that egg production was similar in each treatment (6 replicates of 10 birds each). The layers were fed diets containing 0, 0.1, or 0.2% hydrolyzed yeast for eight weeks. No significant difference was observed in egg production during the first half of the experiment. Egg production and daily egg mass in groups fed diets containing hydrolyzed yeast were significantly higher (<i>P</i>&lt;0.05) than those of the control groups during the second half of the experiment. Egg weight was not affected by the dietary treatment. Eggshell strength and thickness in groups fed diets containing hydrolyzed yeast were significantly higher than those of the control groups during the overall experimental period (<i>P</i>&lt;0.05). Although no significant differences were observed in the Haugh units, yolk color in the group fed diets containing 0.1% hydrolyzed yeast was significantly higher than that in the control group (<i>P</i>&lt;0.05). The mammillary layer thickness increased in a linear manner and significantly following treatment with dietary hydrolyzed yeast (<i>P</i>&lt;0.05). Antibody titer against avian influenza virus in the group fed diets containing 0.2% hydrolyzed yeast was significantly higher (<i>P</i>&lt;0.05) than that in the control group. In conclusion, dietary hydrolyzed yeast improved egg production and eggshell quality of laying hens in the late stages of production.

Genetic variations for the eggshell crystal structure revealed by genome-wide association study in chickens

BackgroundEggshell is a bio-ceramic material comprising columnar calcite (CaCO3) crystals and organic proteinaceous matrix. The size, shape and orientation of the CaCO3 crystals influence the microstructural properties of chicken eggshells. However, the genetic architecture underlying eggshell crystal polymorphism remains to be elucidated.ResultsThe integral intensity of the nine major diffraction peaks, total integral intensity and degree of orientation of the crystals were measured followed by a genome-wide association study in 839 F2 hens. The results showed that the total integral intensity was positively correlated with the eggshell strength, eggshell thickness, eggshell weight, mammillary layer thickness and effective layer thickness. The SNP-based heritabilities of total integral intensity and degree of orientation were 0.23 and 0.06, respectively. The 621 SNPs located in the range from 55.6 to 69.1 Mb in GGA1 were significantly associated with TA. PLCZ1, ABCC9, ITPR2, KCNJ8, CACNA1C and IAPP, which are involved in the biological process of regulating cytosolic calcium ion concentration, can be suggested as key genes regulating the total integral intensity.ConclusionsThe findings greatly advance the understanding of the genetic basis underlying the crystal ultrastructure of eggshell quality and thus will have practical significance in breeding programs for improving eggshell quality.

The partial replacement of sodium chloride with sodium bicarbonate or sodium sulfate in laying hen diets improved laying performance, and eggshell quality and ultrastructure

This study investigated the effects of dietary chloride (Cl) reduction on laying performance and eggshell quality by substitution of sodium bicarbonate (NaHCO3) or sodium sulfate (Na2SO4) for part of dietary sodium chloride (NaCl), and further explored its mechanism for improving eggshell quality. A total of 360 29-wk-old Hy-line Brown laying hens were randomly allocated to 5 dietary treatments, including a basal diet contained 0.33% NaCl (control group, 0.27% dietary Cl), and 4 experimental diets that contained 0.21% and 0.15% dietary Cl by substituting Na2SO4 or NaHCO3 for part of NaCl in the basal diet. No significant differences were observed in blood Na+, Cl−, K+ and Ca2+ levels and pH value as well as serum creatinine and uric acid contents among 5 treatments (P > 0.05). Dietary Cl reduction increased egg production and ADFI during wk 33 to 36, 37 to 40 and 29 to 40 of age and decreased feed conversion ratio during wk 37 to 40 of age (P < 0.05). The hens fed with diets containing 0.15% Cl increased eggshell breaking strength, thickness and weight ratio in wk 40 of age (P < 0.05). Birds fed with dietary 0.21% and 0.15% Cl exhibited higher effective layer thickness and lower mammillary layer thickness of eggshell than those fed with dietary 0.27% Cl (P < 0.05). Apparent Ca metabolizability of hens was increased with dietary Cl reduction (P < 0.05). Total Ca of eggshell of dietary 0.15% Cl group was higher than that of dietary 0.27% Cl group (P < 0.05). No significant differences in laying performance, eggshell quality and Ca metabolism of layers were observed between Na2SO4 and NaHCO3 replacement groups (P > 0.05). Overall, dietary Cl reductions could improve laying performance and eggshell quality by substitution of NaHCO3 or Na2SO4 for part of NaCl, and there were no differences in the improvements between these two substitutes. The improved eggshell quality may be attributed to improved eggshell ultrastructure and increased supply of eggshell Ca2CO3.

