Many types of endocrine cells have been identified in the gastroenteropancreatic system of vertebrates, which have subsequently been named with alphabet (s). L cells which secrete the glucagon-like peptide (GLP)-1 are scattered in the intestinal epithelium. This review discusses the morphological features of chicken L cells and GLP-1 secretion from intestinal L cells. L cells, identified using GLP-1 immunohistochemistry, are open-type endocrine cells that are distributed in the jejunum and ileum of chickens. GLP-1 co-localizes with GLP-2 and neurotensin in the same cells of the chicken ileum. Intestinal L cells secrete GLP-1 in response to food ingestion. Proteins and amino acids, such as lysine and methionine, in the diet trigger GLP-1 secretion from the chicken intestinal L cells. The receptor that specifically binds chicken GLP-1 is expressed in pancreatic D cells, implying that the physiological functions of chicken GLP-1 differ from its functions as an incretin in mammals.

MicroRNAs (miRNAs) are small, non-coding RNA molecules that inhibit protein translation from target mRNAs. Accumulating evidence suggests that miRNAs can regulate a broad range of biological pathways, including cell differentiation, apoptosis, and carcinogenesis. With the development of miRNAs, the investigation of miRNA functions has emerged as a hot research field. Due to the intensive farming in recent decades, chickens are easily influenced by various pathogen transmissions, and this has resulted in large economic losses. Recent reports have shown that miRNAs can play critical roles in the regulation of chicken diseases. Therefore, the aim of this review is to briefly discuss the current knowledge regarding the effects of miRNAs on chickens suffering from common viral diseases, mycoplasmosis, necrotic enteritis, and ovarian tumors. Additionally, the detailed targets of miRNAs and their possible functions are also summarized. This review intends to highlight the key role of miRNAs in regard to chickens and presents the possibility of improving chicken disease resistance through the regulation of miRNAs.

D-Amino acids occur in modest amounts in bacterial proteins and the bacterial cell wall, as well as in peptide antibiotics. Therefore, D-amino acids present in terrestrial vertebrates were believed to be derived from bacteria present in the gastrointestinal tract or fermented food. However, both exogenous and endogenous origins of D-amino acids have been confirmed. Terrestrial vertebrates possess an enzyme for converting certain L-isomers to D-isomers. D-Amino acids have nutritional aspects and functions, some are similar to, and others are different from those of L-isomers. Here, we describe the nutritional characteristics and functions of D-amino acids and also discuss the future perspectives of D-amino acid nutrition in the chicken.

This study was designed to determine the effect of phytase extracted from Aspergillus niger (Natuphos^® E) on growth performance, bone mineralization, phosphorous excretion, and meat quality parameters in broilers fed available phosphorous (aP)-deficient diet. In total, 810 one-day-old Indian River broilers were randomly allotted into one of three dietary treatments, with six replicates per treatment. The three dietary treatments were 1) control group (CON: basal diet with sufficient aP), 2) low phytase (LPY: available phosphorus-deficient diet supplemented with 0.01% phytase), and 3) high phytase (HPY: available phosphorus-deficient diet supplemented with 0.02% phytase). Average daily gain and, feed intake, and feed conversion ratio were measured for 35 days. Excreta were collected from each pen on day 35. One broiler from each cage was euthanized to collect visceral organs and tibia samples. Broiler chickens fed LPY and HPY showed improved (P<0.05) growth performance compared to broilers fed CON on day 35. The tibia length of HPY-fed broilers was more than those of broilers fed other diets on day 35 (P<0.05). However, tibia calcium and phosphorous contents in LPY-fed broilers was higher (P<0.05) than in CON and HPY-fed broilers. Tibia length and calcium and phosphorous content showed a positive correlation (P<0.05) with the weight gain of broilers on day 35. Phosphorous level in the excreta of LPY- and HPY-fed broilers was lesser than those of CON broilers on day 35 (P<0.05). Furthermore, HPY-fed broilers showed lower (P<0.05) phosphorous content in the excreta than LPY-fed broilers. LPY- and HPY-fed broilers showed higher (P<0.05) liver weight than the CON broilers. In conclusion, broilers fed aP-deficient diet supplemented with phytase from Aspergillus excreted less phosphorus, which enhanced growth performance and tibia development from time of hatching to day 35 post-hatching.

Oral administration of l-citrulline (l-Cit) caused hypothermia, but l-Cit is not recommended in poultry diets in Japan. Watermelon is a natural source of l-Cit. The objective of this study is to examine the effect of watermelon waste, i.e., watermelon rind (WR) on the body temperature and plasma free amino acids of chicks. In Experiment 1, 14-day-old chicks were subjected to acute oral administration of WR extract (WRE) (2 ml) under control thermoneutral temperature (CT). In Experiment 2, 15-day-old chicks were orally administered 1.6 ml of either WRE, lowdose l-Cit (7.5 mmol/10 ml), or high-dose l-Cit (15 mmol/10 ml) under CT. In both experiments, rectal temperature (RT) and plasma free amino acids were analyzed. In Experiment 3, after dual oral administration of (1.6 ml) WRE or l-Cit (15 mmol/10 ml), 15-day-old chicks were exposed to high ambient temperature (HT; 35±1°C, 2 h) to monitor changes in RT. Acute oral administration of WRE significantly reduced RT under CT. The degree of RT reduction by WRE was similar to that by high l-Cit. Moreover, RT was significantly low at HT owing to the oral administration of WRE. However, the reduced RT was difficult to explain by the content of Cit in WRE alone. In conclusion, WRE could be used as a dietary ingredient to reduce body temperature for imparting thermotolerance in chicks.

