Transportation is a common stressor in sheep production that is capable of inducing oxidative stress and impairing sheep health and production performance. This study aimed to investigate the alleviating effects of the traditional formula Siji Antiviral Mixture residue after water extraction, which still contains active ingredients, including fiber, polyphenols, and flavonoids, on short-distance transport stress in sheep, as well as its mechanism of action in regulating oxidative stress through the rumen microbiota-metabolism axis.

Twenty first-lambing East Friesian × Hu sheep hybrids weighing 54.49 ± 7.94 kg were randomly assigned to a control group (CON, basal diet) or a Chinese herbal residue group (CMR, basal diet + 50 g/d CMR) feeding at 4 h after approximately 300 km of short-distance transport. Results indicated that 4 h of short-distance transport significantly elevated serum reactive oxygen species (ROS) levels in sheep. Supplementation with Chinese herbal medicine residues markedly reduced serum ROS and lactate dehydrogenase levels while increasing glutathione peroxidase and immunoglobulin G levels. Metagenomic results revealed significantly increased abundance of bacteria such as Selenomonas ruminantium in the rumen of the CMR group, along with substantial increases in CAZymes, including AA7, GH113, and GH84. Metabolomic analysis revealed differentially expressed metabolites in plasma and rumen fluid that were enriched in metabolic pathways such as glycerophospholipid metabolism, α-linolenic acid metabolism, and drug metabolism-cytochrome P450. Correlation network analysis further revealed that Selenomonas ruminantium was significantly negatively correlated with ROS and positively correlated with ruminal LysoPC (16:1(9Z)/0:0), plasma phosphatidylcholine, and key glycerophospholipid metabolism enzymes (e.g., EC 3.1.4.3, PLC). Glycerophospholipid metabolism exhibited synergistic regulatory interactions with arachidonic acid metabolism and drug metabolism-cytochrome P450 pathways.

This study confirmed that 4 h of short-distance transport can induce oxidative stress in sheep. Supplementing feed with Siji Antiviral Mixture herbal residue effectively alleviated transport stress and enhanced immune function. The mechanism of action involved rumen microbial conversion of the herbal residue, which substantially increased the abundance of Selenomonas ruminantium. Related metabolites then regulated host arachidonic acid metabolism and cytochrome P450 drug metabolism indirectly through the glycerophospholipid metabolic pathway and the rumen microbiota-metabolism axis, thereby synergistically exerting antioxidant effects.

Scutellaria baicalensis, a traditional Chinese medicine (TCM), has demonstrated significant therapeutic efficacy in treating respiratory diseases caused by Mycoplasma gallisepticum (MG). However, the effective components of Scutellaria baicalensis are complex, and the material basis for its efficacy anti-MG infection remains unclear. This study aims to elucidate the molecular mechanism by which Scutellaria baicalensis exosome-like nanoparticles (SBELNs) and the key effector molecule, miR159a, regulate inflammation-induced injury caused by MG infection.

SBELNs were isolated from Scutellaria baicalensis root by ultracentrifugation. The in vivo and in vitro transport of SBELNs was investigated through live imaging and laser confocal microscopy after staining with DIR fluorescent dye. Key miRNAs were screened via RNA sequencing, and target genes were predicted using online databases. The interaction between miR159a and its target gene, cyclic nucleotide-gated channel alpha 1 (CNGA1), was validated using a dual-luciferase reporter assay. Furthermore, the regulatory network of the miR159a/CNGA1 axis was systematically analyzed.

SBELNs can specifically target lung tissue. Subsequently, SBELNs release bioactive components that alleviate the lung inflammatory damage caused by MG infection. This beneficial effect stems from two aspects. Firstly, the flavonoid metabolites encapsulated in SBELNs directly suppress the inflammatory damage caused by MG infection. Secondly, the microRNA in SBELNs regulates calcium ion homeostasis via the miR159a/CNGA1 axis. This relieves the intracellular calcium overload induced by MG and participates in the regulation of the immune system by modulating calcium ions. The microRNA in SBELNs regulates calcium ion homeostasis through the miR159a/CNGA1 axis, thereby alleviating MG-induced intracellular calcium overload, mitochondrial damage, excessive ROS, and overactivation of the NF-κB inflammatory pathway.

