Effects of microalgae, with or without xylanase supplementation, on serum immunoglobulins, cecal short-chain fatty acids, microbial diversity, and metabolic pathways of broiler chickens.

Modern broilers are highly susceptible to environmental and pathogenic threats, leading to gut disorders and poor nutrient utilization if not managed properly. Nutritional programming using several feedstuffs and coproducts to manage gut health has been studied. This study used microalgae as a functional compound and xylanase enzyme in broilers' diets as a strategy to manage gut health. A total of 162 one-day-old unsexed Cobb 500 broiler chicks were randomly assigned to 1 of the 3 dietary treatments: a) corn-soybean meal-based control diet (CON), b) 3% microalgae (MAG), and c) MAG with xylanase enzyme (MAG+XYN). The chicks were reared for 35 days (d) on a floor pen system maintaining standard environment conditions to evaluate the effects of microalgae, with or without xylanase supplementation, on serum immunoglobulins, cecal short-chain fatty acids (SCFA) production, cecal microbial diversity, and metabolic pathways. No significant differences were found for serum immunoglobulin and cecal SCFA among the treatment groups (P > 0.05). Relative microbial abundance at the genus level showed that MAG and MAG+XYN groups had a diverse microbial community on d 3 and d 35. However, no bacterial genus had a significant difference (P > 0.05) in their relative abundance on d 3, but 16 genera showed significant differences (P < 0.05) in their relative abundance among the dietary treatments on d 35. Most of these bacteria were SCFA-producing bacteria. Moreover, MAG and MAG+XYN-fed broilers had better responses than CON groups for metabolic pathways (D-mannose degradation, pectin degradation I and II, ß-1-4-mannan degradation, tetrahydrofolate biosynthesis, glutathione biosynthesis, glutathione-peroxide redox reactions, lactate fermentation to propionate, acetate, and hydrogen, etc.) both on d 3 and d 35. The results suggest that using microalgae, with or without xylanase, had no statistical impact on serum immunoglobulins and cecal SCFA production in broilers. However, an improvement in the cecal microbial diversity and metabolic pathways, which are essential indicators of gut health and nutrient utilization, was observed. Most of the improved metabolic pathways were related to fiber utilization and oxidative stress reduction.

Short-chain fatty acids ameliorate necrotizing enterocolitis-like intestinal injury through enhancing Notch1-mediated single immunoglobulin interleukin-1-related receptor, toll-interacting protein, and A20 induction.

Single immunoglobulin interleukin-1-related receptor (SIGIRR), toll-interacting protein (TOLLIP), and A20 are major inhibitors of toll-like receptor (TLR) signaling induced postnatally in the neonatal intestine. Short-chain fatty acids (SCFAs), fermentation products of indigestible carbohydrates produced by symbiotic bacteria, inhibit intestinal inflammation. Herein, we investigated the mechanisms by which SCFAs regulate SIGIRR, A20, and TOLLIP expression and mitigate experimental necrotizing enterocolitis (NEC). Butyrate induced NOTCH activation by repressing sirtuin 1 (SIRT1)-mediated deacetylation of the Notch intracellular domain (NICD) in human intestinal epithelial cells (HIECs). Overexpression of NICD induced SIGIRR, A20, and TOLLIP expression. Chromatin immunoprecipitation revealed that butyrate-induced NICD binds to the SIGIRR, A20, and TOLLIP gene promoters. Notch1-shRNA suppressed butyrate-induced SIGIRR/A20 upregulation in mouse enteroids and HIEC. Flagellin (TLR5 agonist)-induced inflammation in HIEC was inhibited by butyrate in a SIGIRR-dependent manner. Neonatal mice fed butyrate had increased NICD, A20, SIGIRR, and TOLLIP expression in the ileal epithelium. Butyrate inhibited experimental NEC-induced intestinal apoptosis, cytokine expression, and histological injury. Our data suggest that SCFAs can regulate the expression of the major negative regulators of TLR signaling in the neonatal intestine through Notch1 and ameliorate experimental NEC. Enteral SCFAs supplementation in preterm infants provides a promising bacteria-free, therapeutic option for NEC.NEW & NOTEWORTHY Short-chain fatty acids (SCFAs), such as propionate and butyrate, metabolites produced by symbiotic gut bacteria are known to be anti-inflammatory, but the mechanisms by which they protect against NEC are not fully understood. In this study, we reveal that SCFAs regulate intestinal inflammation by inducing the key TLR and IL1R inhibitors, SIGIRR and A20, through activation of the pluripotent transcriptional factor NOTCH1. Butyrate-mediated SIGIRR and A20 induction represses experimental NEC in the neonatal intestine.

