Exploring the expression and preliminary function of chicken regulator of G protein signalling 3 (RGS3) gene in follicular development.

1. The following study explored the expression and preliminary function of the RGS3 gene. The spatial and temporal expression patterns of the RGS3 gene were analysed in the ovarian stroma of Shendan No. 6 Green shell hens and Hy-line Brown hens at four time points (6, 28, 40, and 52 weeks old), as well as in various organs and follicles of Hy-line Brown hens.2. Based on the genomic and protein sequences of RGS3 in NCBI database, phylogenetic trees were constructed using MEGA-X. The protein interaction network was analysed using STRING. According to the results of protein-protein interaction network and pathways, the mRNA expression levels of RGS3 and three interaction proteins were explored by qRT-PCR in vitro.3. Spatio-temporal expression data revealed that RGS3 mRNA was expressed in all the organs tested, being highest in the hypothalamus. In different follicles, RGS3 mRNA was highly expressed in post-ovulatory follicles, followed by ovarian stroma and large white follicles. The expression levels of RGS3 mRNA in the ovarian stroma were significantly higher in Shendan No. 6 Green shell hens than that in the Hy-line Brown hens at all egg-laying stages.4. The phylogenetic tree results showed that ducks, geese and chickens had higher homology based on the genomic and protein sequence of RGS3. Moreover, chicken RGS3 interacted with GSK3B, RAF1 and BRAF based on STRING prediction. In vitro follicle stimulating hormone (FSH) treatment showed that mRNA expression levels of RGS3 and those of its predicted interacting proteins BRAF and GSK3B decreased with increasing FSH concentration. The results suggested that RGS3 responds to FSH and may play an important role in the regulation of follicular development in chicken.

Combined analysis of metagenomic data revealed consistent changes of gut microbiome structure and function in inflammatory bowel disease.

AIMS: To reveal the consistency and discrepancy in the gut microbial structure and function in inflammatory bowel disease (IBD) patients from different regions. METHODS AND RESULTS: Gut microbes, antibiotic resistance genes (ARGs) and virulence factors genes (VFGs) were analysed using metagenome data from three cohorts. The abundance of Escherichia coli extensively increased in IBD patients, whereas Subdoligranulum unclassified decreased dramatically in IBD patients from three countries. Escherichia coli showed a positive correlation with multiple ARGs and VFGs in cohorts from China and the United States, including multidrug-related resistance genes and Capsule and LOS-related virulence factors genes. Escherichia coli biofilm synthesis pathways significantly enriched in IBD patients from three different regions. Notably, Subdoligranulum unclassified and Eubacterium hallii were negatively related to ARGs and VFGs. CONCLUSIONS: Consistent changes of microbiome structure and function were observed in IBD patients from three different regions. As pathogenic bacteria, E. coli may accelerate IBD progression through encapsulation in biofilms by upregulating antibiotic resistance in Crohn's disease patients. Subdoligranulum unclassified and E. hallii may be beneficial for IBD patients and could serve as potential probiotics for IBD treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: This work dispels worries about the regional differences in gut microbial changes in IBD patients and provides useful guidance for more rational microbiome-based therapies.

Endocrine and genetic factors affecting egg laying performance in chickens: a review.

1. Egg-laying performance reflects the overall reproductive performance of breeding hens. The genetic traits for egg-laying performance have low or medium heritability, and, depending on the period involved, usually ranges from 0.16 to 0.64. Egg-laying in chickens is regulated by a combination of environmental, endocrine and genetic factors. 2. The main endocrine factors that regulate egg-laying are gonadotropin-releasing hormone (GnRH), prolactin (PRL), follicle-stimulating hormone (FSH) and luteinising hormone (LH). 3. In the last three decades, many studies have explored this aspect at a molecular genetic level. Recent studies identified 31 reproductive hormone-based candidate genes that were significantly associated with egg-laying performance. With the development of genome-sequencing technology, 64 new candidate genes and 108 single nucleotide polymorphisms (SNPs) related to egg-laying performance have been found using genome-wide association studies (GWAS), providing novel insights into the molecular genetic mechanisms governing egg production. At the same time, microRNAs that regulate genes responsible for egg-laying in chickens were reviewed. 4. Research on endocrinological and genetic factors affecting egg-laying performance will greatly improve the reproductive performance of chickens and promote the protection, development, and utilisation of poultry. This review summarises studies on the endocrine and genetic factors of egg-laying performance in chickens from 1972 to 2019.

