Effects of organic trace minerals chelated with oligosaccharides on growth performance, blood parameters, slaughter performance and meat quality in sheep.

The present study assessed the effects of oligosaccharide-chelated organic trace minerals (OTM) on the growth performance, digestive enzyme activity, blood parameters, slaughter performance, and meat quality indexes of mutton sheep. A total of 60 East Ujumuqin × small-tailed Han crossbred mutton sheep were assigned to two groups (10 duplicates per group) by body weight (26.12 ± 3.22 kg) according to a completely randomized design. Compared to the CON group, the results of the OTM group showed: (1) no significant changes in the initial body weight, final body weight, dry matter intake, average daily gain, and feed conversion ratio (p > 0.05); (2) the activities of trypsin, lipase, and amylase in the jejunum were significantly increased (p < 0.05); (3) serum total protein, albumin, and globulin of the blood were significantly increased (p < 0.05), and the growth factor interleukin IL-10 was significantly higher (p < 0.05), while IL-2, IL-6, and γ-interferon were significantly lower (p < 0.05). Immunoglobulins A, M, and G were significantly higher (p < 0.05); (4) the live weight before slaughter, carcass weights, dressing percentage, eye muscle areas, and GR values did not differ significantly (p > 0.05); (5) shear force of mutton was significantly lower (p < 0.05), while the pH45min, pH24h, drip loss, and cooking loss did not show a significant difference (p > 0.05). The content of crude protein was significantly higher (p < 0.05), while the ether extract content was significantly reduced (p < 0.05), but no significant difference was detected between moisture and ash content; (6) the total amino acids, essential amino acids, semi-essential amino acids, and umami amino acids were significantly increased (p < 0.05). Although umami amino acids were not significant, the total volume increased (p > 0.05). Among these, the essential amino acids, threonine, valine, leucine, lysine in essential amino acids and arginine were significantly increased (p < 0.05). Also, non-essential amino acids, glycine, serine, proline, tyrosine, cysteine, and aspartic acid, were significantly higher (p < 0.05). The content of alanine, aspartate, glutamic acid, phenylalanine, and tyrosine in umami amino acids was significantly higher (p < 0.05).

Trans-anethole exerts protective effects on lipopolysaccharide-induced acute jejunal inflammation of broilers via repressing NF-κB signaling pathway.

This study aimed to explore the effects of trans-anethole (TA) on lipopolysaccharide (LPS)-induced acute jejunal inflammation model of broilers. A total of 160 one-day-old broilers (male; Arbor Acres) were randomly allocated into four treatment groups with 8 replicates of 5 birds each. On d 20, the dose of 5 mg/kg body weight LPS solution and the equal amount of sterile saline were intraperitoneally injected into LPS-challenged and unchallenged broilers, respectively. Compared with the control group, LPS decreased (P < 0.05) the villus height (VH) and the ratio of villus height to crypt depth (VCR) but increased (P < 0.05) the crypt depth (CD), meanwhile, enhanced (P < 0.01) the levels of interleukin-6 (IL-6), interleukin-1beta (IL-1ß) and tumor necrosis factor-alpha (TNF-α) but decreased (P < 0.01) the level of interleukin-10 (IL-10). The group supplemented with 600 mg/kg of TA had lower (P < 0.01) CD and higher (P < 0.01) VCR than the LPS group. TA increased (P < 0.01) the level of IL-10 and decreased (P < 0.01) the level of IL-1ß. The mRNA expression levels of IL-6, nuclear factor kappa B (NF-κB), TNF-α were up-regulated (P < 0.05) and the levels of IL-10 and inhibitor of NF-κB alpha (IκBα) were down-regulated (P < 0.05) by LPS as compared with the control group. TA down-regulated (P < 0.05) the increased mRNA expression levels of genes caused by LPS, as well as up-regulated (P < 0.05) the levels of IL-10 and IκBα. Furthermore, LPS down-regulated (P < 0.05) and up-regulated (P < 0.05) the protein expression levels of IκBα and NF-κB p65, respectively. TA up-regulated (P < 0.05) the level of IκBα and down-regulated (P < 0.05) the level of NF-κB p65. The conclusion of this study is that TA could exert protective effect on the LPS-induced acute jejunal inflammation of broilers via repressing the activation of NF-κB and the 600 mg/kg is the optimal dose against LPS-induced acute jejunal inflammation of broilers.

Pharmacological Effects of Polyphenol Phytochemicals on the Intestinal Inflammation via Targeting TLR4/NF-κB Signaling Pathway.

