This study evaluated the antioxidant capacity of Ricinus communis L. (RC) leaves and powder when used as a feed additive for laying hens. Results showed that the total phenolic content of the aqueous leaf extract of Ricinus communis L. (RCE) was 48.39mg gallic acid equivalent (GAE) per gram dry weight (DW). The flavonoid content was 9.76mg quercetin dihydrate equivalent (QE)/g DW. Ferrous chelating activity was approximately 56.2% with an RCE concentration of 1mg/mL; the highest chelating activity was 91.2% with 4mg/mL extract. The reducing power of 1mg/mL RC was 1.17 times better than 1mg/mL butylated hydroxytoluene (BHT). The Trolox equivalent antioxidant capacity (TEAC) value of 12.5mg/mL RCE was equivalent to 3.09mg/mL Trolox. RCE (10mg/mL) had a lipid oxidative inhibition capacity of 35.3%. A total of 80 ISA brown laying hens at twenty-nine weeks of age were randomly allocated into the control or 1 of 3 treatment groups; the latter received 0.5%, 1% or 2% of RC, respectively, for 12 weeks. Results showed that the RC supplementation improved the feed conversion rate and 0.5% RC generated the best results. Additionally, the egg yolk score was significantly increased in all RC-supplemented groups. Moreover, there was no significant difference in serum characteristics between the treatment groups. Serum antioxidant enzyme activity showed that superoxide dismutase (SOD) activity increased in the RC-supplemented groups relative to the control but was not significantly different. mRNA expression levels of the antioxidant regulatory genes GCLC, GST, HO-1, SOD1, and SOD2 were significantly increased with 2% RC supplementation. In summary, RC is a suitable feed additive for laying hens and the addition of 0.5% RC leaf powder resulted in the greatest benefits.
Neurotensin is secreted from intestinal N cells in response to the food ingestion. Influences of different dietary protein levels on neurotensin-immunoreactive cells in the chicken ileum were examined by using immunohistochemical and morphometrical techniques. The results showed that dietary protein had an obvious influence on neurotensin-immunoreactive cells in the chicken ileum. Four experimental groups were used, with dietary crude protein (CP) levels of 18% (control), 9%, 4.5% and 0%. Enteroendocrine cells showing neurotensin-immunoreactivity were located in crypts and villous epithelium in all groups. Most of the neurotensin-immunoreactive cells in the villous epithelium showed pyramidal or spindle-like shape with a long cytoplasmic process reaching the intestinal lumen, but cells with round or oval shape were found in the CP4.5% and 0% groups. Frequencies of occurrence of neurotensin-immunoreactive cells in the CP18%, 9%, 4.5% and 0% groups were 42.4 ±3.3, 36.6 ±2.2, 30.8 ±2.6 and 25.4 ±3.8 (cell count per mucosal area: cells/mm2, mean ± SD), respectively. There were significant differences in neurotensin-immunoreactive cell frequency between the control and lower CP level, 4.5% and 0%, groups. A significant correlation was found between frequency of occurrence of neurotensin-immunoreactive cells and daily protein intake. These results indicate that ingested protein is likely to be a potential signal for neurotensin production and secretion of N cells in the chicken ileum.
The nasal mucosa plays an important role in the immune system, with nasal mucous cells secreting mucin that, along with pili, exclude foreign substances from intervening. Nasal mucosal-associated lymphoid tissue (NALT), present in the nasal lamina propria, acts as a local immune system. In birds, the Harderian gland in the orbit also plays an important role in the local immune system. In this study, we analyzed the pathway from the nasolacrimal duct to the nasal cavity in chickens and the distribution of the nasal mucous cells responsible for defense mechanisms against pathogens. To determine the three-dimensional structure of the pathway from the nasolacrimal duct to the nasal cavity, we made casts of the anatomy by injecting an acrylic resin into the area. We then prepared paraffin sections to determine the distribution of the NALT and mucous cells. The mucous gland was clearly seen in the mucosal epithelium of the nasal cavity, suggesting that the pathway along the nasal cavity develops a nonspecific immune system to deal with large foreign substances, such as bacteria, using mucins that are secreted from the mucous glands. Hence, there is not only a physical barrier but also an antibacterial activity. Unlike in other animals, morphologically, the nasolacrimal duct in chicken becomes the ventral nasal meatus and opens into the choanae in the caudal portion of the nasal cavity. NALT was prominently present in the lamina propria of the ventral nasal meatus, suggesting the presence of a specific immune system protecting against avian viruses. Thus, responses to vaccine stimulation could be developed from tissues along the pathway of the ventral nasal meatus via the nasolacrimal duct running from the punctum. These morphological studies suggest that the instillation of eye drops could be used as an efficient vaccination method for avoiding respiratory diseases.