Myostatin mRNA expression and its association with body weight and carcass traits in Yunnan Wuding chicken.

Myostatin (MSTN) is expressed in the myotome and developing skeletal muscles, and acts to regulate the number of muscle fibers. Wuding chicken large body, developed muscle, high disease resistance, and tender, delicious meat, and are not selected for fast growth. Broiler chickens (Avian broiler) are selected for fast growth and have a large body size and high muscle mass. Here, 240 one-day-old chickens (120 Wuding chickens and 120 broilers) were examined. Twenty chickens from each breed were sacrificed at days 1, 30, 60, 90, 120, and 150. Breast and leg muscle samples were collected within 20 min of sacrifice to investigate the effects of MSTN gene expression on growth performance and carcass traits. Body weight, carcass traits, and skeletal muscle mass in Wuding chickens were significantly (P < 0.05) lower than those in broiler chickens at all time points. Breast muscle MSTN mRNA was lower in Wuding chickens than in broilers before day 30 (P < 0.05). After day 30, breast muscle MSTN expression was higher in Wuding chicken than in broilers (P < 0.05). Leg muscle MSTN mRNA expression was higher in Wuding chicken than in broilers at all ages except for day 60 (P < 0.05). Correlation analysis revealed that breast muscle MSTN expression has a greater effect in slow growing Wuding chickens than in the fast growing broilers. In contract, leg muscle MSTN mRNA level has a greater effect in broilers than in Wuding chickens. MSTN regulates growth performance and carcass traits in chickens.

Comparison and analysis of Wuding and avian chicken skeletal muscle satellite cells.

Chicken skeletal muscle satellite cells are located between the basement membrane and the sarcolemma of mature muscle fibers. Avian broilers have been genetically selected based on their high growth velocity and large muscle mass. The Wuding chicken is a famous local chicken in Yunnan Province that undergoes non-selection breeding and is slow growing. In this study, we aimed to explore differences in the proliferation and differentiation properties of satellite cells isolated from the two chicken breeds. Using immunofluorescence, hematoxylin-eosin staining and real-time polymerase chain reaction analysis, we analyzed the in vitro characteristics of proliferating and differentiating satellite cells isolated from the two chicken breeds. The growth curve of satellite cells was S-shaped, and cells from Wuding chickens entered the logarithmic phase and plateau phase 1 day later than those from Avian chicken. The results also showed that the two skeletal muscle satellite cell lines were positive for Pax7, MyoD and IGF-1. The expression of Pax7 followed a downward trend, whereas that of MyoD and IGF-1 first increased and subsequently decreased in cells isolated from the two chickens. These data indicated that the skeletal muscle satellite cells of Avian chicken grow and differentiate faster than did those of Wuding chickens. We suggest that the methods of breeding selection applied to these breeds regulate the characteristics of skeletal muscle satellite cells to influence muscle growth.

[The effect of paraquat on autophagy in human embryonic neural progenitor cells].

