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Egg‐laying rate is mainly determined by ovarian function and regulated by the hypothalamic‐pituitary‐gonadal axis; however, the mechanism by which the ovary regulates the egg‐laying rate is still poorly understood. The purpose of this study was to compare the differences in the transcriptomes of the ovary of Lingyun black‐bone chickens with relatively high and low egg‐laying rates and screen candidate genes related to the egg‐laying rate. RNA‐sequencing (RNA‐Seq) was conducted to explore the chicken transcriptome from the ovarian tissue of six Lingyun black‐bone chickens with high (group G, n = 3) and low (group D, n = 3) egg‐laying rates. The results showed that 235 differentially expressed genes (DEGs) were identified between the chickens with high and low egg‐laying rates; among them, 209 DEGs were up‐regulated and 26 DEGs were down‐regulated. Gene Ontology analysis showed that the up‐regulated 209 DEGs were enriched in 50 GO terms and the down‐regulated 26 DEGs were enriched in 40 GO terms. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that up‐regulated DEGs were significantly enriched in 25 pathways and down‐regulated DEGs were significantly enriched in three pathways. Among the pathways, we found the longevity regulating pathway‐multiple species pathway, Estrogen signalling pathway and PPAR signalling pathway may have an essential function in regulating the egg‐laying rate. The results highlighted DEGs in the ovarian tissues of relatively high and low laying Lingyun black‐bone chicken and identified essential candidate genes related to the egg‐laying rate, thereby providing a theoretical basis for improving the egg‐laying rate of Lingyun black‐bone chicken., The purpose of this study was to compare the di?erences in the transcriptomes of ovaries of hens with relatively high and low laying performances and screen candidate genes related to laying performance. RNA sequencing was performed to analyze and compare the mRNA in ovarian tissues of high and low laying performance hens.

In this study, the effects of 5 graded dietary levels (0.025, 0.05, 0.075, 0.1, and 0.125%) of dimethylglycine (DMG) were studied in laying hens during the late laying period (71–78 wk). Graded doses of DMG from 0.025 to 0.125% in the diet produced quadratic positive (P < 0.05) responses in the laying rate, egg-feed ratio, yolk color, grade follicular weight, and the number of large white follicles and linear positive (P < 0.05) responses in average egg weight, and the number of large white follicles. With 0.1% DMG, the laying rate and egg-feed ratio improved (P < 0.05), and the abdominal fat percentage decreased. Considering the laying performance under the conditions used in this study, the best-fit model for the DMG requirements of laying hens was estimated to range from 0.049 to 0.065% DMG during the late laying period based on a regression analysis. The addition of DMG did not affect the total cholesterol (TCH) and triglyceride (TG) contents in the plasma of laying hens; however, it significantly reduced the abdominal fat rate. DMG may change the course of lipid deposition in laying hens during the late laying period. In conclusion, supplementation with DMG can improve the laying rate and follicles development of laying hens.

This study was conducted to compare the infection heterogeneity and cecal microbiota in chicks infected by S. enteritidis. Forty-eight 8-d-old female Arbor Acres chicks were challenged with S. enteritidis and euthanized 24 h later. The eight chicks with the highest Salmonella tissue loads were assigned to group S (S. enteritidis-susceptible), and the eight chicks with the lowest Salmonella tissue loads were assigned to group R (S. enteritidis-resistant). Chicks in group S showed a higher liver index (p &lt, 0.05), obvious liver lesions, and an decreasing trend for the villus height-to-crypt depth ratio (p &lt, 0.10), compared with those in group R. Gene expression of occludin, MUC2, and IL10 was higher, whereas that of iNOS and IL6 was lower (p &lt, 0.05), in chicks of group R relative to those in group S. Separation of the cecal microbial community structure has been found between the two groups. The S. enteritidis-susceptible chicks showed higher abundance of pathogenic bacteria (Fusobacterium and Helicobacter) in their cecal, while Desulfovibrio_piger was enriched in the cecal of S. enteritidis-resistant chicks. In summary, chicks showed heterogeneous responses to S. enteritidis infection. Enhanced intestinal barrier function and cecal microbiota structure, especially a higher abundance of Desulfovibrio_piger, may help chicks resist S. enteritidis invasion.

