Epigenetic Modification of TLRs in Leukocytes Is Associated with Increased Susceptibility to Salmonella enteritidis in Chickens

Toll-like receptors (TLRs) signaling pathways are the first lines in defense against Salmonella enteritidis (S. enteritidis) infection but the molecular mechanism underlying susceptibility to S. enteritidis infection in chicken remains unclear. SPF chickens injected with S. enteritidis were partitioned into two groups, one consisted of those from Salmonella-susceptible chickens (died within 5 d after injection, n = 6), the other consisted of six Salmonella-resistant chickens that survived for 15 d after injection. The present study shows that the bacterial load in susceptible chickens was significantly higher than that in resistant chickens and TLR4, TLR2-1 and TLR21 expression was strongly diminished in the leukocytes of susceptible chickens compared with those of resistant chickens. The induction of expression of pro-inflammatory cytokine genes, IL-6 and IFN-β, was greatly enhanced in the resistant but not in susceptible chickens. Contrasting with the reduced expression of TLR genes, those of the zinc finger protein 493 (ZNF493) gene and Toll-interacting protein (TOLLIP) gene were enhanced in the susceptible chickens. Finally, the expression of TLR4 in peripheral blood mononuclear cells (PBMCs) infected in vitro with S. enteritidis increased significantly as a result of treatment with 5-Aza-2-deoxycytidine (5-Aza-dc) while either 5-Aza-dc or trichostatin A was effective in up-regulating the expression of TLR21 and TLR2-1. DNA methylation, in the predicted promoter region of TLR4 and TLR21 genes, and an exonic CpG island of the TLR2-1 gene was significantly higher in the susceptible chickens than in resistant chickens. Taken together, the results demonstrate that ZNF493-related epigenetic modification in leukocytes probably accounts for increased susceptibility to S. enteritidis in chickens by diminishing the expression and response of TLR4, TLR21 and TLR2-1

Equol Inhibits LPS-Induced Oxidative Stress and Enhances the Immune Response in Chicken HD11 Macrophages

Background/Aims: There has been increasing recent attention on the antioxidative capacity of equol. This study tested the effect of equol on oxidative stress induced by lipopolysaccharide (LPS) and regulation of immunity in chicken macrophages. Methods: Chicken HD11 macrophages were challenged with LPS (100 ng/mL) alone or with LPS (100 ng/mL) and (±)equol (10, 20, 40, 80, 160 μmol/L) together for 24h. Evaluated responses included the contents of malondialdehyde (MDA) and reduced glutathione (GSH), activities of total superoxide dismutase (T-SOD) and inducible nitric oxide synthase (iNOS), transcript abundance of superoxide dismutase 2 (SOD2), catalase (CAT), glutathione transferase (GST), Toll-like receptor 4 (TLR4), tumor necrosis factor alpha (TNFα) and interleukin-1 beta (IL-1β), and contents of the cytokines TNFα, IL-1β, interleukin-2 (IL-2) and interferon beta (IFNβ). Results: Exposure to LPS induced oxidative stress as contents of MDA increased and GSH decreased in LPS-treated cells (P SOD2 and GST transcripts (P TLR4, TNFα and IL-1β (P < 0.05). And there were similar changes in contents of IL-1β, IL-2, IFNβ and TNFα in the cells (P < 0.05). Conclusions: It concluded that equol can protect chicken HD11 macrophages from oxidative stress induced by LPS through reducing lipid peroxidation products and enhancing contents of antioxidants, and activities of relevant antioxidase enzymes; effects were also seen in gene expression related to the immune response and increased contents of cytokines. The optimal concentration of equol on antioxidation and immune enhancement in chicken macrophages was 40 μmol/L

Effects of metabolic energy intervention on lipid content and liver transcriptome in finisher yellow-feathered chickens