OBJECTIVE: To explore the effect of paraquat (PQ) on autophagy in human embryonic neural progenitor cells. METHODS: Using ReNcell CX cell model. After treatment with various concentration (0.00, 1.00, 10.00 and 100.00 µmol/L) of PQ, CCK8 assay was used to detect the cell viability, the transmission electron microscope was used to observe the the cell ultrastructure, the real-time polymerase chain reaction (RT-PCR) was adopted to detect mRNA expression of autophagy related genes which including LC3, Atg12, Atg5, beclin1, Atg7 and mTOR and apoptosis related genes Bax and Bcl-2. RESULTS: The cell viability was significantly inhibited after administered with 100.00 µmol/L of PQ (P<0.01). The mRNA expression of Beclin1 was significantly up-regulated and the emergency of autophagosome were observed at the concentration of 1.00 µmol/L group, while mild cell apoptosis, significantly up-regulated Atg5, Atg8, Atg7 and Atg12 mRNA expression as well as down-regulated expression of mTOR and Bax were detected at the 10.00 µmol/L of PQ group, howere, the obvious apoptosis and the up-regulated mRNA expression of mTOR and Bax were observed at the 100.00 µmol/L of PQ group, the mRNA expression of Bcl-2 were all down-regulated after administered with 1.00, 10.00 and 100.00 µmol/L of PQ and reached the lowest level at the concentration of 10.00 µmol/L. CONCLUSION: PQ can induced autophagy at the low concentration in ReNcell CX cell and autophagy might serve as a protective mechanism to ameliorate PQ-induced cytotoxic effects but apoptosis will be induced at the 100 µmol/L concentration.

Haploinsufficiency caused by a nonsense mutation in NCSTN underlying hidradenitis suppurativa in a Chinese family.

Hidradenitis suppurativa (HS) is a chronic disease of follicular occlusion. It involves the axilla, groin, perianal and perineal regions, and is characterized by recurrent draining sinuses, skin abscesses and disfiguring scars. Loss-of-function mutations in the genes encoding γ-secretase have been identified as a cause of HS. We collected skin samples from three patients with HS from a Chinese family carrying a NCSTN mutation (c.1258C>T (p.Q420X)) and three unrelated healthy controls (HCs). Expression level of nicastrin in skin tissue and cultured keratinocytes and fibroblasts of patients and HCs was determined by real-time quantitative PCR and western blotting. We found that the mRNA and protein levels of nicastrin were significantly reduced in the whole skin, epidermis, dermis, and cultured keratinocytes and fibroblasts compared with HCs. Therefore, we conclude that haploinsufficiency of the NCSTN gene caused by the nonsense mutation c.1258C>T (p.Q420X) contributes to the occurrence of HS in this family.

Estimation of genetic parameters related to eggshell strength using random regression models.

This study examined the changes in eggshell strength and the genetic parameters related to this trait throughout a hen's laying life using random regression. The data were collected from a crossbred population between 2011 and 2014, where the eggshell strength was determined repeatedly for 2260 hens. Using random regression models (RRMs), several Legendre polynomials were employed to estimate the fixed, direct genetic and permanent environment effects. The residual effects were treated as independently distributed with heterogeneous variance for each test week. The direct genetic variance was included with second-order Legendre polynomials and the permanent environment with third-order Legendre polynomials. The heritability of eggshell strength ranged from 0.26 to 0.43, the repeatability ranged between 0.47 and 0.69, and the estimated genetic correlations between test weeks was high at > 0.67. The first eigenvalue of the genetic covariance matrix accounted for about 97% of the sum of all the eigenvalues. The flexibility and statistical power of RRM suggest that this model could be an effective method to improve eggshell quality and to reduce losses due to cracked eggs in a breeding plan.

Genome-wide association analysis of eggshell color of an F2 generation population reveals candidate genes in chickens.

