Alginate Oligosaccharide Alleviates Lipopolysaccharide-Induced Apoptosis and Inflammatory Response of Rumen Epithelial Cells through NF-κB Signaling Pathway.

AOS alleviates inflammatory responses; however, whether it exerts an effect on the rumen or regulates rumen inflammatory reaction remains unknown. In this study, firstly, the ovine ruminal epithelial cells (ORECs) were treated with 0, 200, 400, 600, and 800 µg/mL AOS, hoping to explore whether AOS hurt cell health. The results showed that compared with the AOS-0 group, the AOS-400 group could significantly increase (p < 0.05) cell viability, reduce (p < 0.05) reactive oxygen species (ROS) and interleukin (IL)-6 content, and have no adverse effect on cells. Secondly, we used LPS to construct an in vitro inflammatory model of rumen epithelial cells and then explored the protective role of AOS on rumen epithelial cells. The study was divided into three groups: the control group (CON), LPS, and LPS + AOS. The results demonstrated that the LPS + AOS group significantly increased the cell viability and reduced the ROS level in comparison with the LPS group (p < 0.05). Pretreatment with AOS also repressed (p < 0.05) the secretion of IL-1ß, IL-6, IL-8, and immunoglobulin (Ig)A from ORECs in the culture medium following LPS. In terms of tight junction (TJ) proteins, AOS treatment also significantly increased (p < 0.05) the zonula occludens 1 (ZO-1) and Occludin expression. The apoptosis rate, Caspase3, Caspase9, BAD, and BCL-2/BAX were decreased (p < 0.05) after AOS treatment, and the expression of BCL-2 was increased (p < 0.05). In addition, the expressions of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), and nuclear factor-κB (NF-κB) were inhibited (p < 0.05) with the addition of AOS. At the protein level, pretreatment of AOS decreased (p < 0.05) the expression of MyD88 and the phosphorylation level of inhibitor κB α (IκBα) after the LPS challenge. Taken together, our results indicated that AOS could alleviate the LPS-induced apoptosis and inflammatory response of rumen epithelial cells through the NF-κB signaling pathway, which may be a promising strategy for treating apoptosis and inflammation in sheep breeding.

Effect of tea polyphenols on intestinal barrier and immune function in weaned lambs.

Introduction: The purpose of this study was to explore the effects of tea polyphenols on growth performance, cytokine content, intestinal antioxidant status and intestinal barrier function of lambs, in order to provide reference for intestinal health of ruminants. Methods: Thirty weaned lambs (average initial weight 9.32 ± 1.72 kg) were randomly divided into five groups with six lambs in each group. The control group did not add anything but the basic diet mainly composed of Pennisetum and Corn, and the other four groups added 2, 4, 6 g/kg tea polyphenols and 50 mg/kg chlortetracycline to the basic diet, respectively. The experiment lasted for 42 days. Results: Dietary tea polyphenols improved the growth and stress response and reduced intestinal permeability of lambs (p > 0.05), while CTC did not affect the final lamb weight (p > 0.05). Both tea phenols and CTC significantly reduced inflammatory factors and enhanced the immune system (p > 0.05). Dietary tea polyphenols increased villus height, villus height/crypt depth, secretory immunoglobulin A (p > 0.05), and antioxidant enzymes, while decreasing MDA and apoptosis in the intestine (p > 0.05). However, compared with other groups, the content of T-AOC in jejunum did not change significantly (p > 0.05). Tea polyphenols also increased claudin-1 levels in the duodenum, jejunum, and ileum more than CTC (p > 0.05). CTC had a limited effect on the mRNA expression of Occludin and ZO-1, while tea polyphenols increased these in both the duodenum and ileum (p > 0.05). Conclusion: This study demonstrated that tea polyphenols can effectively improve the intestinal barrier of weaned lambs, and that they have anti-inflammatory and antioxidant effects similar to those of antibiotics. Thus, tea polyphenols could be used to replace antibiotics in ensuring safety of livestock products and in achieving the sustainable development of modern animal husbandry.

Replacing alfalfa hay with amaranth hay: effects on production performance, rumen fermentation, nutrient digestibility and antioxidant ability in dairy cow.