Antibiotic growth promoters (AGPs) have been used for many years as supplements in various livestock diets, including those for poultry. However, the use of AGPs in feed was also associated with an increasing number of antibiotic-resistant bacteria in livestock. In this study, the in vitro antibacterial efficacies of eight commercially available non-AGPs suitable for use in poultry were investigated. Assessments included a combination of antibacterial activity assays and estimations of the minimal inhibitory and bactericidal concentrations along with scanning electron microscopy analysis. The results showed that the probiotic, CloStat^® exerted a bacteriostatic effect against all tested bacteria, namely Salmonella Typhimurium, Escherichia coli, Staphylococcus aureus, and Clostridium perfringens, whereas Gallipro Tect^® and Bacillus Blend^® demonstrated bacteriostatic activity towards most of the pathogens tested. Other commercial non-AGPs, Sangrovit^®, Fysal^®, and Mix oil blend^® showed a stronger or equal antibacterial activity compared to the positive control (AGP Maxus^® G100) againsts all bacteria tested, except C. perfringens. Nor-Spice AB^® and Varium™ did not show any significant effect against the tested bacteria. Several of the tested AGP substitutes exhibited good antibacterial efficiency against pathogenic bacteria and thus may be good candidates for second-stage in vivo investigations into reducing pathogen colonization in broilers.

We evaluated the effects of supplementing an emulsifier blend (sodium stearoyl-2-lactylate and 1, 3-diacylglycerol) in diets with different energy content (normal and 100 kcal/kg reduced) on the growth performance, meat quality, apparent total tract digestibility (ATTD), and blood lipid profile of broiler chickens. Male broiler chickens (n = 1024), with an initial body weight (BW) of 43.60±0.2 g, were used in a 35-day trial. Broiler chickens of similar body weight were randomly allocated to one of four treatment groups in a 2 × 2 factorial arrangement with two levels of dietary energy content and with or without emulsifier blend. Broiler chickens fed on emulsifier blend supplemented diet had a higher body weight gain (BWG) during d 7–21, d 21–35, and overall period (P<0.05), higher BW during overall period (P<0.05), and lower feed conversion ratio (FCR) during d 7–21, d 21–35, and overall period (P<0.05) compared with broilers fed on diets without emulsifier supplementation. Broiler chickens fed on the diet with low energy content had a lower BWG during d 1–7, d 21–35, and overall period (P<0.05), lower BW during overall period, and higher FCR during d 1–7, d 21–35, and overall period (P<0.05). The ATTD of energy tended to decrease in response to low-energy content diet (P<0.10). Drip loss at 7 d post slaughter tended to decrease in response to dietary emulsifier blend supplementation (P<0.10). However, no interactive effects of dietary energy content and emulsifier blend supplementation (P>0.10) were observed on the growth performance, ATTD, blood lipid profiles, meat quality and relative organ weight. In conclusion, dietary emulsifier blend supplementation could improve growth performance, while low dietary energy content would decrease growth performance and ATTD of energy.

Eighty 14-d-old single-comb White Leghorn male chicks were divided into 16 groups with five birds each. Fructosyl-valine, which is a valine-glucose-Amadori product, was intravenously (2,250 nmol/kg body weight) or orally (300 µmol/kg body weight) administered to chicks. Blood samples were collected 15, 30, 60, 120, 180, 360, 720 and 1440 min after administration. Plasma concentrations of fructosyl-valine were measured by using a liquid chromatography / mass spectrometry (LC/MS). The time course change in plasma fructosyl-valine concentration showed an exponential curve, as y=a+be^−λt. The half-life of plasma fructosyl-valine was calculated by the following equation: (log_e2)/λ. When fructosyl-valine was injected intravenously, the highest value for plasma fructosyl-valine concentration was observed 15 min after administration. When injected intravenously, the half-life of plasma fructosyl-valine was calculated to be 231 min. When fructosyl-valine was administered orally to chicks, the highest value for plasma fructosyl-valine concentration was observed 180 min after administration. When administered orally, the half-life of plasma fructosyl-valine was calculated to be 277 min. We conclude that the half-life of fructosyl-valine in plasma was approximately 4 h, which is longer than that of glycated tryptophan.