The effects of N-carbamylglutamate (NCG) on the growth performance, tissue development, and blood parameters of broilers are unknown. In this study, 2 linked experiments were conducted to investigate the effects of 4 graded dietary levels and 3 dietary stages of NCG in a Chinese indigenous yellow-feather broiler breed during 2 growth phases: 1 to 18 d and 19 to 36 d. The dietary levels of NCG were 0.05%, 0.10%, 0.15%, and 0.20%, and dietary stages were designed to add NCG during the starter stage or grower stage or throughout the experimental period. At the age of 18 d, graded doses of NCG from 0.05 to 0.20% in the diet produced quadratic (P < 0.05) positive responses in body weight, width of intermuscular fat cingulum, liver weight, serum blood urea nitrogen, and serum low-density lipoprotein as well as linear (P < 0.05) positive responses in albumin serum concentration. The average feed per gain and mortality were unaffected by dietary NCG levels. Among 3 dietary treatments, only NCG dietary treatments throughout the experimental period improved the body weight and daily weight gain linearly (P < 0.05). The daily weight gain under the 3 dietary treatments used indicated that the most fitting dose is 0.1% NCG among the 4 dietary levels of NCG (P < 0.05). At this dose, muscle weight increased, whereas subcutaneous adipose as well as the serum contents of uric acid, triglyceride, and albumin decreased. Considering the growth performance and tissue development under the conditions used in this study, the best-fit model for NCG requirements of Chinese yellow-feather broilers was estimated from regression analysis to be 0.09 to 0.12% dietary NCG treatments during the grower stage. The modified blood parameters indicated that NCG dietary effects on broiler growth may be accompanied by modified homeostasis of arginine metabolism, lipid deposition, protein synthesis, and immune response.

This experiment was to study the effects of newly harvested corn on growth performance, intestine development and metabolism of nutrients in broilers. A total of 380 one-day-old Arbor Acres male broilers were assigned to ageing corn group (AC group), 1/3 newly harvested corn group (1/3 NC group), 2/3 newly harvested corn group (2/3 NC group), and newly harvested corn group (NC group), which according to the proportion of newly harvested corn in the total amount of corn of basal diets. Each group included six replicates with 16 birds. Digestion and metabolism trials were conducted at 39 to 42 days of age. Results showed that feed: gain ratio (F/G) of NC group was higher than that of 1/3 NC group and 2/3 NC group at 22 to 42 and 1 to 42 days of age (p

An experiment was conducted to evaluate the effects of cyclic heat stress on phenotypic response, meat quality and muscle glycolysis of broiler. One hundred and eighty 35-day-old male Xueshan broilers with similar BW were randomly selected and exposed to either thermal neutral (TN; 23 °C, 55–60% RH, n = 6) or heat stress conditions (CHS; cyclic temperatures of 35 °C from 10:00 to 18:00, 32 °C from 18:00 to 20:00, 28 °C from 20:00 to 08:00, and 32 °C from 08:00 to 10:00 at 40–45% RH, n = 6) for 7 days. Relative to broilers under thermoneutral conditions, broilers subjected to cyclic heat stress had an overall increase in rectal temperature and respiration rate (p

Rational: Salmonella Enteritidis (S. Enteritidis) is a globally significant zoonotic foodborne pathogen which has led to large numbers of deaths in humans and caused economic losses in animal husbandry. S. Enteritidis invades host cells and survives within the cells, causing resistance to antibiotic treatment. Effective methods of elimination and eradication of intracellular S. Enteritidis are still very limited. Here we evaluated whether a new intracellular antibacterial strategy using iron oxide nanozymes (IONzymes) exerted highly antibacterial efficacy via its intrinsic peroxidase-like activity in vitro and in vivo. Methods: The antibacterial activities of IONzymes against planktonic S. Enteritidis, intracellular S. Enteritidis in Leghorn Male Hepatoma-derived cells (LMH), and liver from specific pathogen free (SPF) chicks were investigated by spread-plate colony count method and cell viability assay. Changes in levels of microtubule-associated protein light chain 3 (LC3), a widely used marker for autophagosomes, were analyzed by immunoblotting, immunofluorescence, and electron microscopy. Reactive oxygen species (ROS) production was also assessed in vitro. High-throughput RNA sequencing was used to investigate the effects of IONzymes on liver transcriptome of S. Enteritidis-infected chicks. Results: We demonstrated that IONzymes had high biocompatibility with cultured LMH cells and chickens, which significantly inhibited intracellular S. Enteritidis survival in vitro and in vivo. In addition, co-localization of IONzymes with S. Enteritidis were observed in autophagic vacuoles of LMH cells and liver of chickens infected by S. Enteritidis, indicating that IONzymes mediated antibacterial reaction of S. Enteritidis with autophagic pathway. We found ROS level was significantly increased in infected LMH cells treated with IONzymes, which might enhance the autophagic elimination of intracellular S. Enteritidis. Moreover, orally administered IONzymes decreased S. Enteritidis organ invasion of the liver and prevented pathological lesions in a chicken-infection model. Non-target transcriptomic profiling also discovered IONzymes could change hepatic oxidation-reduction and autophagy related gene expressions in the S. Enteritidis infected chickens. Conclusion: These data suggest that IONzymes can increase ROS levels to promote the antibacterial effects of acid autophagic vacuoles, and thus suppress the establishment and survival of invading intracellular S. Enteritidis. As a result, IONzymes may be a novel alternative to current antibiotics for the control of intractable S. Enteritidis infections.