TLR4/NF-κB is a key inflammatory signaling transduction pathway, closely involved in cell differentiation, proliferation, apoptosis, and pro-inflammatory response. Toll like receptor 4 (TLR4), the first mammalian TLR to be characterized, is the innate immune receptor that plays a key role in inflammatory signal transductions. Nuclear factor kappa B (NF-κB), the TLR4 downstream, is the key to accounting for the expression of multiple genes involved in inflammatory responses, such as pro-inflammatory cytokines. Inflammatory bowel disease (IBD) in humans is a chronic inflammatory disease with high incidence and prevalence worldwide. Targeting the TLR4/NF-κB signaling pathway might be an effective strategy to alleviate intestinal inflammation. Polyphenol phytochemicals have shown noticeable alleviative effects by acting on the TLR4/NF-κB signaling pathway in intestinal inflammation. This review summarizes the pharmacological effects of more than 20 kinds of polyphenols on intestinal inflammation via targeting the TLR4/NF-κB signaling pathway. We expected that polyphenol phytochemicals targeting the TLR4/NF-κB signaling pathway might be an effective approach to treat IBD in future clinical research applications.

Trans-anethole ameliorates lipopolysaccharide-induced acute liver inflammation in broilers via inhibiting NF-κB signaling pathway.

The aim of this study was to investigate the protective effect of trans-anethole (TA) on lipopolysaccharide-induced acute liver inflammation model of chickens by determining the levels of inflammatory mediators in serum and liver, relative mRNA expression and protein expression of inflammation-related genes in NF-κB signaling pathway. A total of 160 one-day-old male chickens (Arbor Acres) were assigned into 4 treatments with 8 replicates of 5 birds each. On d 20, the control group was intraperitoneally injected with sterile saline and the other groups were injected with lipopolysaccharide (LPS; 5 mg/kg body weight). There were no significant differences in average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) among groups. However, compared with the control group, the LPS group significantly increased (P < 0.01) the serum levels of interleukin-6 (IL-6), interleukin-1beta (IL-1ß), tumor necrosis factor-alpha (TNF-α), alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and decreased (P < 0.01) the interleukin-10 (IL-10) level. TA attenuated (P < 0.01) these increases in IL-1ß, TNF-α, ALT, and AST levels and improved (P < 0.01) the IL-10 level. In liver, the groups fed with TA had lower (P < 0.01) concentrations of IL-6 and TNF-α as well as higher (P < 0.05) concentration of IL-10. Furthermore, TA downregulated (P < 0.05) the mRNA expression levels of nuclear factor kappa B p65 (NF-κB p65) and TNF-α, also upregulated (P < 0.05) IL-10 and inhibitor of NF-κB alpha (IκBα) upon LPS challenge. In protein level, supplementation of 600 mg/kg of TA downregulated (P < 0.05) and upregulated (P < 0.05) the protein expression of NF-κB p65 and IκBα, respectively. The present findings suggest that TA could alleviate the acute liver inflammation induced by LPS via blocking the activation of NF-κB and the 600 mg/kg of TA plays more fruitful role in protecting broilers against LPS stimulus.

Trans-anethole ameliorates LPS-induced inflammation via suppression of TLR4/NF-κB pathway in IEC-6 cells.

This study was undertaken to investigate the protective role of trans-anethole (TA) in lipopolysaccharide (LPS)-induced rat intestinal epithelial cells (IEC-6) injury and the potential mechanisms. The cells were pretreated with TA (0 and 1 mM) for 24 h, prior to stimulation by LPS (1 mg/mL) for 24 h. Compared with the control group (CON), LPS stimulus resulted in decreased cell viability, intestinal barrier injury, increased cell apoptosis and cell cycle arrest at the G2/M phase. These effects triggered by LPS were reversed by TA. In order to reveal the main genes and pathways involved among the groups, transcriptome analysis was performed to identify the differential expression genes (DEGs) among the treatment groups. There were a total of 493 DEGs (275 upregulated and 218 downregulated) that were identified between the LPS and CON group. Meanwhile, a total of 361 DEGs (103 regulated and 258 downregulated) were identified in the LPS+TA group compared with the LPS group. The results showed that the DEGs were mostly enriched in immune related pathways, such as tumor necrosis factor (TNF) signaling pathway, cytokine-cytokine receptor interaction, complement and coagulation cascades, interleukin-17 (IL-17) signaling pathway, NF-kappa B (NF-κB) signaling pathway, antigen processing and presentation, and NOD-like receptor signaling pathway. Based on the results of RNA-sequencing, further investigation of the signaling pathway involved revealed that TA could inhibit the activation of toll like receptor 4 (TLR4)/NF-κB signaling pathway and NLR family pyrin domain containing 3 (NLRP3) inflammasome in LPS-induced IEC-6 cells. In conclusion, this finding demonstrated a functional role of TA in intestinal epithelial cells injury and indicated that TA may be a potential strategy for treatment of inflammatory intestinal diseases.

Trans-Anethole Alleviates Subclinical Necro-Haemorrhagic Enteritis-Induced Intestinal Barrier Dysfunction and Intestinal Inflammation in Broilers.

