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Simple Summary: n-3 PUFA, selenium and lutein cannot be synthesized in the human body, but these substances are very important for human health. Consuming functional eggs enriched with nutrients can increase the body's intake of these substances. To efficiently produce functional eggs, selecting breeds that can deposit higher levels of these nutrients is necessary. Therefore, we investigated the types, dosages, and cycles of additives used in the production of functional eggs. In Trial 1, we investigated the differences in n-3 PUFA deposition among five breeds of eggs. The results showed significant variations in the ability of different breeds to deposit n-3 PUFA in eggs, with Dwarf Layer demonstrating a significant advantage. In Trial 2, we investigated the difference of n-3 PUFA, selenium and lutein deposition in eggs from two reeds in terms of their ability to deposit selenium and lutein in eggs. Additionally, the n-3 PUFA content in eggs from the two breeds was similar to the findings in Trial 1. Therefore, this experiment provides valuable insights for selecting different breeds in the production of functional eggs, ultimately improving the quality and economic benefits of the production process. The purpose of this study was to compare the differences in the content of functional nutrients in eggs, performance parameters, and egg quality parameters of different chicken breeds. In Trial 1, 150 41-week-old hens of each breed, including the Dwarf Layer, White Leghorn, Silky fowl, Beijing-you chicken, and Shouguang chicken, were randomly assigned to the control (CON) and 2.5% flaxseed oil (FSO) groups to compare the difference in n-3 polyunsaturated fatty acid (PUFA) content in eggs. The contents of α-linolenic acid (ALA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and total n-3 PUFA in eggs were increased (p < 0.05) in the FSO groups. The ALA (4.28%), DHA (2.03%), and total n-3 PUFA (6.46%) contents in eggs of Dwarf Layer were the highest among the five breeds (p < 0.05). To further verify if other functional nutrients also have such differences, 600 24-week-old White Leghorn and Dwarf Layer were allocated to four groups: CON, FSO, 0.02% selenium-enriched yeast (SEY), and 0.20% marigold flower extract (MFE), in Trial 2. The content of functional nutrients in eggs was significantly increased (p < 0.05) after feeding these additions. After feeding FSO, the eggs of the Dwarf Layer had a higher n-3 PUFA content than the White Leghorn (p < 0.05). However, no significant differences were found in selenium and lutein content in different breeds. Performance and egg quality were not negatively impacted by FSO, SEY, or MFE. [ABSTRACT FROM AUTHOR]

Simple Summary: Autosexing based on sex-linked phenotypes of late feathering (LF) and early feathering (EF) was widely used in poultry production. Previous studies found that tandem duplication on the Z chromosome is responsible for LF. In this study, based on tandem duplication, we developed a multiplex PCR test for genotyping through reaction optimization. The genotyping result for feathering rate with the multiplex PCR test was consistent with the sex and phenotype records, and the result of homozygote and heterozygote tests of LF males were verified by test cross and tallied with offspring's phenotypes. The multiplex PCR test is stable and accurate for feathering rate genotyping, applicable to all LF chickens associated with tandem duplication, and has great significance for poultry breeding. Sex-linked phenotypes of late feathering (LF) and early feathering (EF) are controlled by a pair of alleles K and k+. Autosexing based on the feathering rate is widely used in poultry production. It is reported that a tandem duplication of 176,324 base pairs linked to the K locus is responsible for LF expression and could be used as a molecular marker to detect LF chicken. So far, there is no genotyping method that can accurately and stably identify the LF homozygote and heterozygote in all chicken breeds. In the present study, a multiplex PCR test was developed to identify EF, LF homozygote, and heterozygote according to electrophoretic bands and the relative height of the peaks by Sanger sequencing. We tested 413 chickens of six native Chinese breeds with this method. The identification was consistent with the sex and phenotype records of the chickens. Band density analysis was performed, and the results supported our genotyping using the new assay. In order to further verify the accuracy of this test in distinguishing homozygote and heterozygote males, 152 LF males were mated with EF females, and the results of the offspring's phenotypes were consistent with our expectations. Our results support tandem duplication as molecular markers of LF, and this new test is applicable to all LF chickens associated with tandem duplication. [ABSTRACT FROM AUTHOR]

