Eggshell Quality Traits Research Articles

Ovomucoid gene polymorphism and its influence on quality changes at various storage timepoint of eggs from two strains of Japanese quail

During storage, irreversible changes occur in eggs, resulting in a decline in their quality, predominantly affecting the albumen. Ovomucoid, a major protein found in egg white, belongs to the Kazal-type serine proteinase inhibitors and serves to protect the embryo from microorganisms. Notably, in chicken eggs, it is a significant allergen. There is a possibility that its polymorphism also influences the quality and stability of table eggs. Hence, this study aimed to evaluate the potential effect of polymorphism in the ovomucoid gene and protein on quality changes during the storage of eggs derived from 2 strains of Japanese quail, encompassing various utility types. Eggs from selected females of laying and meat-type breeds were stored for 14 wk, with egg quality traits assessed 10 times during this duration. DNA was isolated from each female, and sequencing was conducted on all exons of the ovomucoid gene. In total, 5 SNPs were identified in exons and adjacent intronic sequences, with SNP1 (13:12355585), SNP4 (13:12356594), and SNP5 (13:12358538) leading to amino acid substitutions in the ovomucoid protein. Notably, all SNPs except SNP5 were identified in the ovomucoid gene of Japanese quail for the first time. The results demonstrated that in the F33 strain, SNP1, SNP3, and SNP4 exhibited significant associations with egg weight, whereas in the S22 strain, SNP5 significantly affected yolk color and various eggshell quality traits, including eggshell weight, eggshell thickness, and breaking strength, throughout the storage period. Furthermore, a haplotype block containing 2 SNPs (3 and 4) was identified, exhibiting 2 distinct haplotypes that significantly affected egg weight, eggshell weight, and breaking strength at various storage time points during egg quality analyses. These findings provide novel insights into the genetic basis of egg quality during storage and have the potential to be integrated into breeding programs for these strains.

CYP24A1 and TRPC3 Gene Expression in Kidneys and Their Involvement in Calcium and Phosphate Metabolism in Laying Hens.

Ca and P homeostasis across the egg-laying cycle is a complex process involving absorption in the small intestine, reabsorption/excretion in the kidneys, and eggshell gland secretion. Diets with inadequate calcium and phosphorus can interfere with their absorption and digestibility, resulting in eggshell quality losses and reduced productive life, affecting egg production and welfare. A better understanding of gene expression profiles in the kidneys of laying hens during the late egg-laying period could clarify the renal role in mineral metabolism at this late stage. Therefore, the performance, egg quality and bone integrity-related traits, and expression profiles of kidney candidate genes were evaluated in 73-week-old laying hens receiving different Ca and P ratios in their diet: a high Ca/P ratio (HR, 22.43), a low ratio (LR, 6.71), and a medium ratio (MR, 11.43). The laying hens receiving the HR diet had improved egg production and eggshell quality traits compared to the other two groups. Humerus length was shorter in the HR than in the other groups. The CYP24A1 and TRPC3 genes were differentially expressed (p.adj ≤ 0.05) among the groups. Therefore, their expression profiles could be involved in calcium and phosphate transcellular transport in 73-week-old laying hens as a way to keep mineral absorption at adequate levels.

Genetic patterns and genome-wide association analysis of eggshell quality traits of egg-type chicken across an extended laying period