20S-ginsenoside Rg3 inhibits the biofilm formation and haemolytic activity of Staphylococcus aureus by inhibiting the SaeR/SaeS two-component system.

Introduction. Staphylococcus aureus is a major cause of chronic diseases and biofilm formation is a contributing factor. 20S-ginsenoside Rg3 (Rg3) is a natural product extracted from the traditional Chinese medicine red ginseng.Gap statement. The effects of Rg3 on biofilm formation and haemolytic activity as well as its antibacterial mechanism against S. aureus have not been reported.Aim. This study aimed to investigate the effects of Rg3 on biofilm formation and haemolytic activity as well as its antibacterial action against clinical S. aureus isolates.Methodology. The effect of Rg3 on biofilm formation of clinical S. aureus isolates was studied by crystal violet staining. Haemolytic activity analysis was carried out. Furthermore, the influence of Rg3 on the proteome profile of S. aureus was studied by quantitative proteomics to clarify the mechanism underlying its antibacterial action and further verified by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR).Results. Rg3 significantly inhibited biofilm formation and haemolytic activity in clinical S. aureus isolates. A total of 63 with >1.5-fold changes in expression were identified, including 34 upregulated proteins and 29 downregulated proteins. Based on bioinformatics analysis, the expression of several virulence factors and biofilm-related proteins, containing CopZ, CspA, SasG, SaeR/SaeS two-component system and SaeR/SaeS-regulated proteins, including leukocidin-like protein 2, immunoglobulin-binding protein G (Sbi) and fibrinogen-binding protein, in the S. aureus of the Rg3-treated group was downregulated. RT-qPCR confirmed that Rg3 inhibited the regulation of SaeR/SaeS and decreased the transcriptional levels of the biofilm-related genes CopZ, CspA and SasG.Conclusions. Rg3 reduces the formation of biofilm by reducing cell adhesion and aggregation. Further, Rg3 can inhibit the SaeR/SaeS two-component system, which acts as a crucial signal transduction system for the anti-virulence activity of Rg3 against clinical S. aureus isolates.

The soluble CD83 protein prevents bone destruction by inhibiting the formation of osteoclasts and inducing resolution of inflammation in arthritis.

Here we show that soluble CD83 induces the resolution of inflammation in an antigen-induced arthritis (AIA) model. Joint swelling and the arthritis-related expression levels of IL-1ß, IL-6, RANKL, MMP9, and OC-Stamp were strongly reduced, while Foxp3 was induced. In addition, we observed a significant inhibition of TRAP+ osteoclast formation, correlating with the reduced arthritic disease score. In contrast, cell-specific deletion of CD83 in human and murine precursor cells resulted in an enhanced formation of mature osteoclasts. RNA sequencing analyses, comparing sCD83- with mock treated cells, revealed a strong downregulation of osteoclastogenic factors, such as Oc-Stamp, Mmp9 and Nfatc1, Ctsk, and Trap. Concomitantly, transcripts typical for pro-resolving macrophages, e.g., Mrc1/2, Marco, Klf4, and Mertk, were upregulated. Interestingly, members of the metallothionein (MT) family, which have been associated with a reduced arthritic disease severity, were also highly induced by sCD83 in samples derived from RA patients. Finally, we elucidated the sCD83-induced signaling cascade downstream to its binding to the Toll-like receptor 4/(TLR4/MD2) receptor complex using CRISPR/Cas9-induced knockdowns of TLR4/MyD88/TRIF and MTs, revealing that sCD83 acts via the TRIF-signaling cascade. In conclusion, sCD83 represents a promising therapeutic approach to induce the resolution of inflammation and to prevent bone erosion in autoimmune arthritis.

Transfer and persistence of bovine immunoglobulins in lambs fed a colostrum replacer.

BACKGROUND: Colostrum-derived antibodies are crucial for the protection of newborn lambs from infectious diseases. Several colostrum replacer products that contain bovine antibodies are on the market. We investigated the absorption and persistence of bovine antibodies from a powdered colostrum replacer in newborn lambs. METHODS: We tested a lamb colostrum replacer containing bovine serum in lambs that were separated from their dams at birth. Immunoglobulin G (IgG) uptake was analysed by ELISA, and the persistence of antigen-specific antibodies was analysed by parainfluenza 3 virus (PI-3) neutralisation assay. RESULTS: Serum antibody ELISA performed on days 1 and 14 revealed IgG levels of 17.9 ± 2.8 and 27.5 ± 2.5 mg/ml, respectively. PI-3 antibodies derived from the colostrum replacer were present for 86.3 ± 10.6 days. CONCLUSIONS: Antibodies derived from bovine serum protein delivered to lambs via a commercial colostrum replacer are readily absorbed and persist for months, suggesting that these products may offer adequate protection.

