Several conventional traits, including eggshell thickness, are commonly being improved genetically as a means to increase eggshell strength. At the same time, researchers have come to recognize that factors related to egg geometry, such as egg shape, are important determinants of the variability remaining in eggshell strength, after conventional traits have been considered. Therefore, given that the value of the egg shape index -the egg’s width to length ratio- depends highly on the hen strain, it is necessary to examine the relationship between eggshell strength and shape index more closely in a variety of breeds. From this perspective, by using REML methodology under a five-trait animal model, we analyzed a two-way selection experiment for non-destructive eggshell deformation in 31 generations of White Leghorns, to evaluate the effect of selection for eggshell strength on egg shape. In the strong line, which refers to the line that was selected for decreased non-destructive deformation value, the genetic correlation between eggshell breaking strength and shape index was 0.285±0.055, whereas that between non-destructive deformation and shape index was -0.021±0.063. In the weak line, these values were 0.244±0.055 and -0.093±0.060, respectively. The heritability estimates were 0.381±0.033 for non-destructive deformation, 0.349±0.029 for eggshell breaking strength, and 0.544±0.027 for shape index in the strong line, and 0.408±0.031, 0.468±0.032, and 0.484±0.028, respectively, in the weak line. The genetic correlation between eggshell breaking strength and shape index suggests that rounder eggs are somewhat more resistant to breakage than more elongated eggs. The moderately high heritability estimates for shape index indicate the potential to improve egg shape through genetic gain.

Chicken egg white ovotransferrin (OTF) has been reported to exist in three electrophoretic variants (OTF^A, OTF^B and OTF^C). In this report, we identified a causal polymorphism between the OTF^B and OTF^C variants in Japanese and Taiwanese native chickens and compared the antibacterial activity between these two variants. The cDNA sequence analyses from Satsumadori oviducts revealed that three non-synonymous SNPs T1809G (Ser52Ala), A2258G (Ile96Val) and G7823A (Asp500Asn) corresponded to the OTF electrophoretic phenotypes. Of the three SNPs, the G7823A mutation perfectly corresponded to the electrophoretic phenotypes OTF^B (G/G, Asp500Asp), OTF^B/C (G/A, Asp500Asn) and OTF^C (A/A, Asn500Asn) in three chicken populations. The variants OTF^B and OTF^C exhibited similar antibacterial potency against Gram-positive and Gram-negative bacteria. This study provides, for the first time, molecular information on polymorphism of OTF^B and OTF^C variants of chicken ovotransferrin and its effect on the antimicrobial activity of the respective variants.

The aim of this study was to investigate the effect of reduced dietary energy (ME) and protein (CP) levels along with administration of a phytogenic feed additive (PFA) based on oregano, anise and citrus essential oils, on broiler growth performance, nutrient digestibility, meat and blood biochemical parameters and total antioxidant capacity (TAC).

Depending on dietary ME and CP level down regulation compared to a corn-soybean meal basal diet A used as positive control, three diet types [A, B(＝A-3%) and C(＝A-6%)] were implemented. Depending on the inclusion or not of PFA at 125 mg/kg diet, 450 1-d old, male Cobb broilers were randomly allocated in six treatments according to a 3×2 factorial arrangement with 5 replicates of 15 broilers; A: diets formulated optimally to meet broiler nutrient requirements for maximizing protein content of meat for starter, grower and finisher growth periods; APh: A+PFA; B: suboptimal in ME and CP levels by 3%; BPh: B+PFA; C: suboptimal in ME and CP levels by 6%; CPh: C+PFA.

Feed conversion ratio (FCR) was improved in birds fed diet A compared to diet C during the grower period (P_D＝0.021) and overall (P_D＝0.010). Phytogenic supplementation resulted in higher (P_D×Ph＝0.020) total tract apparent digestibility of fat in birds fed diet C compared to diet A. Birds fed diet A had higher (P_D＝0.001) plasma cholesterol, compared to birds fed diet C. In addition, birds fed diets A and B had higher (P_D＝0.002) breast protein content compared to C. Overall, PFA inclusion reduced cholesterol (P_Ph＝0.002) and increased plasma TAC (P_Ph<0.001). Moreover, PFA increased breast (P_Ph＝0.001) and thigh (P_Ph＝0.01) TAC. In conclusion, a reduction in dietary ME and CP levels, adversely affected the FCR, whereas PFA supplementation tended (P_Ph＝0.089) to compensate these effects. Moreover, the addition of PFA reduced plasma cholesterol and improved plasma and meat TAC.