This study investigated the alleviative potential of trans-anethole (TA) on the impaired intestinal barrier and intestinal inflammation and its regulatory effects on gut microbiota in broilers with subclinical necro-hemorrhagic enteritis (NE) challenge. Subclinical NE challenge led to a severe decline in the 21-day body weight (BW) and average daily gain (ADG), but an increase in feed conversion ratio (FCR) and intestinal lesion score of birds compared with controls (P < 0.05). Compared with the subclinical NE group, the TA administration group exhibited lower (P < 0.05) intestinal lesion score and crypt depth (CD), serum diamine oxidase activity, and D-lactate concentration, but higher (P < 0.05) intestinal tight junction protein expressions, villus height (VH), VH/CD, and numbers of proliferating cell nuclear antigen (PCNA)-positive cells. The administration of TA also inhibited (P < 0.05) the expression of intestinal pro-inflammatory cytokines including interleukin (IL)-1ß, IL-8, interferon-gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) but increased (P < 0.05) jejunal IL-10 and secretory immunoglobulin A (sIgA) concentration. TA inclusion also led to a remarkable reduction of intestinal NF-kappa-B inhibitor alpha (IκBα) degradation and nuclear factor kappa beta (NF-κB) translocation. Moreover, TA modulated the cecal microbiota abundance and diversity of NE birds, as confirmed by reducing the phylum Firmicutes and genera Ruminococcaceae_UCG-014, Eubacterium_coprostanoligenes_group, and Ruminococcaceae_NK4A214_group when supplemented at 600 mg/kg and reducing genera Butyricicoccus, Oscillibacter, and Flavonifractor when supplemented at 400 mg/kg (P < 0.05). Supplementation of TA in broiler diets could alleviate subclinical NE infection by restoring intestinal barrier integrity, inhibiting NF-κB signaling pathway, and modulating gut microbiota. A 600-mg/kg dose of TA is the optimum concentration for ameliorating subclinical NE in broilers.

Effects of yeast hydrolysate on growth performance, serum parameters, carcass traits, meat quality and antioxidant status of broiler chickens.

BACKGROUND: Yeast hydrolysate (YH) has multiple salutary biological activities. Nevertheless, the application of YH in broiler production is limited. This study was conducted to evaluate the protective effects of YH derived from Saccharomyces cerevisiae by exploring growth performance, serum parameters, organs relative weight, carcass traits, meat quality and antioxidant status of broilers. RESULTS: Supplementing YH linearly and quadratically improved (P < 0.05) body weight gain and gain-to-feed ratio compared to that in the control group. Triglycerides, low-density lipoprotein cholesterol and total cholesterol in serum, the decline in pH and cooking loss of breast muscle, and malonaldehyde concentration in serum and liver were decreased linearly and/or quadratically by YH (P < 0.05), whereas high-density lipoprotein cholesterol in serum, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in serum, GSH-Px activity in liver, glutathione content in serum and liver, eviscerated yield rate and chest muscle yield, and the relative weight of spleen and liver were linearly and/or quadratically increased (P < 0.05). Moreover, YH enhanced the mRNA levels of nuclear factor erythroid 2-related factor 2, heme oxygennase-1 (HO-1), GSH-Px1 and SOD1 (linear and/or quadratic, P < 0.05). CONCLUSION: Dietary YH beneficially affected growth performance, serum parameters, organ relative weight, carcass traits, meat quality and antioxidant status in broilers, indicating its potential application as a promising feed additive in broiler production. © 2021 Society of Chemical Industry.

The Effects of Zearalenone on the Localization and Expression of Reproductive Hormones in the Ovaries of Weaned Gilts.

This study aims to investigate the effects of zearalenone (ZEA) on the localizations and expressions of follicle stimulating hormone receptor (FSHR), luteinizing hormone receptor (LHR), gonadotropin releasing hormone (GnRH) and gonadotropin releasing hormone receptor (GnRHR) in the ovaries of weaned gilts. Twenty 42-day-old weaned gilts were randomly allocated into two groups, and treated with a control diet and a ZEA-contaminated diet (ZEA 1.04 mg/kg), respectively. After 7-day adjustment, gilts were fed individually for 35 days and euthanized for blood and ovarian samples collection before morning feeding on the 36th day. Serum hormones of E2, PRG, FSH, LH and GnRH were determined using radioimmunoassay kits. The ovaries were collected for relative mRNA and protein expression, and immunohistochemical analysis of FSHR, LHR, GnRH and GnRHR. The results revealed that ZEA exposure significantly increased the final vulva area (p < 0.05), significantly elevated the serum concentrations of estradiol, follicle stimulating hormone and GnRH (p < 0.05), and markedly up-regulated the mRNA and protein expressions of FSHR, LHR, GnRH and GnRHR (p < 0.05). Besides, the results of immunohistochemistry showed that the immunoreactive substances of ovarian FSHR, LHR, GnRH and GnRHR in the gilts fed the ZEA-contaminated diet were stronger than the gilts fed the control diet. Our findings indicated that dietary ZEA (1.04 mg/kg) could cause follicular proliferation by interfering with the localization and expression of FSHR, LHR, GnRH and GnRHR, and then affect the follicular development of weaned gilts.

Illicium verum and Eucommia ulmoides leaf extracts promote nutrient availability and antioxidant capacity in piglets by upregulating duodenal and jejunal Nrf2/TNF-α.