Zinc and L-Ascorbic Acid Dietary Supplementation Affected Antioxidant Status, Heat Shock Protein 70, and Some Immunoglobulins in Japanese Quail Under Heat Stress Conditions.

This study assessed the impact of the dietary inclusion of L-ascorbic acid and organic zinc (Availa-Zn) on heat-stressed Japanese quails. Growth performance, antioxidant status, immune status, heat shock protein 70 (HSP70), and some blood biochemical parameters were assessed. One-day-old, unsexed Japanese quail chicks (n = 240) were randomly allocated into 4 dietary treatments (6 replicates per treatment; 10 birds per replicate). Birds were fed a basal corn-soybean meal diet (control treatment) with different supplemental levels of L-ascorbic acid and/or Availa-Zn (200 mg L-ascorbic acid/kg diet, 62 mg Availa-Zn/kg diet, and 200 mg L-ascorbic acid + 62 mg Availa-Zn/kg diet) from July to August 2020 for 35 days. The average minimum and maximum ambient temperatures varied from 85.4 to 98.8 °F, and the relative humidity was between 69 and 74%. Supplemented L-ascorbic acid and Availa-Zn, either as separate supplements or as combined supplements, increased bird growth performance, blood hemoglobin, thyroid hormones, total protein, globulin, total antioxidant capacity, HSP70, catalase, superoxide dismutase enzyme activity, and immunoglobulin A and G (P < 0.05), while heterophil/lymphocytes decreased (P < 0.01) during the entire rearing period (1-35 days). Most of the assessed parameters showed stronger responses when L-ascorbic acid and Availa-Zn were added together, which may suggest a synergistic effect. In conclusion, the combined supplementation of L-ascorbic acid and Availa-Zn at 200 and 62 mg/kg, respectively, could be considered an efficient dietary supplement to enhance Japanese quail growth performance, antioxidant capacity, immune status, and general health under heat stress conditions.

Nile tilapia bones as eco-friendly alternative in water treatment and enhancing performance and immunity in broiler chickens.

Background: Water pollution has become a major threat to the environment and the living so an eco-friendly bio-filter was chosen for its merits over conventional techniques. Aim: Investigating the purifying activities of the Tilapia bone powder against inorganics, heavy metals, and microbial water pollutants and its impacts on performance, biochemical and antioxidant levels, cortisol and immunoglobulin concentrations, and intestinal microbiota in challenged broiler chickens. Methods: The in-vitro activity of Tilapia bone powder was evaluated against magnesium chloride and lead nitrate using tube minimal inhibitory concentration (MIC), as well as against Escherichia coli O1527:H7, Salmonella Typhimurium, Mycoplasma gallisepticum, Aspergillus niger, Trichophyton mentagrophytes, and Candida albicans using a 96-micro-well MIC. A total of 250 1-day-old Hubbard chicks were divided into five groups on a deep litter system. Chicks were supplemented daily with Tilapia bone powder (1 g/l) for 4-6 hours from the 3rd day. Challenges were served on the 7th, 14th, 21st, 28th, and 35th days for four broiler groups using magnesium chloride (100 mg/l), lead nitrate (350 mg/l), E. coli (2.4 × 1012 CFU/ml), S. Typhimurium (1.8 × 108 CFU/ml), respectively, and the 5th group was assigned as a control. A total of 2,250 samples (90 Tilapia-pollutants mixes, 480 Tilapia-microbial mixes, 240 sera, 240 intestinal swabs, and 1,200 tissue samples) were collected. Results: Tilapia bone powder 1% reveals a 100% reduction in the lead after 1 hour, total and calcium hardness after 0.5 hours, as well as 100% killing efficacy against E. coli O1527:H7, S. Typhimurium, M. gallisepticum, A. niger, T. mentagrophytes, and C. albicans after 0.5, 1, 1, 1, 1, and 1 hour, respectively. Tilapia bone powder 1% treated water reveals highly significant (p < 0.01) increases in dissolved oxygen and declines in physicochemical and microbial parameters compared with tap water. Challenged treated broilers revealed highly significant (p < 0.01) increases in weight gains, performance index, body weights, carcasses, and organs weights, immunoglobulin concentrations, and antioxidant levels, as well as highly significant (p < 0.01) improvements in feed conversions, feed and water intakes, biochemical profile, cortisol hormone, and intestinal microbiota. Conclusion: Tilapia bone powder provided significant in-vitro adsorptive and antimicrobial actions, as well as supported the broiler chickens to mitigate the polluted water stress accompanied by enhanced performance, carcass quality, immunity, and intestinal microbiota.