High-energy diets improve growth rates and shorten the growth period while resulting in excessive fat deposition in the liver of birds. The effect of different energy diets on lipid content and liver transcriptome in finisher Huxu chickens was examined here. A total of 200 90-d-old Huxu chickens with an average BW of 1.08 kg were randomly divided into two dietary treatments. The birds were fed diets either containing 12.06 MJ ME/kg (low metabolic energy, LME) or 13.31 MJ ME/kg (high metabolic energy, HME) for 45 days. Chickens fed the HME diet had significantly higher total triglyceride and total cholesterol concentrations in both plasma and liver than did those fed the LME diet. Transcriptomic profiling showed 480 known differentially expressed genes (DEG), with 180 down-regulated and 300 up-regulated chickens from the HME diet compared with the LME diet. Notably, these genes were mostly enriched in lipid metabolism including the PPAR signalling pathway, fatty acid elongation, and glycerophospholipid metabolism. The relative mRNA expression level of 14 lipid metabolism-related genes, included 13 up-regulated (MBOAT2, PLA2G2E, AGPAT2, AGPAT3, PCYT2, PEMT, ELOVL2, HADHA, ACOT1L, FABP3, FABP1, PLIN1, RXRG) and one down-regulated (LCAT) gene. In conclusion, the present findings indicate that a high-energy diet likely regulates gene expression of the 14 lipid metabolism-related genes through the PPAR signalling pathway, fatty acid elongation, and glycerophospholipid metabolism resulting in excessive fat accumulation in the liver of Huxu chickens. These results provide a better understanding of the biological mechanisms by which the increased dietary energy induces fat accumulation in the liver.Highlights A high-energy diet without change in protein level induced excessive fat accumulation in the Huxu chickens. Chickens fed the high-energy diet had induced higher total triglyceride and total cholesterol concentrations in both plasma and liver. This study showed a comprehensive profile of hepatic gene expression in chickens with fatty liver induced by high energy

Potential Effects of Acidifier and Amylase as Substitutes for Antibiotic on the Growth Performance, Nutrient Digestion and Gut Microbiota in Yellow-Feathered Broilers

This study was conducted to evaluate the effects of acidifier (benzoic acid, BA), amylase (AL) and their combination as substitutes for antibiotics on growth performance, antioxidation, nutrient digestion and gut microbiota of yellow-feathered broilers. A total of 1440 twenty-one-day-old broilers were randomly allocated to six treatments. Broilers in the control group (CON) were fed a basal diet, whereas birds in the other five groups were fed the basal diet supplemented with antibiotic (zinc bacitracin, AT, 40 mg/kg), BA (2000 mg/kg), low level AL (AL-L, 300 mg/kg), high level AL (AL-H, 500 mg/kg) and the combination of AL-H and BA (BA+AL-H). The experimental animals were killed at the end of the trial (21 day-63 day) then blood samples were collected from two birds per pen. Bird weight, feed intake and survival rate were recorded on pen basis. Growth performance was not significantly influenced by AT, BA, AL-L, AL-H or BA+AL-H. Plasma uric acid (UA) was decreased from CON by all treatments; the activity of AKP in plasma was also lowered by AT, BA, AL-H and BA+AL-H. Plasma activity of LDH was reduced by BA. In the jejunal mucosa, Na+K+-ATP activity was increased by BA, AL-L, AL-H and BA+AL-H. Mucosal activities of T-AOC and CAT were increased with AL-L and AT supplementation, respectively. Additionally, the relative abundance of Escherichia coli (E. coli) in the cecal contents was reduced by BA+AL-H and, with the exception of AL-H, all treatments increased the relative abundance of Lactobacillus. In conclusion, dietary AT, BA, AL-L, AL-H or BA+AL were effective in improving the antioxidant capacity, nutrient digestion and gut microbiota composition. No significant differences were observed in the tested variables between AT and other treatments, indicating that BA, AL and their combination may be alternatives to dietary inclusion of zinc bacitracin. Dietary addition of 500 mg/kg AL and 2000 mg/kg BA was an optimum supplementation dose

Dietary supplementation of bilberry anthocyanin on growth performance, intestinal mucosal barrier and cecal microbes of chickens challenged with Salmonella Typhimurium