The effect of Illicium verum extracts (IVE) or Eucommia ulmoides leaf extracts (ELE) on nutrient availability, duodenal and jejunal antioxidant ability of Duroc ×Landrace × Yorkshire (DLY) and Chinese native Licha-black (LCB) piglets was investigated. Ninety-six piglets (48 DLY and 48 LCB respectively) without significant difference in body weight (11.22 ± 0.32 kg) were used in a 2 × 4 factorial design. Animals were randomly allocated to four treatments, and each had four replicates with three DLY and three LCB piglets. Treatments were basal diet (CON) and basal diet with 500 mg/kg IVE, 250 mg/kg ELE and 50 mg/kg chlortetracycline (CHL) respectively. Animals were placed individually for 7-days adaptation following 42-days test. Results showed the significant interaction (p < 0.05) between dietary treatments and pig species in activity of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px), content of malondialdehyde (MDA), and α-tumour necrosis factor (TNF-α), nuclear factor erythroid 2-related factor 2 (Nrf2) and Nrf2/TNF-α at mRNA and protein level in duodenum and jejunum of DLY and LCB piglets. The IVE and ELE increased (p < 0.05) activity of GSH-Px and T-SOD, and the Nrf2/TNF-α at mRNA and protein level, however, the decreased (p < 0.05) MDA content, and TNF-α at mRNA and protein level in duodenum and jejunum were observed. The CHL decreased (p < 0.05) activity of GSH-Px and T-SOD, TNF-α and Nrf2 at mRNA and protein level in duodenum, but increased (p < 0.05) MDA content and Nrf2/TNF-α in jejunum. DLY piglets had higher (p < 0.05) nutrient digestibility (organic matter, crude protein and gross energy), availability (biological value and net protein utilization), MDA content, and TNF-α at mRNA and protein level in jejunum, and had lower (p < 0.05) activity of GSH-Px and T-SOD, and Nrf2 and Nrf2/TNF-α at mRNA and protein level in duodenum and jejunum than LCB. In conclusion, the 500 mg/kg IVE and 250 mg/kg ELE improved the nutrient availability, and the improvement of antioxidant capacity is realized by activating the Nrf2/TNF-α of duodenum and jejunum. The CHL had adverse effects on antioxidant ability of DLY and LCB piglets. The results showed that the nutrient digestion and absorption capacity of DLY were stronger than that of LCB piglets, but the antioxidant capacity was lower than that of LCB piglets. Therefore, the IVE and ELE are recommended as a new potential alternative to antibiotics in piglets' diets.

Effects of star anise (Illicium verum Hook.f.) essential oil administration under three different dietary energy levels on growth performance, nutrient, and energy utilization in broilers.

This experiment was conducted to investigate the effect of star anise essential oil (SAO) supplementation in diets with different energy levels on growth performance, nutrient metabolic efficiency of broilers. One hundred and ninety-two Arbor Acres male broiler chicks at 28 days of age were divided into a 3 × 4 factorial arrangement design with three dietary energy levels (13.41, 12.82, 12.23 MJ/kg) and 4 levels of SAO supplementation (0, 200, 400, and 600 mg/kg of diet). Dietary supplementation with SAO increased (p < .05) apparent metabolic efficiency of CP, EE, GE, and all of the amino acids except Trp. Significant interactions were noted between energy level and SAO administration for metabolic efficiency of CP, all of the amino acids except Cys. Inclusion of SAO enhanced apparent nutrient metabolic efficiency of broilers in a dose-dependent manner, birds supplemented with 400 mg/kg of SAO in high-energy diets appeared to contain highest nutrient metabolic efficiency, moreover, the metabolic efficiency of nutrients in low-energy diets along with 200 or 400 mg/kg of SAO was similar with that in high-energy diets without SAO, which indicated that the SAO might ameliorate the negative effects of reduced dietary metabolic energy on nutrient utilization in broilers.

Effects of star anise (Illicium verum Hook.f) oil on the nuclear factor E2-related factor 2 signaling pathway of chickens during subclinical Escherichia coli challenge.

We characterized the mechanism underlying star anise (Illicium verum Hook.f) oil (SAO)-mediated antioxidant status during subclinical Escherichia coli (E. coli) challenge. A total of 512 male birds (White Leghorn) at 30 wk of age with similar body weight (2.14 ± 0.02 kg) were randomly divided into 2 groups with 1 group being orally challenged with E. coli (every other day from day 15 to day 27) during the experiment. Each group of birds was then randomly allocated to dietary treatment of SAO supplementation at 0, 200, 400, or 600 mg/kg of basal diet (8 replicate cages during each treatment). The treatments were arranged a 4 × 2 factorial arrangement. The experiment comprised 1 wk of adaptation and 3 wks of data collection. There was no interaction (P > 0.05) between SAO supplementation and E. coli challenge for final body weight and average daily feed intake of birds. However, E. coli challenge resulted in a significant decrease (P < 0.001) in final body weight of birds as compared with unchallenged birds. There were interactions between SAO supplementation and E. coli challenge for the activity of glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) concentration in serum and for the activity of GSH-Px in the liver of birds. Supplementation of SAO enhanced the activities of antioxidant enzymes but decreased the MDA content in the serum and liver of birds, and it also enhanced the expression of genes including superoxide dismutase, catalase, and nuclear factor E2-related factor 2 (Nrf2) in the liver of the birds. Meanwhile, supplementation of SAO can also reduce E. coli challenge-induced oxidative stress in the serum and liver of birds, and the efficacy of SAO in birds during subclinical E. coli challenge is dose-dependent. In conclusion, the enhancement of antioxidant capacity by star anise or its effective compounds is through upregulation of Nrf2 signaling pathway. The optimum supplementation dose of SAO for protecting birds against E. coli challenge is 400 mg/kg.

