Effects of High-Concentrate-Induced SARA on Antioxidant Capacity, Immune Levels and Rumen Microbiota and Function in Goats.

This study aims to explore the antioxidant, immune, and enzyme metabolism aspects in goats experiencing subacute ruminal acidosis (SARA). Furthermore, we seek to elucidate the relationship between the symbiotic microbiota of goats and their metabolic function. Sixteen goats were equally divided into two groups and fed a normal-concentrate diet (NC, 55% concentrate) or a high-concentrate diet (HC, 90% concentrate) for five weeks. We found that the HC diet reduced the total antioxidant capacity (T-AOC) (p = 0.022) and increased interleukin-1ß (IL-1ß) (p = 0.015), interleukin-4 (IL-4) (p = 0.008) and interleukin-6 (IL-6) (p = 0.002) concentration of goats. Simultaneously, the HC diet significantly increased the concentrations of alkaline phosphatase (ALP) and amylase (AMY) in the blood and rumen fluid of goats (p < 0.05). Microbial analysis in the rumen of goats revealed that the HC diet decreased bacterial richness and diversity, as evidenced by the changed observed species, Chao 1, PD whole tree and Shannon when compared to the NC diet (p < 0.01). The proportion of Proteobacteria increased while that of Spirochaetes and Fibrobacteres significantly decreased with the HC diet (p < 0.05). The Christensenellaceae_R-7_group and Ruminococcaceae_UCG-010 in rumen was notably decreased when a diet was switched from 55% concentrate diet to 90% concentrate diet (p < 0.05). Additionally, microbial functional potentials deduced that the HC diet significantly increased the abundance of the citrate cycle (TCA cycle) (ko00020) associated with carbohydrate metabolism (p = 0.028). Furthermore, the HC diet significantly increased the glutathione metabolism (ko00480) associated with the metabolism of other amino acids (p = 0.008). Our findings suggested that SARA reduced the total antioxidant capacity and increased levels of inflammatory factors in goats, as well as decreased rumen bacterial species and abundance.

Alterations in nutrient digestibility and performance of heat-stressed dairy cows by dietary L-theanine supplementation.

The purpose of this study was to investigate the effects of dietary L-theanine supplementation on apparent nutrient digestibility, milk yield, milk composition, and blood biochemical indices of dairy cows under heat stress. Thirty Chinese Holstein cows (19.84 ± 2.42 kg milk/d, 192.36 ± 40.77 d in milk and 2 ± 0.93 parities) were divided into 3 groups of 10 animals each. The control group was fed a basal total mixed ration (TMR) diet, while treatment 1 (LTA16) and treatment 2 (LTA32) groups were fed a basal TMR diet supplemented with L-theanine at 16 and 32 g/cow per day, respectively. The results showed that feeding the dairy cows with LTA16 treatment decreased (P < 0.05) their rectal temperature, whereas feeding with LTA32 treatment decreased (P < 0.05) their rumen fluid ammonia nitrogen content. In comparison to the control group, the supplementation of L-theanine had no significant effect (P > 0.05) on the dry matter intake, nutrient digestibility, total volatile fatty acid (TVFA) concentration and molar proportion of volatile fatty acid, milk yield, milk composition, feed efficiency and antioxidant capacity of the dairy cows. The triglyceride (TG) content of the LTA32 group was significantly greater (P = 0.014) than that of the control group. With the increase in L-theanine dosage, the serum cholesterol (CHOL) content significantly increased (P = 0.013). The serum albumin (ALB; P = 0.067), low-density lipoprotein cholesterol (LDL-C; P = 0.053), and high-density lipoprotein cholesterol (HDL-C; P = 0.067) contents showed an upward trend as L-theanine dosage increased. Ultimately, the results of this study show that supplementing dairy cow diet with L-theanine could decrease dairy cow rectal temperature, affect lipid metabolism, and potentially relieve the heat stress of dairy cows to some extent.

Oral administration of Lactobacillus delbrueckii enhances intestinal immunity through inducing dendritic cell activation in suckling piglets.