The basic units of skeletal muscle in all vertebrates are multinucleate myofibers, which are formed from the fusion of mononuclear myoblasts during the embryonic period. In order to understand the regulation of embryonic muscle development, we selected four chicken breeds, namely, Cornish (CN), White Plymouth Rock (WPR), White Leghorn (WL), and Beijing-You Chicken (BYC), for evaluation of their temporal expression patterns of known key regulatory genes (Myomaker, MYOD, and MSTN) during pectoral muscle (PM) and thigh muscle (TM) development. The highest expression level of Myomaker occurred from embryonic days E13 to E15 for all breeds, indicating that it was the crucial stage of myoblast fusion. Interestingly, the fast-growing CN showed the highest gene expression level of Myomaker during the crucial stage. The MYOD gene expression at D1 was much higher, implying that MYOD might have an important role after hatching. Histomorphology of PM and TM suggested that the myofibers was largely complete at E17, which was speculated to have occurred because of the expression increase in MSTN and the expression decrease in Myomaker. Our research contributes to lay a foundation for the study of myofiber development during the embryonic period in different chicken breeds. [ABSTRACT FROM AUTHOR]

Background: Duration of fertility (DF) is an important economic trait in poultry production because it has a strong effect on chick output. Various criteria or traits to assess DF on individual hens have been reported but they are affected by many nongenetic factors. Thus, a reliable definition and associated genetic parameters are needed. Because egg production is also vital in chicken breeding, knowledge of the relationship between DF and laying performance is needed for designing selection programs. Methods: We used five traits that consider both fertility and embryonic livability to delineate DF. Phenotypic and genetic analyses were completed for 2094 hens, with measurements of DF at 35 and 60 weeks of age and hatching egg production at 400 days of age (HEP400). The selection differentials for DF and HEP400 were evaluated. Results: DF is largely independent of the number of oviposited eggs in the peak laying period but both egg production and DF naturally decline with age. The heritability of the five DF traits ranged from 0.11 to 0.13 at 35 weeks of age and increased slightly in the later laying period, ranging from 0.14 to 0.17 (except for efficient duration, time between insemination and the first unhatched egg). Estimates of the genetic correlation for a given trait measured at the two ages were moderate (0.37–0.44), except for efficient duration. However, number of viable embryos depends strongly on egg production. Estimates of genetic correlations of fertility duration day (FDD) at both ages with HEP were weak. Selection for FDD improved DF but without a significant change in laying performance. Selection for increased HEP400 did not contribute to DF improvement. Conclusions: Although estimates of heritability of the five traits related to DF were low, selection to improve DF based on any one of them is possible. Among these, FDD is an effective selection criterion when the eggs are collected for approximately two weeks after insemination. The best selection procedure for DF improvement would involve multiple measurements at various ages. FDD is independent of laying performance and can be incorporated into a breeding program with egg production to improve reproductive efficiency. [ABSTRACT FROM AUTHOR]

Beijing You Chicken, a valuable local chicken breed from Beijing, China, was once listed as an endangered breed. From the point of view of conservation, the preservation of this breed is an important task for the local researchers. Semen cryopreservation is a popular method to maintain valuable species. However, during cryopreservation, semen is susceptible to oxidative damage. Melatonin is a potent antioxidant and free radical scavenger, so it has been selected to improve the efficiency of sperm cryopreservation. In this study, the chicken semen was treated with different concentrations of melatonin in the cryopreservation solution. The results showed that melatonin at concentrations of 10−3 M and 10−5 M significantly improved sperm progressive motility and total motility, respectively, compared to the control (P < 0.05). Melatonin at 10−3 M also significantly improved the plasma membrane and acrosome integrity of spermatozoa compared to the control. The mechanisms are that melatonin significantly reduces the level of ROS and preserves sperm mitochondrial membrane potential. Most importantly, the melatonin-treated cryopreserved chicken sperm after artificial insemination significantly increased the hatching rate of chicks compared to the control (p < 0.05). The results show that melatonin has a positive effect on the quality of the cryopreserved spermatozoa. These results provide the theoretical and practical basis for using melatonin to improve Beijing You Chicken conservation, and they may also be applicable to poultry as a whole. [ABSTRACT FROM AUTHOR]

