Ex ovo culture of avian embryos can be applied not only to embryology but also to various fields of basic research such as embryo manipulation, toxicology, and regenerative medicine. The windowing method, which facilitates various manipulations and observations by opening a hole in one part of the eggshell, and culture systems using surrogate eggshells, are widely used. Despite this, biology lessons in high schools cover shell-less culture systems, which involve the development of avian embryos in artificial vessels, such as rice bowls, without using surrogate eggshells. However, as embryo development stops at its early stages in this method, it is not possible to continuously observe the development of the embryo. This led to attempts to develop an embryo culture method using a complete artificial culture vessel that does not use surrogate eggshells, and Kamihira et al. (1998) succeeded in hatching quail embryos in an artificial culture vessel using polytetrafluoroethylene membranes. In addition, Tahara succeeded in hatching chick embryos in artificial culture vessels that used cling film made of polymethylpentene and reported their detailed methodology (Tahara and Obara, 2014). These technologies are being applied not only to school education but also to various fields of research.

The aim of this study was to identify genes involved in comb development to provide insights into the molecular mechanism of chickens' comb formation. Fixation index (F_ST) and average number of base differences (π) of males with large and small combs were calculated based on whole-genome resequencing data. Chromosome regions with larger F_ST values and smaller π were considered candidate selection regions. Through further annotation of gene functions and pathways, we sought to screen possible selected genes associated with comb development. By screening whole genome resequencing data, F_ST and π were calculated using a 40 Kb sliding window strategy and eight regions were identified. Quantitative trait loci (QTL; FOX1 gene) related to comb length were found on chromosome 1. QTL (GLP1R, BTBD9, MIR6633, and MDGA1 genes) related to comb weight were found on chromosome 3. QTL (ALDH1A1, TMC1, and ANXA1 genes) associated with comb area were found on the Z chromosome. Nineteen genes, Wnt signaling pathway and neuroactive ligand-receptor interaction signaling pathway directly or indirectly related to comb growth and development were found through functional annotation and GO analysis. Among the selected genes LYN, GLP1R, FOX1, TBK1, STRAP, ST6GALNAC, and Wnt signaling pathways were related to immunity. MDGA1, BTBD9, MTSS1, SrGAPs, and neuroactive ligand receptor interaction signaling pathways related to neural function were screened. ALDH1A1, ANXAl, THBS, HIF-1α, and ACTN1 genes were related to heat dissipation. Among the selected genes FOX1, MDGAl, and ANXAl associated with immunity, neurological function, and heat dissipation function coincided with genes affecting the length, weight, and area of the comb. Comprehensive analysis suggested that comb development was due to multiple genes and signaling pathways.

This study aimed to examine 1) whether selection for a superior allele at a single nucleotide polymorphism site (SNP; AB604331, g.420 C>A) of the chicken cholecystokinin type A receptor (CCKAR) gene in Miyazaki Jitokko chickens is detectable in commercial poultry farms, and 2) whether the reproductive traits of the Kyushu Rhode hens, as a maternal stock line of the Miyazaki Jitokko chickens, are affected by SNP selection. Conventional and A-allele fixed (improved) Miyazaki Jitokko chicks were hatched on the same day and raised in a battery cage until 7 days of age. The chicks were then deposited at two commercial poultry farms and reared until slaughter at 126 and 163 days for cockerels and pullets, respectively. Body weight on the day of hatching (day 0), at 5 days of age, and at slaughter were measured. The differences in the body weights of the farm and test groups at slaughter were analyzed using the generalized linear model. A-allele fixation increased the body weight at slaughter by approximately +123.5 g and +131.9 g in cockerels and pullets, respectively. No significant differences between the conventional and improved hens were detected in terms of egg-laying rate, fertilization rate, and hatchability in the Kyushu Rhode hens. The data suggest that fattening chicks can be supplied as usual, even if Kyushu Rhode hens are switched from the conventional to improved type. In conclusion, genetic improvements using the CCKAR SNP site as a marker were effectively established in terms of the growth of the Miyazaki Jitokko chickens in commercial farms and the reproductive traits of the Kyushu Rhode hens.