Puerarin improved growth performance and postmortem meat quality by regulating lipid metabolism of cattle under hot environment.

This study aims to evaluate the effect of puerarin on performance, meat quality, and serum indexes of beef cattle under hot environment. Thirty-two bulls were divided into four groups and fed diet supplemented with puerarin at 0, 200, 400, or 800 mg/kg. Results showed that heat stress was employed for 54 out of 60 days, 400 mg/kg group declined serum cortisol (COR) contents, all treatments increased the contents of total cholesterol, high density lipoprotein cholesterol, and total superoxide dismutase activity; in addition, glutathione peroxidase activity of 200 mg/kg group were enhanced, only 800 mg/kg group enhanced immunoglobulin (IgA, IgM, and IgG) and low density lipoprotein cholesterol contents compared with the control (p < .05). Moreover, 400-mg/kg puerarin increased serum growth hormone levels compared with 200 mg/kg group but declined COR concentrations compared with 200 mg/kg and 800 mg/kg groups (p < .05). More importantly, average daily gain and daily matter intake, and intramuscular fat contents of 400 mg/kg group were enhanced, but the shear force of beef in 400 mg/kg and 800 mg/kg groups were declined compared with the control (p < .05). These findings indicated that supplemental with puerarin enhanced immune and antioxidant, and 400 mg/kg of puerarin improved performance and meat quality by normalizing levels of stress hormones and increasing intramuscular fat deposition of beef cattle under hot environment.

Effects of dietary resveratrol supplementation on immunity, antioxidative capacity and intestinal barrier function in weaning piglets.

The aim of this study was to investigate effects of dietary supplementation of resveratrol on immunity, antioxidative capacity, intestinal barrier function in weaning piglets. Here, fifty-four 28-day-old Duroc × Landrace × Yorkshire weaning piglets were randomly divided into three dietary treatments and fed with a basal diet or a basal diet supplemented with 150 and 300 mg/kg resveratrol, respectively, for 42 days. The results indicated that resveratrol increased serum immunoglobulin G content. In serum, resveratrol increased glutathione peroxidase enzyme activity and decreased malondialdehyde (MDA) content. In liver, resveratrol not only increased T-AOC and total superoxide dismutase enzyme activities but also decreased MDA content. Meanwhile, the results showed that resveratrol had significantly increased the jejunum villus height and villus height/crypt depth, and decreased the crypt depth in jejunum. Furthermore, the mRNA expressions of IL-10 and ZO-1 were significantly increased in jejunal mucosa. However, there was no significant difference in the mRNA expressions of TNF-α, IL-1ß, IL-6, Occludin and Claudin1 between the treatment groups and the control group. Taken together, these results indicated that dietary supplementation of resveratrol could increase antioxidant activity, promote the integrity of intestinal barrier and increase the production of anti-inflammatory cytokines in weaning piglets.

Siglec-15 as an Emerging Target for Next-generation Cancer Immunotherapy.

Immunomodulatory agents blocking the PD-1/PD-L1 pathway have shown a new way to treat cancer. The explanation underlying the success of these agents may be the selective expression of PD-L1 with dominant immune-suppressive activities in the tumor microenvironment (TME), supporting a more favorable tumor response-to-toxicity ratio. However, despite the big success of these drugs, most patients with cancer show primary or acquired resistance, calling for the identification of new immune modulators in the TME. Using a genome-scale T-cell activity array in combination with bioinformatic analysis of human cancer databases, we identified Siglec-15 as a critical immune suppressor with broad upregulation on various cancer types and a potential target for cancer immunotherapy. Siglec-15 has unique molecular features compared with many other known checkpoint inhibitory ligands. It shows prominent expression on macrophages and cancer cells and a mutually exclusive expression with PD-L1, suggesting that it may be a critical immune evasion mechanism in PD-L1-negative patients. Interestingly, Siglec-15 has also been identified as a key regulator for osteoclast differentiation and may have potential implications in bone disorders not limited to osteoporosis. Here, we provide an overview of Siglec-15 biology, its role in cancer immune regulation, the preliminary and encouraging clinical data related to the first-in-class Siglec-15 targeting mAb, as well as many unsolved questions in this pathway. As a new player in the cancer immunotherapeutic arena, Siglec-15 may represent a novel class of immune inhibitors with tumor-associated expression and divergent mechanisms of action to PD-L1, with potential implications in anti-PD-1/PD-L1-resistant patients.