OBJECTIVE: The aim of this research was to explore the effects of dietary substitution of alfalfa hay by amaranth hay on production performance, rumen fermentation, nutrient digestibility, serum biochemical parameters and antioxidant ability in dairy cows. METHODS: A total of 45 healthy Holstein cows with same parity and similar milk yield and body weight were randomly divided into 3 groups: control diet without amaranth hay (CON) or 50% and 100% alfalfa hay replaced by an equal amount of amaranth hay (dry matter basis, AH1 and AH2, respectively). All the cows were fed regularly 3 times a day at 06:30, 14:30, and 22:30 and had free access to water. The experiment lasted for 60 d. RESULTS: The dry matter intake of CON and AH1 groups was higher (p<0.05) than that of AH2 group. Compared with AH1 group, the milk yield of AH2 group was reduced (p<0.05). Moreover, dietary substitution of alfalfa hay by amaranth hay increased (p<0.05) milk fat, ammonia nitrogen and acetate concentrations. However, the crude protein digestibility of AH2 group was lower (p<0.05) than that of CON group, while an opposite tendency of serum urea nitrogen was found between two groups. The neutral detergent fiber digestibility of AH1 group was increased (p<0.05) when compared to AH2 group. Amaranth hay treatment increased (p<0.05) the serum concentration of glutathione peroxidase in dairy cows. Compared with CON group, the malonaldehyde activity of AH1 group was decreased (p<0.05). CONCLUSION: Dietary replacing alfalfa hay with amaranth hay (50% ratio) in dairy cows did not affect production performance but improved their antioxidant ability.

Grape Seed Extract as a Feed Additive Improves the Growth Performance, Ruminal Fermentation and Immunity of Weaned Beef Calves.

The purpose of this research was to evaluate effects of grape seed extract (Gse) supplementation on the growth performance; ruminal fermentation; nutrient digestibility; and serum biochemical, antioxidative, and immune parameters of weaned beef calves. A total of 30 Simmental crossbred male calves with similar age and body weight were randomly allocated to two groups: a control group with no Gse (CON) and a Gse supplementation group (GSE) (4 g/d Gse per animal). The results show that, compared with the CON group, the average daily gain significantly increased (p = 0.043) in the GSE group. The ruminal contents of microbial protein and butyrate in GSE group were higher (p < 0.05) than those in the CON group. Additionally, calves fed Gse displayed increased (p < 0.05) dry matter and neutral detergent fiber digestibility. Moreover, the serum concentrations of triglyceride, catalase, superoxide dismutase, immunoglobulin G and immunoglobulin M were higher (p < 0.05) in the GSE group than those in the CON group. However, opposite tendencies of non-esterified fatty acid, malondialdehyde, tumor necrosis factor-α and interleukin-6 were found between the two groups. Overall, the supplementation of Gse can improve ruminal fermentation, nutrient digestibility, antioxidant ability, and immunity, as well as promoting the healthy growth of weaned cross-breed beef calves.

Silage additives improve fermentation quality, aerobic stability and rumen degradation in mixed silage composed of amaranth and corn straw.

The objective of this research was to investigate effects of different additives on the fermentation quality, aerobic stability and rumen degradation of mixed silage composed of amaranth and corn straw. The mixture ratio of amaranth to corn straw was 78%: 22%. Three additives were selected in this study and five groups were as follows: control group (CON, without additive), lactic acid bacteria group (LAB, 5 mg/kg, Lactobacillus plantarum ≥ 1.6×1010 CFU/g and L. buchneri ≥ 4.0×109 CFU/g), glucose group (GLU, 30 g/kg), cellulase group (CEL, 2 mg/kg) and lactic acid bacteria, glucose and cellulase group (LGC, added at the same levels as in individual group). The period of ensiling was 60 days. Fermentation quality, chemical composition and aerobic stability of mixed silage were analyzed. Four cows with permanent ruminal fistula were selected as experimental animals. Nylon bag technique was used to study rumen degradation characteristic of dry matter (DM), crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber (ADF) of mixed silage. Compared with CON group, the addition of different silage additives could improve mixed silage quality of amaranth and corn straw to some extent. Combining three additives significantly increased (P < 0.05) the DM, CP and lactic acid contents, whereas decreased (P < 0.05) the ADF and NDF contents as well as pH and ammonia nitrogen/total nitrogen. Moreover, the aerobic stability and rumen degradation of DM, CP and NDF were significantly improved (P < 0.05) in LGC group when compared to other groups. In conclusion, the combined addition of lactic acid bacteria, glucose and cellulase increased DM, CP and lactic acid contents as well as lactic acid bacteria count, decreased NDF and ADF contents and aerobic bacteria and mold counts, improved aerobic stability and rumen degradation of amaranth and corn straw mixed silage.