The industry of egg-type chicken has shown a trend of extending the rearing period, with the goal of breeding chicken breeds capable of producing 500 qualified eggs by 700 d of age. However, the rapid decline in eggshell quality during the late laying period is one of the major challenges. In this study, a total of 3,261 Rhode Island Red chickens were used to measure eggshell quality traits including eggshell strength (ESS), eggshell thickness (EST), eggshell color (ESC) and eggshell gloss (ESG) at seven age points ranging from 36 to 90 wk of age. Phenotypic variations increased with the aging process, especially during the late laying period (> 55 wk), and the heritability during this period decreased by 22.7 to 81.4% compared to the initial and peak laying periods. Then we performed genome-wide association study (GWAS) to identify the genomic variants that associated with eggshell quality, with a custom Illumina 50K BeadChip, named PhenoixChip-I. The results indicated that 2 genomic regions on GGA1(23.24-25.15Mb; 175.95-176.05 Mb) were significantly (P < 4.48E-06) or suggestively (P < 8.97E-05) associated with ESS, which can explain 9.59% and 0.48% of the phenotypic variations of ESS46 and ESS36, respectively. Three genes, FRY, PCNX2, and ENSGALG00000052468, were considered to be the candidate genes for ESS. For other traits, the genome-wide suggestive SNPs were identified at each age point, exhibiting a certain trend with aging process. Additionally, SNP enrichment analysis and functional annotation of cross-tissue regulatory elements to ESS36 revealed a high concentration of enhancer elements specific to shell gland and kidney tissues. This study, deepened our knowledge of eggshells and laying a valued scientific foundation for chicken molecular breeding.

Genome-wide association study exploring the genetic architecture of eggshell speckles in laying hens

BackgroundEggshell speckle phenotype is an important trait in poultry production because they affect eggshell quality. However, the genetic architecture of speckled eggshells remains unclear. In this study, we determined the heritability of eggshell speckles and conducted a genome-wide association study (GWAS) on purebred Rhode Island Red (RIR) hens at 28 weeks to detect potential genomic loci and candidate genes associated with eggshell speckles.ResultsThe heritability of eggshell speckles was 0.35 at 28 weeks, and the speckle level is not related to other eggshell quality traits in terms of phenotypic correlation. We detected 311 SNPs (6 significantly, and 305 suggestively associated) and 39 candidate genes associated with eggshell speckles. Based on the pathway analysis, the 39 candidate genes were mainly involved in alpha-linolenic acid metabolism, linoleic acid metabolism, ether lipid metabolism, GnRH signaling pathway, vascular smooth muscle contraction, and MAPK signaling pathway. Ultimately, ten genes, LOC423226, SPTBN5, EHD4, LOC77155, TYRO3, ITPKA, DLL4, PLA2G4B, PLA2G4EL5, and PLA2G4EL6 were considered the most promising genes associated with eggshell speckles that were implicated in immunoregulation, calcium transport, and phospholipid metabolism, while its function in laying hens requires further studies.ConclusionsThis study provides new insights into understanding the genetic basis of eggshell speckles and has practical application value for the genetic improvement of eggshell quality.

Genetic parameters for production, quality, and colors from eggs in Brazilian lineages of chickens.

Genetic parameters were estimated for egg production, egg quality, and eggshell colors in eight lineages of Brazilian laying hens. Age at first egg (AFE), total egg production up to the 45th week (PROD), egg weight (EW), albumen height (AH), yolk color (YC), the Haugh units (HU), eggshell strength (ESS), eggshell thickness (EST), yolk weight (YW), eggshell weight (ESW), and eggshell color (L*, a*, and b*) were measured in 2030 eggs obtained from 645 laying hens. Variance components were estimated from a mixed animal model, which included the fixed effects of contemporary groups, cage location, and hen line, and the additive genetic, permanent environmental, and residual as random effects. In general, heritabilities were low to moderate (h2 = 0.11 to 0.48). Genetic correlations among eggshell quality traits were moderate to high (0.36 and 0.69). High genetic correlations were obtained between the eggshell color traits [rg = -0.90 (L* and a*); rg = -0.64 (L* and b*); and rg = 0.65 (a* and b*)]. Results suggest that EW is strongly correlated with ESW, but the genetic correlations between EW and ESS and between EW and EST were low. Genetic correlations between L* and eggshell quality traits were low to moderate, suggesting that L* has little or no relation with external egg quality. However, genetic correlations between a* and b* values and eggshell quality traits were high. The genetic correlations between eggshell color and eggshell quality traits were low, suggesting that the eggshell color does not influence external egg quality. Genetic correlations between PROD and egg quality traits were negative and varied between -0.42 and -0.05. This antagonistic relationship emphasizes the importance of adopting breeding schemes that allow the simultaneous genetic progress of these traits by considering their genetic correlation and economic relevance, such as the selection index.