Autophagy is an important component of the innate immune system in mammals. Low levels of basic autophagy are sustained in normal cells, to help with the clearance of aging organelles and misfolded proteins, thus maintaining their structural and functional stability. However, when cells are faced with challenges, such as starvation or pathogenic infection, their level of autophagy increases significantly. Salmonella is a facultative intracellular pathogen, which imposes an economic burden on the poultry farming industry and human public health. Previous studies have shown that Salmonella can induce the autophagy of cells following invasion, which to a certain extent helps to protect the cells from bacterial colonization. This review summarizes the latest research in the field of Salmonella-induced autophagy, including: (i) the autophagy induction and escape mechanisms employed by Salmonella during the infection of host cells; (ii) the effect of autophagy on intracellular Salmonella; (iii) the important autophagy adaptors that recognize intracellular Salmonella in host cells; and (iv) the effect of autophagy-modulating drugs on Salmonella infection.

Magnetic Fe 3 O 4 nanoparticles (Fe 3 O 4 -NPs) have been widely investigated for their biomedical applications. The main purpose of this study was to evaluate the cytotoxic effects of different sizes of Fe 3 O 4 -NPs in chicken macrophage cells (HD11). Experimental groups based on three sizes of Fe 3 O 4 -NPs (60, 120 and 250 nm) were created, and the Fe 3 O 4 -NPs were added to the cells at different doses according to the experimental group. The cell activity, oxidative index (malondialdehyde (MDA), superoxide dismutase (SOD) and reactive oxygen species (ROS)), apoptosis and pro-inflammatory cytokine secretion level were detected to analyse the cytotoxic effects of Fe 3 O 4 -NPs of different sizes in HD11 cells. The results revealed that the cell viability of the 60 nm Fe 3 O 4 -NPs group was lower than those of the 120 and 250 nm groups when the same concentration of Fe 3 O 4 -NPs was added. No significant difference in MDA was observed among the three Fe 3 O 4 -NP groups. The SOD level and ROS production of the 60 nm group were significantly greater than those of the 120 and 250 nm groups. Furthermore, the highest levels of apoptosis and pro-inflammatory cytokine secretion were caused by the 60 nm Fe 3 O 4 -NPs. In conclusion, the smaller Fe 3 O 4 -NPs produced stronger cytotoxicity in chicken macrophage cells, and the cytotoxic effects may be related to the oxidative stress and apoptosis induced by increased ROS production as well as the increased expression of pro-inflammatory cytokines.

Understanding the differences in microbial communities shaped by different food selective forces, especially during early post-hatch period, is critical to gain insight into how to select, evaluate, and improve antibiotic growth promoters (AGPs) alternatives in food animals. As a model system, commercial diet-administered OAs (DOAs) and water-administered OAs (WOAs) were used separately or in combination as Virginiamycin alternatives for broiler feeding during two growth phases: 1–21 days and 22–42 days. Among these three OA-treated groups, the DOA group was most similar to the AGP group in the composition and the proportion of these dominant bacterial communities at the level of phylum, family, and genus in cecal chyme of broilers. Sub-therapeutic Virginiamycin decreased the richness, homogenization, and species diversity of gut microbiota, especially in the early growth stage from days 1 to 21. Among these three OA supplementation schemes, it was clear that DOA supplementation was more likely to increase or maintain the richness, homogenization, species diversity, and predicted gene functions of cecal microbiota in treated broilers than either no supplementation or AGP supplementation during two experimental stages. The interference of DOA treatment with early colonization of probiotics and pathogens in broiler cecum was the most similar to AGP treatment, and OAs did not cause the occurrence of Virginiamycin-resistant strains of Enterococcus at the end of this trial. In terms of the predicted gene functions of the microbiota, AGP and DOA treatments provided a similar selective force for microbial metabolism functions in the cecum of broiler chickens, especially in the early growth stage. Noticeably, the relative abundance of some microbiome that was modified by Virginiamycin or DOA supplementation was significantly correlated with body weight gain and KEGG pathway analysis-annotated gene functions such as replication and repair, translation, nucleotide metabolism, and so on. With the comprehensive analysis of these results and practical application, shortened DOA supplementation, after optimization of the amount of addition, would be a suitable alternative to sub-therapeutic Virginiamycin. It was suggested that the programed intestinal microecology under such early selection forces and the effective addition time may be the key elements to focus on the designed alternate strategies of AGPs in food animals.