Eggshell color is an important visual characteristic that affects consumer preferences for eggs. Eggshell color, which has moderate to high heritability, can be effectively enhanced through molecular marker selection. Various studies have been conducted on eggshell color at specific time points. However, few longitudinal data are available on eggshell color. Therefore, the objective of this study was to investigate eggshell color using the Commission International de L'Eclairage L*a*b* system with multiple measurements at different ages (age at the first egg and at 32, 36, 40, 44, 48, 52, 56, 60, 66, and 72 weeks) within the same individuals from an F2 resource population produced by crossing White Leghorn and Dongxiang Blue chicken. Using an Affymetrix 600 single nucleotide polymorphism (SNP) array, we estimated the genetic parameters of the eggshell color trait, performed genome-wide association studies (GWASs), and screened for the potential candidate genes. The results showed that pink-shelled eggs displayed a significant negative correlation between L* values and both a* and b* values. Genetic heritability based on SNPs showed that the heritability of L*, a*, and b* values ranged from 0.32 to 0.82 for pink-shelled eggs, indicating a moderate to high level of genetic control. The genetic correlations at each time point were mostly above 0.5. The major-effect regions affecting the pink eggshell color were identified in the 10.3-13.0 Mb interval on Gallus gallus chromosome 20, and candidate genes were selected, including SLC35C2, PCIF1, and SLC12A5. Minor effect polygenic regions were identified on chromosomes 1, 6, 9, 12, and 15, revealing 11 candidate genes, including MTMR3 and SLC35E4. Members of the solute carrier family play an important role in influencing eggshell color. Overall, our findings provide valuable insights into the phenotypic and genetic aspects underlying the variation in eggshell color. Using GWAS analysis, we identified multiple quantitative trait loci (QTLs) for pink eggshell color, including a major QTL on chromosome 20. Genetic variants associated with eggshell color may be used in genomic breeding programs.

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.

Effects of exogenous energy on synthesis of steroid hormones and expression characteristics of the CREB/StAR signaling pathway in theca cells of laying hen.

Energy and the cAMP-response element binding protein (CREB)/steroidogenic acute regulatory protein (StAR) signaling pathway play important roles in steroid hormone production and follicular development in hens. This present study aimed to investigate the effects of exogenous energy on the synthesis of steroid hormones and the expression characteristics of the CREB/StAR signaling pathway in theca cells of laying hen. The primary theca cells of small yellow follicles were randomly divided into 6 treatments and cultured in medium with glucose concentrations of 1, 1.5, 3, 4.5, 6, and 7.5 mg/mL for 48 h. It was found that growth was robust and cell outlines were clear when cells were cultured with 1, 1.5, 3, and 4.5 mg/mL glucose, but cell viability was diminished and cell density decreased after exposure to glucose at 6 and 7.5 mg/mL for 48 h. Cell viability showed an increasing and then decreasing quadratic response to increasing glucose concentration in culture (r2 = 0.688, P < 0.001). The cell viability of theca cells cultured with 4.5 mg/mL glucose was greater than those cultured with 1, 1.5, 6, and 7.5 mg/mL glucose (P < 0.05). The concentration of estradiol in the medium containing 3 mg/mL glucose was higher than in medium containing 1, 1.5, and 6 mg/mL glucose (P < 0.05). There was an increasing and then decreasing quadratic correlation between progesterone concentrations and glucose concentrations (r2 = 0.522, P = 0.002). The concentration of progesterone in medium with 4.5 mg/mL glucose was higher than in medium with 1 and 7.5 mg/mL glucose (P < 0.05). There was an increasing and then decreasing quadratic correlation between the relative expression of CREB1 (r2 = 0.752, P < 0.001), StAR (r2 = 0.456, P = 0.002), CYP1B1 (r2 = 0.568, P < 0.001), and 3ß-HSD (r2 = 0.319, P = 0.018) in theca cells of laying hens and glucose concentrations after treatment with different glucose concentrations for 48 h. After treatment with 4.5 mg/mL glucose, the expression of StAR, CYP1B1, and 3ß-HSD genes were increased compared to treatment with 1, 1.5, 3, 6, and 7.5 mg/mL glucose (P < 0.001). There was an increasing and then decreasing quadratic correlation between glucose concentrations and protein expression of CREB1 (r2 = 0.819, P < 0.001), StAR (r2 = 0.844, P < 0.001), 3ß-HSD (r2 = 0.801, P < 0.001), and CYP11A1 (r2 = 0.800, P < 0.001) in theca cells of laying hens. The protein expression of CREB1, StAR, and 3ß-HSD in theca cells cultured with 4.5 mg/mL glucose was higher than in other groups (P < 0.001). The results indicate that the appropriate glucose concentration (4.5 mg/mL) can improve the synthesis of steroid hormones in theca cells of laying hens through the upregulation of key genes and proteins in the CREB/StAR signaling pathway.