The effects of dietary supplementation of zinc (Zn) sources and concentrations were investigated on growth performance, absorption into tissues, fecal excretion, nutrient retention, and intestinal morphology in broilers fed a corn-soybean meal basal diet. A total of 525 one-day-old chicks (Ross 308) were assigned based on body weight to seven dietary treatments. There were five replicate pens for each treatment and 15 broilers per replicate pen. The dietary treatments included a basal diet (control, without supplementing Zn), and basal diet supplemented with Zn, as inorganic zinc sulfate (ZnS; 110 mg/kg); organic Zn-methionine (ZnM; 110 mg/kg); hot-melt extruded (HME) 25 zinc sulfate (27.5 mg/kg); HME50 zinc sulfate (55 mg/kg); HME75 zinc sulfate (82.5 mg/kg); or HME100 zinc sulfate (110 mg/kg) for 35 days in two phases (d 1–21, phase I and d 22–35, phase II). Bodyweight and feed efficiency of broiler chicks fed diets supplemented with increasing dietary concentrations of HME-Zn improved linearly during the study period (P<0.05). Compared to the control treatment, the ZnS, ZnM, and HME diets increased Zn concentrations in the serum and liver. Inorganic ZnS supply resulted in the highest Zn concentration in excreta. Increasing supplemented Zn content in diets as HME linearly increased Zn concentration in the excreta, serum, liver, and tibia. Broiler chicks fed diets supplemented with increasing concentrations of HME increased villus height (VH; linear and quadratic) of the jejunum and VH of the ileum (linear). Increasing concentrations of dietary Zn supplied as HME resulted in linearly enhanced dry matter, gross energy, and nitrogen retention of broilers on day 21. These results suggest that dietary HME-Zn at a lower level (55 ppm) shows the same growth performance as common ZnSO₄ at 110 ppm.

Necrotic enteritis caused by Clostridium perfringens has reemerged as a severe poultry disease worldwide since the ban on the routine use of antibiotics in animal feed. Probiotics are considered alternatives to antibiotic growth promoters. Bacillus species are the most common microorganisms used as probiotics in the feed industry. The current study investigated the growth and surfactin levels of Bacillus licheniformis in solid-state fermentation using different substrates and evaluated the effects of B. licheniformis-fermented products on growth performance and intestinal morphology in broilers exposed to C. perfringens. The highest viable biomass and surfactin concentration of B. licheniformis-fermented products was observed at 2% molasses and 20% soybean meal supplementation during fermentation (P<0.05). B. licheniformis-fermented product-derived surfactin inhibited the growth of C. perfringens in vitro in a dose dependent manner (P<0.05). B. licheniformis-fermented product supplementation (2 g/kg) significantly improved the body weight and average daily gain weight of broilers challenged with C. perfringens (P<0.05). B. licheniformis-fermented products significantly alleviated necrotic lesions and ameliorated intestinal morphology in broilers exposed to C. perfringens (P<0.05). Collectively, these observations demonstrate that B. licheniformis-fermented products improve growth performance and gut morphology in broilers under C. perfringens challenge. B. licheniformis-fermented products may have the potential to be used as alternatives to antibiotic growth promoters for preventive treatments against C. perfringens in broilers.

The aim of this study was to study the regulation of abdominal fat deposition by DL-α-tocopherol acetate (vitamin E) in broilers. Diets supplemented with 50 IU vitamin E significantly diminished abdominal fat deposition in broilers at day 35. Transcriptome sequencing results for abdominal fat tissues of the control (FC) and 50 IU vitamin E-supplemented (FT) groups identified 602 differentially expressed genes (DEGs), which were enriched in cellular process, cell and cell part, and binding Gene Ontology terms. Pathway functional analysis revealed that the DEGs were enriched in 42 metabolic pathways. Notably, the most enriched pathway, fatty acid biosynthesis, was found to play a key role in lipid metabolism. Further, the key regulators of lipid metabolism, including fatty acid synthase, acetyl-CoA carboxylase alpha, and acyl-CoA synthetase long-chain family member 1, demonstrated decreased expression following vitamin E supplementation. Herein, we have identified pathways and genes regulated by vitamin E, thereby providing novel insights into the nutrients regulating abdominal fat deposition in broilers.