Effect of organic and inorganic manganese supplementation on performance and eggshell quality in aged laying hens.

Manganese (Mn) is an important trace element for laying hen's nutrition, which is required in small amounts in the diet. Its deficiency results in lowered production performance and eggshell quality. This experiment was conducted to investigate the influence of different sources and levels of Mn on egg production performance and eggshell quality in aged laying hens. A total of 720, 83-week-old Hy-Line W-36 laying hens were fed a non-Mn supplemented basal diet for 4-week (to ime Mn-exhaustion of body) and then were allocated to a completely randomized design with 10 treatments, six replicates and 12 birds each. Concentration of Mn in the non-Mn supplemented basal diet was 10.34 mg/kg (treatment 1), the added doses of dietary Mn were included 30, 60, and 90mg/kg of three different sources (Mn-oxide, Mn-sulphate, and Mn-organic) for treatments 2-10, respectively. The experiment lasted for 12 week. Dietary supplementation with either organic or inorganic Mn sources significantly enhanced egg production (EP), egg mass (EM), feed conversion ratio (FCR), and relative eggshell weight (RESW) compared with the non-Mn supplemented diet. However, the experimental diets did not influence feed intake (FI), egg weight, and other eggshell quality traits. Based on the broken line regression models, the performance traits were optimized at 30-40mg/kg Mn concentration when supplemented by Mn-sulphate or Mn-organic. Although, it was 80-90mg/kg when supplemented by Mn-oxide. The relative bio-efficacy of inorganic Mn sources include Mn-oxide and Mn-sulphate in compare with Mn-organic were estimated 45% and 87% (for EP trait), 30% and 94% (for EM trait), 36% and 99% (for FCR trait), and 37% and 78% (for RESW trait), respectively. In the aged laying hens, Mn requirement is higher than the NRC's recommendation. Sulphate and organic sources of Mn are more effective than Mn-oxide.

Guinea fowl (Numida meleagris) eggs and free-range housing: a convenient alternative to laying hens' eggs in terms of food safety?

The aim of this study was to evaluate the impact of the genotype (guinea fowl, native breed Leghorn, and commercial hybrid hens), storage time (0, 14, 28 d) and storage temperature (fresh, 5, 20°C) on eggshell quality traits and microbiological contamination of eggshell, eggshell membranes, and albumen. A total of 150 hens (50 hens per genotype—divided into 2 equal groups because of the results replication) were used. There were 150 eggs (50 per genotype) used for microbial analysis and 600 eggs used for the analysis of eggshell quality. The effects of genotype, storage time, and storage temperature were observed. Moreover, interactions between these factors were calculated. The significant effect of genotype (P = 0.0001) was found in egg weight, in all observed parameters of eggshell quality (proportion, thickness, strength, surface, and index), eggshell contamination of Escherichia coli (EC) and total number of micro-organisms (TNM), penetration of TNM into eggshell membranes (P = 0.0014), and penetration of TNM into albumen (P = 0.0019). Storage time significantly affected egg weight and all parameters of eggshell quality except the eggshell strength and index. It also significantly affected count of Enterococcus (ENT) on eggshell, TNM in eggshell membranes and TNM in albumen. Storage temperature significantly influenced egg weight (P = 0.0001) and all parameters but eggshell thickness and surface. Regarding the microbial contamination, storage temperature significantly affected a count of ENT on shell, TNM in shell membranes, and TNM in albumen. Concerning significant interactions, the interaction among genotype and storage time was found significant (P = 0.0148). Fresh and 28-day-old commercial hybrid eggs were the most contaminated, whereas guinea fowl eggs (fresh and 14 d old) and Leghorn hen eggs (fresh, 14, 28 d old) had the lowest level of contamination by EC. When looking for an alternative to laying hens, guinea fowls should be taken into consideration due to their higher resistance to diseases, ability of adaptation to different environmental conditions, and especially in terms of eggshell quality and therefore egg safety.