Lactobacillus delbrueckii (LAB) has been demonstrated to exert versatile beneficial effects on modulating intestinal immunity, increasing gut microbial diversity, promoting growth performance, and even preventing disease onset in pigs. However, the underlying mechanism of LAB-mediated gut immunity regulation in piglets remains unclear. In this study, we found that supplementation of LAB significantly increases serum TNF-α, ileum IL-4, and IL-10 levels compared with the control group. Meanwhile, oral supplementation of LAB-modified gut microbial communities was evidenced by the increased abundance of the Lactobacillus genus in the colon. Mechanistically, LAB induced dendritic cell (DC) maturation and activation, which may be relevant to the activation of NF-κB and MAPK signaling pathways. Moreover, we found that oral administration of LAB during the suckling period shows long-lasting immunomodulatory impacts on intestinal immunity after weaning. Collectively, this study uncovers the mechanism of LAB in regulating the intestinal immunity of piglets, suggesting that LAB can be developed as an immunoenhancing biological agent during the suckling period.

Alterations of endotoxin distribution across different biofluids and relevant inflammatory responses by supplementing L-theanine in dairy cows during heat stress.

The present trial was performed to reveal the regulatory effects of L-theanine on the levels of lipopolysaccharide (LPS) endotoxin within different biofluids, as well as relevant inflammatory responses of dairy cattle under heat stress conditions. Thirty lactating Chinese Holstein dairy cattle (189 ± 47 days in milk, and 2 ± 1 parities) were allocated in a completely randomized design to each of 3 dietary treatments: the control (CON, 0 g/d per cow L-theanine), the low L-theanine dosage treatment (LL, 16 g/d per cow L-theanine), and the high L-theanine dosage treatment (HL, 32 g/d per cow L-theanine). This trial consisted of 38 d (7 d for adaption and 31 d for data and sample collection), and sample collection for rumen liquid, blood plasma or serum, and milk were conducted on the d 27 and 38, respectively. Dairy cattle were constantly exposed to environmental heat stress during this experiment according to the recorded temperature-humidity index (THI). In the LL treatment, LPS concentration in rumen liquid was higher (P < 0.05), whilst LPS densities in plasma and milk were lower (P < 0.05) than those of the CON. Supplementing L-theanine at 2 dosages both significantly lowered (P < 0.05) the level of interleukin (IL)-1ß in the serum. Results of the present study suggested that L-theanine could be a promising additive in reducing the detrimental effects of heat stress on dairy cows, and L-theanine supplementation at 16 g/d per cow is preferred because it reduced the LPS translocation into the peripheral blood and LPS accumulation in the milk, as well as mitigated LPS-induced inflammatory reactions in dairy cows during heat stress. Further studies are necessitated to investigate the underlying mechanisms of L-theanine in LPS alteration and inflammation alleviation.

Effects of Dietary Supplementation of Lactobacillus delbrueckii on Gut Microbiome and Intestinal Morphology in Weaned Piglets.

Lactobacillus delbrueckii is a Gram-positive bacterium mostly used in the dairy industry for yogurt and cheese. The present study was designed to evaluate the effects of Lactobacillus delbrueckii on serum biochemical parameters, intestinal morphology, and performance by supplementing at a dietary level of 0.1% in diets for weaned piglets. Eighty healthy weaned piglets (initial body weight: 7.56 ± 0.2 kg) were randomly divided into two feeding groups with four replicates in each group (n = 10 animals per replicate); piglets were fed with basal diet (CON) or basal diet containing 0.1% Lactobacillus delbrueckii (LAC). The results showed that dietary supplementation of Lactobacillus delbrueckii improved growth performance and increased serum HDL and insulin levels in piglets on the 28th day of the experimental time (p < 0.05). The gut microbe analysis revealed that Lactobacillus delbrueckii significantly decreased the relative abundance of the phyla Bacteroidetes, but increased the relative abundance of the phyla Firmicutes. The Lactobacillus delbrueckii also significantly increased the relative abundance of Bifidobacterium and Lactobacillus at the genus level of the bacterial community in the ileum, but decreased the relative abundance of unclassified Clostridiales. Moreover, Lactobacillus delbrueckii improved mucosal morphology by obtaining higher intestinal villus height (p < 0.05), significantly increasing the concentrations of butyrate, isobutyric acid, and isovaleric acid in colonic chyme of piglets, but decreasing the intestinal pH at the duodenum and ileum on the 28th day of the experimental time. In conclusion, dietary supplementation of Lactobacillus delbrueckii in the diet of weaned piglets can improve intestinal morphology and modulate the microbiota community to promote growth performance.