An experiment was conducted to evaluate the effects of crude protein (CP) levels on growth performance, carcase quality, meat quality, immune indices, litter and faeces quality, as well as welfare quality, in free-range yellow broilers using balanced amino acid technology. In total, 600 one-d-old Suqin yellow broilers were raised for 21 d. On d 22, 540 birds with similar BW (390 ± 7 g SD) were randomly selected and placed into 9 pens (3 groups with 3 replicates per group and 60 birds each replicate) and then reared to 56 d of age. The birds were fed a diet containing CP 19% (control), 18% or 17% (measured value: 18.878%, 17.881%, 17.052%, respectively), in three groups for 35 d. The results revealed that decreasing dietary CP levels from 19% to 17% had no impact on broiler growth performance, carcase quality and meat quality (p > .05). The relative immune organ weights also showed no change to the lowering of dietary CP levels (p > .05), whereas serum albumin was significantly influenced by decreasing dietary CP levels (p = .042). Furthermore, nitrogen content in litter and faeces, as well as the footpad dermatitis score for welfare indices, were decreased with decreased dietary CP levels (p = .009, p = .014, p = .045, respectively). The results of this study suggest that low-protein diets with balanced amino acids reduce nitrogen excretion and footpad dermatitis without effecting the performance, carcase quality, and meat quality of free-range yellow broilers.

To investigate the effects of Clostridium butyricum (C. butyricum) on breast muscle lipid metabolism of broilers, 192 one-day-old male Arbor Acres broilers were randomly allocated into 2 treatments with 6 replicates in a completely randomised design. The broilers were fed corn-soybean meal-based diets and supplemented with 0 or 1 × 109 cfu of C. butyricum/kg of diet for 42 days. The birds in the C. butyricum-supplemented group showed higher (p

The present study was conducted to evaluate the effects of lecithin on the performance, meat quality, lipid metabolism, and cecum microbiota of broilers. One hundred and ninety-two one-day-old AA broilers with similar body weights (38 ± 1.0 g) were randomly assigned to two groups with six replicates of sixteen birds each and were supplemented with 0 and 1 g/kg of lecithin for forty-two days. Performance and clinical observations were measured and recorded throughout the study. Relative organ weight, meat quality, lipid-related biochemical parameters and enzyme activities were also measured. Compared with broilers in the control group, broilers in the lecithin treatment group showed a significant increase in L* value and tenderness (p &lt, 0.05). Meanwhile, the abdominal adipose index of broilers was markedly decreased in lecithin treatment after 42 days (p &lt, 0.05). In the lipid metabolism, broilers in the lecithin treatment group showed a significant increase in hepatic lipase and general esterase values at 21 days compared with the control group (p &lt, 0.05). Lower Firmicutes and higher Bacteroidetes levels in phylum levels were observed in the lecithin treatment group after 21 and 42 days. The distribution of lactobacillus, clostridia, and rikenella in genus levels were higher in the lecithin treatment group after 21 and 42 days. No statistically significant changes were observed in performance, relative organ weight, or other serum parameters (p &gt, 0.05). These results indicate that supplementation with lecithin significantly influence the lipid metabolism in broilers at 21 and 42 days, which resulted in the positive effect on the meat color, tenderness, and abdominal adipose in broilers.

Simple Summary Normally, researchers use feces or rectal swabs to characterize a gut microbiome, but because there is significant spatiotemporal variation across different intestinal segments there are marked differences in composition and function of microbiomes among various gut sites. Hence, a consensus has not been reached on the location for sampling for gut microbial metagenome sequencing. This study provides a comparative perspective on gut microbial function that takes into account different sampling resources when conducting a metagenomic sequence analysis, highlighting the differences in the choice of a gut microbiome sampling site, and investigating whether feces and rectal swab samples are efficient proxies for gut microbiome sampling. Abstract When conducting metagenomic analysis on gut microbiomes, there is no general consensus concerning the mode of sampling: non-contact (feces), noninvasive (rectal swabs), or cecal. This study aimed to determine the feasibility and comparative merits and disadvantages of using fecal samples or rectal swabs as a proxy for the cecal microbiome. Using broiler as a model, gut microbiomes were obtained from cecal, cloacal, and fecal samples and were characterized according to an analysis of the microbial community, function, and resistome. Cecal samples had higher microbial diversity than feces, while the cecum and cloaca exhibited higher levels of microbial community structure similarity compared with fecal samples. Cecal microbiota possessed higher levels of DNA replicative viability than feces, while fecal microbiota were correlated with increased metabolic activity. When feces were excreted, the abundance of antibiotic resistance genes like tet and ErmG decreased, but some antibiotic genes became more prevalent, such as fexA, tetL, and vatE. Interestingly, Lactobacillus was a dominant bacterial genus in feces that led to differences in microbial community structure, metabolism, and resistome. In conclusion, fecal microbiota have limited potential as a proxy in chicken gut microbial community studies. Thus, feces should be used with caution for characterizing gut microbiomes by metagenomic analysis.