The effect of breed and age on the gloss of chicken eggshells

The monitoring of eggshell quality is important mainly in terms of production economy. Eggshell appearance is one of the most characteristics, influencing the purchasing behavior of consumers. Besides numerous eggshell appearance quality (color, shape, etc.), gloss is an important trait to reflect the eggshell appearance. In this study, 2 experiments were conducted to investigate the effect of breed and age on the gloss of eggshells. In experiment 1, we compared the eggshell gloss of 7 chicken breeds. In experiment 2, 105 Wanan (WA) chickens were raised, and 1 egg was collected from each individual at 26, 32, 40, and 50 wks of age. Eggshell gloss, color (L*, a*, b*), cuticle coverage (ΔE*ab), and thickness were measured. The results of experiment 1 showed that the average gloss values were highly variable among different breeds, and the highest was found in WA (gloss unit [GU] = 8.12), almost 2.5 folds as many as the lowest in Rhode Island Red (GU = 3.23). Also, the eggshell gloss of the local chicken breeds was significantly higher than the highly selected lines of egg-type chicken breeds (P < 0.001). In experiment 2, the results showed that gloss ranged from 9.08 GU to 12.12 GU with a variation of 28.38 to 39.71%. It fluctuated with the increasing age of hens and had the peak value at 26 wk. But, the correlation analysis between eggshell gloss and other eggshell quality traits were very low (−0.07 to 0.25). This study laid a foundation for improving the uniformity and intensity of eggshell gloss for breeders.

Genetic improvement of egg laying traits in Fayoumi chickens bred under conditions of heat stress through selection and gene expression studies.

Thermal stress has been shown to result in decreased egg production, decreased eggshell quality, and ultimately millions of dollars in losses to the industry. Therefore, there are many factors to consider when implementing genetic selection programs aimed at improving egg production under tropical conditions. So, trial is trying to improve the productivity and eggshell quality traits of the Fayoumi chicken under high ambient temperatures via selection programs and gene expression. In the present study, day-old Fayoumi chicks were raised either under normal temperature (control) or conditions of thermal stress (the heated group). At 35 weeks, male and female chickens from the control group were mated randomly and females selected for higher egg production and eggshell strength were mated to male siblings to obtain the progeny of the first generation (F1). F1 birds were further selected and mated to obtain the progeny of the second generation. Our results show that egg production and eggshell strength traits improved over successive generations via selection under conditions of heat stress. Furthermore, the reduction in egg production and eggshell strength as a result of heat stress declined from one generation to the next in birds selected for good heat tolerance, and an inverse relationship was observed between the OC-17 and eggshell strength. Additionally, levels of HSP90 and gene expression increased in the two successive generations, indicating that both productivity and heat tolerance were enhanced due to selection in birds raised under conditions of thermal stress. Moreover, generation exerted an important effect on this trait. Thus, desirable traits such as improved heat tolerance in producing lines were observed in Fayoumi chickens exposed to conditions of thermal stress via selection. Therefore, modern advances in studies of poultry breeding and genetics, such as gene expression studies, should be examined.

Long term effect of dietary lipid saturation on eggshell quality and bone characteristics of laying hens.