Illicium verum extracts and probiotics with added glucose oxidase promote antioxidant capacity through upregulating hepatic and jejunal Nrf2/Keap1 of weaned piglets.

Accumulating evidences indicate that plant extracts and probiotics are effective antioxidant substitutes which play important roles in animal production. However, the comparative study of the mechanism underlying the antioxidant property of Illicium verum extracts (IVE) and probiotics with added glucose oxidase (PGO) on piglets remains to be explored. This study evaluated the difference and the interaction effect of IVE and PGO on serum, liver, and jejunum antioxidant capacity of weaned piglets. A total of 32 weaned piglets (Duroc × Landrace × Yorkshire) at the age of 28 d with an average body weight of 14.96 ± 0.32 kg were randomly divided into four treatments with eight replicates per treatment in a 2 × 2 factorial arrangement. Treatments included basal diet (IVE-PGO-), basal diet + 1,000 mg/kg PGO (IVE-PGO+), basal diet + 500 mg/kg IVE (IVE+PGO-), and basal diet + 500 mg/kg IVE + 1,000 mg/kg PGO (IVE+PGO+). All the piglets were housed individually for the 42-d trial period after 7-d adaptation. The piglets were euthanized at the end of the experiment and the liver and jejunum samples were taken and subjected to immunohistochemistry, Western blotting, as well as antioxidant and qRT-PCR analysis. Significant interactions were observed between IVE and PGO for total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) in serum (42 d), liver, and jejunum; malondialdehyde (MDA) in serum (21 d); and mRNA and protein expression of kelch sample related protein-1 (Keap1) and nuclear factor erythroid-2 related factor (Nrf2)/Keap1 in the liver and jejunum (P < 0.05). Both IVE and PGO improved (P < 0.05) T-SOD and GSH-Px in the serum (42 d), liver, and jejunum, and the mRNA and protein expression of Nrf2 and Nrf2/Keap1 in the liver and jejunum, but decreased (P < 0.05) MDA in the serum (21 d) and the mRNA and protein expression of Keap1 in the liver and jejunum. Immunohistochemical results confirmed that IVE and PGO enhanced the positive reactions of Nrf2 but weakened Keap1 in both the liver and jejunum. In conclusion, the results confirmed that IVE (500 mg/kg) and PGO (1,000 mg/kg) can improve the antioxidant capacity of weaned piglets and that the interaction effect between IVE and PGO is significant. At the same time, the fact that IVE and PGO activate the Nrf2/Keap1 in the liver and jejunum signaling pathway suggests that they play an important role in the ameliorative antioxidant capacity of weaned piglets. Therefore, the combination of IVE and PGO could be recommended as a new potential alternative to antibiotics in piglets' diets.

Effect of Illicium verum or Eucommia ulmoides leaf extracts on the anti-stress ability, and mRNA and protein expression of Nrf2 and TNF-α in Duroc × Landrace × Yorkshire and Chinese native Licha-black nursery piglets.

Plant extracts are considered to be an effective alternative to antibiotics in response to weaning stress in piglets. This study evaluated the effect of Illicium verum extracts (IVE) or Eucommia ulmoides leaf extracts (ELE) on growth performance, serum and liver antioxidant ability of nursery piglets, as well as the difference of IVE and ELE on Duroc × Landrace × Yorkshire (DLY) and Chinese native Licha-black (LCB) piglets. A total of 96 nursery piglets (48 DLY and 48 LCB piglets) with an average body weight of 11.22 ± 0.32 kg were randomly divided into four treatments in a 2 × 4 factorial design. Each treatment had four replicates with 3 DLY and 3 LCB piglets per replicate respectively. Treatments included: basal diet, basal diet + 500 mg/kg IVE, basal diet + 250 mg/kg ELE and basal diet + 50 mg/kg chlortetracycline (CHL). All piglets were housed individually for the 42 days trial period after 7 days adaptation. Results showed that there were significant interactions (p < .05) between piglets species and dietary treatments in average daily gain (ADG) and feed efficiency, serum and hepatic glutathione peroxidase (GSH-Px) and malondialdehyde (MDA), hepatic integral optical density (IOD) of α-tumour necrosis factor (TNF-α), hepatic relative mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2)/TNF-α and protein expression of TNF-α. Regardless of piglets species, supplementation with IVE and ELE increased (p < .05) ADG and feed efficiency, T-SOD and GSH-Px in serum and liver, hepatic IOD of Nrf2, hepatic mRNA and protein expression of Nrf2/TNF-α. However, CHL treatment resulted in lower (p < .05) serum GSH-Px and hepatic mRNA and protein expression of Nrf2/TNF-α, and higher hepatic MDA and IOD of TNF-α. Compared to LCB, DLY piglets had higher (p < .05) ADG and feed efficiency, serum and hepatic MDA, and protein expression of TNF-α, but lower (p < .05) ADFI, liver index, serum and hepatic GSH-Px, hepatic IOD of TNF-α, mRNA expressions of Nrf2/TNF-α were observed. In conclusion, Illicium verum (500 mg/kg) and Eucommia ulmoides leaf (250 mg/kg) extracts can increase the growth performance and antioxidant ability of DLY and LCB piglets, while chlortetracycline produces undesirable side-effects on the antioxidant ability of DLY and LCB piglets. Illicium verum and Eucommia ulmoides leaf extracts produced different antioxidant effects in DLY and LCB piglets with the Chinese native Licha-black pig responding better than Duroc × Landrace × Yorkshire.