Sophoricoside from Sophora japonica ameliorates allergic asthma by preventing mast cell activation and CD4+ T cell differentiation in ovalbumin-induced mice.

Asthma is a chronic inflammatory lung disorder with continuously increasing prevalence worldwide. Novel strategies are needed to prevent or improve asthma. The aim of this study was to investigate the effects of sophoricoside from Sophora japonica on allergic asthma. The mature seeds of S. japonica contain a large amount of sophoricoside. Sophoricoside reduced allergic and asthmatic symptoms by suppressing airway inflammation and antibody-antigen reaction in mouse models. In particular, sophoricoside suppressed immune cell recruitment into the airway lumens of the lungs and production of pro-inflammatory cytokines in the bronchoalveolar lavage fluid (BALF) of ovalbumin (OVA)-induced mice. It also decreased the amounts of histamine and arachidonic acid metabolites released in OVA-induced mice and antibody-antigen stimulated mast cells. In addition, sophoricoside decreased differentiation of naïve CD4+ T cells into T helper type 1 (Th1), Th2, and Th17 cells. Overall, we demonstrated that sophoricoside improved allergic asthma by suppressing mast cell activation and CD4+ T cell differentiation.

Effects of dietary supplementation of compound enzymes on performance, nutrient digestibility, serum antioxidant status, immunoglobulins, intestinal morphology and microbiota community in weaned pigs.

The objective of this study was to evaluate the effects of compound enzymes (CE) (containing per g 375 U amylase, 2500 U protease, 4000 U xylanase and 150 U ß-glucanase) on performance, nutrient digestibility, serum antioxidant status, immunoglobulins, intestinal morphology, volatile fatty acids contents and microbiota community in weaned pigs. Seventy-two pigs (Duroc × Landrace × Yorkshire, weaned at d 28) with an average body weight of 8.49 ± 0.87 kg were allotted into two treatments with six replicate pens per treatment (three barrows and three gilts per pen) according to sex and body weight in a randomised complete block design. The treatments contained a corn-soybean meal-barley basal diet (CON) or a basal diet supplemented with 1000 mg CE/kg (CE). The study was divided into phase 1 (d 1 to 14) and 2 (d 15 to 35). The average daily gain was increased (p < 0.05) in pigs fed CE in phase 2 and overall (d 1 to 35) compared with CON. These pigs had greater (p ≤ 0.05) serum IgA, IgG, superoxide dismutase and catalase contents, as well as tended to increase serum IgM content and apparent total tract digestibility (ATTD) of organic matter in phase 1 compared with CON. In phase 2, pigs supplemented with CE showed greater (p < 0.01) ATTD of dry matter, organic matter, crude protein and gross energy compared with CON. These pigs also had increased (p < 0.05) IgA, IgG, IgM, superoxide dismutase contents, and decreased (p < 0.05) malondialdehyde content in serum compared with CON. Moreover, pigs fed CE had higher (p < 0.05) villus height and villus height to crypt depth ratio in ileum, and tended to increased acetic acid content in colon compared with CON. Furthermore, pigs fed CE had increased (p < 0.05) relative abundance of Firmicutes at phylum level, Lactobacillales at order level, Lactobacillaceae at family level, Bacilli at class level, Lactobacillus at genus level in caecum and colon, as well as lower (p < 0.05) relative abundance of Bacteroidetes at phylum level, Bacteroidales at the order level, Bacteroidia at class level, Clostridium_sensu_stricto_6 at genus level in colon compared with CON. In conclusion, dietary inclusion of compound enzymes could effectively improve nutrient digestibility, serum antioxidant status, immunoglobulin, gut morphology, microbiota community, and therefore improve performance in weaned pigs.

MiR-196-5p involvement in selenium deficiency-induced immune damage via targeting of NFκBIA in the chicken trachea.