The effects of fucoidan as a dairy substitute on diarrhea rate and intestinal barrier function of the large intestine in weaned lambs.

This paper explores the effects of fucoidan on the frequency of diarrhea, colon morphology, colon antioxidant status, cytokine content, short-chain fatty acids, and microflora of cecal contents in early weaned lambs in order to provide a reference for the intestinal health of young ruminants. Fucoidan is a natural active polysaccharide extracted from kelp and other large brown algae. It has many biological effects, such as improving immunity, nourishing the stomach and intestines, and anti-tumor properties. This study investigated the effects of fucoidan supplementation in milk replacer on the large intestine's ability to act as an intestinal barrier in weaned lambs. With six duplicate pens and one lamb per pen, a total of 24 weaned lambs (average starting body weight of 7.32 ± 0.37 kg) were randomly assigned to one of four milk replacer treatments. Four concentrations of fucoidan supplementation (0, 0.1, 0.3, and 0.6% dry matter intake) were employed to investigate the effects of fucoidan on cecal fermentation and colon microbial organization. The test period lasted 37 days (1 week before the test and 1 month after the test), and lamb cecal contents and colon organization were collected for examination. In addition, the fecal status of all lambs was observed and recorded daily, allowing us to calculate the incidence of diarrhea in weaned lambs. The findings demonstrated that fucoidan may significantly increase the concentration of short-chain fatty acids (propionic acid and butyric acid) in the cecal digesta of weaned lambs. In weaned lambs, 16S rDNA testing showed that fucoidan at 0.3-0.6% (dry matter intake) was beneficial for boosting the variety of the intestinal bacteria and modifying the relative abundance of a few bacterial strains. In addition, fucoidan enhanced colon antioxidant and immune functions and decreased the diarrhea rate to relieve weaning stress. This result demonstrates that milk replacer supplementation with fucoidan contributes to the improvement in the large intestinal health of weaned lambs.

Chitosan Oligosaccharide Inhibits the Synthesis of Milk Fat in Bovine Mammary Epithelial Cells through AMPK-Mediated Downstream Signaling Pathway.

Chitosan oligosaccharide (COS) is a variety of oligosaccharides, and it is also the only abundant basic amino oligosaccharide in natural polysaccharides. Chitosan oligosaccharide is a low molecular weight product of chitosan after enzymatic degradation. It has many biological effects, such as lipid-lowering, antioxidant and immune regulation. Previous studies have shown that chitosan oligosaccharide has a certain effect on fat synthesis, but the effect of chitosan oligosaccharide on milk fat synthesis of bovine mammary epithelial cells (BMECs) has not been studied. Therefore, this study aimed to investigate chitosan oligosaccharide's effect on milk fat synthesis in bovine mammary epithelial cells and explore the underlying mechanism. We treated bovine mammary epithelial cells with different concentrations of chitosan oligosaccharide (0, 100, 150, 200, 400 and 800 µg/mL) for 24 h, 36 h and 48 h respectively. To assess the effect of chitosan oligosaccharide on bovine mammary epithelial cells and determine the concentration and time for chitosan oligosaccharide treatment on cells, several in vitro cellular experiments, including on cell viability, cycle and proliferation were carried out. The results highlighted that chitosan oligosaccharide (100, 150 µg/mL) significantly promoted cell viability, cycle and proliferation, increased intracellular cholesterol content, and reduced intracellular triglyceride and non-esterified fatty acids content. Under the stimulation of chitosan oligosaccharide, the expression of genes downstream of Phosphorylated AMP-activated protein kinase (P-AMPK) and AMP-activated protein kinase (AMPK) signaling pathway changed, increasing the expression of peroxisome proliferator-activated receptor alpha (PPARα) and hormone-sensitive lipase (HSL), but the expression of sterol regulatory element-binding protein 1c (SREBP1) and its downstream target gene stearoyl-CoA desaturase (SCD1) decreased. In conclusion, these results suggest that chitosan oligosaccharide may inhibit milk fat synthesis in bovine mammary epithelial cells by activating the AMP-activated protein kinase signaling pathway, promoting the oxidative decomposition of fatty acids and inhibiting fatty acid synthesis.