Lactobacillus delbrueckii Protected Intestinal Integrity, Alleviated Intestinal Oxidative Damage, and Activated Toll-Like Receptor-Bruton's Tyrosine Kinase-Nuclear Factor Erythroid 2-Related Factor 2 Pathway in Weaned Piglets Challenged with Lipopolysaccharide.

Oxidative stress is increasingly being recognized as a player in the pathogenesis of intestinal pathologies, and probiotics are becoming an attractive means of addressing it. The present study investigated the effects of dietary supplementation with Lactobacillus delbrueckii (LAB) on intestinal integrity and oxidative damage in lipopolysaccharide (LPS)-challenged piglets. A total of 36 crossbred weaned piglets (Duroc × Landrace × Large Yorkshire) were randomly divided into three groups: (1) non-challenged controls (CON), (2) LPS-challenged controls (LPS), and (3) 0.2% LAB (2.01 × 1010 CFU/g) + LPS treatment (LAB + LPS). On the 29th day of the experiment, the LPS and CON groups were injected intraperitoneally with LPS and saline at 100 ug/kg body weight, respectively. The results show that the LPS-induced elevation of the serum diamine oxidase (DAO) level and small intestinal crypt depth (CD) were reversed by the dietary addition of LAB, which also markedly increased the ileal expression of tight junction proteins (occludin, ZO-1, and claudin-1) in the LPS-challenged piglets. Furthermore, LAB supplementation normalized other LPS-induced changes, such as by decreasing malondialdehyde (MDA) in both the serum and intestinal mucosa and 8-hydroxy-2-deoxyguanosine (8-OHdG) in the jejunal mucosa, increasing glutathione reductase (GR) and glutathione peroxidase (GSH-Px) in both the serum and intestinal mucosa, and increasing glutathione (GSH) and superoxide dismutase (SOD) in the jejunal mucosa. LAB also activated Toll-like receptor (TLR)-Bruton's tyrosine kinase (Btk)-nuclear factor erythroid 2-related factor 2(Nrf2) signaling pathways in the intestine, suggesting that it plays a vital role in the ameliorative antioxidant capacity of weaned piglets. In summary, LAB increased intestinal integrity by improving the intestinal structure and tight junctions while enhancing antioxidant functions via the activation of the TLR-Btk-Nrf2 signaling pathway.

Natural Polyphenols as Targeted Modulators in Colon Cancer: Molecular Mechanisms and Applications.

Colon cancer commonly develops from long-term chronic inflammation in the intestine and seriously threatens human health. Natural polyphenols have been valued as a crucial regulator of nutrient metabolism and metabolic diseases, owing to their anti-inflammatory and antioxidant functions and the ability to maintain a balance between gut microbes and their hosts. Notably, experimental and clinical evidence has shown that natural polyphenols could act as a targeted modulator to play a key role in the prevention or treatment of colon cancer. Thus, in this review, we summarized recent advances in the possible regulatory mechanism and the potential application of natural polyphenols in colon cancer, which might be regarded as a novel platform for the colon cancer management.

The Associated Regulatory Mechanisms of Zinc Lactate in Redox Balance and Mitochondrial Function of Intestinal Porcine Epithelial Cells.