Daidzein (DA) is a kind of isoflavone that is extracted primarily from soy plants and that has become increasingly popular as a dietary supplement. The objective of this study was to evaluate the effects of dietary DA supplementation for laying breeder hens on laying performance, reproductive organ development, hatching performance of seed eggs, and growth performance of offspring and to investigate the underlying molecular mechanisms. A total of 180 55-week-old laying breeder hens were randomly divided into 2 treatment groups and, after 3 weeks of acclimation, were fed either a control diet (CON) or a DA-supplemented diet (DAS, CON+30 mg/kg DA) for a total of 12 weeks. DAS treatment improved the laying rate, luteinizing hormone (LH) levels, and small yellow follicle (SYF) numbers without negative effects on the hatchability of breeder eggs or the growth performance of offspring. High-throughput RNA sequencing was utilized to identify differentially expressed genes in the SYF granulosa layer in the two groups. Transcriptome analysis showed that 161 genes (fold change ≥2 or ≤0.5; P-value

The purpose of this study was to investigate the effects of Bacillus subtilis on growth performance, intestinal morphology, and cecal microbial composition of broilers. A total of 270 healthy one-day-old Arbor Acres male broiler chicks were randomly divided into 3 dietary treatment groups, with 6 replicates per group and 15 chickens per replicate. The dietary treatment groups were as follows: 1) basal diet, negative control group; 2) basal diet +250 g/t of zinc bacitracin, positive control group; and 3) basal diet +750 g/t of B. subtilis, B. subtilis group. Results of this experiment showed that compared with the negative control group, body weight at 42 d, average daily gain and European Production Efficiency Factor over the 42 d phase in the B. subtilis group and positive control group were significantly increased (P 0.05). The villus height to crypt depth ratio in the ileum of the B. subtilis group was significantly higher (P

Soybean meal (SBM) is high in antinutritional factors (ANFs), which is not conducive to the starter growth of broilers. The purpose of this study was to investigate the effects of soy protein concentrate (SPC) in starter diet on growth performance, carcass traits, meat quality, immune organ indices and blood biochemical indices of broilers. A total of 384 1-day-old Arbor Acres (AA) male broilers (46.05 &plusmn, 0.37 g) with similar body weight were randomly divided into 4 groups with 8 replicates in each group and 12 broilers in each replicate. The experiment was divided into three phases: in starter phase (1&ndash, 10 d), birds were fed a corn-SBM-based basal mash diet (control) and the basal diet was supplemented with SPC at 4% (SPC4), 8% (SPC8), 12% (SPC12). In the grower phase (11&ndash, 21 d) and the finisher phase (22&ndash, 42 d), the birds in all four treatment groups were fed the same diets. The results showed that the body weight was significantly increased in the SPC8 and SPC12 groups of broilers at 10 d and 42 d (p &lt, 0.05). The average daily gain was significantly increased in the SPC12 group of broilers at 1&ndash, 10 d and 1&ndash, 42 d (p &lt, 0.05). The average daily feed intake was significantly increased in the SPC8 and SPC12 groups of broilers at 1-10 d (p &lt, 0.05). The feed conversion rates at 1&ndash, 42 d (p = 0.055) tended to decline in the SPC12 group. The carcass yield and the thymus indices were significantly increased in the SPC12 group of broilers at 42 d (p &lt, 0.05). Alanine aminotransferase (ALT)/aspartate aminotransferase (AST) tended to decline in SPC12 group at 10 d (p = 0.055) and total protein (TP) tended to increase in the SPC12 group at 42 d (p = 0.080). The contents of total cholesterol (T-CHO) and high-density lipoprotein (HDL) were significantly elevated in the SPC12 group of broilers at 42 d (p &lt, 0.05). In conclusion, dietary inclusion of 12% SPC as a starter diet can be recommended due to the positive effects on broilers.