The present study aims to investigate the maternal origin and genetic diversity of laying-type Japanese quail lines based on partial sequences (453 base pairs) of a mitochondrial DNA (mtDNA) control region. A total of 478 individuals from 12 lines were sequenced and six different haplotypes with eight variable sites were identified. All haplotypes, two of which were identical to previously reported sequences, were typical for the Japanese quail (Coturnix japonica) and were distinct from those of the common quail (Coturnix coturnix) in a phylogenetic analysis including other published haplotypes. One haplotype was distributed in the majority of individuals (84.9%, 406/478) across all lines. Within each line, 72.5–100% of individuals had this predominant haplotype. The second most common haplotype was detected in 12.8% (61/478) individuals. These two haplotypes accounted for 97.7% of all individuals. The remaining four haplotypes were distributed with a low frequency; these were observed in five, three, two, and one individuals across all lines, respectively. All lines showed a low degree of haplotype diversity ranging from 0.0000 to 0.4321. Genetic differentiation indexes (FST) were not significant in approximately 80% pairwise comparisons of lines. The results suggest limited maternal origin and low mtDNA diversity of laying-type quail lines and may reflect their breeding history where the present gene pool was rooted in a small number of founders.
A study using pair-feeding technique was conducted to determine whether heat exposure directly or indirectly (via reduced feed intake) increases intestinal mucosal damage and permeability to endotoxin in broiler chickens. Male broiler chickens (Ross 308), 27-d-old, were subjected to one of the three treatments (n=8): 1) thermo-neutral conditions (24°C) with ad libitum feed intake, 2) heat stress conditions (33°C) with ad libitum feed intake, or 3) pair-feeding under thermo-neutral conditions, with the feed intake identical to that of heat-stressed chickens. Using these groups, two experiments were performed to evaluate temporal changes in the intestinal morphology in response to each treatment. In experiment 1, chickens were sacrificed after 24 h of exposure to the treatment conditions, while in experiment 2, chickens were sacrificed after 12 or 72 h of exposure to the treatment conditions. In experiment 1, exposure to heat stress conditions for 24 h significantly decreased both the villus height to crypt depth ratio and number of proliferating cell nuclear antigen (PCNA)-positive cells in the duodenum and increased the plasma endotoxin concentration. These findings were not observed in pair-fed chickens. In experiment 2, intestinal integrity and function were unaffected by 12 h of heat stress. On the other hand, chickens exposed to heat stress for 72 h exhibited significantly damaged intestinal morphology in the duodenum as well as increased plasma endotoxin concentration; these negative effects were not observed in pair-fed chickens. These findings suggest that the intestinal morphology and permeability changes observed in chickens that are heat-stressed for 24-72 h are due to the heat stress conditions and not due to reduced feed intake.
The objective of the study was to investigate the effect of transportation distance and crating density on broiler meat quality during hot and humid weather. Breast meat samples were collected from broilers (Ross-308) after transportation for 80, 160, and 240km at crating densities of 10, 12, and 15 birds per crate. The broilers were transported in plastic crates of 0.91×0.55×0.3m (length×width×height) in summer at an ambient temperature of 27.2–33.6°C and relative humidity of 52.7–62.9%. The birds were then slaughtered, and the meats’ physicochemical characteristics, marination and cooking attributes were measured. Meat pH at 2hours (h) postmortem (PM) was significantly higher in birds transported for 80 and 160km compared to those transported for up to 240km, while meat pH at 24h PM was higher in birds transported for up to 160km compared to the other treatments; however, no effect of different crating densities on meat pH and drip loss was observed. Meat losses such as drip loss, thaw loss, cooking loss in raw and marinated breast, and meat shear force were significantly higher in birds transported for up to 240km compared to the other treatments. An increase in crating density above 12 birds per crate also increased thaw loss, whereas crating density of 12 birds per crate and higher increased cooking loss and meat shear force. Meat lightness also increased significantly with the increase in transportation distance above 160km and crating density above 12 birds per crate. In conclusion, transportation of broilers for more than 80km with crating density higher than 12 birds per crate during summer was associated with severe losses in yield and deterioration in physical appearance and functional characteristics of meat.