Dietary selenium (Se) deficiency can induce multifarious immune injury in tissues, accompanied by inflammation and a decreased expression of selenoproteins. The results of previous studies indicated that these issues are associated with Se-mediated microRNAs involved in immune regulation, although the specific mechanisms associated with these interactions have not been reported in the trachea of chickens. To explore the effects of Se deficiency in the trachea of chickens and the role of miR-196-5p, we established correlational models of tracheal injury in chickens. One hundred broilers were divided into four groups, including a control group (C group), a Se deficient group (L group), a lipopolysaccharide (LPS)-induced control group (C + LPS group) and a LPS-induced Se deficient group (L + LPS group). Light microscopy observations indicated that the infiltration of inflammatory cells was the major histopathological change caused by Se deficiency. Furthermore, ultrastructural observation of the tracheal epithelium and ciliary showed typical inflammatory signs owing to Se deficiency. We determined the targeting relationship between miR-196-5p and NFκBIA by bioinformatics analysis. In the case of Se deficiency, the changes were detected as follows: 19 selenoproteins showed different degrees of decrease (p < 0.05). Significant inhibition of both antimicrobial peptides and immunoglobulin production were observed (p < 0.05). IκB-α (NFκBIA) expression degraded with the increasing miR-196-5p (p < 0.05), and the NF-κB pathway was activated. Thereafter, we can see a significant increase in the mRNA levels of inflammatory cytokines-related genes (tumor necrosis factor (TNF)-α, inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, prostaglandin E (PTGE), interleukin (IL)-1ß, IL-6) and protein expression of NF-κB/iNOS pathway-related genes (NF-κB, iNOS, TNF-α, COX-2) (p < 0.05). The release of IL-2, interferon (IFN)-γ inhibited (p < 0.05) and the secretion of IL-4, IL-6 increased, suggesting the imbalance of Th1/Th2 (Th, helper T cell) cytokines. Compared to the control, the mRNA and protein expression levels of the anti-inflammatory system components with antioxidant activity (PPAR-γ/HO-1) were in an inhibitory state (p < 0.05). Antioxidases (SOD, CAT, GSH-Px) activities were suppressed. The activities of the peroxide markers (MDA, H2O2) were enhanced (p < 0.05). In addition, Se deficiency had a positive effect on the pathological changes of inflammation and the exceptional immunity in LPS-treated groups (p < 0.05). The results confirmed the relationship between miR-196-5p and NFκBIA in chickens, revealing that Se deficiency causes respiratory mucosal immune dysfunction via the miR-196-5p-NFκBIA axis, oxidative stress and inflammation. Moreover, Se deficiency exacerbates the inflammatory damage stimulated by LPS. Our work provides a theoretical basis for the prevention of tracheal injury owing to Se deficiency and can be used as a reference for comparative medicine. Furthermore, the targeted regulation of miR-196-5p and NFκBIA may contribute to the protection of the tracheal mucosa in chickens.

Effect of Trichoderma reesei Degraded Date Pits Supplementation on Growth Performance, Immunoglobulin Levels, and Intestinal Barrier Functions of Broiler Chickens.

BACKGROUND: Date palm (Phoenix dactylifera L.) is a dominant fruit crop in most Arabian countries. Date pits, as a major byproduct which remained after consumption of date flesh, proved to be a valuable source of energy. OBJECTIVES: The impact of degraded date pits (DDP) on growth performance, intestinal bacterial population, and expression profiles of intestinal genes in broilers was determined. Recent patents have been established on DDP from the European patent office (EP2586318B1), Hong Kong patent registry office (HK1184642) and by the United States patent and trademark office (US8968729B2 and US10265368B2). METHODS: Solid-state degradation system (SSD) was used for the preparation of DDP using Trichoderma reesei. One-day-old Brazilian broiler chicks "Cobb 500" were randomly divided into six treatments with six replicates, which consisted of a normal diet containing only corn-soy (control), diet containing corn-soy + (20%, 50g/100Kg oxytetracycline), diet containing corn-soy + 10% (DDP), diet containing corn-soy + 0.2% mannan oligosaccharides (MOS), diet containing corn-soy + 0.1% mannose, and diet containing corn-soy + 0.2% mannose. RESULTS: There were no significant differences in body weight, feed intake, and feed conversion ratio (FCR) in broilers among the treatments. The bacterial count was significantly decreased in 10% DDP diet-fed broilers, 0.2% MOS and antibiotic diet-fed broilers. Immunoglobulin levels in serum and intestinal contents and expression pattern of genes in jejunum were upregulated in 10% DDP and 0.2% MOS diet-fed broilers. CONCLUSION: DDP can be used as an energy source for replacing part of corn, mannan oligosaccharide and also recommended as a potential alternative to antimicrobials in broilers diet.

Effects of astragalus and ginseng polysaccharides on growth performance, immune function and intestinal barrier in weaned piglets challenged with lipopolysaccharide.