Salmonella spp. pose a threat to both human and animal health, with more than 2600 serovars having been reported to date. Salmonella serovars are usually identified by slide agglutination tests, which are labor intensive and time consuming. In an attempt to develop a more rapid screening method for the major poultry Salmonella serovars, we developed a loop-mediated isothermal amplification (LAMP) assay, which directly detected the sefA gene, a fimbrial operon gene existing in several specific serovars of Salmonella enterica including the major poultry serovars, namely Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) and Salmonella enterica serovar Gallinarum (Salmonella Gallinarum). With the 177 bacterial strains we tested, positive reactions were only observed with 85 strains of serovar Salmonella Enteritidis and Salmonella Gallinarum. The detection limit of the LAMP assay was 4 CFU/reaction with genomic DNAs of Salmonella Enteritidis (ATCC 13076) from pure culture and 400 CFU/ reaction with DNA extracted from spiked chicken feces. The LAMP assay was more sensitive than conventional culture, especially without enrichment, in detecting Salmonella Enteritidis (CMCC 50041) in the spiked fecal samples. The results show the sefA LAMP method is a rapid, sensitive, specific, and practical method for directly detection of Salmonella Enteritidis and Salmonella Gallinarum in chickens. The sefA LAMP assay can potentially serve as new on-site diagnostics in the poultry industry.

Highly drug-resistant Salmonella enterica serovar Indiana became the most common serovar in broilers with diarrhea in China over the course of this study (15% in 2010 to 70% in 2014). While most S . Indiana isolates (87%, 384/440) were resistant to 13 to 16 of the 16 antibiotics tested, 89% of non- S . Indiana isolates (528/595) were resistant to 0 to 6 antibiotics. Class 1 integrons and IncHI2-type plasmids were detected in all S . Indiana isolates, but only in 39% and 1% of non- S . Indiana isolates.

The present study was conducted to investigate the effects of Enterococcus faecium (E. faecium) on the meat quality and antioxidant capacity of muscle in broilers. A total of 600 Arbor Acre broiler chickens (1-day-old, male) were randomly divided into five treatments with six replicates (20 chickens per replicate) for each treatment. The five treatments were the control treatment (CON, basal diet), antibiotic treatment (ANT, basal diet supplemented with 0.1% chlortetracycline) and E. faecium-supplemented treatments (LEF, MEF and HEF, basal diet supplemented with 50, 100 and 200 mg/kg of E. faecium respectively). The experiment lasted 42 days in two periods of 21 days. Results showed that there were no differences in breast meat quality among different treatments (p > 0.05). Compared with the CON and ANT treatments, the yellowness of thigh meat in E. faecium-supplemented treatments was significantly increased (p < 0.05); the shear force of thigh meat in the LEF and MEF treatments was lower than that of the CON treatment (p < 0.05). In addition, the concentration of the inosine monophosphate (IMP) in the breast and thigh meat of the MEF treatment was significantly higher than that of the other treatments (p < 0.05). At 21 days, the total antioxidant capability (T-AOC) level and glutathione peroxidase (GSH-Px) activity of breast meat and superoxide dismutase (SOD) in the thigh meat of the MEF treatment were greatly increased (p < 0.05). At 42 days of age, the catalase (CAT), GSH-Px and T-AOC activities in the breast meat of the MEF treatment were increased (p < 0.05) and the CAT activity of thigh meat in the LEF and MEF treatments was increased (p < 0.05). In conclusion, E. faecium supplementation increased the meat quality of the thigh muscle, increased the IMP content and the activities of CAT, SOD, T-AOC, and GSH-Px of muscle in broilers. Supplementation with 100 mg/kg E. faecium had the greatest effects.