Summary: Chicken plumage color, the genetic basis of which is often affected by epistasis, has long interested scientists. In the current study, a population of complex epistasis was constructed by crossing dominant White Leghorn chickens with recessive white feather chickens. Through a genome‐wide association study, we identified single nucleotide polymorphisms and genes significantly associated with white and colored plumage in hens at different developmental stages. Interestingly, white plumage in adulthood was associated with the recessive white feather gene (TYR), whereas white feathers at birth stage were associated with the dominant white feather gene (PMEL), indicating age‐related roles for these genes. TYR was shown to exert an epistatic effect on PMEL in adult hens. Additionally, TYR had an epistatic effect on barred plumage, while barred plumage had an epistatic effect on black plumage. TYR had no epistatic effect on the yellow plumage. We confirmed that the barred plumage gene is CDKN2A, as reported in previous studies. Golgb1 and REEP3, which play important roles in the Golgi network and affect the formation of feather pigments, are important candidate genes for yellow plumage. The candidate genes for black plumage are CAMKK1 and IFT22. Further research is warranted to elucidate the molecular mechanisms underlying these traits. [ABSTRACT FROM AUTHOR]

The chicken major histocompatibility complex (MHC) on chromosome 16 is the most polymorphic region across the whole genome, and also an ideal model for genetic diversity investigation. The MHC B-F/B-L region is 92 kb in length with high GC content consisting of 18 genes and one pseudogene (Blec4), which plays important roles in immune response. To evaluate polymorphism of the Chinese indigenous chickens as well as to analyze the effect of selection to genetic diversity, we used WaferGen platform to identify sequence variants of the B-F/B-L region in 21 chicken populations, including the Red Jungle Fowl (RJF), Cornish (CS), White Leghorns (WLs), 16 Chinese domestic breeds, and two well-known inbred lines 63 and 72. A total of 3,319 single nucleotide polymorphism (SNPs) and 181 INDELs in the B-F/B-L region were identified among 21 populations, of which 2,057 SNPs (62%) and 159 INDELs (88%) were novel. Most of the variants were within the intron and the flanking regions. The average variation density was 36 SNPs and 2 INDELs per kb, indicating dramatical high diversity of this region. Furthermore, BF2 was identified as the hypervariable genes with 67 SNPs per kb. Chinese domestic populations showed higher diversity than the WLs and CS. The indigenous breeds, Nandan Yao (NY), Xishuangbanna Game (XG), Gushi (GS), and Xiayan (XY) chickens, were the top four with the highest density of SNPs and INDELs. The highly inbred lines 63 and 72 have the lowest diversity, which might be resulted from a long-term intense selection for decades. Collectively, we refined the genetic map of chicken MHC B-F/B-L region, and illustrated genetic diversity of 21 chicken populations. Abundant genetic variants were identified, which not only strikingly expanded the current Ensembl SNP database, but also provided comprehensive data for researchers to further investigate association between variants in MHC and immune traits. [ABSTRACT FROM AUTHOR]