This study investigated the long-term effect of dietary fatty acid saturation on eggshell quality and bone characteristics of laying hens at end of lay. Five isoenergetic (3,011.5 kcal AME/kg DM) and isonitrogenous (169 g CP/kg DM) diets were formulated using different lipid sources at a constant 30 g/kg inclusion level. The control, polyunsaturated omega-3 (PUFA n-3), polyunsaturated omega-6 (PUFA n-6), monounsaturated omega-9 (MUFA n-9), and saturated fatty acid (SFA) diets consisted of linseed- and fish oil (50:50 blend), fish oil, sunflower oil, high oleic acid sunflower oil, and tallow, respectively. A total of 200 individually caged Hy-Line Silver-Brown hens (20 wk of age) were randomly allocated to the 5 treatments (n = 40 replicates/treatment) and received the experimental diets for 54 wk. During weeks 58, 62, 66, 70, and 74 of age (end-of-lay period), 20 eggs/treatment per day (n = 140 eggs/treatment per week) were selected for determining eggshell quality traits. At 74 wk of age, 10 birds per treatment (n = 10 birds/treatment) were randomly selected for the determination of bone quality characteristics. Data were statistically analyzed (P < 0.05) using a fully randomized 1-way ANOVA. Dietary treatment had no effect (P > 0.05) on eggshell quality traits. The MUFA n-9 treatment with the highest unsaturated to SFA ratio (UFA: SFA) resulted in a higher (P < 0.05) femur weight (10.34 g) as well as femur- (52.99%) and tibia ash (51.07%) content than the SFA treatment. Also, the PUFA n-3 diet resulted in a higher (P < 0.05) femur weight (10.21 g), femur ash (51.82%), and percentage femur (0.57%) compared to the SFA diet. Results suggested that prolonged feeding of diets varying in FA profile had no negative effect on eggshell quality, whereas UFA: SFA and long-chain n-3 PUFA affected the bone quality (especially the femur) of hens at the end of lay.

Validated quantitative trait loci for eggshell quality in experimental and commercial laying hens.

Compromised eggshell quality causes considerable economic losses for the egg industry. Breeding for improved eggshell quality has been very challenging. Eggshell quality is a trait that would greatly benefit from marker-assisted selection, which would allow the selection of sires for their direct contribution to the trait and would also allow implementation of measurements integrating a number of shell parameters that are difficult to measure. In this study, we selected the most promising autosomal quantitative trait loci (QTL) affecting eggshell quality on chromosomes 2, 3, 6 and 14 from earlier experiments and we extended the F2 population to include 1599 F2 females. The study was repeated on two commercial populations: Lohmann Tierzucht Rhode Island Red line (n=692 females) and a Hy-Line White Plymouth Rock line (n=290 progeny tested males). We analyzed the selected autosomal QTL regions on the three populations with SNP markers at 4-13 SNPs/Mb density. QTL for eggshell quality were replicated on all studied regions in the F2 population. New QTL were detected for eggshell color on chromosomes 3 and 6. Marker associations with eggshell quality traits were validated in the tested commercial lines on chromosomes 2, 3 and 6, thus paving the way for marker-assisted selection for improved eggshell quality.

Nutritional Quality of Eggs of Amberlink and Hyline Layers Fed on Different Levels of Provitamin A-Biofortified Maize

ABStRACt The study was conducted to determine the interaction of types of maize (Provitamin A-biofortified maize (PABM) versus white maize) and strain of laying birds (Amberlink and Hyline) on nutritional quality of eggs. Twenty-one of each of Amberlink and Hyline laying hens were fed on three diets for 30 days. Birds were distributed in a 3 × 2 factorial arrangement constituting three diets (0, 50 or 100 % of PABM) and two strains (Amberlink & Hyline). There was a diet × strain interaction on egg shell weight, average daily feed intake, egg production, egg weight and egg shell thickness. Eggs produced by layers under 100 % PABM had lighter eggshell weight and lower eggshell percentage. In contrast, eggs produced by layers under 50 % PABM diet had signi-ficantly higher eggshell percentage, heavier egg shell weight and thicker eggshell. As the level of PABM increased, the yellow and red hue (Hunter a* and b* values) significantly increased while the lightness values (Hunter L* values) decreased. As the level of PABM in the ration increased, the vitamin A content of the egg yolk also significantly increased. Assimilation of vitamin A from feed to egg yolk in Amberlink and Hyline hens was similar. Hence, egg enrichment with vitamin A can be achieved by using PABM in layers ration. The use of high level of PABM had a negative effect on the eggshell quality traits.