To select metabolic biomarkers and differentially expressed genes (DEGs) associated with resistant-ascites syndrome (resistant-AS), we used innovative techniques such as metabolomics and transcriptomics to comparatively examine resistant-AS chickens and AS controls. Metabolomic evaluation of chicken serum using ultra-performance liquid chromatography-quadruple time-of-flight high-sensitivity mass spectrometry (UPLC-QTOF/HSMS) showed significantly altered lysoPC(18:1), PE(18:3/16:0), PC(20:1/18:3), DG(24:1/22:6/0:0), PS(18:2/18:0), PI(16:0/16:0), PS(18:0/18:1), PS(14:1/14:0), dihydroxyacetone, ursodeoxycholic acid, tryptophan, L-valine, cycloserine, hypoxanthine, and 4-O-Methylmelleolide concentrations on day 21 and LysoPC(18:0), LysoPE(20:1/0:0), LysoPC(16:0), LysoPE(16:0/0:0), hypoxanthine, dihydroxyacetone, 4-O-Methylmelleolide, LysoPC(18:2), and PC(14:1/22:1) concentrations on day 35, between the susceptible and resistant groups. Compared to the susceptible group, transcriptomic analysis of liver samples using RNA-seq revealed 413 DEGs on day 21 and 214 DEGs on day 35 in the resistant group. Additional evaluations using gene ontology (GO) indicate that significant enrichment occurred in the oxygen transportation, defensive reactions, and protein modifications of the decreased DEGs as well as in the cell morphological formation, neural development, and transforming growth factor (TGF)-beta signalling of the increased DEGs on day 21. Oxygen transportation was also significantly enriched for downregulated DEGs on day 35. The combinatory evaluation of the metabolome and the transcriptome suggests the possible involvement of glycerophospholipid metabolism in the development of resistant-AS in broilers.

Ascites syndrome (AS) still has an unacceptably high incidence rate in both humans and animals although there have been many studies on AS. To continue our previous pathological and biochemical investigation on the underlying mechanisms of AS incidence in broiler chickens, cutting-edge technologies including RNA-seq and metabolimics were used by directly comparing AS chickens and healthy controls. The RNA-seq analysis in the liver identified 390 differentially expressed genes (DEGs), among which 212 genes were up-regulated and 178 genes were down-regulated in the AS group compared to the control. For the down-regulated DEGs, further gene ontology (GO) analysis suggested that lipid metabolism, cell differentiation, enzyme linked receptor protein signaling pathway and steroid biosynthesis pathway were significantly enriched. For up-regulated DEGs, the cholesterol metabolic process has the lowest p value (0.000966) of fold enrichment while the cholesterol biosynthetic process has the highest fold enrichment (46.67). The metabolomic analysis of serum revealed statistically significant changes in the concentrations of LysoPC(20 : 4), LysoPC(16 : 0), LysoPC(18 : 0), LysoPC(18 : 1), LysoPC(18 : 2), PC(14 : 1/20 : 1), PC(20 : 4/18 : 0), PC(14 : 1/22 : 1), dihydroxyacetone, indoleacrylic acid, ursodeoxycholic acid, l-valine, and l-tryptophan. The integrative analysis of transcriptome and metabolome indicated that two biological pathways of tryptophan biosynthesis and metabolism, and glycerophospholipid metabolism may contribute to the induction of AS in broilers. These findings have provided novel insights into our understanding of molecular mechanisms of AS incidence in broilers.

Phytosterols are intended for use as a novel food ingredient with plasma cholesterol-lowering activity. Although phytosterols are naturally present in the normal diet, daily consumption is insufficient to ensure plasma cholesterol-lowering levels. Therefore, phytosterols may be added to the diets to achieve the desired cholesterol-lowering activity. A subchronic laying hen safety study was conducted to examine if high-dose phytosterols could affect the safety of hens. Three hundred sixty 21-wk-old Hy-Line Brown laying hens were randomly assigned to 5 groups with 6 replicates of 12 birds each; after 3 wk, birds were fed diets supplemented with 0, 20, 80, 400, and 800 mg/kg of phytosterols for 12 wk. Throughout the study, clinical observations and laying performance were measured. At the end of the study, birds were subjected to a full postmortem examination: blood samples were taken for clinical pathology, selected organs were weighed, and specified tissues were taken for subsequent histological examination. No treatment-related changes that were considered to be of toxicological significance were observed. Therefore, a nominal phytosterol concentration of 800 mg/kg was considered to be the no-observed-adverse-effect level.