Zinc lactate (ZnLA) is a new organic zinc salt which has antioxidant properties in mammals and can improve intestinal function. This study explored the effects of ZnLA and ZnSO4 on cell proliferation, Zn transport, antioxidant capacity, mitochondrial function, and their underlying molecular mechanisms in intestinal porcine epithelial cells (IPEC-J2). The results showed that addition of ZnLA promoted cell proliferation, inhibited cell apoptosis and IL-6 secretion, and upregulated the mRNA expression and concentration of MT-2B, ZNT-1, and CRIP, as well as affected the gene expression and activity of oxidation or antioxidant enzymes (e.g., CuZnSOD, CAT, and Gpx1, GSH-PX, LDH, and MDA), compared to ZnSO4 or control. Compared with the control, ZnLA treatment had no significant effect on mitochondrial membrane potential, whereas it markedly increased the mitochondrial basal OCR, nonmitochondrial respiratory capacity, and mitochondrial proton leakage and reduced spare respiratory capacity and mitochondrial reactive oxygen (ROS) production in IPEC-J2 cells. Furthermore, ZnLA treatment increased the protein expression of Nrf2 and phosphorylated AMPK, but reduced Keap1 and p62 protein expression and autophagy-related genes LC3B-1 and Beclin mRNA abundance. Under H2O2-induced oxidative stress conditions, ZnLA supplementation markedly reduced cell apoptosis and mitochondrial ROS levels in IPEC-J2 cells. Moreover, ZnLA administration increased the protein expression of Nrf2 and decreased the protein expression of caspase-3, Keap1, and p62 in H2O2-induced IPEC-J2 cells. In addition, when the activity of AMPK was inhibited by Compound C, ZnLA supplementation did not increase the protein expression of nuclear Nrf2, but when Compound C was removed, the activities of AMPK and Nfr2 were both increased by ZnLA treatment. Our results indicated that ZnLA could improve the antioxidant capacity and mitochondrial function in IPEC-J2 cells by activating the AMPK-Nrf2-p62 pathway under normal or oxidative stress conditions. Our novel finding also suggested that ZnLA, as a new feed additive for piglets, has the potential to be an alternative for ZnSO4.

Effects of Combined Supplementation of Conjugated Linoleic Acid, Methionine Chromium, Betaine, and Cysteamine on Meat Tenderness of Rats.

A systemic design was carried out to investigate the optimal combination of BET, Met-Cr, CLA, and CS for improving the meat tenderness in rats. A total of 104 six-week old male Sprague-Dawley rats were randomly assigned to 13 treatments with 4 replicates of 2 rats each. The experiments lasted for 5 weeks. The results showed that inclusion of Met-Cr decreased the contents of intramuscular fat (IMF), fat among muscle cells, and lipid droplets inside muscle cells (P < 0.05), and inclusion of CLA or Met-Cr increased the contents of IMF, fat among muscle cells, and lipid droplets inside muscle cells (P < 0.05). CS increased the contents of total collagen (TC) and soluble collagen (SC), and CLA decreased the contents of TC and SC (P < 0.05). The combination of BET and CLA increased IMF and SC contents and decreased TC contents (P < 0.05). The combination of BET and CS could increase fat contents among muscle cells and decrease TC and SC contents (P < 0.05). The combination of CLA and Met-Cr decreased IMF contents (P < 0.05). The combination of CLA and CS, as well as Met-Cr and CS, decreased fat contents among muscle cells (P < 0.05). These combinations may regulate lipogenesis and decrease the deposition of fat in muscles. There existed a significant positive correlation between IMF and SC content, which might indicate that IMF content improves meat's tenderness partly by increasing SC content in muscle.

Oral Administration of Lactobacillus delbrueckii during the Suckling Phase Improves Antioxidant Activities and Immune Responses after the Weaning Event in a Piglet Model.