Genetic parameters of body weight, egg production, and shell quality traits in the Shan Ma laying duck (Anas platyrhynchos)

Three hundred and four female ducks of the Chinese indigenous Shan Ma breed, progeny of 11 sires and 104 dams, were used to study laying traits. Among them, 264 ducks were used to study the egg shell quality traits of eggs laid at 300days of age. The mean age at first egg was 109days with an average egg weight of 49.6±3.7g. Between 210 and 300days of age, egg weight increased from 65.0±3.9g to 67.0±4.2g and the mean of the number of eggs laid up to 300days was 161±15.0. Egg length was 59.57±3.01mm and egg width was 45.02±1.98mm, leading to a shape index of 1.32±0.08. Egg shell thickness was about 0.31mm whatever the shell region, and the breaking strength was 28.80±8.29N. The heritability's estimated using restricted maximum likelihood ( REML: ) methodology were high for egg weights (ranging from 0.43 to 0.61), intermediate for the number of eggs laid (ranging from 0.38 to 0.43), and low for the age at first egg (0.13). Heritability's for egg shell quality traits varied from 0.20 for the breaking strength to 0.44 for egg length, with in-between values of 0.28 for shell thickness and 0.34 for the shape index. The number of eggs laid was not genetically correlated with the age at first egg or egg weight, but was correlated with body weight (rg = +0.54±0.23). High positive correlations were found between egg weight and body weight traits, and both of these traits (except egg weight at first egg) were highly and positively correlated with egg length and width. Breaking strength was genetically correlated with egg shell thickness (rg = +0.54±0.19) and the shape index (rg = +0.71±0.23). These results suggest that an efficient selection strategy could be implemented to improve the egg production of the pure Shan Ma duck line.

Genome-wide association study revealed a promising region and candidate genes for eggshell quality in an F2 resource population.

BackgroundEggshell 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).ResultsESWs 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.ConclusionsIdentification 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1795-7) contains supplementary material, which is available to authorized users.

Egg shell quality in Japanese quail: characteristics, heritabilities and genetic and phenotypic relationships

The objective of the present study was to estimate heritabilities as well as genetic and phenotypic correlations for egg weight, specific gravity, shape index, shell ratio, egg shell strength, egg length, egg width and shell weight in Japanese quail eggs. External egg quality traits were measured on 5864 eggs of 934 female quails from a dam line selected for two generations. Within the Bayesian framework, using Gibbs Sampling algorithm, a multivariate animal model was applied to estimate heritabilities and genetic correlations for external egg quality traits. The heritability estimates for external egg quality traits were moderate to high and ranged from 0.29 to 0.81. The heritability estimates for egg and shell weight of 0.81 and 0.76 were fairly high. The genetic and phenotypic correlations between egg shell strength with specific gravity, shell ratio and shell weight ranging from 0.55 to 0.79 were relatively high. It can be concluded that it is possible to determine egg shell quality using the egg specific gravity values utilizing its high heritability and fairly high positive correlation with most of the egg shell quality traits. As a result, egg specific gravity may be the choice of selection criterion rather than other external egg traits for genetic improvement of egg shell quality in Japanese quails.

The sodium channel gene family is specifically expressed in hen uterus and associated with eggshell quality traits