This article expounds that SBELNs alleviate lung injury caused by MG infection by regulating calcium homeostasis. This discovery demonstrates the anti-infective capability SBELNs, but also supports the development of natural drug delivery systems.

Citrulline (Cit), an effective precursor of arginine (Arg), escapes hepatic catabolism to be almost completely absorbed into the systemic circulation, thereby being efficiently converted to Arg in the kidneys to enhance its systemic bioavailability. This study investigated the effects of dietary Cit supplementation on lactation performance in sows, as well as the underlying mechanisms related to intestinal health in their suckling piglets, using multi-omics analyses.

Dietary Arg and Cit supplementation significantly increased average daily feed intake of lactating sows. Milk fat content and plasma nitric oxide (NO) concentration increased significantly in the Arg group and the 40%Cit group (P < 0.05), while milk threonine content increased slightly (P = 0.084). Consequently, the average daily gain of suckling piglets over the 21-day lactation period was also significantly improved. Furthermore, maternal 40%Cit supplementation improved the intestinal health of offspring by enhancing jejunal morphology and upregulating the expression of the tight junction protein occludin (P < 0.05), indicating a strengthened intestinal barrier. Mechanistically, this was achieved by activating the mTOR/S6 pathway in the piglets' jejunum. Maternal 40%Cit supplementation upregulated the expression of proteins related to mitochondrial fusion and fission (MFN2 and MFF, P < 0.05), and the protein expression of OPA1 showed an increasing trend (P = 0.097), indicating the structural and functional status of mitochondria was improved. Maternal 40%Cit supplementation also modulated the gut microbiota of piglets, increasing the abundance of beneficial bacteria (Lachnoclostridium). Metabolomic analysis of sow milk identified 58 differential metabolites. Among these metabolites, palmitic acid levels were significantly increased and positively correlated with the abundance of Lachnoclostridium in the intestine (P < 0.05).

Dietary Cit supplementation enhanced sow lactation performance and improved intestinal barrier function in their offspring via activation of the jejunal mTOR/S6 pathway and improved mitochondrial structure and function in the piglet jejunum. These benefits were further supported by modulation of the gut microbiota and alterations in the milk fat and metabolome, ultimately promoting piglet growth.

Chronic exposure to low-dose lipopolysaccharide (LPS) in poultry farming environments induces persistent respiratory inflammation, resulting in lung injury and impaired growth performance in broilers. Bacillus velezensis (BV) is a probiotic with known antibacterial and immunomodulatory activities, yet its role in respiratory health remains poorly understood. This study aimed to assess the potential benefits of BV in alleviating chronic pneumonia triggered by LPS in broilers and to clarify its mechanistic pathways.

A chronic LPS intratracheal instillation model was established, comprising control, LPS, and BV + LPS groups. BV supplementation significantly ameliorated LPS-induced growth impairment (P < 0.05), inhibited the synthesis of key inflammatory mediators, and mitigated oxidative stress in serum and bronchoalveolar lavage fluid (P < 0.05). Integrated multi-omics analyses revealed that BV remodeled the pulmonary microbiota, enriching isoflavone-metabolizing taxa including Blautia and unclassified Lachnospiraceae (P < 0.05), which was associated with elevated pulmonary concentrations of daidzein, genistein, and glycitein (P < 0.05). Transcriptomic together with molecular analyses revealed that BV enhanced the activation of PPAR-γ while attenuating NF-κB pathway activity, thereby reducing the expression of genes associated with inflammation (P < 0.05). In vitro, experiments showed that daidzein and genistein inhibited cellular inflammatory responses through PPAR-γ signaling. BV culture supernatant directly suppressed NF-κB/NLRP3 inflammasome activation in chicken HD11 macrophages, reduced intracellular reactive oxygen species (ROS) generation, and shifted macrophage polarization toward an anti-inflammatory phenotype (P < 0.05).