The influencing factor of in vitro fertilization and embryonic transfer in the domestic fowl (Gallus domesticus).

This study was conducted to explore the influencing factors of ova in vitro fertilization (IVF) and transfer of the fertilized ova into the oviduct of recipient hens. The efficiency of fertilization was compared using three aspects: (i) the different time of ova collection and transfer, (ii) egg-laying period of recipient hen; and (iii) semen volume. The following results are observed: 72%, 40% and 0% of ova were found in ovarian sac in 30∼40 min, 50∼60 min and more than 90 min post-oviposition, respectively; 20%, 18%, 14% and 5.8% of ova were fertilized with 0.1, 0.2, 0.5 and 1.0 ml semen, respectively; and 33% and 100% of healthy chickens were hatched from fertile ova with 0.1 and 0.5 ml of semen, respectively. All oocytes obtained from ovary and mid-oviduct were unfertilized. Embryos were transferred into recipient hens 30 min ± 10 min post-oviposition, and 70% of shelled eggs were produced. There were no eggs produced in the other transfer times. This demonstrated that live chicken can be obtained by IVF of ova collected shortly after oviposition. It was important that the ovum was transferred into the oviduct infundibulum of recipient hens immediately or shortly after oviposition.

Effects of energy restriction during growing phase on the productive performance of Hyline Brown laying hens aged 6 to 72 wk.

The aim of this study was to assess the effects of energy-restricted feeding during growing phase on the productive performance of Hyline Brown laying hens aged 6 to 72 wk. A total of 720 six-week-old layer chicks were allocated equally to 3 groups with 6 replicates of 40 pullets each, and were fed 1 of 3 diets that were nutritionally similar except for the apparent metabolizable energy corrected for nitrogen (AMEn) content. At the age of 6 to 17 wk, the pullets in the control group were given diet with 2,850 kcal/kg AMEn, and were fed ad libitum. The levels of AMEn in diet of pullets in the experimental groups were 90% (2,565 [2,850 × 90%] kcal/kg) and 80% (2,280 [2,850 × 80%] kcal/kg) of that in control group, and the daily amount of feed was restricted to the absolute quantity of the diet consumed by pullets in control group. At the age of 18 to 72 wk, all the hens were fed with the same diets ad libitum. As energy restriction increased in the growing phase, body weight (BW) dropped at the ages of 12 and 15 to 23 wk (at 23 wk: P = 0.001; at other ages: P < 0.001), but it showed no significant difference at 24 wk (P = 0.071). At 20 wk, restricting energy induced a delay in the development of sexual organs, including the ovary stroma, oviduct, and small yellow follicle (P < 0.05), as well as a delay in sexual maturity (P < 0.05). Consequently, the laying rate in the first and second periods dropped linearly (P = 0.046, 0.030, and 0.038, P < 0.001, respectively). The coefficient of variation (CV) in the BW at 19, 20, and 21 wk (P = 0.040, 0.023, and 0.042, respectively), the CV of age at first egg (P < 0.001), and CV of individual egg number at age 18 to 72 wk (P < 0.001) decreased linearly. There was a linear increase in the laying rate of hens in the later periods (at age 32-72 wk, P < 0.05), as well as in the average total egg number per hen and average laying rate at the age of 18 to 72 wk (P = 0.006). The average egg mass also showed a linear increase with increasing levels of energy restriction (P < 0.001). In summary, although appropriate energy restriction during growing phase delayed sexual maturity and sexual organ development in early-laying Hyline Brown pullets, it improved uniformity of BW, age at first egg laying, and individual egg number, and increased egg number per hen, laying rate, average egg mass, and number of settable eggs from 18 to 72 wk of age.

Genome-wide association studies for mottled eggs in chickens using a high-density single-nucleotide polymorphism array.