BackgroundEggshell quality is important for the poultry industry. During eggshell formation a mass of inorganic minerals is deposited. The Sodium Channel (SCNN1) gene family plays an essential role in cation transportation. The objective of this study was to investigate the pattern of expression of members of the SCNN1 gene family, their variation and their effects on eggshell quality.ResultThe highest expression of SCNN1a, SCNN1b, and SCNN1g genes were in the active uterus during eggshell mineralization, while SCNN1d showed its highest expression level in the quiescent uterus (no egg present). Nineteen candidate SNPs from the four genes were genotyped in a population of 338 White Leghorn layers. Association analysis between SNPs (haplotypes/diplotypes) and eggshell traits was performed. Among seven significant SNPs, five SNPs were associated with eggshell strength, eggshell thickness, eggshell percentage or/and egg weight, while the other two SNPs within SCNN1d were only associated with eggshell percentage. These SNPs had a 0.25-6.99% contribution to phenotypic variance, depending on the trait. In haplotype analysis, SCNN1b and SCNN1d were associated with egg weight. The SCNN1b and SCNN1g were significantly associated with eggshell weight while only SCNN1g explained 2.04% of phenotypic variance. All the alleles of the members of SCNN1 gene family were associated with eggshell percentage and eggshell thickness, and others members had an association with eggshell strength except for SCNN1a. The contribution of different haplotypes of the SCNN1 gene family to eggshell phenotypic variance ranged from 0.09% to 5.74%.ConclusionsOur study indicated that the SCNN1 gene family showed tissue expression specificity and was significantly associated with eggshell traits in chicken. This study provides evidence that genetic variation in members of the sodium channel can influence eggshell quality.

Variation in the ovocalyxin‐32 gene in commercial egg‐laying chickens and its relationship with egg production and egg quality traits

Avian eggshell quality is an important trait for commercial egg production, as the eggshell is the primary packaging material and antimicrobial barrier for the internal food resource. Strong eggshells are essential to ensure that eggs can reach their final destination without damage. Ovocalyxin-32 (OCX32) is a matrix protein found within the outer layers of the eggshell and in the cuticle. Numerous reports in the literature have identified association between variants in the gene encoding this protein, OCX32, and various eggshell quality traits. Thus, OCX32 is a candidate gene for selection for eggshell traits in commercial poultry populations. Sequencing of exons 2-6 of the OCX32 gene in eight elite brown and white eggshell commercial egg-laying lines revealed 28 SNPs and one SNP/indel. Eighteen of these SNPs were predicted to alter the amino acid sequence of the protein. Clusters of SNPs in complete linkage disequilibrium were found in both exons 2 and 6. A total of 19 different versions or protein-sequence haplotypes of the OCX32 protein were inferred, revealing considerable variation within commercial lines. Genotypes for 13 of the SNPs were determined for 330-1819 individuals per line. Trait association studies revealed a significant effect of OCX32 on shell color in white egg lines and line-specific significant effects on albumen height, early egg weight, puncture score, and yolk weight. Three of the lines showed a significant change in OCX32 frequency over time, indicating selection pressure for certain variants of this gene during the breeding program.

A single nucleotide polymorphism in the parathyroid hormone gene and effects on eggshell quality in chickens

Parathyroid hormone (PTH), released by the parathyroid gland of animals, plays an important role in regulating the metabolism of calcium and phosphate. As a candidate gene for eggshell quality traits, the SNP was screened and its genetic effects on eggshell qualities and levels of serum calcium, phosphate, and PTH were analyzed in this study. Three hundred Houdan hens, an indigenous breed of chicken in France, were used for genotyping and data recording. Of the 3 sets of primers used to amplify the exons, exon 3 was polymorphic and 3 genotypes were identified. Sequencing revealed a nucleotide transition, A2205G (GenBank accession no. NC_006092), which was a synonymous mutation and caused a codon for lysine to change from AAA to AAG. Eggshell percentage and breaking strength for genotypes GG and AG were greater (P<0.05) than for AA, respectively; the total serum calcium for genotype GG was higher than for genotypes AA and AG by 11.8 and 10.1%, respectively; the serum phosphate for genotype GG was greater than genotype AA and AG by 16.7 and 12%, respectively. All genotypes shared the same calcium:phosphate ratio. For serum PTH, genotype GG was approximately 30% higher than genotype AA. Therefore, the SNP A2205G in PTH affected the eggshell percentage and breaking strength, and it may be associated with the variation of serum calcium and PTH level, indicating that the SNP in PTH has the potential for utilization in a MAS program for eggshell quality in chicken.