This experiment was conducted to evaluate the effects of astragalus polysaccharides (Aps) and ginseng polysaccharide (Gps) on growth performance, liver function, immune function, TLR4 signalling pathways and intestinal barrier in weaned piglets challenged with lipopolysaccharide (LPS). In an experiment spanning 28 days, 180 weaned piglets were randomly divided into three treatment groups: basal diet (Con), basal diet supplemented with 800 mg/kg Gps (Gps) and basal diet supplemented with 800 mg/kg Aps (Aps). At the end of the experiment, 12 piglets of each group were selected; half (n = 6) were intraperitoneally injected with LPS and half with normal saline. Dietary supplementation with Aps and Gps significantly increased (p < .05) the average daily gain and feed conversion rate. Lipopolysaccharide challenge increased (p < .05) expression of serum urea nitrogen (BUN), alanine aminotransferase (ALT), aspartate aminotransferase (AST), interleukin-1ß (IL-1ß) and tumour inflammatory factor-α (TNF-α), but decreased (p < .05) serum superoxide dismutase (SOD) level, total antioxidant capacity (T-AOC) and immunoglobulin A (IgA) expression. Lipopolysaccharide-challenged piglets fed with Aps or Gps had lower (p < .05) BUN, ALT, AST, IL-1ß and TNF-α levels and greater (p < .05) SOD, T-AOC and IgA levels. Lipopolysaccharide challenge increased (p < .05) the expression of TLR4, MyD88 and NF-κB, and LPS-challenged piglets fed diets supplemented with Aps or Gps increased TLR4 and MyD88 and decreased NF-κB expression. Lipopolysaccharide challenge reduced (p < .05) the jejunal villus height, and piglets fed with Aps or Gps had increased (p < .05) jejunal villus height. Supplementation with Aps or Gps enhanced the expression of occludin and claudin in challenged or unchallenged piglets. In conclusion, dietary supplementation with Aps or Gps enhanced piglet growth performance, alleviated liver dysfunction and reduced immunological stress caused by LPS, as well as increased the intestinal barrier function.

Short ragweed pollen promotes M2 macrophage polarization via TSLP/TSLPR/OX40L signaling in allergic inflammation.

This study was to explore the role and mechanism of macrophages in pollen-triggered allergic inflammation. A murine model of short ragweed (SRW) pollen-induced experimental allergic conjunctivitis (EAC), and bone marrow (BM)-macrophages cultures were used. Typical allergic manifestations and TSLP-stimulated Th2 hyperresponse were observed in ocular surface of EAC model in wild-type (WT) mice induced by SRW. The M2 phenotype markers, Arg1, Ym1 and FIZZ1, were highly expressed by conjunctiva and draining cervical lymph nodes (CLNs) of WT-EAC mice when compared with controls, as evaluated by RT-qPCR and Immunofluorescent double staining with macrophage marker F4/80. The stimulated expression of TSLPR and OX40L by macrophage was detected in conjunctiva and CLNs by RT-qPCR, double staining, and flow cytometry. M2 macrophages were found to produce TARC and MDC. In contrast, EAC model with TSLPR-/- mice did not show allergic signs and any increase of Th2 cytokines (IL-4, IL-5 and IL-13) and M2 markers. In vitro cultures confirmed that SRW extract stimulates expression of TSLPR, OX40L, TARC, MDC, and three M2 markers by BM-macrophages from WT mice, but not from TSLPR-/- mice. These findings demonstrate that SRW pollen primes macrophage polarization toward to M2 phenotype via TSLP/TSLPR/OX40L signaling to amplify allergic inflammation.

Lubiprostone as a potential therapeutic agent to improve intestinal permeability and prevent the development of atherosclerosis in apolipoprotein E-deficient mice.

The interaction between atherosclerosis and commensal microbes through leaky gut syndrome (LGS), which is characterized by impaired intestinal permeability and the introduction of undesired pathogens into the body, has not been fully elucidated. Our aim was to investigate the potential role of a ClC-2 chloride channel activator, lubiprostone, which is reported to have beneficial effects on LGS, in the development of atherosclerosis in apolipoprotein E-deficient (ApoE-/-) mice. After a 15-week feeding period of a Western diet (WD), ApoE-/- mice were treated with a Western-type diet (WD) alone or WD with oral supplementation of lubiprostone for 10 weeks. This feeding protocol was followed by experimental evaluation of LGS and atherosclerotic lesions in the aorta. In mice with lubiprostone, in vivo translocation of orally administered 4-kDa FITC-dextran was significantly improved, and RNA expression of the epithelial tight junction proteins, Zo-1 and occludin, was significantly up-regulated in the ileum, compared to the WD alone group, suggesting a possible reversal of WD-induced intestinal barrier dysfunction. As a result, WD-induced exacerbation of atherosclerotic lesion formation was reduced by 69% in longitudinally opened aortas and 26% in aortic root regions. In addition, there was a significant decrease in circulating immunoglobulin level, followed by an attenuation of inflammatory responses in the perivascular adipose tissue, as evidenced by reduced expression of pro-inflammatory cytokines and chemokines. Lubiprostone attenuates atherosclerosis by ameliorating LGS-induced inflammation through the restoration of the intestinal barrier. These findings raise the possibility of targeting LGS for the treatment of atherosclerosis.