The aim of this study was to determine the effects of live probiotics Lactobacillus reuteri (LR) and Clostridium butyricum (CB) on the expression of genes of innate immune system, in broiler chick ileum and cecum. Chicks were administered 500μl water with or without LR or CB, daily from day 1 to 6 after hatching. The ileum and cecum were collected on day 7 for analysis of gene expression of Toll-like receptors (TLRs), pro- and anti-inflammatory cytokines, and antimicrobial peptides (AMPs) using real-time PCR. The expression of TLR2-1 was upregulated by CB in the ileum and that of TLR5 was upregulated by both LR and CB. Expression of IL-1β and TGFβ2 in the ileum and of TGFβ3 and TGFβ4 in the cecum was upregulated by both LR and CB. The gene expressions of avian β-defensin (AvBD) 1 and cathelicidin (CATH) 3 were upregulated by CB and that of AvBD4 was upregulated by LR in the cecum. However, the expression of CATH2 in the ileum was downregulated by LR. These results suggest that probiotic LR and CB treatments affect a part of the innate defense system in the ileum and cecum by modulating the expression of innate immune molecules including TLRs, pro- and anti-inflammatory cytokines, and AMPs.
Canola meal (CM) is a commonly used feedstuff; however, it is known to be bitter, and chickens have a low preference for it. The purpose of this study was to seek clarity regarding the taste quality of CM and find methods to increase the preference for CM by chickens. We examined whether CM activates the bitter taste receptors in chickens, whether chickens show aversive responses to CM, and whether an antagonist for bitter taste receptors inhibits the bitterness of CM. Using the Ca2+ imaging technique, we showed that CM contains bitter compounds, which activate the bitter taste receptors in chickens. Further, we showed that 6-methoxyflavanone (6-meth), an antagonist for the bitter taste receptors in chickens, inhibits the activation of these receptors by CM extract. Although chickens showed a low preference for the solution of the CM extract, their preference was improved by adding 6-meth in behavioral tests. These results suggest that the preference for CM could be improved by inhibiting the bitter taste receptors in chickens.
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 (FST) 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 FST 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, FST and π were calculated using a 40Kb 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.
Glucose is a major circulating carbohydrate in birds and its level in the blood is often used as a biometric indicator in clinical diagnosis and various studies. Notably, hypoglycemia is often associated with Spiking Mortality Syndrome in broilers; therefore, blood glucose levels need to be correctly evaluated in clinical diagnosis. In the present study, we investigated the effect of different blood treatment methods after blood collection on chicken blood glucose measurements. The blood glucose level of plasma separated from blood cell components immediately after blood collection was used as a reference and compared with glucose levels in serum and stored plasma. The mean glucose level in plasma separated from blood cell components immediately after blood collection was 236.1 ± 15.9mg/dL and remained stable for at least one week in refrigerated storage (between 2°C and 5°C). However, glucose levels decreased slowly in plasma unseparated from blood cell components in storage with ice water. Mean glucose level in serum separated from blood cell components 1 h after blood collection was 206.4 ± 9.2mg/dL and fell to 108.3 ± 30.0mg/dL after 24 h. Therefore, the chicken blood serum glucose level was significantly lower than the level in plasma immediately after blood collection, regardless of elapsed time after blood collection. For the measurement of glucose in chicken blood, it is necessary to use refrigeration, use plasma from which blood cell components have been removed, and take measurements within at least 30 min.
Feeding behavior and energy metabolism are precisely regulated by humoral and/or neural factors in the central nervous system. In particular, nuclei, such as the arcuate nucleus, ventromedial hypothalamic nucleus, and lateral hypothalamic area located near the third ventricle of the hypothalamus are the centers of feeding and energy metabolism in various vertebrate species, including chickens. In this study, we evaluated the effects of cannulation of the third ventricle on chick growth and feeding behavior in the neonatal stage, to develop a method for local and chronic central nervous system-mediated energy metabolism. Referring to the chick brain atlas, a guide cannula was inserted into the third ventricle of the chick under anesthesia immediately after hatching using a stereotaxic instrument. The chicks that recovered from anesthesia were bred for 11 days under normal feeding management conditions, and then feed intake amount, body weight gain, and metabolic tissue weight were measured. The effects of direct stimulation of the third ventricle with 2-deoxy-D-glucose on the expression level of the immediate-early gene, cFOS, and feed intake in 5-day-old chicks were also evaluated. There were no differences in feed intake, body weight gain, and metabolic tissue weight between 11-day-old cannulated and control chicks. The expression of cFOS mRNA in the ventromedial hypothalamic nucleus was higher than that in the amygdala after the third ventricular administration of 2-deoxy-D-glucose. Additionally, direct third ventricular injection of 2-deoxy-D-glucose attenuated the feeding behavior of chicks for a while. Overall, we speculate that the technique is effective for local and/or chronic stimulation of the nucleus near the third ventricle of the chick hypothalamus, which is important for feed and energy metabolism regulation.