Effects of zearalenone-induced oxidative stress and Keap1-Nrf2 signaling pathway-related gene expression in the ileum and mesenteric lymph nodes of post-weaning gilts.

Zearalenone (ZEA) contamination of feed affects animal husbandry and the human health. Currently, the molecular mechanism underlying small intestine-related diseases caused by ZEA-induced oxidative stress is not well understood. In this study, we aimed to identify the mechanisms involved in ZEA (0.5-1.5 mg/kg)-induced oxidative stress in the ileum and mesenteric lymph nodes (MLNs) and the role of the Kelch-like erythroid cell-derived protein with CNC homology-associated protein 1 (Keap1)-nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway in post-weaning gilts. Forty post-weaning gilts (Landrace × Yorkshire × Duroc) with an average body weight of 14.01 ± 0.86 kg were randomly allocated to four groups and fed a corn-soybean meal basal diet supplemented with < 0.1, 0.5, 1.0, or 1.5 mg/kg ZEA. The results showed that the activity of total superoxide dismutase and glutathione peroxidase decreased (p < 0.05) linearly and quadratically and that the content of malondialdehyde increased (p < 0.05) quadratically in the ileum and MLNs with increasing ZEA in the diet. Immunohistochemical analysis showed that the expression of Nrf2 and glutathione peroxidase 1 (Gpx1) immunoreactive proteins in the ileum and MLNs were significantly enhanced with increasing ZEA. The relative mRNA and protein expression of Nrf2, Gpx1, quinone oxidoreductase 1 (Nqo1), hemeoxygenase 1 (Ho1), modifier subunit of glutamate-cysteine ligase (Gclm), and catalytic subunit of glutamate-cysteine ligase (Gclc) increased (p < 0.05) linearly and quadratically, and the relative mRNA and protein expression of Keap1 decreased (p < 0.05) linearly and quadratically in the ileum with increasing ZEA concentrations in the diet. Further, the relative mRNA and protein expression of Nrf2 and Gpx1 increased (p < 0.05) linearly and quadratically, and the relative mRNA and protein expression of Nqo1, Ho1, and Gclm decreased (p < 0.05) quadratically in the MLNs as ZEA concentrations increased in the diet. Our results provide valuable genetic information on ZEA-induced oxidative stress in the ileum and MLNs of post-weaning gilts and have elucidated the key regulatory genes involved in the Keap1-Nrf2 signaling pathway. Results indicated that the Keap1-Nrf2 signaling pathway might be a key target to further prevent and treat ZEA-induced injury to the ileum in post-weaning gilts.

Comparative study of stress response, growth and development of uteri in post-weaning gilts challenged with zearalenone and estradiol benzoate.

The objective of this study was to evaluate the effects of zearalenone (ZEA) and estradiol benzoate (EB) on stress injury and uterine development in post-weaning gilts. Thirty healthy post-weaning female gilts (Duroc × Landrace × Large White) aged 28-32 days were randomly allocated to three treatments as follows: (a) basal diet (Control), (b) basal diet plus 1.0 mg/kg purified ZEA (ZEA) and (c) basal diet plus 0.75 ml (1.5 mg) EB per pig at 3-days intervals by intramuscular injection (EB). The serum estradiol (E2 ), the final and the increased vulvar area, uterine index, thickness of the myometrium and endometrium, and protein expression of heat shock protein 70 (HSP70) in ZEA group were higher than those in the control group (p < .05), but lower than those in the EB group (p < .05). The serum luteinizing hormone in ZEA group was lower than that of the control group (p < .05), but higher than that in the EB group (p < .05). Higher serum follicle-stimulating hormone and progesterone were observed in the ZEA and control groups than those in the EB group (p < .05). The serum glutathione peroxidase activity in the ZEA group was lower than that in the control and EB groups (p < .001), and the malondialdehyde in the ZEA group was higher than that in the control and EB groups (p < .001). Moreover, the relative mRNA and protein expression of growth hormone receptor (GHR) and relative mRNA expression of HSP70 in the ZEA and EB groups were higher than those in the control group (p < .05). In conclusion, both ZEA (1.0 mg/kg) and EB (1.5 mg at 3 days intervals by intramuscular injection) stimulated vulvar swelling and uterine hypertrophy by disordering serum hormones and up-regulating GHR expression, and induced stress by different mechanisms in this study. Furthermore, the observed up-regulating HSP70 expression challenged by ZEA or EB may be part of the mechanism to resist stress injury.