Association between ovocalyxin‐32 gene haplotypes and eggshell quality traits in an F2 intercross between two chicken lines divergently selected for eggshell strength

Broken and cracked eggshells contribute significantly to economic losses in the egg production industry. We previously identified ovocalyxin-32 as a potential gene influencing eggshell traits, by analysing an intercross between two parent lines developed from the same founder population by a two-way selection for eggshell strength with non-destructive deformation (DEF) conducted over 14 generations. We determined the nucleotide sequences of six ovocalyxin-32 exons in the parent individuals and analysed the association between ovocalyxin-32 and eggshell traits in the F2 individuals. We identified three haplotypes (W, M and S) of ovocalyxin-32 in the parent individuals. A mismatch amplification mutation assay was performed to distinguish six diplotype individuals (WW, MM, SS, WM, MS and WS) inthe F2 population. The egg weight (EW) of SS-diplotype individuals was significantly higher than that of WW-, WM- and WS-diplotypes. Short length of the egg (SLE) of SS-diplotype individuals was significantly higher than that of WW-, WM- and MS-diplotypes. Long length of the egg (LLE) of SS-diplotype individuals was significantly higher than that of WM and WS-diplotypes. DEF of WW-diplotype individuals was significantly higher than that ofSS-, WM, MS and WM-diplotypes. Haplotypic effect analyses showed significant differences between the W-haplotype and the S-haplotypes in the EW, SLE, LLE and DEF. The DEF of M-haplotype was significantly lower than that of W- and S-haplotypes. These results suggest that S- and M-haplotypes are critical for high quality of eggshells in the F2 population. In conclusion, ovocalyxin-32 is a useful marker of eggshell traits and can be used to develop strategies for improving eggshell traits in commercial layer houses.

Using Scanning Electron Microscopy to Detect the Ultrastructural Variations in Eggshell Quality of Fayoumi and Dandarawi Chicken Breeds

An experiment was conducted to evaluate egg quality and ultrastructural measurements of eggshell in two Egyptian local breeds of chicken (Fayoumi and Dandarawi). A total of 162 females representing the two breeds were randomly assigned to the current experiment. They were housed in individual cages placed in an open-sided house under the same environmental, managerial and hygienic conditions. Maturation measurements including body weight and age at sexual maturity were determined. Also, egg production (number and weight) was recorded for the first 3 months of production cycle to calculate egg mass. To as sess egg quality parameters (internal and external), a total of 90 eggs were randomly collected from each breed at 26 weeks of age. In addition, eggshell samples were prepared to investigate ultrastructural variations using scanning electron microscopy (SEM). The current results revealed that Fayoumi breed had a better performance at sexual maturity compared to Dandarawi one. Inferior egg production parameters during the first 3 months of laying cycle were associated with Dandarawi breed. With respect to internal egg quality, there were no significant differences between Fayoumi and Dandarawi breeds for liquid components as a percentage of egg weight. In addition, Haugh unit and yolk index as indicators to the quality of albumen and yolk, did not exhibit a difference between breeds. Regarding eggshell quality traits, it could be noticed that Fayoumi breed significantly increased specific gravity and eggshell strength compared to Dandarawi one. In terms of ultrastructural measurements of eggshell, SEM inspection revealed that eggshell of Fayoumi breed had a higher effective thickness (palisade layer) compared to Dandarawi one. On the other hand, the length of mammillary layer (absolute or relative) was significantly lower in eggshells of Fayoumi hens compared to those of Dandarawi counterparts. Type B bodies, which are rounded and located among mammillary caps, were more frequent in eggshell of Dandarawi breed resulting in poor shell quality. However, Fayoumi breed had good rounded caps and early fusion as compared to Dandarawi one. The incidence of mammillary alignment and erosion were more common in eggshell of Dandarawi breed suggesting poor shell strength. In conclusion, Fayoumi breed had genetically not only better egg production and egg quality but also good ultrastructural formation of eggshell compared to Dandarawi breed.