This study was conducted to determine effects of krill meal supplementation on production performance, egg quality, antioxidant substances, and fatty acid composition of egg yolk from hens fed a low-pigment diet. A total of 640 laying hens (Lohmann Brown), which were each 25 weeks old, were divided into 4 dietary treatment groups. Each treatment consisted of 8 replications with 20 laying hens per replication. The treatments were corn-soybean meal basal diet (CS), CS with 7.5% cassava meal (low-pigment; LP), and LP with 1.5 or 3% krill meal. All dietary treatments were formulated to be isocaloric (2,750 kcal/kg metabolizable energy) and isonitrogenous (17.5% crude protein). Birds were raised in an evaporative cooling system house for 8 weeks (25-33 weeks of age). Water was provided ad libitum and feed was provided according to breed requirement recommendations. The LP diets supplemented with krill meal had no effect on production performance and egg quality compared to those of the CS group (P>0.05). However, the LP diet caused a significant reduction in yolk color score, and astaxanthin, vitamin A, and vitamin E contents of egg yolk (P<0.05). However, the contents of these nutrients increased as the level of krill meal was increased in the diets (P<0.05). The highest yolk color score, and astaxanthin, vitamin A, and vitamin E contents were observed in laying hens fed 3% krill meal (P<0.05). The LP diet had no effect on n-3 fatty acid content; however, a significant reduction in the content of n-6 fatty acids, especially linoleic acid was observed (P<0.05). Further reduction occurred when higher level of krill was used in the diets (P<0.05). An increase in krill meal level significantly increased docosahexaenoic acid but not linolenic acid content of egg yolk. Krill meal, therefore, could be used to produce docosahexaenoic acid and antioxidant enriched eggs.
We conducted a 28-day trial to evaluate the influence of Bacillus cereus and phytase supplementation on the expression of musculoskeletal strength and intestinal histological features in Japanese quail. Two-hundred day-old quail chicks were divided into four groups with five replicates (n = 10): the first group served as a control and was fed only a basal diet (BD); the second group was fed BD + 0.1% B. cereus; the third group was fed BD + 0.01% microbial phytase; and the fourth group was fed BD + 0.01% microbial phytase + 0.1% B. cereus. Compared to the control, individual and combined supplementation of probiotic and phytase increased (P < 0.05) the tibial weight, length, outside diameter, and weight of bone ash, but decreased (P < 0.05) the weight-to-length index. The water-holding capacity, fiber diameter, fiber cross-sectional area, number of fibers per unit area, fascicle diameter, and fascicle cross-sectional area increased significantly (P < 0.05) in birds fed on the combination of phytase and B. cereus. The villus height, width, depth, height-to-crypt depth, and surface area increased significantly (P < 0.05) in the B. cereus and phytase groups on an individual basis. The strength of the musculoskeletal system was fully expressed when B. cereus and phytase were given synergistically. However, the histological features of the intestines improved in birds fed on an individual basis.
Insulin stimulates protein synthesis in skeletal muscles. Protein synthesis is controlled by the mechanistic target of rapamycin (mTOR) signaling in skeletal muscles. This study was conducted to investigate the effect of insulin on protein synthesis and mTOR signaling in chick myotube cultures. Chick myotubes were incubated with insulin (1μg/ml) for 1 h. Protein synthesis, measured using the surface sensing of translation method, was significantly increased by insulin. The phosphorylation of AKT (Thr308 and Ser473), p70 ribosomal S6 kinase 1 (Thr389), S6 ribosomal protein (Ser235/236), and eukaryotic translation initiation factor 4E-binding protein 1 (Thr37/46) was also significantly increased by insulin. These results suggest that insulin stimulates protein synthesis via mTOR signaling (phosphorylation of AKT, S6K1, S6 ribosomal protein, and 4E-BP1) in chick myotube cultures.
Innate fear responses to tonic immobility (TI) and open field (OF) were evaluated in newly hatched chicks of three breeds with distinct breed origin and genetic relationships. The breeds studied were Nagoya (NAG), a native Japanese breed; White Leghorn (WL), a representative of layers; and White Plymouth Rock (WPR), a parental breed of common broilers. The TI test revealed that WL was the most sensitive to extensive fear evoked by the TI test among the three breeds, followed in order by WPR, and NAG. In contrast, the OF test revealed that NAG was the most sensitive to mild fear evoked by the OF test, followed in order by WPR, and WL. The different fear responses between NAG and WL were supported by minimal phenotypic correlations between TI and OF traits in each breed. These results demonstrated that NAG and WL breeds exhibit extreme and opposite responses to TI and OF fears.