Effects of Dietary Fucoidan Supplementation on Serum Biochemical Parameters, Small Intestinal Barrier Function, and Cecal Microbiota of Weaned Goat Kids.

The purpose of this study was to evaluate the effects of fucoidan supplementation on serum biochemical parameters, small intestinal barrier function, and cecal microbiota of weaned goat kids. A total of 60 2-month-old weaned castrated male goat kids (Chuanzhong black goat) were used in this 30-day experiment. The goat kids were randomly divided into four groups: a control group (CON) fed the basal diet, and three other groups supplemented with 0.1%, 0.3%, and 0.5% fucoidan in the basal diet (denoted as F1, F2, and F3 groups, respectively). The results indicated that dietary fucoidan supplementation decreased (p < 0.05) the activity of lactate dehydrogenase (LDH) and the content of glucose (GLU) as measured on day 15. As measured on day 30, dietary fucoidan increased (p < 0.05) the content of total protein (TP) and decreased the activity of aspartate aminotransferase (AST), and supplementation with 0.3% and 0.5% fucoidan decreased (p < 0.05) the activity of LDH. Dietary fucoidan decreased (p < 0.05) the content of D-lactic acid (D-LA) and the activity of diamine oxidase (DAO). Dietary fucoidan increased (p < 0.05) the activity of catalase (CAT) in the duodenum. Dietary 0.3% and 0.5% fucoidan enhanced (p < 0.05) the activity of glutathione peroxidase (GSH-Px) in the ileum, the activity of total superoxide dismutase (T-SOD) in the jejunum and ileum, and the activity of CAT in the ileum. Dietary 0.3% and 0.5% fucoidan reduced the contents of malondialdehyde (MDA) in the duodenum, jejunum, and ileum and the content of hydrogen peroxide (H2O2) in the duodenum. Dietary fucoidan increased (p < 0.05) the content of secretory immunoglobulin A (sIgA) in the duodenum. Supplementation of 0.3% and 0.5% fucoidan upregulated (p < 0.05) the gene expression of ZO-1 and claudin-1 in the duodenum, jejunum, and ileum, and dietary supplementation of 0.3% and 0.5% fucoidan upregulated (p < 0.05) the gene expression of occludin in the jejunum and ileum. The 16S rRNA high-throughput sequencing results showed that at the phylum level, dietary fucoidan increased (p < 0.05) the abundance of Bacteroidetes while decreasing (p < 0.05) the abundance of Firmicutes. At the genus level, dietary 0.3% and 0.5% fucoidan increased (p < 0.05) the abundances of Unspecified_Ruminococcaceae, Unspecified_Bacteroidale, Unspecified_Clostridiales, and Akkermansia. In conclusion, dietary fucoidan supplementation had positive effects on intestinal permeability, antioxidant capacity, immunity function, tight junctions, and the cecal microflora balance in weaned goat kids.

The Effects of Fucoidan Dietary Supplementation on Growth Performance, Serum Antioxidant Capacity, Immune Function Indices and Intestinal Morphology in Weaned Kids.