Effects of star anise (Illicium verum Hook.f.) essential oil on nutrient and energy utilization of laying hens.

An experiment using 96 Hy-Line brown laying hens at 29 weeks of age that were randomly allocated to four treatments with eight replicates was conducted to assess the effects of star anise (Illicium verum Hook.f.) oil (SAO) on nutrient and energy utilization. Dietary treatments were corn-soybean meal-based diets supplemented with 0, 200, 400, and 600 mg/kg SAO. The birds were fed the diets for 14 days. The conventional nutrients and amino acids of feed and excreta samples were measured for calculating apparent metabolic efficiency, and gross energy (GE) was analyzed for computation of apparent metabolic energy (AME) and true metabolic energy (TME). Endogenous losses were obtained from another 16 birds for calculating true metabolic efficiency. Birds supplemented with SAO had higher (p < 0.05) metabolic efficiency of crude protein (CP), organic matter (OM), Lys, Met, Arg, and Thr than those of control group. Quadratic effects on utilization of Lys, Met, and Thr were observed (p < 0.05). Inclusion of SAO to diet improved nutrient utilization, and the efficacy is best at 400 mg/kg.

Polysaccharide from alfalfa activates RAW 264.7 macrophages through MAPK and NF-κB signaling pathways.

Alfalfa polysaccharide (APS), a bioactive compound extracted from alfalfa, has been proposed to exhibit potential growth-promoting and immune-enhancing functions. But, little is known about the cellular immunomodulatory and intrinsic molecular mechanisms. Here we extracted the APS, and performed in vitro experiments to characterize the immunomodulatory functions as well as the molecular mechanisms of APS on RAW 264.7 macrophages cells. Chemical analyses showed that APS was mainly composed of fucose, arabinose, galactose, glucose, xylose, mannose, galacturonic acid and glucuronic acid. The results of in vitro assays demonstrated that 50 and 100 µg/mL APS increased the cell viability of RAW 264.7 cells. The secretion and gene expression of NO/iNOS, IL-6 and TNF-α in APS-induced macrophage cell were significantly enhanced. However, APS-induced TNF-α production was decreased by blocking the MAPK or NF-κB signaling pathways, especially for the blockade of p38. Moreover, APS enhanced the phosphorylation of p38, ERK, and JNK, promoted the degradation of IκBα, and increased the nuclear translocation of NF-κB p65 subunit. Therefore, we demonstrated that APS could improve the immune functions of RAW 264.7 macrophages cells by promoting the cell viability and increasing secretion and gene expressions of NO/iNOS, IL-6 and TNF-α through the MAPK and NF-κB signaling pathways.

Effects of star anise (Illicium verum Hook.f.) essential oil on laying performance and antioxidant status of laying hens.

To investigate the effects of dietary supplementation of star anise oil (SAO) on performance and antioxidant status of laying hens, a total of 864 Hy-Line brown laying hens at 26 wk of age were randomly allocated to 4 treatments with 6 replicates of 36 birds. Dietary treatments were non-star anise oil supplementation and supplemented with SAO at the level of 200, 400, and 600 mg/kg diet. The birds were fed the diets for 56 d. Average egg weight, average daily feed intake (ADFI), egg mass, laying rate, and feed conversion of each replicate were measured. Blood and liver samples from 12 birds were obtained, 72 eggs were picked out, per treatment at day 28 and day 56 of the experiment, and eggs stored for 56 d, to determine antioxidant status in serum, liver, and yolk. All laying hens had similar average egg weight, egg mass, laying rate, and feed conversion in day 29 to day 56 or the entire period of the experiment but significant difference on ADFI in day 1 to day 56. However, increasing diet concentration of SAO tended to improve (P < 0.10) egg mass, average egg weight, and ADFI in day 1 to day 28. Supplementation of SAO linearly increased (P < 0.05) activities of total superoxide dismutase (T-SOD) (day 28 and day 56) and glutathione peroxidase (GSH-PX) (day 56) in serum, GSH-PX (day 28 and day 56) in liver and total antioxidant capacity (T-AOC) (day 56) in serum and liver, but linearly reduced (P < 0.05) concentrations of malondialdehyde (MDA) (day 28 and day 56) in liver. Supplementation of SAO linearly increased (P < 0.05) T-SOD activity at day 14 and day 28, reduced (P < 0.05) MDA concentration at day 42 and day 56 of the experiment in yolk. Increasing content of SAO linearly (P < 0.05) increased T-SOD activity in yolk of eggs stored at day 0, 14, 28, 42, and 56, decreased MDA content of eggs stored at day 42 and 56, whether laying hens fed diets for 28 or 56 d. Dietary supplementation of SAO enhanced laying performance and overall antioxidant status of laying hens in a dose-dependent manner.