Mottled eggs in layer chickens are gaining increasing attention because of the economic impact on the egg industry caused by the reduced sale value of commodity eggs. However, the genetic architecture underlying mottled eggs is not well understood. The genetic architecture underlying the mottled egg trait was investigated using genome-wide association studies (GWAS) by high-density arrays, using a total of 407 pink eggs and 799 blue eggs from an F2 resource population generated by crossing Dongxiang Blue-shelled and White Leghorn chickens. The mottled egg score in blue eggs was found to be higher than that in pink eggs. The single-nucleotide polymorphism heritability of mottled egg at laying day and storage for 7 days was 0.18 and 0.20, respectively. Bivariate GWAS provided 29 significant loci, mainly located on GGA2, GGA3, GGA8, GGA10, GGA15, GGA17, and GGA23, affecting mottled egg on laying day. Candidate genes RIMS2, SLC25A32, RIMBP2, VPS13B, and RGS3 were obtained for mottled eggshell by bivariate GWAS and gene annotation. Our findings provide new insights into the genetic architecture of mottled egg in hens, and demonstrate that a genomic selection method would be profitable for breeding out the mottled egg trait.

Effects of energy-restricted feeding during rearing on sexual maturation and reproductive performance of Rugao layer breeders.

The aim of this study was to assess the effects of energy-restricted feeding during rearing on the sexual maturation and reproductive performance of Rugao layer breeders. A total of 2,400 8-wk-old Rugao layer breeders were randomly assigned to one of 5 groups (480 pullets per group) with eight replicates and were fed one of 5 diets that were nutritionally similar with the exception of apparent metabolizable energy corrected for nitrogen (AMEn) content (2,850, 2,750, 2,650, 2,550, and 2,450 kcal AMEn/kg) from 8 to 18 wks of age. The daily amount of feed was restricted to the absolute quantity of the diet consumed by laying hens fed 2,850 kcal AMEn per kg diet ad libitum (control). From 18 to 52 wks of age, all hens were fed basal diets ad libitum. The body weight of layer breeders at 18 wks of age decreased linearly with increasing energy restriction (P < 0.001), but caught up within 3 wks of ad libitum feeding (P = 0.290). The coefficient of variation of the body weight of the hens at 18, 21, and 24 wks of age decreased linearly (P = 0.010, 0.025, and 0.041, respectively) with increasing energy restriction during rearing. Energy-restricted feeding delayed sexual organ development at 18, 20, and 22 wks of age, including the number of large yellow follicles, oviduct length, oviduct length index, oviduct index, and ovary stroma index (P < 0.05), and delayed sexual maturity, including the age at laying the first egg and the age at 5% and 50% egg production (P = 0.042, 0.004, and 0.029, respectively). Consequently, egg number from 5% to 50% egg production decreased linearly as the degree of energy restriction increased (P = 0.001) and egg production of hens in the energy-restricted feeding groups was lower than that of hens in the ad libitum feeding group (6.36, 6.43, 6.4, and 4.61% vs. 14.29%; P < 0.05) from 18 to 20 wks of age. Furthermore, egg weight increased linearly as energy restriction increased (P < 0.001) and laying hens in the most severe energy-restricted feeding group had more setting eggs (normal eggs weighing >40 g) than hens in the ad libitum feeding and lighter energy-restricted feeding groups (149.57 vs. 144.34, 142.66, 143.63, and 141.78; P < 0.05). No significant differences were observed in fertility, hatchability of fertile eggs, and hatchability of setting eggs (P = 0.381, 0.790, and 0.605, respectively). In conclusion, moderate energy restriction (85.97%, 2,450 vs. 2,850 kcal AMEn/kg) from 8 to 18 wks of age increased egg weight as well as the production of setting eggs in native layer breeders throughout the laying period, without adverse effects on productive performance from 18 to 52 wks of age, or fertility and hatchability at 52 wks of age.

Effect of a free-range raising system on growth performance, carcass yield, and meat quality of slow-growing chicken.