Abstract Background Anthocyanins (AC) showed positive effects on improving the intestinal health and alleviating intestinal pathogen infections, therefore, an experiment was conducted to explore the protective effects of supplemented AC on Salmonella-infected chickens. Methods A total of 240 hatchling chickens were randomly allocated to 4 treatments, each with 6 replicates. Birds were fed a basal diet supplemented with 0 (CON, and ST), 100 (ACL) and 400 (ACH) mg/kg of AC for d 60, and orally challenged with PBS (CON) or 109 CFU/bird (ST, ACL, ACH) Salmonella Typhimurium at d 14 and 16. Results (1) Compared with birds in ST, AC supplementation increased the body weight (BW) at d 18 and the average daily gain (ADG) from d 1 to 18 of the Salmonella-infected chickens (P < 0.05); (2) AC decreased the number of Salmonella cells in the liver and spleen, the contents of NO in plasma and inflammatory cytokines in ileal mucosa of Salmonella-infected chickens (P < 0.05); (3) Salmonella infection decreased the ileal villi height, villi height to crypt depth (V/C), and the expression of zonulaoccludins-1 (ZO-1), claudin-1, occludin, and mucin 2 (MUC2) in ileal mucosa. AC supplementation relieved these adverse effects, and decreased ileal crypt depth (P < 0.05); (4) In cecal microbiota of Salmonella-infected chickens, AC increased (P < 0.05) the alpha-diversity (Chao1, Pd, Shannon and Sobs indexes) and the relative abundance of Firmicutes, and decreased (P < 0.05) the relative abundance of Proteobacteria and Bacteroidota and the enrichment of drug antimicrobial resistance, infectious bacterial disease, and immune disease pathways. Conclusions Dietary AC protected chicken against Salmonella infection via inhibiting the Salmonella colonization in liver and spleen, suppressing secretion of inflammatory cytokines, up-regulating the expression of ileal barrier-related genes, and ameliorating the composition and function of cecal microbes. Under conditions here used, 100 mg/kg bilberry anthocyanin was recommended

Effects of Dietary Iron Level on Growth Performance, Immune Organ Indices and Meat Quality in Chinese Yellow Broilers

The objective of three trials was to investigate the effects of dietary Fe on growth performance, immune organ indices and meat quality of Chinese yellow broilers during the whole growth period. A total of 1440 1-day-old, 1440 22-day-old, and 1080 43-day-old Lingnan yellow male broilers were randomly assigned to one of six dietary treatments with six replicates per treatment (40 birds per replicate for both 1 to 21 d and 22 to 42 d, 30 birds for 43 to 63 d). Additional Fe (0, 20, 40, 60, 80, and 100 mg/kg) was added as FeSO4 &bull; H2O to the three basal diets (calculated Fe 50 mg/kg, analyzed 48.3, 49.1, 48.7 mg/kg, respectively). The calculated final dietary Fe concentrations in Starter, Grower and Finisher phases were 50, 70, 90, 110, 130, and 150 mg/kg. The results showed that average daily gain (ADG), average daily feed intake (ADFI) and feed conversion rate (FCR) of the broilers were not influenced by the different levels of Fe (p&gt; 0.05). Weight indices of the spleen, thymus and bursa of Fabricius were not influenced (p &gt; 0.05) by the different levels of Fe during three 21-day experimental periods. Hematocrit, and Fe contents of the liver and kidney were not affected by different levels of Fe (p&gt; 0.05). The diet with 150 mg/kg of Fe increased the a* (relative redness) value of breast muscle compared to the 50 and 70 mg/kg diets at 24 h post mortem (p&lt; 0.05). The diet with 90 mg/kg Fe increased the pH of breast muscle compared to broilers fed 50 or 150 mg/kg Fe (p &lt; 0.05) 45 min after slaughter. The diet with 90 mg/kg Fe decreased drip loss of breast muscle compared to 150 mg/kg Fe (p&lt; 0.05). These data suggest that feeding yellow-feathered broilers on a conventional corn-soy based diet satisfies their requirements without additional Fe at ages 1 to 21, and 22 to 42 d, while 90 mg/kg in the finisher phase improved meat quality, and from the QP (quadratic polynomial) models of the key meat quality variables, pH of breast muscle and drip loss of breast muscle, the optimal dietary Fe level was 89 to 108 mg/kg, and daily Fe fed allowance was 11 to 13 mg in the finisher phase (43 to 63 d)