With the appeals for antibiotic-free production and the attention to natural feed additives in poultry. The effects of fresh lemon (FM) supplementation on older laying hens (79–96 weeks) and its comparison with citric acid (CA) and flavomycin (FLA) were evaluated in the present study. Female Jingfen chickens (n = 540) were randomly distributed into 5 dietary treatments: 0 (control), 20 g/kg (2%) and 40 g/kg (4%) FM, 0.7 g/kg CA and 0.005 g/kg FLA. Production performances were recorded daily and egg quality traits were measured biweekly. Serum biochemical indices and antibody titres were detected every 4 weeks from 84 to 96 weeks of age. Compared with the control diet, FM, CA and FLA increased the laying rate (p <.05) and average daily feed intake (ADFI), and reduced the feed to egg ratio (FER), indicating positive effects on production performance. Albumen height and HU were increased by 2% and 4% FM groups compared to the control, while albumen height was decreased in CA and FLA groups. FM improved serum antioxidant enzyme activities (p <.05) and reduced serum HDL, LDL, TG and T-CH compared to other groups, indicating favourable effects on antioxidative status and lipid metabolism. Antibody titres against H5N1, H9N2 and NDV were higher in FM groups than that in other groups during most experimental periods (p <.05). In conclusion, FM supplementation enhanced the production performance and antioxidant capacity of laying hens during the late laying period and was superior to the CA and FLA in improving albumen quality, immune status and lipid metabolism. FM supplementation in layers' diets had positive effects on production performance, egg albumen quality and lipid metabolism, and could enhance antioxidant capacity and immune levels. Adding FM had better effects on the albumen quality, immune status and lipid metabolism of laying hens than adding CA and FLA during the late laying period. [ABSTRACT FROM AUTHOR]

Artificial selection is often associated with numerous changes in seemingly unrelated phenotypic traits. The genetic mechanisms of correlated phenotypes probably involve pleiotropy or linkage of genes related to such phenotypes. Dongxiang blue-shelled chicken, an indigenous chicken breed of China, has segregated significantly for the dermal hyperpigmentation phenotype. Two lines of the chicken have been divergently selected with respect to comb color for over 20 generations. The red comb line chicken produces significantly higher number of eggs than the dark comb line chicken. The objective of this study was to explore potential mechanisms involved in the relationship between comb color and egg production among chickens. Based on the genome-wide association study results, we identified a genomic region on chromosome 20 involving EDN3 and BMP7 , which is associated with hyperpigmentation of chicken comb. Further analyses by selection signatures in the two divergent lines revealed that several candidate genes, including EDN3 , BMP7 , BPIFB3 , and PCK1 , closely located on chromosome 20 are involved in the development of neural crest cell and reproductive system. The two genes EDN3 and BMP7 have known roles in regulating both ovarian function and melanogenesis, indicating the pleiotropic effect on hyperpigmentation and egg production in blue-shelled chickens. Association analysis for egg production confirmed the pleiotropic effect of selected loci identified by selection signatures. The study provides insights into phenotypic evolution due to genetic variation across the genome. The information might be useful in the current breeding efforts to develop improved breeds for egg production. [ABSTRACT FROM AUTHOR]

Simple Summary: With the increased interest in animal welfare, poultry housing systems have been a concern for the last decade, and conventional cages have been replaced by non-cage systems or enriched cages. However, the environmental conditions in non-cage housing systems and their association with hens' production performance and health have not been studied extensively. Therefore, the present study compared the indoor environmental microbial content in two different non-caged systems, namely, a plastic-net housing system and floor-litter housing system, and investigated its effects on the production performance, serum parameters and intestinal morphology of hens during the peak laying period. The results indicated that the NRS resulted in better indoor environmental air quality and ground hygiene than the LRS and enhanced the production performance, antioxidant capacity and intestinal health of hens, represented by positive changes in the laying rate, serum parameters and intestinal morphology. This study investigated the effects of plastic-net housing system (NRS) and floor-litter housing system (LRS) on the production performance, serum parameters and intestinal morphology of Shendan laying hens. A total of 1200 30-week-old hens were randomly allocated to the NRS and LRS groups, each of which included five replicates with 120 chickens in each replicate. The experiment was conducted from 32 to 40 weeks of age. Indoor airborne parameters were measured every 2 weeks, and indoor ground contamination was measured monthly. The laying rate and mortality of hens were recorded daily, and egg quality traits and serum parameters were measured every 2 weeks. At 40 weeks of age, four birds per replicate from each experimental group were selected for intestinal morphological observation. The results showed that the airborne bacteria number in the LRS was significantly higher than that in the NRS (p < 0.05) for most of the experimental period (except at 32 and 38 weeks of age), and the bacterial numbers on the surfaces of the floor and floor eggs in the LRS were approximately 10 times higher than those in the NRS (p < 0.05). Compared with the LRS, the NRS improved the laying rate (p < 0.05), reduced serum malondialdehyde (MDA) (p < 0.05) and corticosterone (CORT) concentrations and increased serum glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities, indicating favourable effects on antioxidative status. The NRS was significantly associated with an increased villus height (VH), villus height to crypt depth ratio (VCR) in the small intestine (p < 0.05) and increased VCR in the caecum (p < 0.05). Overall, the lower rate of bacterial contamination in the NRS than in the LRS indicated better environmental hygiene. The NRS enhanced the laying performance and antioxidant capacity of hens and was superior to the LRS in improving intestinal health. The current findings support the advantages of the NRS for the health and welfare of Shendan chickens during the peak laying period. [ABSTRACT FROM AUTHOR]