Uterine transcriptome analysis reveals mRNA expression changes associated with the ultrastructure differences of eggshell in young and aged laying hens

BackgroundLower eggshell quality in the late laying period leads to economic loss. It is a major threat to the quality and safety of egg products. Age-related variations in ultrastructure were thought to induce this deterioration. Eggshell formation is a highly complex process under precise regulation of genes and biological pathways in uterus of laying hens. Herein, we evaluated the physical, mechanical and ultrastructure properties of eggshell and conducted RNA sequencing to learn the transcriptomic differences in uterus between laying hens in the peak (young hens) and late phase (aged hens) of production.ResultsThe declined breaking strength and fracture toughness of eggshell were observed in aged hen group compared to those in young hen group, accompanied with ultrastructure variations including the increased thickness of mammillary layer and the decreased incidence of early fusion. During the initial stage of eggshell formation, a total of 183 differentially expressed genes (DEGs; 125 upregulated and 58 downregulated) were identified in uterus of laying hens in the late phase in relative to those at peak production. The DEGs annotated to Gene Ontology terms related to antigen processing and presentation were downregulated in aged hens compared to young hens. The contents of proinflammatory cytokine IL-1β in uterus were higher in aged hens relative to those in young hens. Besides, the genes of some matrix proteins potentially involved in eggshell mineralization, such as ovalbumin, versican and glypican 3, were also differentially expressed between two groups.ConclusionsAltered gene expression of matrix proteins along with the compromised immune function in uterus of laying hens in the late phase of production may conduce to age-related impairments of eggshell ultrastructure and mechanical properties. The current study enhances our understanding of the age-related deteriorations in eggshell ultrastructure and provides potential targets for improvement of eggshell quality in the late laying period.

EFFECT OF NANO DICALCIUM PHOSPHATE SUPPLEMETATION ON EGGSHELL STRUCTURE AND CALCIUM AND PHOSPHORUS UTILIZATION

This study was conducted to evaluate the efficiency of dietary varying levels of nano di-calcium phosphate (NDCP) on eggshell quality [total thickness (EST), mammillary layer thickness (MT), and mammillary density (MD)] egg mineral contents, bone breaking strength, serum calcium and phosphorus concentrations and calcium (Ca) and phosphorus (P) retention in old laying hen. Therefore, one hundred and eighty brown LSL hens at the age of 52 weeks of age were used for 12 weeks at the end of their production period. Hens were divided into four equal groups, 45 birds/group with nine replicates each. The treatments were as follows: Group (1): basal diet (control); Group (2): basal diet + 200g NDCP/ ton; Group (3): basal diet + 400g NDCP/ ton; and Group (4): basal diet + 800g NDCP/ ton. Results detected that serum Ca and P concentration, tibia fresh weight, tibia ash, tibia Ca, Ca and P retention were significantly increased stuck with the increase in NDCP supplementation level up to 400 g / ton and tend to decrease after that. Furthermore, Yolk mineral content (Ca and p) accumulation was significantly decreased with increasing NDCP levels. There were no significant differences in tibia breaking strength and tibia P with the supplementation in comparison with the control group. Eggshell thickness and breaking strength increased parallel to the increase in NDCP levels due to the increase in palisade layer and mammillary layer. In conclusion supplementation of NDCP up to 400 g/ ton to layers diet may be help in solving the problems of egg shell weakness at the end of egg laying season or old hen production.