Few studies have focused on the regulation of cytokine and avian β-defensin (AvBDs) expression for promoting immune defense in the avian intestine. The aim of this study was to investigate the effects of different Toll-like receptor (TLR) ligands (bacterial patterns) on the expression of proinflammatory cytokines (IL-1β and IL-6) and AvBDs (AvBD1, AvBD4, and AvBD7) in the chick intestine. The ileum and cecum of 3-day-old chicks were collected and examined histologically to identity the cells present in the intestinal mucosa. Other tissues were cultured with or without the TLR2, TLR4, and TLR21 ligands— Pam3CSK4, LPS, and CpG-ODN—for 1 or 3 h. The gene expression profiles of proinflammatory cytokines and AvBDs were determined in these tissues using real-time polymerase chain reaction (PCR). The mucosa of the ileum and cecum contained leukocytes, luminal and crypt epithelial cells, and other enterocytes. Pam3CSK4 tended to downregulate the expression of IL-1β, AvBD1, and AvBD7 in the ileum but upregulated their expression in the cecum. LPS downregulated the expression of IL-1β and IL-6 in both the ileum and the cecum, whereas it upregulated the expression of AvBD1, AvBD4, and AvBD7 in the cecum. CpG-ODN upregulated the expression of IL-6 and AvBD7 in the ileum and IL-1β in the cecum, and downregulated the expression of IL-1β and AvBDs in the ileum. We suggested that the expression levels of proinflammatory cytokines and AvBDs in the chick intestine are affected by TLR2, TLR4, and TLR21 ligands. Thus, these innate immune factors may be modulated by the luminal microbe complex in the intestine.
Testosterone (T) is known to induce aggressive behavior, particularly in male animals. However, our recent results showed that a certain kind of aggressive behavior is T-independent; moreover, the role of T in chicken territorial and isolation-induced aggressive behavior has not yet been investigated. In addition, castration alone is insufficient to evaluate the role of T in aggressive behavior because we found that non-testicular T concentration, probably derived from the adrenal gland, in the blood of castrated chicks was low, but not zero. In the present study, therefore, the role of testicular T in chicken aggressive behavior was evaluated through castration, and the role of non-testicular T was assessed using the subcutaneous implantation of flutamide, a non-steroidal antiandrogen, in the castrated male layer chicks. Resident-intruder (R-I) and social interaction (SI) tests were used to quantitatively monitor territorial and isolation-induced aggressive behavior, respectively. Castration and drug implantation of the chicks were performed at 14 days of age. The R-I test was performed at 29 and 30 days of age, and the SI test was performed at 31 and 32 days of age. The total aggression frequencies (TAFs) and aggression establishment rate (AER) were used as indices of chick aggressive behavior. In the R-I test, castration significantly decreased the TAFs but the AER was not affected by castration or flutamide implantation. In the SI test, on the other hand, there were no significant differences in the TAFs, but the AER tended to increase in the intact chicks and decrease in the flutamide-implanted, castrated male chicks. These results suggest that the role of T in chicken aggression depends on the differences in social context of the behavior, and that both testicular and non-testicular T play an important role in the occurrence of isolation-induced aggression in male layer chicks.
Adjuvants are common vaccine components. Novel adjuvants may improve the protective immunity conferred by vaccines against poultry diseases. Here, a less-hemolytic saponin, platycodin D (PD), isolated from the root of Platycodon grandiflorum was investigated as a potential alternative adjuvant. PD was tested as an adjuvant in the infectious bronchitis (IB) vaccine, because the existing IB vaccine has often failed to induce effective immune responses. The adjuvant activity of PD in conjunction with IB vaccine was evaluated in this study. Compared to control treatment, PD treatment significantly increased the proliferation of chicken peripheral blood mononuclear cells, concentration of interferon-γ in culture supernatants, and anti-IB antibody titer. In chickens pre-challenged with the Mass 41 infectious bronchitis virus (IBV), PD administration resulted in fewer and less severe clinical signs, lower mortality rate, and higher protection compared to control treatment. Histopathological examination showed that the lungs and kidneys of PD-treated chickens displayed fewer pathological lesions than those of control chickens. Our results also demonstrated that this new vaccine adjuvant improved chicken humoral and cellular immune responses without any side effects. Hence, our findings suggest that PD might serve as an effective adjuvant in IBV vaccines.