Probiotic bacteria are known for their beneficial effects on the intestinal immune function of the host animal. However, their effects on mucosal barrier function in chicks are not completely understood. The aim of this study was to determine the effects of the probiotic bacterium, Lactobacillus reuteri (LR), on the gastrointestinal mucosal barrier function of broiler chicks. One day-old male broiler chicks were orally injected water (300 µL) with or without 1 × 10⁸ cfu of LR (5 mg FINELACT, Asahi Calpis Wellness Co. Ltd.) every morning for 7 days (day 0 to 6). The crop, duodenum, ileum, and cecum were collected on day 7 and were used for histological analysis and RNA extraction. Then, the thickness of the mucosal structures and the number of goblet cells in the digestive tract were assessed using histological analysis. The expression of Mucin 2, factors related to the formation of tight junctions (Claudin1, 5, and 16, ZO2, and JAM2), cytokines (IL-6, CXCLi2, and IL-10), and avian β-defensin 10 (AvBDs) (AvBD2, 10, and 12) in the crop, duodenum, ileum, and cecum were analyzed using real-time polymerase chain reaction (PCR). Results showed that oral administration of LR increased ileal villus height and crypt depth, decreased Claudin16 level in the crop and increased JAM2 level in the crop and ileum, and decreased the expression of AvBD10 in the ileum and cecum and that of AvBD12 in the crop. It did not affect goblet cell number and Mucin 2 expression. These results suggested that LR used in this study may enhance mucosal barrier function by regulating tight junctions in the upper gastrointestinal tract.

Autophagy in the skeletal muscle increases under catabolic conditions resulting in muscle atrophy. This study investigated the effect of inhibition of mechanistic target of rapamycin (mTOR) on autophagy in chick skeletal muscle. We examined the effects of Torin1, an mTOR inhibitor, on autophagy. Chick myotubes were incubated with Torin1 (100 nM) for 3 h. It was observed that Torin1 inhibited the phosphorylation of AKT (Ser473), p70 ribosomal S6 kinase 1 (S6K1, Thr389), S6 ribosomal protein (Ser235/236), and eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1, Thr37/46), which are used for measurement of mTOR activity. Torin1 significantly (P< 0.01) increased the LC3-II/LC3-I ratio, an index for autophagosome formation, while it did not influence the expression of autophagy-related genes (LC3B, GABARAPL1, and ATG12). In addition, Torin1 increased atrogin-1/MAFbx (a muscle-specific ubiquitin ligase) mRNA expression. Fasting for 24 h inhibited the phosphorylation of AKT (Ser473), S6K1 (Ther389), S6 ribosomal protein (Ser235/236), and 4E-BP1 (Thr37/46) in chick skeletal muscle and significantly (P<0.01) increased the LC3-II/LC3-I ratio. Fasting also increased GABARAPL1 and atrogin-1/MAFbx mRNA expression but not LC3B or ATG12 mRNA expression. These results indicate that mTOR signaling regulates autophagy and the ubiquitin-proteasome proteolytic pathway in chick skeletal muscle.

L-Pipecolic acid is an intermediate of L-lysine catabolism. Its central injection exerted a hypnotic effect on the brain, which was partially mediated by the activation of γ-aminobutyric acid-A and γ-aminobutyric acid-B receptors. L-Proline has also been shown to exert a similar effect on N-methyl-D-aspartate receptors. Furthermore, L-pipecolic acid is known as L-homoproline, and both L-pipecolic acid and L-proline belong to the imino acid group; therefore, it is plausible that they share certain commonalities, including similar functions. However, the role of N-methyl-D-aspartate receptors with respect to the effects of L-pipecolic acid has not been examined yet. In the present study, the relationship between N-methyl-D-aspartate receptors and the central function of L-pipecolic acid was investigated in neonatal chicks. The behavioral postures for active wakefulness and standing/sitting motionless with eyes opened were significantly affected after intracerebroventricular injection of L-pipecolic acid; whereas, sitting motionless with head drooped (sleeping posture) was significantly enhanced. However, the N-methyl-D-aspartate receptor antagonist, MK-801, did not affect these changes. In conclusion, the central administration of L-pipecolic acid did not exert hypnotic effects through the activation of N-methyl-D-aspartate receptors in neonatal chicks. These results suggest that the imino group is not a determinant for activating N-methyl-D-aspartate receptors.

Because of the presence of sperm storage tubules (SSTs) in the utero-vaginal junction (UVJ) in the oviduct, once ejaculated sperm enter the female reproductive tract, they can survive for a prolonged period in domestic birds; however, the specific mechanisms involved in sperm maintenance within the SST remain to be elucidated. In this study, we showed that transferrin (TF) and albumin (ALB) are expressed in SSTs. When UVJ extracts were subjected to size-exclusion column chromatography, we obtained fractions that extend sperm longevity in vitro. LC-MS/MS analysis of the two major proteins in the fractions identified these proteins as TF and ALB. Immunohistochemical analysis using specific antisera against TF and ALB indicated that both proteins were localized not only in the SSTs, but also in the surface epithelium of the UVJ. When the ejaculated sperm were incubated with either purified TF or ALB, sperm viability increased after 24 h. These results indicated that oviductal TF and ALB are involved in the process of sperm storage in SSTs and may open a new approach for technological improvement to prolong sperm longevity in vitro.