Feed Restriction Improves Lipid Metabolism by Changing the Structure of the Cecal Microbial Community and Enhances the Meat Quality and Flavor of Bearded Chickens

Excessive fat deposition in full-fed Bearded chickens does not only reduce carcass yield but also causes consumer rejection of meat. Feed restriction (FR) is an effective method to save on feed cost, reduce carcass fat deposition, and improve meat quality. A total of 560 150-d Bearded chickens were randomly divided into seven groups (each with eight replicates of ten birds) for 40 days. The control group was fed with the basal diet ad libitum (CON), and the other six groups were fed with 90% of the feed intake (90% FI), 80% FI, 70% FI, 90% metabolizable energy (90% ME), 80% ME, and 70% ME of the CON, respectively. Compared to the CON group, FR increased meat yield, but the total weight of the Bearded chickens was slighter; 80% FI and 70% ME improved the relative lipid metabolism indices of chickens, especially the levels of triglycerides and total cholesterol in the plasma and liver (p &lt; 0.05), and decreased calpastatin activity in the breast muscle (p &lt; 0.05). Additionally, 16S rRNA sequencing of cecal microbial community indicated that an increase in the abundance of Hydrogenoanaerobacterium and Bacteroides plebeius was observed in the 80% FI group (p &lt; 0.05), and an enrichment in Olsenella, Catabacter, and Lachnospiraceae were observed in the 70% ME group (p &lt; 0.05) compared to the CON group. Moreover, compared to the CON group, the L * value of the breast muscle significantly decreased, and a * value significantly increased in the 80% FI group (p &lt; 0.05). Notably, the concentrations of threonine, lysine, aspartic acid, glutamic acid, proline, and arginine and the activity of calpain in breast muscle increased in the 80% FI group more than in the CON group (p &lt; 0.05), while valine, isoleucine, leucine, phenylalanine, lysine, alanine, tyrosine and proline decreased in ME restriction groups (p &lt; 0.05). Taken together, our results indicated that 80% FI could improve lipid metabolism by changing the structure of the cecal microbial community, and the meat quality and flavor of the Bearded chickens in 80% FI group was improved with a promoted meat color score, flavor substances, and the calproteinase system

Feed Restriction Improves Lipid Metabolism by Changing the Structure of the Cecal Microbial Community and Enhances the Meat Quality and Flavor of Bearded Chickens

Excessive fat deposition in full-fed Bearded chickens does not only reduce carcass yield but also causes consumer rejection of meat. Feed restriction (FR) is an effective method to save on feed cost, reduce carcass fat deposition, and improve meat quality. A total of 560 150-d Bearded chickens were randomly divided into seven groups (each with eight replicates of ten birds) for 40 days. The control group was fed with the basal diet ad libitum (CON), and the other six groups were fed with 90% of the feed intake (90% FI), 80% FI, 70% FI, 90% metabolizable energy (90% ME), 80% ME, and 70% ME of the CON, respectively. Compared to the CON group, FR increased meat yield, but the total weight of the Bearded chickens was slighter; 80% FI and 70% ME improved the relative lipid metabolism indices of chickens, especially the levels of triglycerides and total cholesterol in the plasma and liver (p p Hydrogenoanaerobacterium and Bacteroides plebeius was observed in the 80% FI group (p Olsenella, Catabacter, and Lachnospiraceae were observed in the 70% ME group (p p p p < 0.05). Taken together, our results indicated that 80% FI could improve lipid metabolism by changing the structure of the cecal microbial community, and the meat quality and flavor of the Bearded chickens in 80% FI group was improved with a promoted meat color score, flavor substances, and the calproteinase system