The purpose of this study was to evaluate the effects of fucoidan dietary supplementation on growth performance, organs' relative weight, serum anti-oxidation markers, immune function indices and intestinal morphology in weaned kids. A total of 60 2-month-old weaned castrated male kids (Chuanzhong black goat) were used for this 30-day experiment and randomly allocated to four groups. The control group (CON) fed a basal diet, while the other three groups were provided with the same diet further supplemented with fucoidan at 0.1%, 0.3% or 0.5%, namely, F1, F2 and F3 groups, respectively. The results indicated that dietary fucoidan supplementation significantly increased (p < 0.05) the activity of catalase (CAT) when compared to the CON group on day 15. Moreover, the addition of fucoidan at 0.3% and 0.5% significantly increased (p < 0.05) the activities of glutathione peroxidase (GSH-Px) and total superoxide dismutase (T-SOD). On day 30, dietary fucoidan supplementation significantly reduced (p < 0.05) the feed conversion rate (FCR), contents of tumor necrosis-α (TNF-α), interleukin-1ß (IL-1ß) and interleukin-6 (IL-6), while it significantly increased (p < 0.05) the activity of total superoxide dismutase (T-SOD), the content of immunoglobulin G (IgG) and the villus height (VH) of the duodenum. Moreover, dietary 0.3% and 0.5% fucoidan supplementation significantly increased (p < 0.05) the villus height (VH) of the jejunum and ileum and significantly reduced (p < 0.05) the crypt depth (CD) of ileum. In conclusion, dietary fucoidan had positive effects on growth performance, serum anti-oxidation, immune function and intestinal morphology of weaned kids.

Effects of Dietary Supplementation of Algae-Derived Polysaccharides on Morphology, Tight Junctions, Antioxidant Capacity and Immune Response of Duodenum in Broilers under Heat Stress.

To evaluate the ameliorative effect of algae-derived polysaccharide (ADP) supplementation on duodenal injury caused by heat stress (HS) in broilers, a total of 144 male yellow-feathered broilers (56-day-old) were randomly allocated into three groups: The TN group (thermoneutral zone, broilers were raised at 23.6 ± 1.8 °C); HS group (heat stress, broilers were exposed to 33.2 ± 1.5 °C 10 h/day, 8:00 a.m.-18:00 p.m., the temperature in the remaining period was consistent with the TN group); HSA group (heat-stressed broilers were fed with ADP supplemented diet at 1000 mg/kg). There were six replications in each treatment, and eight broilers in each replication. The feeding trial lasted four weeks. The results showed that dietary ADP supplementation tended to increase the villus height (p = 0.077) and villus width (p = 0.062), and decrease the apoptosis rate (p = 0.081) in the duodenum of broilers under HS. Furthermore, dietary ADP increased the relative mRNA and protein (based on immunofluorescence) expression levels of occludin and zonula occludens-1 (ZO-1) in the duodenum of broilers under HS (p < 0.05). In addition, dietary ADP enhanced the total antioxidation capacity (T-AOC) and activity of glutathione-S transferase (GST), while reducing the malondialdehyde (MDA) concentration of the duodenum in broilers under HS (p < 0.05). Moreover, dietary ADP supplementation upregulated the duodenal nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), glutathione peroxidase 1 (GPx1) and glutathione S-transferase theta 1 (GSTT1) mRNA expression levels in heat-stressed broilers (p < 0.05). Furthermore, compared with the HS group, broilers fed with an ADP supplemented diet had a higher relative mRNA expression of inhibitor kappa B alpha (IκBα) (p < 0.05) and a lower relative mRNA expression of tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) in the duodenum (p < 0.05). In summary, dietary ADP supplementation had an ameliorative effect on HS-induced impairment of tight junctions, antioxidant capacity and the immune response of the duodenum in broilers. These beneficial effects might be related to the modulation of Nrf2 and NF-κB signaling pathways.

MiR-485 targets the DTX4 gene to regulate milk fat synthesis in bovine mammary epithelial cells.

MicroRNAs (miRNAs) are mRNA suppressors that regulate a variety of cellular and physiological processes, including cell proliferation, apoptosis, triglyceride synthesis, fat formation, and lipolysis, by post-transcriptional processing. In previous studies, we isolated and sequenced miRNAs from mammary epithelial cells from Chinese Holstein cows with high and low milk fat percentages. MiR-485 was one of the significantly differentially expressed miRNAs that were identified. In the present study, the relationship between the candidate target gene DTX4 and miR-485 was validated by bioinformatics and real-time fluorescent quantitative PCR (qRT-PCR) and Western blot (WB) analyses in bovine mammary epithelial cells (bMECs). The results indicated that miR-485 negatively regulated the mRNA expression of the target gene DTX4. Furthermore, an shRNA interference vector for the target gene DTX4 was constructed successfully, and it increased the triglyceride content and reduced the cholesterol content of transfected cells. These results suggest that miR-485 may affect the contents of triglycerides (TGs) and cholesterol (CHOL) by targeting the DTX4 gene. This study indicates that miR-485 has a role in regulating milk fat synthesis and that miR-485 targets the DTX4 gene to regulate lipid metabolism in bMECs. These findings contribute to the understanding of the functional significance of miR-485 in milk fat synthesis.