The present study was conducted to analyze the chemical properties of persimmon peel (PP) and the in vitro digestibility of shrimp meal (SM) diets containing PP. Discussions whether PP can be used as a feed additive to promote digestion of SM in chickens are also included. The chemical composition and chitinase activity of dried PP was studied. SM diets containing PP were formulated according to the 4 by 6 factorial design: 4 levels of SM (0%, 10%, 15%, and 20%) × 6 levels of PP (0%, 2%, 4%, 6%, 8%, and 10%). The in vitro digestibility of dry matter (IVDMD), crude protein (IVCPD), and chitin (IVCD) was also studied. PP was rich in nitrogen-free extract (NFE, about 74%) and tannin (2.8%), and the highest chitinase activity of PP was observed at pH 4.5. Approximately 50% of chitinase activity was also observed at acidic (3.0) and alkaline (8.0) pH. Its activity was slightly affected by pepsin treatment. IVDMD increased upon addition of up to 8% PP, but decreased with an increase in the level of SM. When PP level was increased up to 6%, IVCPD in the group containing 0% SM, changed slightly; however, an increasing trend was observed in the other groups. When PP level was more than 6%, IVCPD decreased in all the groups. IVCD increased dose-dependently with increasing level of PP and decreased with increasing level of SM. In conclusion, PP was rich in NFE, had high chitinase activity, and improved all digestibility parameters, such as IVDMD, IVCPD, and IVCD, in SM diets where the PP level was under 6%. Thus, up to 6% of PP can be safely included in SM diets as a digestion promoter.

The experiment reported herein was conducted to estimate the ileal and excreta endogenous phosphorus (P) losses in broiler chickens. Three purified diets, namely a P-free diet, a gelatin-based diet containing negligible amounts of P, and a casein-based diet with 100% available P, were formulated. Test diets were offered ad libitum from day 25 to 28 post hatch and ileal digesta were collected. Excreta samples were also collected to estimate total tract endogenous P losses. Ileal endogenous P losses in birds fed the casein-based diet were higher (P<0.05) than those in birds fed P-free and gelatin-based diets. The ileal endogenous losses of P in birds fed P-free, gelatin-based, and casein-based diets were 25, 104 and 438 mg/kg dry matter intake, respectively. The endogenous P loss values estimated at the excreta level were 830, 560 and 372 mg/kg dry matter intake, respectively. Ileal and excreta endogenous losses of P in birds fed a casein-based diet were similar (P>0.05), but ileal losses were lower (P<0.05) than the excreta values in birds fed P-free and gelatin-based diets, resulting in a significant (P<0.001) assay diet by site of measurement interaction. The present data demonstrate that values determined for endogenous P losses in broiler chickens vary widely depending on the assay diet used.

The molecular mechanism underlying photoperiodic response in seasonal breeding animals such as the Japanese quail, red jungle fowl, sheep, mouse, and hamster involves thyroid-stimulating hormone beta subunit (TSHβ) mRNA expression in the pars tuberalis stimulated by the extension in day length. Furthermore, this mechanism is regulated by eyes absent 3 (Eya3) in mammals. Even in birds, the expression of both TSHβ and EYA3 is induced in the pars tuberalis by the extension in day length; however, the relationship between the two genes is unknown. To clarify the function of EYA3 in quail photoperiodism, in the present study, we performed mRNA structure analysis of the Japanese quail EYA3 mRNA using reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analysis. The results revealed that there are four types of splice variants within regions of exons 7, 8, and 9 of quail EYA3 mRNA. Among the four splice variants of quail EYA3, the splice variant containing exon 7 was expressed in the pars tuberalis on the first long day, when quails were transferred from the short-day condition to the long-day condition. The results indicate that EYA3 splice variant containing exon 7 is involved in the photoperiodic response of the pars tuberalis in the Japanese quail.

Effects of a large-sized cage with a low metabolizable energy and low crude protein (LME-LCP) diet on growth performance, feed cost, blood biochemistry, and antibody response of growing layers were evaluated. A total of 668 one-day-old female Gushi chicks were randomly allocated into three different cages, namely, large, medium, and small cages, referred to as Group A, Group B, and Group C, respectively, and fed LME-LCP diets. A fourth group of birds raised in small cages and fed a standard diet (STD) was designated Group D i.e. the control. Equal stocking densities were maintained among the four groups throughout the experiment, from 50–20 birds/m². Large cages with LME-LCP diet (Group A) increased the shank length and girth as chicks grew, improved the activities of serum creatine kinase, and reduced serum triglyceride and cholesterol concentrations. The total feed intake in birds from Group A was higher than those from the other groups at every stage. The total cost (rmb/bird) of feed was 6.70% lower in Group A than that in Group D, which indicated the cost-effectiveness of large cages. In conclusion, large cages with LME-LCP diets have positive effects on body weight, shank growth, and serum biochemical indices of growing Gushi chicks, and can reduce feed costs.