The aim of this study was to investigate the effect of starch granules enriched with carvacrol and mixed with straw pellets (as poultry litter) on the mortality of larvae and adults of the lesser mealworm, Alphitobius diaperinus Panzer, a cosmopolitan pest inhabiting chicken houses in vast numbers worldwide. Additionally, the effect of starch granules on the growth parameters and survival of broiler chickens exposed to treated litter was examined. In this study, granules containing 3, 5, and 10% carvacrol was used. In a simulated chicken house bioassay, this material was mixed with pellets in three different proportions: 30/70%, 40/60%, and 50/50% (granules/pellets, respectively). On this medium, young larvae (approximately 10 days old), older larvae (last stage before pupa), and unsexed 7–10 days old adults of the lesser mealworm, with access to food, were colonized. Experiments were performed at 29 °C in the dark. The study shows that poultry litter with the addition of starch granules enriched with 10% of carvacrol in the proportion of 40:60% (granules:pellets) appears to be the optimal medium applicable to broiler houses for A. diaperinus control. In this environment, all larvae and adults died within 3–4 days and the overall development of the experimental chickens was similar to that of the control. However, the feed conversion rate was slightly higher in the treated group (1.72) than in the control group (1.56). The average final body weight in the treated group was 100 g lower than that in the control group (the differences were not statistically significant).
This study aimed to modify the feed mixtures of laying hens to enrich the consumer eggs with n-3 polyunsaturated fatty acids (PUFA): α-linolenic acid (ALA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). One hundred and twenty Tetra-SL laying hens used in the study were divided into three groups of 40 laying hens arranged in five repetitions: C, control with 5% soybean oil; E1, 0.5% fish oil + 0.5% microalgae Schizochytrium limacinum; and E2, 0.75% fish oil + 0.75% microalgae. The composition of the mixtures was balanced at the level of 17.5% raw protein and 11.81 MJ/kg metabolic energy (ME). Feed and water were provided ad libitum, and the experiment lasted for 21 days. In this study, the different physical and chemical properties of eggs, the fatty acid profile and lipid oxidation of fat in egg yolks were analyzed. The results of the study showed that the weight of the egg yolk and that of the shell depended on the feeding treatments (P = 0.014 and P < 0.001), and the weight of eggs and basic parts, as well as the thickness of the shell depended on the storage duration (P < 0.001). The storage time affected the pH of egg yolks and albumen and the reduction in Haugh units and albumen height (P < 0.001). Significant differences were observed in the content of ALA, DHA, Σn-3 PUFA (mg/100 g) and the n-6/n-3 PUFA ratio between the C and E1/E2 egg groups (P < 0.001). The results of the study indicate that it is sufficient to use a lower level of fish oil and the microalgae Schizochytrium limacinum in hens’ feed to achieve a satisfactory increase in n-3 PUFA in eggs, while maintaining optimal values of egg quality and freshness indicators.
A significant association was reported between a single nucleotide polymorphism (SNP; AB604331, g. 420 C > A) in the cholecystokinin type A receptor gene and growth traits in some Japanese slow-growing chickens. Demonstration tests of the genetic improvement effect by comparing the superior allele-A fixed chickens with conventional ones were carried out considering the effect of different seasons on growth traits in other slow-growing chickens. Meat-type Okumino-kojidori chickens from Gifu Prefecture are a three-way cross of Gifu-jidori improved, White Plymouth Rock, and Rhode Island Red breeds. We used a total of 468 meat-type Okumino-kojidori: 264 individuals from a private hatchery as conventional chickens and 204 A-allele fixed individuals from the Gifu Prefectural Livestock Research Institute as improved chickens. We performed fattening experiments over two seasons: summer and winter. In each season, experimental birds of both sexes were hatched on the same day, raised in the same chicken house, and fed the same diet adlibitum for 12 weeks. Body weight was recorded at 3, 6, 9, and 12 weeks of age. SNP genotypes were determined using the mismatch amplification mutation assay. Association between the SNP and growth traits was analyzed using generalized linear models built on sex-based, seasonal, additive, and dominance genetic effects. The observed AA, AC, and CC genotype frequencies in the conventional chickens were 0.158, 0.479, and 0.363, respectively; body weight at 12 weeks and average daily gain from 3 to 12 weeks was superior for the A allele compared to the C allele. The improved chickens were heavier than the conventional ones at 12 weeks. Body weight at 12 weeks in allele-A fixed chickens increased by 3.2% compared to the conventional chickens. We concluded that g. 420 C > A is a good selective marker that increases slaughter weight in the meat-type Okumino-kojidori chickens.