Gene Polymorphisms are Associated with Eggshell Ultrastructure Organization in Hens

ABSTRACT Background: Eggshell ultrastructure organization, including effective layer thickness, mammillary layer thickness, and average size of mammillary cones, is important for breeding and significantly influences eggshell mechanical properties. Several matrix proteins were known to be important in eggshell formation. However, the proteins and variations that determine eggshell ultrastructure organization are not known. Results: In this study, 17 single-nucleotide polymorphisms of three major genes in a hen population using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Five single-nucleotide polymorphisms with a very low minor allele frequency (< 1%) were excluded from further analysis. The remaining 12 single-nucleotide polymorphisms in Hardy-Weinberg equilibrium were used for analysis of associations with eggshell ultrastructure organization. Associations were found for (i) ovocleidin-116 with effective layer thickness (EFF), mammillary layer thickness (MAM), and average size of mammillary cones (SMAM); (ii) ovalbumin with eggshell thickness (ESH), effective layer thickness, and density of the mammillary cone (DMAM); and (iii) calmodulin1 with density of the mammillary cone. Conclusions: The single-nucleotide polymorphisms identified in the present study may be used as potential markers to improve eggshell quality.

Genetic architecture dissection by genome-wide association analysis reveals avian eggshell ultrastructure traits.

The ultrastructure of an eggshell is considered the major determinant of eggshell quality, which has biological and economic significance for the avian and poultry industries. However, the interrelationships and genome-wide architecture of eggshell ultrastructure remain to be elucidated. Herein, we measured eggshell thickness (EST), effective layer thickness (ET), mammillary layer thickness (MT), and mammillary density (MD) and conducted genome-wide association studies in 927 F2 hens. The SNP-based heritabilities of eggshell ultrastructure traits were estimated to be 0.39, 0.36, 0.17 and 0.19 for EST, ET, MT and MD, respectively, and a total of 719, 784, 1 and 10 genome-wide significant SNPs were associated with EST, ET, MT and MD, respectively. ABCC9, ITPR2, KCNJ8 and WNK1, which are involved in ion transport, were suggested to be the key genes regulating EST and ET. ITM2C and KNDC1 likely affect MT and MD, respectively. Additionally, there were linear relationships between the chromosome lengths and the variance explained per chromosome for EST (R2 = 0.57) and ET (R2 = 0.67). In conclusion, the interrelationships and genetic architecture of eggshell ultrastructure traits revealed in this study are valuable for our understanding of the avian eggshell and contribute to research on a variety of other calcified shells.

Revisiting Sabath's "Larger Avian Eggs" from the Gobi Cretaceous

In 1991, Sabath described “larger avian eggs” from the Upper Cretaceous Barun Goyot and Djadokhta Formations of Mongolia. These were later included in the ootaxon Gobioolithus major. Here we recognize the larger avian eggs of Sabath as a distinct ootaxon, Styloolithus sabathi, oogen. et oosp. nov. These eggs differ from those of Gobioolithus in being larger (70 by 32 mm) and more elongate. Microscopically, the shell bears a third layer (possible external zone) thicker than the mammillary layer and nearly as thick as the second layer (possible squamatic zone); the continuous layer (including layers two and three) to mammillary layer thickness ratio is 3.1:1. Within the clutch, the tightly spaced eggs stand with their long axes steeply inclined. Adult remains are associated with two clutches, suggesting an incubation mode similar to that of troodontid maniraptorans, where adults sat atop largely buried eggs. S. sabathi provides evidence that relative egg size in Mesozoic non-ornithuromorph birds had increased markedly from the non-avian theropod condition in oviraptorids and troodontids, but had not yet reached the modern egg-adult proportions of Neornithes. Sediment-bound upright eggs appear common to Enantiornithes and more basal avians, suggesting that like non-avian theropods, these birds lacked chalazae, the chords of albumen allowing egg rotation in modern birds. Absence of this simple structure may have restricted these basal birds to ground nesting in areas with appropriate substrates and not permitted the type of nesting diversity found in Neornithes. Neornithes are the only Mesozoic clade of Dinosauria to nest completely free of sediment; this may have played a crucial role in their surviving the K-Pg mass extinction event.

Influence of eggshell ultrastructural organization on hatchability

Eggshell quality is associated with hatchability, and ultrastructural organization is an important eggshell quality parameter. To the determine the relationship of shell ultrastructural properties with hatchability, we measured the effect of eggshell thickness, mammillary layer thickness, and the average size of mammillary cones on hatchability. Eggs with thick shells and thick mammillary layers had the highest incidence of hatching. As such, hatchability correlated positively with both eggshell thickness (r = 0.30; P < 0.05) and mammillary layer thickness (r = 0.28; P < 0.05). In addition, the correlation between the average size of mammillary cones and hatchability was not statistically significant. However, a significant positive correlation was observed between the average size of mammillary cones and mammillary layer thickness. It can be concluded that eggshell thickness and mammillary layer thickness affect hatchability.