The objective of the present study was to examine the effects of nano-selenium on growth performance, antioxidative status, and immune function in broiler chickens reared under thermoneutral (22±1°C) or high ambient temperature (35±1°C) conditions. Thirty-six broiler chicks at 15d old were randomly divided into 6 treatments in a 3×2 factorial design. The main factors included the dietary supplementation (basal diet without Se supplementation [control], basal diet with 0.3 mg of nano-elemental Se per kilogram of diet [nano-Se], and basal diet with 0.3 mg of sodium selenite per kilogram of diet [SSe]) and the ambient temperature challenge (22±1°C or 35±1°C). The birds were given the experimental diets from 15 to 30 d of age. High ambient temperature significantly depressed body weight gain, feed intake, feed conversion ratio, breast muscle weight, and abdominal fat weight, while feeding nano-Se clearly alleviated these negative effects of high ambient temperature. In addition, feeding nano-Se increased glutathione peroxidase mRNA expression in liver and alleviated the negative effects of high ambient temperature via reducing the malondialdehyde content in liver and breast muscle. Furthermore, feeding nano-Se increased mRNA expression of cytokine genes (interleukins 2 and 6) under both thermoneutral and high ambient temperature conditions. Under both thermoneutral and high-temperature conditions, broiler chickens fed nano-Se had higher Se and vitamin E concentrations in breast muscle than broiler chickens fed the control diet. In contrast, feeding SSe at the same dose as nano-Se did not alleviate the negative effects of high ambient temperature on broiler chickens. In conclusion, dietary supplementation with nano-Se at 0.3 mg/kg diet might enhance growth performance by improving antioxidative or immune properties in broilers reared under high ambient temperature.

The aim of the study was to test the effect of high moisture ear corn (HMEC) and high moisture shelled corn (HMSC) feeds on laying hen performance. A total of 108 Rhode Island Red laying hens were divided by body weight and performance into 12 blocks (9 in each). Each block was assigned to one of three dietary treatments. The hens were fed one of three experimental diets containing 48.0% commercial laying hen diet (CON), 55.7% HMEC (EC) or 48.5% HMSC (SC) on a dry matter (DM) basis. All diets were isocaloric (2.80 mega calorie (Mcal) of (metabolizable energy (ME)/kg of DM) and isonitrogenous (15.5% CP of DM). DM Feed intake (139 to 148 g DM/d per hen), egg production rate (79 to 85%), egg mass (47.6 to 51.2 g/d per hen) and feed utilization (2.7 to 3.1 g of feed DM/g of egg) were not affected by diet. Body weight for the SC diet was significantly less than that for the CON diet (P<0.05). This was possibly due to the low feed intake during weeks 1 and 2 because of the short adaptation period to the experimental diet. The body weight loss of hens fed the SC diet may not have occurred when there had been a sufficient adaptation period. Eggshell strength (3.27 to 3.52 kg/cm²) and Haugh unit (80.0 to 83.6) were not affected by diet. In conclusion, HMEC and HMSC diets do not significantly affect laying hen performance and can be used as a main ingredient of the laying hen diet.

Remarkably hardened pectoralis major muscles are increasingly found in broiler chickens in some slaughterhouses in Japan. Based on physical examinations, we selected three affected birds and three control birds from a 43-day-old Ross 308 commercial broiler flock, and examined them biochemically, pathologically, and microbiologically. We found that the presence of degenerative lesions in the pectoralis major muscles was associated with an inability to lift the wings to achieve back-to-back wing contact. As the pectoralis major muscle is a major adductor of the humerus, these degenerative lesions may inhibit the extensibility of the muscle, thereby limiting the range of motion of the humerus. Testing the ability to lift the wing is proposed as a new diagnostic method for detecting affected birds.