Simple Summary: Although there is an established understanding of the nutritional requirements for poultry production, the evaluation of the feed form for chickens still needs to be further examined. It is generally believed that a pellet diet has a positive effect on chicken growth, but there are some studies that have reported no difference between pellet and mash on chicken's performance. The present study indicated that pellet diets improved the production performance and nutrition metabolism through positive changes in the laying rate, feed intake, egg albumen quality and apparent digestibility of laying hens. One of the most important factors that determine feed utilization by chickens is the feed form. Although it is generally believed that pellet diets have a positive effect on chicken growth, there are some studies that have indicated no difference between pellet and mash on chickens performance. This study was conducted to assess the effects of feed form on production performance, egg quality, nutrient metabolism and intestinal morphology in two breed laying hens. Two hundred and sixteen 25-week-old Hy-Line brown (n = 108) and Hy-Line grey (n = 108) hens were selected. Each breed was randomly allocated into two treatments with 6 replications (9 birds in each replication), which were fed mash and pellet diets, respectively. Production performances were recorded daily and egg quality traits were measured every two weeks. At 42 weeks of age, one bird per replication from each experimental group was selected for metabolism determination and intestine morphology observation. Compared with mash diets, pellet diets improved laying rate (p < 0.05), ADFI (average daily feed intake, p < 0.05), egg weight, shell strength, yolk proportion and Haugh unit (p < 0.05) in both breeds and reduced the FCR (feed conversion ratio, p < 0.05) in Hy-Line grey. The apparent digestibility of DM% (dry matter) and CP% (crude protein) were significantly higher (p < 0.05) in both breed laying hens fed pellet than those fed mash. The apparent digestibility of P% (phosphorus) and Ca% (calcium) was higher in Hy-Line grey fed pellet and was higher in Hy-Line brown fed mash. Compared to mash diets, pellet diets increased the VH (villus height), CD (crypt depth) and VCR (ratio of villus height to crypt depth) of the small intestine of Hy-Line grey, and increased the VH and CD of duodenum and ileum of Hy-Line brown. Overall, pellet diets improved production performance and nutrition metabolism through positive changes in the laying rate, feed intake, egg albumen quality and apparent digestibility of laying hens. The current findings provided support for the advantages of feeding pellets during the peak egg laying period for the two popular laying hen strains, Hy-Line brown and Hy-Line grey. [ABSTRACT FROM AUTHOR]