The objective of this study was to examine the effect of Gynura divaricata (Jakr-Na-Rai, JNR) on the growth performance, hematology, and carcass fat deposition of broilers. A total of 240 male Cobb-500 birds, 22 d old, were randomly allocated into five treatment groups of six replicates. Each group was raised at a high stocking density of 28kg of body weight/m2 until day 43. The treatments consisted of (i) a basal diet, or the basal diet supplemented with (ii) 2.5mg/kg avilamycin (T2), (iii) JNR crude ethanol extract at a flavonoid level of 1.3g/kg (T3), and (iv and v) JNR powder at a flavonoid level of 1.3 (T4) or 2.6g/kg (T5). Dietary supplementation with JNR powder at both flavonoid levels decreased the heterophil/lymphocyte ratio compared to the other groups (P < 0.001). Birds in the T5 group presented decreased blood glucose (P < 0.005) and cholesterol (P < 0.002) levels and a numerically (P = 0.056) decreased triglyceride level. The total bile acid concentration increased (P < 0.001) in all the JNR-fed groups, but there was no significant effect on the digestibility of ileal protein or fat. At 1.3g/kg of diet, JNR increased the final body weight and feed intake (P < 0.05), but the average daily gain and feed conversion ratio were not different among groups. The carcass and abdominal fat percentages were lowest in chicks fed on the diets supplemented with 2.6g/kg JNR powder (P < 0.007 and P < 0.025, respectively). Drip loss and malondialdehyde concentrations in the breast meat did not change. In conclusion, JNR powder improved several hematological parameters, increased total bile acid concentrations, and decreased the percentage of abdominal fat. The powder form of JNR elicited better results than the ethanol extract form at the same flavonoid inclusion level.
1α-Hydroxycholecalciferol (1α-OH-D3) is an active vitamin D derivative. In this study, three experiments were conducted to evaluate the optimal dietary levels of 1α-OH-D3 in broiler chickens from 1 to 42 days of age. 1α-OH-D3 levels used were 0, 1.25, 2.5, 5, and 10μg/kg in experiment 1, 0.625, 1.25, 2.5, 5, 7.5, and 10μg/kg in experiment 2, and 2, 2.5, 3, 3.5, 4, 4.5, and 5 μg/kg in experiment 3. In experiment 1, the addition of 0 to 10μg/kg of 1α-OH-D3 quadratically improved growth performance, tibia development, and mRNA expression levels of nuclear vitamin D receptor (nVDR), membrane vitamin D receptor (mVDR), and type IIb sodium-phosphate cotransporter (NaPi-IIb) in the duodenum of broiler chickens from 1 to 12 days of age. Body weight gain (BWG), the weight and ash weight of the tibia, and mRNA expression levels of mVDR and NaPi-IIb of broilers fed with 0 and 10μg/kg of 1α-OH-D3 were lower than those of birds fed with 2.5μg/kg of 1α-OH-D3. In experiment 2, 1α-OH-D3 levels were quadratically related to BWG and to weight and ash weight of the femur and the tibia of broiler chickens at 42 days of age. The highest values of growth performance and bone mineralization were recorded in broilers fed with 2.5 to 5μg/kg of 1α-OH-D3. In experiment 3, there was no difference observed in BWG and the weight and ash weight of the femur and the tibia of the 42-day-old broilers fed with 2 to 5μg/kg of 1α-OH-D3. These data suggest that the optimal dietary levels of 1α-OH-D3 were 2 to 5μg/kg for broiler chickens from 1 to 42 days of age.
Broiler chickens grow rapidly within a short period; in this regard, our group had previously reported a decrease in the active transport of glucose in the intestines of broiler chickens with their growth. Therefore, in this study, we compared the active transport process of amino acids in the intestines between 1- and 5-week-old broilers using everted sac, Ussing chamber techniques, and real-time quantitative polymerase chain reaction (RT-PCR). The everted sac experiment showed that amino acids were absorbed from all segments of the small intestine in both age groups. There were no significant differences in the serosal to mucosal ratio between 1- and 5-week-old broilers. The Ussing chamber experiment showed that amino acid-induced short-circuit current (ΔIsc) in the ileal epithelium was significantly greater in the 5-week-old chickens than in the 1-week-old chicks (P = 0.035). Membrane conductance, an indicator of ion permeability, showed no significant difference between the two groups. Moreover, the mRNA expression levels of amino acid transporters (ASCT1, EAAT3, B0AT1, and y+LAT1) were significantly elevated in the distal ileum of the 5-week-old broilers compared to those in the 1-week-old broilers (P < 0.05), while no significant differences were observed in the mRNA levels of ATB0’+, B0/+AT, rBAT, CAT1, and CAT2 in both groups. Our study provides clear evidence that age-dependent increase in the active transport of amino acid across the ileal epithelium is caused by the high expression of Na+-dependent amino acid transporters in broiler chickens.