Experiments were conducted to evaluate the effect of free-range raising systems on growth performance, carcass yield, and meat quality of slow-growing chickens. Slow-growing female chickens, Gushi chickens, were selected as the experimental birds. Two hundred 1-d-old female chicks were raised in a pen for 35 d. On d 36, ninety healthy birds, with similar BW (353.7+/-32.1g), were selected and randomly assigned to 2 treatments (indoor treatment and free-range treatment, P>0.05). Each treatment was represented by 3 groups containing 15 birds (45 birds per treatment). During the indoor treatment, the chickens were raised in floor pens in a conventional poultry research house (7 birds/m2). In the free-range treatment, the chickens were housed in a similar indoor house (7 birds/m2); in addition, they also had a free-range grass paddock (1 bird/m2). All birds were provided with the same starter and finisher diets and were raised for 112 d. Results showed that the BW and weight gain of the chickens in the free-range treatment were much lower than that of the chickens in the indoor floor treatments (P<0.05). There was no effect of the free-range raising system on eviscerated carcass, breast, thigh, and wing yield (P>0.05). However, the abdominal fat yield and tibia strength (P<0.05) significantly declined. The nutrient composition (water, protein, and fat), water-holding capacity, shear force, and pH of the muscle were largely unaffected (P>0.05) by the free-range raising system. The data indicated that the free-range raising system could significantly reduce growth performance, abdominal fat, and tibia strength, but with no effect on carcass traits and meat quality in slow-growing chickens.

Making global river ecosystem health assessments objective, quantitative and comparable.

Assessing and comparing global river ecosystem health in an objective and quantitative way remains a major challenge. In this study the widely-used semi-quantitative methods Rapid Biological assessment Protocols (RBPs) was used to determine the health of rivers. The findings were then compared to the results derived from our new UAV (Unmanned aerial vehicles) orthophotographic imagery method. This method quantitatively and objectively assesses river ecosystem health. As a comparison, our method was used to quantitatively measure distance and areas of a range of hydrological and biological attributes thus improving the accuracy of distance- and area-related indices, consequently avoiding subjective errors in these estimations that is fraught in methods like the RBPs. To strengthen the objectivity of the assessment the weights of these indices were objectively determined using the entropy weighting method. This new method was then tested using 9551 UAV orthophotographs taken over six field campaigns. It performed satisfactorily, showing that in our study area the health status of mountain rivers was the best with the highest score of 0.94 out of 1.0. Temporally, the health of the river was better in summer (0.65) compared with that in autumn (0.40). Changes in river ecosystem health were driven by variations in biology and water quality. In contrast the outputs of RBPs, especially in relation to distance and area indices, had ~ 20% uncertainty due to visual errors and subjectivity in estimations by observers. The UAV orthophotographic imaging method proposed in this study can improve the ability to compare the health of rivers across different periods and regions throughout the globe.

Effects of high-dose selenium-enriched yeast on laying performance, egg quality, clinical blood parameters, organ development, and selenium deposition in laying hens.

Organic selenium (Se) supplementation from Se-enriched yeast (SY) has been advocated and approved for use in animal feeds by some nutritionists and researchers rather than inorganic Se from sodium selenite. However, there is little available safety data of SY in laying hens. A subchronic study was conducted to determine if high-dose SY affects the safety of hens. A total of 768, 30-wk-old, Hy-Line Brown hens were randomly assigned to 1 of 4 groups (192 laying hens per group) with 6 replicates of 32 birds each. After a 2-wk acclimation period, the birds were fed diets supplemented with 0, 0.3, 1.5, or 3.0 mg/kg Se from SY for 12 wk. Throughout the study period, clinical observations and laying performance were measured. The hematological and chemical parameters of blood samples and the Se concentration in eggs were examined after SY supplementation for 4, 8, and 12 wk, and the egg quality was measured after 12 wk. At the end of the study, full post-mortem examinations were conducted: breast Se concentrations were measured, visceral, and reproductive organs were weighed, and specified tissues were collected for subsequent histological examinations. Although the Se concentrations in the eggs and breast meat from hens fed 3.0 mg/kg of Se from SY were 1036.73% and 2127.93% higher (P < 0.001) than those from hens fed a basal diet after 12 wk, no treatment-related changes of toxicological significance were observed. Therefore, up to 3 mg/kg organic Se from SY can be used to supplement the diets for laying hens without adverse effects following 84-d administration.