An experiment was conducted to evaluate the effect of stocking density on growth performance, carcass yield, and immune status of a local chicken breed. In total, 840 one-day-old male Suqin yellow chickens were placed into 4-m(2) cages in groups of 50 (low), 70 (medium), or 90 (high) birds. Each treatment was represented by 4 replicates (cages). The cages measured 2.84 × 1.42 m; half of the area of the cage (2 m(2)) was used from 1 to 28 d and the whole cage was used from 29 to 42 d. Stocking densities were 25, 35, and 45 birds/m(2) from 1 to 28 d and 12.5, 17.5, and 22.5 birds/m(2) from 29 to 42 d (low, medium, and high, respectively). Final production (live bird mass after fasting) per unit area was 14.46, 19.46, and 24.23 kg/m(2), respectively, at 42 d of age. Several immune parameters were evaluated, and the growth performance, carcass yield, and meat quality were determined. Body weight at 28 and 42 d of age was significantly reduced as the stocking density increased (P0.05). A depression in daily weight gain was noticed from 1 to 28 d and 1 to 42 d of age, and daily feed intake decreased significantly in each period as density increased (P0.05). The feed/gain from 29 to 42 d and from 1 to 42 d of age decreased as density increased (P0.05). At 42 d, there was no effect of the stocking density on carcass, eviscerated carcass, breast, and abdominal fat yields (P0.05). The thigh yield of chickens in the medium-density group improved significantly (P0.05) compared with those of the other 2 groups. The water-loss rate, shear force, and meat color of the muscle were unaffected (P0.05) by the stocking density, but pH values increased slightly as density increased. No significant difference was noted in the immunological parameters, but the blood total protein and potassium were significantly affected by stocking density (P0.05). The findings of this study suggest that increasing the stocking density advantageously affected feed/gain and decreased the final BW, whereas no evidence was found that stocking density caused changes in any of the measured immune parameters.

The experiment was conducted to investigate the effects of starter feeding time on body growth and viscera development of newly hatched chicks. A total of 1080 Yangzhou chicks with similar weight derived from 1800 eggs were selected in this experiment. These chicks were randomly assigned to six groups (1, 2, 3, 4, 5 and 6) according to six feeding schedules (fasting for 12, 24, 36, 48, 60, 72 hours posthatch), with 3 replicates per group and 60 chicks (30 males and 30 females) per replicate. After fasting, all birds were fed a crumbled starter feedstuff ad libitum until 6 days of age. At 0, 1, 2, 3, 4, 5 and 6d posthatch, eight chicks (4 males and 4 females) per replicate of each experimental group were randomly selected and individually weighed. Yolk sac, liver, heart, pancreas, bursal and small intestine of each chick were taken out and weighed; the length of small intestine, moreover, was measured. Results showed that food deprivation reduced body weight (BW) of experimental chicks; furthermore BW decreased more and more with a prolonged fasting time. At 6 d of age, the BW of the former five groups was higher than that of the Group 6 (P

Experiments were conducted to compare endogenous amino acid losses and the true amino acid availability (TAAA) of 3 feedstuffs by using methods involving a short-term fasting and an N-free diet with cecectomized ganders. Diets were formulated to contain soybean meal, fish meal, and cottonseed meal as the sole source of protein. A precision-fed assay was used in which each feed sample was precise-fed (60 g) to geese and excreta were collected for 48 h. A N-free diet and fasting methods were used to evaluate the endogenous amino acid losses. Endogenous losses of 3 amino acids were significantly different (P0.01) with the N-free diet and fasting methods. The TAAA of soybean meal, fish meal, and cottonseed meal determined by N-free diet method ranged from 84.49 to 97.09%, 89.18 to 98.16%, and 77.09 to 98.32%, respectively. The TAAA of these 3 diets determined by the fasting method ranged from 83.50 to 97.77%, 88.08 to 99.60%, and 76.09 to 98.09%, respectively. However, there were only a few small differences (P0.05) between methods in each amino acid. In conclusion, there was no difference in determination of the amino acid availability of these feedstuffs using cecectomized ganders between the N-free diet and fasting methods.

Ascites is a major problem for both human health and animal production, due to its association with high rates of morbidity and mortality, low efficiency of nutrient utilization, and permanent adverse effects on performance. Although it is one of the three major metabolic diseases in poultry production, the underlying mechanisms are largely unknown. In this study, six ascites syndrome (AS) chickens and six normal chickens were obtained from each group (108 chickens) at 21 and 35 days. A liver metabolomics method based on ultra-performance liquid chromatography/quadruple time-of-flight mass spectrometry (UPLC/Q-TOF/MS) was used to explore the metabolic pattern of low molecular mass metabolites in chickens with low-temperature-induced AS. Coupled with blood biochemistry and histopathology results, the significant difference in metabolic profiling between the AS group and the control group, as determined through pattern recognition analysis, indicated changes in global tissue metabolites. The results showed that a primary bile acid synthesis disorder and inflammation had occurred by 21 days and that lysophospholipid metabolism was disrupted by 35 days with the continuation of low temperatures. Several metabolites, including taurodeoxycholic acid, cholic acid glucuronide, glycocholic acid, LysoPC(15 : 0) and taurocholic acid, were identified as the potential and proper biomarkers of AS. These biochemical changes in tissue metabolites are related to perturbations of lipid metabolism, which may be helpful to further understand the AS mechanisms. This work shows that the metabolomics is a valuable tool for studying metabolic diseases.