Early colonization in the gut by probiotics influences the progressive development and maturity of antioxidant and immune system functionality in the future. This study investigated the impact of orally administrated Lactobacillus delbrueckii (LAB) during the suckling phase on future antioxidant and immune responses of the host, using a piglet model. One hundred neonatal piglets received saline (CON) or LAB at the amounts of 1, 2, 3, and 4 mL at 1, 3, 7, and 14 d of age, respectively. The piglets were weaned at the age of 21 d and fed until the age of 49 d. Serum, liver, and intestinal samples were obtained at 21, 28, and 49 d of age. The results showed that LAB tended to decrease serum 8-hydroxy-2-deoxyguanosine concentration and decreased the concentration of serum and hepatic malondialdehyde, but increased the activity of hepatic glutathione peroxidase on days 21, 28, and 49. The concentrations of secretory immunoglobulin A and some inflammatory cytokines and chemokines were increased (P < 0.05) in the intestinal mucosa of LAB-treated piglets on days 21, 28, and 49 compared to that of CON piglets. Likewise, protein expression of cyclooxygenase 2 and inducible nitric oxide synthase in the intestine of LAB-treated piglets was increased (P < 0.05) during the whole period. These results indicate that administration of LAB to the suckling piglet could improve antioxidant capacity and stimulate intestinal immune response, and these long-lasting effects are also observed up to 4 weeks after weaning. A proper utilization of LAB to neonates would be beneficial to human and animal's future health.

[Accuracy of predicting in vitro ruminal methane production in goats using volatile fatty acids stoichiometric models].

This study was conducted to investigate the accuracy of predicting in vitro ruminal methane (CH4) production using volatile fatty acids (VFA) stoichiometric models [CH4 = 0.5Ace-0.25Pro + 0.5But-0.25Val] (model 1), where CH4, Ace, Pro, But and Val are the production amounts of CH4, acetate, propionate, butyrate and valerate, respectively. Ten common feedstuffs, including four concentrates and six roughages with a wide range of chemical composition were incubated in serum bottles, and VFAs and CH4 production at 72 h were determined. The differences between the predicted and measured CH4 production were quantified using the model accuracy analysis. The results showed that the predicted CH4 production amounts were generally greater than the measured values obtained using the model 1, and the bias, slope and random error were 62.6%, 11.7% and 25.7%, respectively, indicating that fixed error exceeded 70%. By assuming 80% of total hydrogen being used for CH4 synthesis, the VFA stoichiometric model could be re-expressed as [CH4 = 0.8 (0.5Ace-0.25Pro + 0.5But-0.25Val)] (model 2). The root mean square prediction error (rMSPE = 0.18) for model 2 was less than for model 1 (rMSPE = 0.60). In addition, the bias, slope and random error of the model 2 were 2.1%, 5.7%, 92.3%, respectively, indicating that fixed error was less than 10%. In model 1, hydrogen formation resulting from VFA production were assumed to be totally consumed by methanogens for CH4 synthesis, without considering other pathways of hydrogen metabolism, which was the main factor resulting in the higher predicted values than the measured values.

Determination of GABA(Aα1) and GABA (B1) receptor subunits expression in tissues of gilts during the late gestation.

GABA(Aα1) and GABA(B1) receptor subunits are responsible for most behavioral, physiological and pharmacological effects of GABA receptors. We investigated the expression of GABA(Aα1) and GABA(B1) receptor subunits in different tissues of gilts during late pregnancy in hot summer. The mRNA abundance of GABA(Aα1) receptor subunit in different tissues of gilts at d 90 and d 110 of gestation was as follows: d 90: brain > lung > liver > ovary > spleen > kidney > heart; d 110: brain > lung > spleen > liver > ovary > kidney > heart. And, the mRNA abundance of GABA(B1) receptor subunit was as follows: d 90: spleen > lung > brain > kidney > ovary > liver > heart; d 110: spleen > lung > kidney > brain > ovary > liver > heart. The results in this trial indicated that the GABA(Aα1) receptor subunit was abundantly expressed in brain, while GABA(B1) receptor subunit was abundant in spleen and lung of gilts during late gestation. There were no gestation stage-dependent effects on GABA(Aα1) and GABA(B1) receptor subunits expression in all tissues.