Comparison of dynamic change of egg selenium deposition after feeding sodium selenite or selenium-enriched yeast.

The aim of this study was to compare the dynamic change of egg selenium (Se) deposition after sodium selenite (SS) or selenium-enriched yeast (SY) supplementation for 1, 3, 5, 7, 14, 21, 28, 56, and 84 d. A total of 576 32-wk-old Hy-Line Brown laying hens were randomly assigned to 3 groups (192 laying hens per group) with 6 replicates, and fed a basal diet (without Se supplementation) or basal diets with 0.3 mg/kg of Se from SS or 0.3 mg/kg of Se from SY, respectively. The results showed that the Se concentrations in the eggs from hens fed a SY-supplemented diet were significantly higher (P < 0.001) than those from hens fed a SS-supplemented diet or a basal diet after 3 d. And the Se concentrations in the eggs from hens fed a SS-supplemented diet were significantly higher (P < 0.001) than those from hens fed a basal diet after 14 d. There was a positive linear and quadratic correlation between Se concentrations in the eggs from hens fed a SY-supplemented diet (r2 = 0.782, P < 0.001; r2 = 0.837, P < 0.001) or SS-supplemented diet (r2 = 0.355, P < 0.001; r2 = 0.413, P < 0.001) and number of feeding days. The Se concentrations in the breasts from hens fed a SY-supplemented diet were 126.98% higher (P < 0.001) than those from hens fed a SS-supplemented diet, and were 299.44% higher (P < 0.001) than those from hens fed a basal diet after the 84-d feeding period. In conclusion, the dietary Se was gradually transferred into eggs with the extension of the experimental duration. The deposition rate of Se in the eggs from hens fed a SY-supplemented diet was much more rapid than that from hens fed a SS-supplemented diet, and the organic Se from SY had higher bioavailability as compared to inorganic Se from SS.

A prospective gating method to acquire a diverse set of free-breathing CT images for model-based 4DCT.

Breathing motion modeling requires observation of tissues at sufficiently distinct respiratory states for proper 4D characterization. This work proposes a method to improve sampling of the breathing cycle with limited imaging dose. We designed and tested a prospective free-breathing acquisition protocol with a simulation using datasets from five patients imaged with a model-based 4DCT technique. Each dataset contained 25 free-breathing fast helical CT scans with simultaneous breathing surrogate measurements. Tissue displacements were measured using deformable image registration. A correspondence model related tissue displacement to the surrogate. Model residual was computed by comparing predicted displacements to image registration results. To determine a stopping criteria for the prospective protocol, i.e. when the breathing cycle had been sufficiently sampled, subsets of N scans where 5 ⩽ N ⩽ 9 were used to fit reduced models for each patient. A previously published metric was employed to describe the phase coverage, or 'spread', of the respiratory trajectories of each subset. Minimum phase coverage necessary to achieve mean model residual within 0.5 mm of the full 25-scan model was determined and used as the stopping criteria. Using the patient breathing traces, a prospective acquisition protocol was simulated. In all patients, phase coverage greater than the threshold necessary for model accuracy within 0.5 mm of the 25 scan model was achieved in six or fewer scans. The prospectively selected respiratory trajectories ranked in the (97.5 ± 4.2)th percentile among subsets of the originally sampled scans on average. Simulation results suggest that the proposed prospective method provides an effective means to sample the breathing cycle with limited free-breathing scans. One application of the method is to reduce the imaging dose of a previously published model-based 4DCT protocol to 25% of its original value while achieving mean model residual within 0.5 mm.