Background: Gene expression variation is a key underlying factor influencing phenotypic variation, and can occur via cis- or trans-regulation. To understand the role of cis- and trans-regulatory variation on population divergence in chicken, we developed reciprocal crosses of two chicken breeds, White Leghorn and Cornish Game, which exhibit major differences in body size and reproductive traits, and used them to determine the degree of cis versus trans variation in the brain, liver, and muscle tissue of male and female 1-day-old specimens. Results: We provided an overview of how transcriptomes are regulated in hybrid progenies of two contrasting breeds based on allele specific expression analysis. Compared with cis-regulatory divergence, trans-acting genes were more extensive in the chicken genome. In addition, considerable compensatory cis- and trans-regulatory changes exist in the chicken genome. Most importantly, stronger purifying selection was observed on genes regulated by trans-variations than in genes regulated by the cis elements. Conclusions: We present a pipeline to explore allele-specific expression in hybrid progenies of inbred lines without a specific reference genome. Our research is the first study to describe the regulatory divergence between two contrasting breeds. The results suggest that artificial selection associated with domestication in chicken could have acted more on trans-regulatory divergence than on cis-regulatory divergence. [ABSTRACT FROM AUTHOR]

The Huiyang beard chicken is a well-known Chinese local breed known for its elongated feathers gathered from both sides of the face (muffs) and below the beak (beard), as well as short wattles (SW). The muff and beard (Mb) mutation is caused by ectopic upregulation of the homeobox B8 ( HOXB8 ) gene in the mandibular skin; and the chi-square test showed a significant correlation between SW and Mb genotypes. However, the underlying molecular mechanisms that regulate Mb and SW variations remain unclear. In this study, we investigated the transcriptomes of the mandibular skin and wattles of chickens with and without the Mb genotype to elucidate the molecular basis of these traits. Our results show that HOXB8 is expressed at significantly higher levels in both the mandibular skin and wattles of Mb chickens than in those of wild-type chickens, indicating that HOXB8 regulates both the Mb and SW phenotypes. Key genes for keratin synthesis were highly expressed in the mandibular skin of Mb chickens, suggesting that HOXB8 may play a role in feather development. In wattles, changes in the expression of extracellular matrix synthesis genes may contribute to SW traits. DNA-binding motif analyses revealed that differentially expressed genes were likely to be directly regulated by HOXB8 binding, indicating that HOXB8 may directly or indirectly regulate feather follicle development and wattle growth. Our study identified both known and novel targets, including several genes not previously implicated in feather development and mesenchymal formation. These findings provide insights into the molecular mechanisms of skin appendage variation in birds and offer potential applications in breeding poultry breeds with unique phenotypes. [ABSTRACT FROM AUTHOR]

Piao chicken, a Chinese indigenous rumpless chicken breed, lacks pygostyle, caudal vertebra, uropygial gland and tail feathers. The rumplessness in Piao chicken presents an autosomal dominant inheritance pattern. However, the molecular genetic mechanisms underlying the rumplessness in Piao chicken remains unclear. In this study, whole-genome resequencing was performed for 146 individuals from 10 chicken breeds, including 9 tailed chicken breeds and Piao rumpless breed. Tailbone CT scan for Piao chickens and WL chickens, revealed that some Piao chicken tails were normal in number, and for a few Piao chickens tail length and tail bone numbers were between the rumpless and the normal tailed chickens. The results showed that the rumpless phenotype has not been completely fixed in Piao chicken breed. Using selection signature analysis and structural variation detection, we found a 4174 bp deletion located in the upstream region of IRX1 gene on chromosome 2 related to rumpless phenotype. Structural variation genotyping showed that the deletion was present in all 32 rumpless Piao chickens (del/del, wild/del) and absent from all 112 tailed chickens included in the dataset for the other 9 breeds and 2 tailed Piao chickens (wild/wild). In summary, all rumpless Piao chickens tested here carry this deletion mutation, to show a complete linkage association with rumplessness trait. We suggested that the 4174 bp deletion could be causative for rumpless phenotype in Piao chicken since this is the only mutation to show the complete linkage disequilibrium with rumplessness on whole genome level across all of 146 chickens from the 10 breeds. This study could facilitate a better understanding of the genetic characteristics of Piao chicken. • Piao chicken structural variation detection. • In Piao Chicken, a deletion variant was found to be associated with rumpless. • This study could facilitate a better understanding of tail degeneration in chicken. [ABSTRACT FROM AUTHOR]