The aim of this study was to determine whether probiotic-feeding affected the expression of cathelicidins (CATHs), a major family of antimicrobial peptides, in response to lipopolysaccharides (LPS) challenge in the proventriculus and cecum of broiler chicks. One-day-old male Chunky broiler chicks were fed with or without 0.4% probiotics for 7 days (P-group and non-P-group, respectively). Then, they were orally challenged with no LPS (0-LPS), 1 μg LPS (1-LPS), or 100 μg LPS (100-LPS) (n＝5 in all groups) in Experiment 1, and with no LPS or 1 μg LPS (n＝6 in all groups) in Experiment 2. Five hours after LPS challenge, the proventriculi and ceca were collected to analyze CATHs expression. Expression of CATHs was examined at first by reverse transcription-polymerase chain reaction (RT-PCR) using the 0-LPS chicks of non-P-group. The differences in CATHs expression upon probiotics-feeding and LPS were analyzed by real time-PCR. All four CATHs (CATH1, 2, 3 and 4) were expressed in the proventriculus and cecum of chicks. In the proventriculus, the expression of CATHs after LPS challenge did not show significant differences between non-P and P-groups in Experiment 1 and 2. In the cecum, the interactions of the effects of probiotics and LPS on the expression of CATH2 in Experiment 1 and CATH1 and 2 in Experiment 2 were significant, and their expression in 1-LPS chicks was higher in P-group than in non-P-group. However, CATH3 and 4 did not show any significant differences between non-P- and P-groups challenged with LPS. These results suggest that probiotics-feeding may stimulate the immunodefense system mediated by CATH2 and possibly CATH1 against infection by Gram-negative bacteria in the cecum.

Cryopreserved sperm undergoes serious damage which affects its fertilizing ability. Despite progress in understanding the nature of functional deterioration in mammalian sperm, little is known about the mechanism involved in the induction of functional damage in avian sperm. Cellular membranes are considered the primary site of cryodamage to sperm. Membrane rafts are specific membrane regions enriched in sterols, ganglioside G_M1, and functional proteins and they play important roles in the regulation of diverse functions exerted in mammalian sperm during fertilization. Several reports investigating cryopreservation-induced membrane changes in mammalian sperm have suggested that cryopreservation induces a compositional alteration of membrane rafts via a loss of membrane sterols, leading to impaired fertilizing ability. Recently, we demonstrated that membrane rafts are present in chicken sperm. Therefore, we investigated a possible mechanism for the induction of functional damage in cryopreserved chicken sperm, with particular attention to cryopreservation-induced compositional changes in membrane rafts. Sterol quantification showed that loss of sterols from sperm membranes occurred following cryopreservation. Biochemical analyses of detergent-insoluble membranes showed that the lipid and protein compositions of membrane rafts were altered dramatically by cryopreservation. To determine the physiological role of these changes, we examined external translocation of phosphatidylserine (PS), representing an early apoptotic change, and found that cryopreservation induced apoptotic changes in chicken sperm. Furthermore, methyl-β-cyclodextrin-induced loss of sterols from the plasma membranes stimulated PS translocation that was not accompanied with caspase-3 activation, which plays an important role downstream of the apoptotic cascade. Based on the results obtained in this study, we discuss a new mechanism for reduction of the fertilizing ability in avian sperm after cryopreservation.