Dietary l-arginine supplementation influences growth performance and B-cell secretion of immunoglobulin in broiler chickens.

Our previous study has shown that high levels of l-arginine (ARG) have reduced serum and mucosal antibody concentrations. In order to provide a better understanding in the application of ARG supplementation in the poultry industry, the study was conducted to investigate the effect of high levels of ARG on performance and B-cell secretion of immunoglobulin M (IgM) and IgG development in broiler chickens. A total of 192 1-day-old male Arbor Acres Plus broilers were randomly allocated into 4 groups (8 replicates per group, 6 birds per replicate) fed diets containing one of four ARG concentrations (analysed): 9.8, 14.7, 19.1 and 23.4 g/kg respectively. Growth performance was measured based on body weight gain (BWG), feed intake (FI) and feed conversion ratio (FCR). Increasing ARG quadratically increased (p < 0.05) BWG and FI with reaching plateau at 14.7 g/kg, while linearly decreased (p < 0.05) FCR, indicating that maximal performance required ARG no more than 14.7 g/kg in diets. Serum IgG and IgM concentrations were linearly reduced (p < 0.05) with increasing ARG. Chickens fed 19.1 g/kg or 23.4 g/kg ARG had lower (p < 0.05) serum IgG or IgM than chickens fed 9.8 g/kg ARG. As for the mRNA expression of bursal IgG and IgM, they were significantly downregulated with increasing ARG (p < 0.05). Chickens on ARG (>19.1 g/kg) had a lower (p < 0.05) IgG and IgM mRNA expression than chickens fed 9.8 g/kg. Activator of transcription 3 (STAT3) mRNA expression was linearly reduced with increasing ARG (p < 0.05), the transcriptional repressor B-cell lymphoma 6 (BCL6) mRNA expression was quadratically (p < 0.05) responded, and these cytokines had the lowest expression at 19.1 g/kg. ARG supplementation (>14.7 g/kg) did not significantly improve the growth performance, while it may have a potential negative regulatory effect on B-cell-mediated humoral immunity in chickens associated with suppression of the STAT3 expression associated with the JAK/STAT3 pathway.

Glutamine protects against LPS-induced inflammation via adjusted NODs signaling and enhanced immunoglobulins secretion in rainbow trout leukocytes.

To evaluate effects of glutamine (GLN) on fish immune responses, leukocytes were isolated from head kidney of rainbow trout and cultured in GLN-free DMEM media supplemented with different combinations of lipopolysaccharide (LPS) and GLN. LPS significantly increased expression of pro-inflammatory cytokines, while GLN supplementation alleviated LPS-induced inflammation. Leukocytes in +GLN + LPS group showed more active GLN anabolism and catabolism, which signals could be sensed by O-GlcNAcylation, and then affected LPS binding to cell surface (LBP) and adjusted NODs signaling. The mRNA expression of immunoglobulins (Igs) and their receptor (pIgR) was also significantly increased after GLN supplementation. Further analysis showed that GLN increased the percentage of IgM+ B cells and IgT+ B cells, accompanied with the increased IgM and IgT secretion in culture media, which further increased complement C3 expression to perform effector functions. All these results illustrated the regulating mechanism of GLN against LPS-induced inflammation both via adjusted NODs signaling and increased Igs+ B cells to secrete Igs.

Enhancement of immune maturation in suckling rats by leptin and adiponectin supplementation.

Leptin and adiponectin, adipokines present in breast milk, have shown immunomodulatory properties. The current study aimed to ascertain whether a nutritional supplementation with leptin or adiponectin in neonatal rats was able to influence the maturation of the systemic immune response in early life. To achieve this, suckling Wistar rats were supplemented with either leptin (0.7 µg/kg/day) or adiponectin (35 µg/kg/day) during the whole suckling period. Plasmatic immunoglobulins were quantified, and spleen lymphocyte composition and their ability to proliferate and release cytokines were evaluated during (day 14) and at the end (day 21) of the suckling period. Rats fed with either adipokine showed higher plasma IgM and IgG1 concentrations and adiponectin supplementation also increased IgG2a at both studied days (P < 0.05). With regard to the lymphocyte composition, both adipokine supplementations increased T cell proportion and both CD4+ and CD8+ T cell subsets after two weeks of supplementation (P < 0.05). Moreover, only leptin administration increased NK and NKT cell proportions at the end of the suckling period. Finally, both adipokines influenced the cytokine secretion pattern by splenocytes. In conclusion, these results suggest that leptin and adiponectin play a role in the maturation of the systemic immune response during the suckling period.