A study was conducted to determine the effect of transportation distance and crating density on meat quality traits during winter. ROSS-308 broilers aged 35 days were divided into 10 treatment groups based on three transportation distances (80, 160, and 240km) with three crating densities (10, 12, and 15 birds per crate) along with birds slaughtered on farm without crating as a control group. Each treatment was replicated 10 times and placed at various locations in the truck. The birds were transported in plastic crates of 3 ft × 1.8 ft × 1 ft at ambient temperature and relative humidity of 3.6–9.5°C and 63.3–78.8%, respectively. The breast meat quality parameters including pH, color, drip loss, shear force, marinade uptake and retention, cooking losses, and yield were determined. The results showed significantly higher pH 15 min post-slaughter (PS), drip loss, lightness, marinade uptake, raw meat cooking yield, and shear force in birds slaughtered on farm without any transportation. The birds transported for 240km had significantly higher pH at 2 hours (h) PS, marinade retention, and cooking yield. Whereas, pH at 24h PS, meat redness, yellowness, chroma, and marinated meat cooking yield were significantly higher in all transported treatments compared to the control group. Among different crating densities, the birds transported with 10 birds per crate showed significantly higher meat redness, marinade retention, and cooking yield. The crating density of 15 birds per crate had significantly higher lightness, hue, cooking loss, and shear force. However, no effect of crating density was observed on meat pH, drip loss, and marinade uptake. It can be concluded that an increase in transportation distance and low crating density during winter can negatively affect physical quality of meat with significant improvement in marination and cooking characteristics.
We have previously reported a significant association between the single-nucleotide polymorphism (SNP; g.420 C>A) in the cholecystokinin type A receptor gene (CCKAR) and the growth traits of Hinai-dori, a breed of chicken that is indigenous to Japan. Moreover, we have demonstrated that the minor allele of this SNP improved the growth rate in a low-growth line of the Hinai-dori breed. Hence, in the present study, we verified the association between this SNP and the growth traits of the Hinai-jidori chicken: a cross between a Hinai-dori sire and Rhode Island Red dam. In addition, we verified whether the growth rate was improved in Hinai-jidori chickens produced from the parent stocks in which the SNP A/A genotype was fixed by selection (improved Hinai-jidori chickens). The Hinai-jidori female chicks at 4 weeks of age, were subdivided into three genotypic groups (A/A, A/C, and C/C), with 20 chicks in each group, and reared in an open-sided poultry shed until 23 weeks of age. The results showed that the body weight at 23 weeks of age and the average daily gain after 14 weeks of age were significantly higher in group A/A than in group C/C. Subsequently, the improved and the conventional Hinai-jidori chickens were reared until they reached 22 weeks of age to verify the effects on their growth traits. The body weight of the improved Hinai-jidori chickens at 22 weeks was significantly greater than the conventional Hinai-jidori chickens. Moreover, the association between the SNP and body weights of Hinai-jidori chickens at market age (24 weeks) on the production farms showed that the A allele was significantly superior to the C allele. In conclusion, the CCKAR g.420 C>A SNP improves the growth rate of commercial Hinai-jidori chickens and could be a candidate marker for improving the growth performance in selective breeding of Hinai-jidori chickens.
Salmonella Typhimurium (ST) infection in chickens inhibits their growth and can lead to food-borne diseases in humans. Probiotics are expected to enhance the function of host intestinal barrier against pathogen infection. The aim of our study was to determine the effect of viable Lactobacillus reuteri (LR) on the response of the mucosal barrier function to antigen stimulation in broiler chicks. Day-old male (n = 8) and female (n = 4) broiler chicks were orally administered either 1 × 108 LR or a water-only control, every day for 7 days. After 7 days, either 1 × 108 heat-killed ST (k-ST), or a buffer-only control, was administered via intra-cardiac injection. The ileum and cecum were collected 3 h post-injection, and paraffin sections were prepared for either mRNA extraction (males), or gut permeability tests (females). Villus and crypt lengths were determined via histological analysis. Real-time PCR was used to calculate expression levels of Toll-like receptors (TLRs), pro-inflammatory cytokines, anti-inflammatory cytokines, avian β-defensins, and tight-junction-associated molecules. Gut permeability was assessed using the inverted intestine method. We found that (1) expression of TLR2-1, TLR21,TGF-β2 and TGF-β3 were reduced following k-ST stimulation, but were unaffected by LR-treatment; (2) oral administration of LR led to increased Claudin1, Claudin5, ZO2, and JAM2 expression following k-ST stimulation; (3) cecal permeability was reduced by co-treatment with LR and k-ST, but not by treatment with LR or k-ST alone. These results suggest that LR, as used in this study, may enhance the intestinal mucosal physical barrier function, but not the expression of other immune-related factors in newly hatched chicks.