Vasoactive intestinal peptide (VIP) treatment induced mRNA expression of Prolactin (PRL) in the chicken anterior pituitary gland. VIP responsive element (VRE) of the PRL promoter was identified in the various bird species. However, transcription factor, which binds to VRE, has not yet been identified. Prolactin regulatory element-binding protein (PREB) gene cloned as a candidate transcription factor binds to VRE. Increases of mRNA levels of PRL and PREB during embryogenesis were identified. However, whether VIP affects levels of PRL and PREB mRNA during embryogenesis remains unknown. The effects of VIP and forskolin on mRNA expression of PRL and PREB in the embryonic anterior pituitary gland were assessed. Furthermore, administration of VIP to laying hens was conducted to examine the relationship between VIP and PREB mRNA expression. At day 14 of the embryonic growth stage, VIP treatment did not affect mRNA levels of either PRL or PREB, whereas forskolin treatment induced the increase of these mRNA levels. At day 20, both VIP and forskolin induced an increase of PRL and PREB mRNA levels. The administration of VIP significantly increased mRNA levels of PRL and PREB in the anterior pituitary gland of White Leghorn and Nagoya. These results indicate that the effects of VIP on PRL and PREB mRNA expression levels of VIP receptor may in turn affect PRL and PREB mRNA levels in the chicken anterior pituitary gland.

Salmonella Enteritidis, S. Gallinarum and S. Pullorum are common serovars to infect poultry and cause diseases differently. The antibody production and cellular immune responses of male and female layers were evaluated before and after inoculation. Before inoculation, S. Gallinarum and S. Pullorum could survive and grow in 10% sera from 6-week-old layers, and S.Enteritidis and E. coli were completely eliminated. The weights of the male and female layers were increased the lowest by inoculation with S. Gallinarum, followed by S. Pullorum, and S. Enteritidis. Inoculation with S. Enteritidis, S. Gallinarum and S. Pullorum increased the antibody titer in the males depending on the serovars and maintained same higher antibody level in females. Furthermore, an increased anti-Salmonella IgG titer was associated with bactericidal ability and the level was reduced by serovars and complemente. Despite the vaccination and serovars, the male layers expressed more IgG2a than IgG1, indicating preferential activation of the Th1 pathway. The inoculation number affected the expression level of IFN-γ and IL-12 in the blood not in the secretion of the peripheral blood mononucleated cells (PBMCs) and more inoculations increased the expression of both cytokines. Inoculation increased more reactive oxygen species (ROS) production in polymorphonuclear (PMN) cells, not the PBMCs. ROS production was greater in cells from the males than from the females and greater in the cells treated with S. Enteritidis than S. Gallinarum and S. Pullorum. These three serovars and their vaccinations differed in sera killing and immune responses.

Necrotic enteritis (NE) is a poultry disease caused by Clostridium perfringens and characterized by severe intestinal necrosis. The incidence of avian NE has been progressively increasing following the removal of antibiotics from poultry feed. We evaluated the effect of diets supplemented with the thermally-processed clays, calcium montmorillonite (CaMM) on clinical signs, immunopathology, and cytokine responses in broiler chickens using an experimental model of NE consisting of co-infection with Eimeria maxima and C. perfringens. In Trial 1, Ross/Ross chickens were fed from hatch with a normal basal diet or a CaMM-supplemented diet with or without a fermentable fiber, an organic acid, and/or a plant extract, and co-infected with E. maxima and C. perfringens under conditions simulating clinical infection in the field. Chickens fed a diet supplemented with CaMM plus a fermentable fiber and an organic acid had increased body weight gain, reduced gut lesions, and increased serum antibody levels to C. perfringens α-toxin and NetB toxin compared with chickens fed the basal diet alone. Levels of transcripts for interleukin-1β (IL-1β), IL-6, inducible nitric oxide synthase, and tumor necrosis factor-α superfamily-15 were significantly altered in the intestine and spleen of CaMM-supplemented chickens compared with unsupplemented controls (p<0.05). In Trial 2, Cobb/Cobb chickens were fed an unsupplemented diet or a diet supplemented with CaMM or Varium^®, each with a fermentable fiber and an organic acid, and co-infected with E. maxima and C. perfringens under subclinical infection conditions. Compared with unsupplemented controls, broilers fed with CaMM plus a fermentable fiber and an organic acid had increased body weight gain, and reduced feed conversion ratio, mortality, and intestinal lesions, compared with chickens fed an unsupplemented diet (p<0.05). Dietary supplementation of broiler chickens with CaMM plus a fermentable fiber and an organic acid might be useful to control avian NE in the field.