Yupingfeng polysaccharides enhances growth performance in Qingyuan partridge chicken by up-regulating the mRNA expression of SGLT1, GLUT2 and GLUT5.

The ban on the use of antibiotic in feed encouraged nutritionists to using alternatives to maintain growth performance and intestinal function of broilers. This study was conducted to evaluate the effects of Yupingfeng polysaccharides (YP) supplementation on growth performance and expression of SGLT1, GLUT2 and GLUT5 in Qingyuan partridge chicken. Experiment 1: a total of 540 chickens were randomly allocated to five groups with six replication. Dietary treatments were: (1) CON (control group), basal diet; (2) T1, CON + 0.5 g kg-1 YP; (3) T2, CON + 1 g kg-1 YP; (4) T3, CON + 2 g kg-1 YP; (5) T4, CON + 4 g kg-1 YP. Experiment 2, a total of 162 were randomly allocated to three groups with three replication. Dietary treatments were: (1) CON, basal diet; (2) T1, CON + 0.5 g kg-1 YP; (3) T2, CON + 1 g kg-1 YP. From days 1 to 14 and overall, chicken fed T1 diet had higher ADG. On day 42, there was increased villus height of jejunum in T1 group. On days 14 and 28, there was decreased villus height of duodenum and jejunum in T2 group. In duodenum, the expression of SGLT1 (days 21, 35 and 42), GLUT2 (days 7, 14, 21, 28, 35 and 42) and GLUT5 (days 7, 14, 21 and 28) was increased with YP supplementation. In jejunum, the expression of SGLT1 (days 7, 14, 21, 28 and 35), GLUT2 (days 14, 21, 28, 35 and 42) and GLUT5 (days 7, 14, 21, 28, 35 and 42) was increased with YP supplementation. In ileum, the expression of SGLT1 (days 7, 21, 35 and 42), GLUT2 (days 7, 14, 21 and 42) and GLUT5 (days 7, 14, 21, 28, 35 and 42) was increased with YP supplementation. Dietary YP supplementation improves growth performance and expression of SGLT1, GLUT2 and GLUT5 in intestine.

Study of Bacillus subtilis on growth performance, nutrition metabolism and intestinal microflora of 1 to 42 d broiler chickens.

To study the influence of different levels of Bacillus subtilis on growth performance, nutrition metabolism and intestinal microflora of 1 to 42 d Arbor Acres (AA) broilers, a total of 800 one-day-old healthy broilers were randomly divided into 5 groups with 4 replicates per group and 40 broilers per replicate. Broilers were fed a basic diet (group 1) which acted as the control group, and 4 other groups (2 to 5) were fed the basal diet with B. subtilis added at concentrations of 100, 150, 200 and 250 mg/kg, respectively for 42 days. The results showed as follow: the average daily gain (ADG) of group 4 was significantly higher than (P < 0.05) that of group 1, and the average daily feed intake (ADFI) of group 5 was the highest but the differences between groups were not significant (P > 0.05). The feed to gain ratio (F/G) of all the experimental groups was lower than that of the control and the difference was significant in group 4 (P < 0.05). In addition, supplementation of B. subtilis increased the apparent metabolism of crude protein (P > 0.05), crude fat (P > 0.05), dry matter (P > 0.05) and organic matter (P < 0.05). B. subtilis decreased the Escherichia coli and Salmonella populations in the cecum. This shows that adding B. subtilis to the broiler diet can improve the growth performance, increase feed efficiency, regulate serum index and reduce harmful bacteria in the intestinal tract. Based on our study, it could be recommended that addition of B. subtilis at 200 mg/kg could improve the growth performance of broilers.