Polymorphisms in eggshell organic matrix genes are associated with eggshell quality measurements in pedigree Rhode Island Red hens

Novel and traditional eggshell quality measurements were made from up to 2000 commercial pedigree hens for a candidate gene association analysis with organic eggshell matrix genes: ovocleidin-116, osteopontin (SPP1), ovocalyxin-32 (RARRES1), ovotransferrin (LTF), ovalbumin and ovocalyxin-36, as well as key genes in the maintenance and function of the shell gland [estrogen receptor (ESR1) and carbonic anhydrase II (CAII)]. Associations were found for (i) ovalbumin with breaking strength and shell thickness; (ii) ovocleidin-116 with elastic modulus, shell thickness and egg shape; (iii) RARRES1 with mammillary layer thickness; (iv) ESR1 with dynamic stiffness; (v) SPP1 with fracture toughness and (vi) CAII with egg shape. The marker effects are as large as 17% of trait standard deviations and could be used to improve eggshell quality.

Changes in egg shell characteristics of the Sparrowhawk (Accipiter nisus) and Peregrine (Falco peregrinus) associated with exposure to environmental pollutants during recent decades

In Britain since the late 1940s, the Sparrowhawk (Accipiter nisus) and Peregrine (Falco peregrinus) have laid eggs with unusually thin shells. DDT and its metabolites have been blamed. This paper reports how various properties of modern shells differ from those of older shells of normal thickness.For the shells of both species, there were roughly proportional reductions in thickness of the two main component layers, the mammillary and palisade layers, suggesting that thin shells resulted from a decreased rate of deposition. In addition, in modern Sparrowhawk shells, the resistant surface layer was especially deficient perhaps due to a further reduction in deposition rate or to slight premature termination. Shell thinning was associated with a proportionately greater decrease in shell strength. For the Peregrine, porosity, measured as the rate of passage of water vapour, was lower for the thin shells than for normal shells. This difference could not be explained by a change in the area of the pore channels, since the modern shells did not have fewer pores and the pore channels were the same size and shape in the two samples of shells. However, in the thin shells of both species, mammillae height increased relative to mammillary layer thickness, and in the Peregrine shells the extent of this increase was related to the change in porosity.

Egg shell characteristics and incidence of shell breakage for grey herons Ardea cinerea exposed to environmental pollutants

Egg shell characteristics and incidences of shell breakage have been studied for the grey heron Ardea cinerea at several colonies in eastern England. Eggs collected in the late 1960s and in the 1970s had shells which were significantly thinner than shells collected before 1946. For a sample taken in 1973, shell thickness, shell strength, mammillary layer thickness and palisade layer thickness were all inversely related to the concentration of pp'-DDE(+ pp'-DDT + pp'-TDE) in the egg contents. Shell thinning was considered to be due to a reduced availability of shell components leading to a depressed rate of shell formation. A separate mechanism resulted in increased mammillae height relative to mammillary layer thickness. For comparison, the structure of other types of thin shells was also examined. Incidence of egg shell breakage was positively related to the residue levels of pp′-DDE or dieldrin in intact eggs. Eggs with shells thinner than 240 μm were unlikely to survive incubation. Since such eggs tend to contain higher-than-average residues, sampling intact eggs leads to a mean egg residue figure that is lower than the real mean for the colony. Measuring shell thickness is suggested as a rapid and economical bioassay of DDE levels. Shell thinning leads to a proportionally greater reduction in shell strength, and breakage of shells less than 240 μm in thickness was probably due to a combination of shell fragility and aberrant behaviour by the brooding adults. Breakage of thicker shells was believed to be mainly due to deliberate destruction. Despite more than half of the pairs breaking their shells during some years, heron populations have not declined overall perhaps because of the ability of this species to lay repeat clutches. There are indications that egg residues of DDE and other organochlorines are decreasing, and shell thickness and the incidence of shell breakage are returning to normal.