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.

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.

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.

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 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.

Safety evaluation of daidzein in laying hens: Effects on laying performance, hatchability, egg quality, clinical blood parameters, and organ development.

Daidzein has become increasingly popular as a dietary supplement, particularly for postpeak-estrus animals, as a safe and natural alternative estrogen-like compound. However, there is little available safety data of daidzein in laying hens. A study was conducted to examine if high-dose daidzein affected the safety of hens, including mortality, laying performance, egg quality, hematological parameters, clinical chemical parameters, organ development parameters, and hatchability. A total of 2,448 42-wk-old Rugao laying hens were randomly assigned to 4 groups with 6 replicates of 102 birds each (612 laying hens per group). After a 2-wk acclimation period, the birds were fed diets supplemented with 0, 10, 100, or 200 mg/kg of daidzein for 12 wk. The hatchability of setting eggs increased linearly with increasing dietary daidzein supplementation (P = 0.034), while the hatchability of fertile eggs also tended to increase linearly (P = 0.069). The red cell distribution width (RCDW) and coefficient variation of RCDW showed an increasing and then decreasing quadratic response to increasing dietary daidzein supplementation (P = 0.001 and 0.002, respectively). No statistically significant changes were observed in mortality, laying performance, egg quality, clinical chemistry parameters, or organ development parameters (P > 0.05). The magnitude of these hematological changes was such that they were considered to be of no toxicological significance. Therefore, a nominal daidzein concentration of 200 mg/kg is not expected to cause adverse effects following daily administration to laying hens for 84 d.

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.

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.