Effect of different sources and levels of iron in the diet of sows on iron status in neonatal pigs.

This study was conducted to determine the effects of maternal dietary supplementation of ferrous glycine chelate (Fe-Gly) and ferrous sulfate monohydrate (FeSO4·H2O) on the relative organ weight, tissue iron contents, red blood cells (RBC), hemoglobin concentration (HGB) and hematocrit (HCT) in blood, as well as ferritin (Fn), serum iron (SI), and total iron binding capacity (TIBC) in serum of newborn piglets. Forty-five sows (Landrace × Large white, mean parity 3 to 4, no significant differences in BW) were randomly allotted to 9 treatments (n = 5 sows/treatment): control (basal diet with no Fe supplementation), the basal diet supplemented with 50, 80, 110 or 140 mg Fe/kg as Fe-Gly, and the basal diet supplemented with 50, 80, 110 or 140 mg Fe/kg as FeSO4·H2O. The neonatal piglets (n = 45) were used to determine the relative organ weight, tissue iron contents and blood biochemical indices. Compared with the control, the relative weight of spleen and kidney were significantly increased (P < 0.05) in the Fe-Gly groups. The iron contents in liver, spleen, kidney and femur were also found increased (P < 0.05) in the Fe-Gly groups. The RBC (d 1 and 21), HGB (d 1 and 21) and HCT (d 1 and 21) in blood and Fn (d 1) and SI (d 1 and 21) significantly increased (P < 0.05), but the TIBC (d 1 and 21) in serum decreased (P < 0.05) in the Fe-Gly groups. Moreover, the kidney relative weight, iron content in liver, spleen, kidney and femur, RBC (d 1) and HGB (d 21) in blood, and SI (d 1) in the Fe-Gly groups increased (P < 0.05) compared with the FeSO4·H2O treatment. Linear and quadratic responses of the kidney relative weight, the iron content in liver, spleen, kidney and femur, RBC (d 1 and 21), HGB (d 1 and 21) and HCT (d 1 and 21) in whole blood, SI (d 1) and TIBC (d 1 and 21) in the Fe-Gly groups were observed (P < 0.05). Linear responses of Fn (d 1 and 21) and SI (d 21) in the Fe-Gly groups, and spleen relative weight, HCT (d 1), Fn (d 1) and TIBC (d 1 and 21) in the FeSO4·H2O groups were observed (P < 0.05). These finding suggest that Fe-Gly supplemented at the level of 110 mg/kg in the diet of sows in this experiment is superior to other forms of supplementation, based on HGB concentration, the relative organ weight, tissue iron contents and blood biochemical indices of piglets.

Zearalenone-Promoted Follicle Growth through Modulation of Wnt-1/ß-Catenin Signaling Pathway and Expression of Estrogen Receptor Genes in Ovaries of Postweaning Piglets.

Feedstuffs are severely contaminated by zearalenone (ZEA) worldwide. A specific dietary level of ZEA could cause malformations of the reproductive organs of sows, false estrus, decreased litter size, and abortion. However, the underlying mechanisms are still not clear. The objectives of the present study were to assess the effects of ZEA on morphology, distribution, and expression of estrogen receptors (ERα and ERß) in the ovaries of postweaning piglets. Furthermore, the relationship between ERs/glycogen synthase kinase (GSK)-3ß-dependent pathways mediated by ZEA and the Wnt-1/ß-catenin signaling pathway was examined. Forty healthy weaning piglets were allocated to the following four treatment groups: piglets fed with basal diet only (control), and ZEA0.5, ZEA1.0, and ZEA1.5, which were fed basal diets supplemented with ZEA at 0.5, 1.0, and 1.5 mg·kg-1, respectively. Then, the expression of GSK-3ß, ERα, ERß, and Wnt-1/ß-catenin were examined histomorphologically and immunohistochemically. Results showed that the proportion of primordial follicles (PrF's) decreased ( p < 0.001) but that of atretic primordial follicles (APFs) increased ( p < 0.001) with increasing dietary ZEA levels. More interestingly, the immunopositivity of ERß in the ovaries was stronger than that of ERα with the same treatment. The relative mRNA and protein expression levels of ERα, ERß, Wnt-1, ß-catenin, and GSK-3ß in the ovaries of postweaning gilts increased linearly ( p < 0.05) as dietary ZEA concentrations increased. Moreover, the accumulation of Wnt-1 and ß-catenin in the ovaries indicated that ZEA activated the Wnt-1/ß-catenin pathway, mediated by ERs/GSK-3ß. Our results strongly suggested that ovarian follicles in the ZEA (0.5-1.5 mg·kg-1)-treated groups were highly proliferative state, indicating that ZEA promoted ovarian development. The results also suggested that ZEA activates the ERs/GSK-3ß-dependent Wnt-1/ß-catenin signaling pathway, indicating its important role in accelerating development of the ovaries.