These findings demonstrate that BV alleviates LPS-induced chronic pneumonia through two complementary pathways, as it remodels the pulmonary microbiota to enhance isoflavone metabolism and thereby suppress inflammation, while its own metabolites also directly inhibit inflammatory signaling. This study provides new insight into probiotic-based interventions for respiratory health in livestock.

Mastitis, one of the most prevalent inflammatory diseases in female mammals, causes significant economic losses in livestock farming. Notably, the natural flavonoid compound baicalin exhibits potent anti-inflammatory activity. However, its efficacy in alleviating mastitis severity and the underlying molecular mechanisms remain unexplored. Therefore, this study aims to investigate the protective effects of baicalin in alleviating mastitis and its key molecular mechanisms.

This study demonstrated in vivo that baicalin effectively alleviates mastitis symptoms in dairy cows and mice, primarily manifested by reduced tissue pathological damage, decreased levels of pro-inflammatory cytokines, and maintain the integrity of the blood-milk barrier (BMB). Multi-omics sequencing analysis indicated that IL-17 and TNF signaling pathways play crucial roles in this process. Further studies demonstrated that IL-17RA^-/- mice exhibited a phenotype similar to that observed with baicalin treatment, confirming the importance of this pathway. Notably, network pharmacology screening combined with molecular dynamics simulations revealed stable binding of baicalin to IL-17RA, suggesting that baicalin exerts its protective effect to alleviate mastitis by targeting IL-17RA. Mechanistically, both baicalin treatment and IL-17RA deletion block activation of key downstream pathways of the IL-17 signaling pathway, including MAPK, ERK and NF-κB, thereby suppressing excessive activation of the TNF signaling pathway, preventing exacerbation of the inflammatory response and barrier damage.

In conclusion, this study demonstrates that baicalin inhibits excessive activation of the IL-17/TNF signaling pathway by targeting IL-17RA, thereby reducing inflammatory responses and BMB damage within the mammary gland and alleviating mastitis severity.

Under high-concentrate feeding conditions, ruminants often experience rumen microecological imbalance and dysfunction, which can impair growth performance and increase the risk of antibiotic resistance gene (ARG) dissemination.

To evaluate the ameliorative effects of Aspergillus niger (A. niger) cultures, fattening sheep were randomly allocated into the following five groups: a control group (CON), a control diet supplemented with 250, 500, or 1,000 mg/kg A. niger cultures (designated as LA, MA, and HA, respectively); and an antibiotic group supplemented with 5,000 mg/kg chlortetracycline premix (AN). Microbial community analysis indicated that several bacterial taxa, including Succinivibrio sp900317105, Prevotella sp002353485, Quinella sp017515635, Quinella sp015206805, and Prevotella sp900320255, were significantly enriched in the A. niger culture-supplemented groups (P < 0.05). ARG profiling showed that the abundance of tetracycline resistance genes was significantly lower in all A. niger groups compared with the CON and AN groups (P < 0.05), while β-lactam resistance genes were significantly reduced in the HA group (P < 0.05). Furthermore, the abundances of Rank I and Rank II ARGs were significantly higher in the AN group than in the other groups, whereas the abundances of Rank II and Rank IV ARGs were significantly lower in the A. niger culture groups than in the CON and AN groups. Metabolomic analysis further demonstrated that supplementation with A. niger cultures significantly decreased the concentration of N-decanoyl-L-homoserine lactone (P < 0.05) while increasing the levels of N-3-oxotetradec-7Z-enoyl-L-homoserine lactone, indole-3-methyl acetate, and indole-3-propionic acid (P < 0.05).

These findings suggest that A. niger cultures can reduce the abundance of ARGs and mitigate the risk of ARG dissemination by modulating the rumen microbial community and associated metabolites.

The overuse of antibiotics in both veterinary and human medicine has resulted in the emergence of antibiotic-resistant bacteria, prompting a search for effective alternatives. Antimicrobial peptides (AMPs) are short, often cationic, peptide-based molecules with antimicrobial and immunomodulatory activity, which makes them promising alternatives to conventional antibiotics in poultry production.

From a prior machine-learning-guided screen of 875 candidate AMPs against a wide bacterial panel, 62 exhibited activity against avian pathogenic Escherichia coli (APEC) and low in vitro hemolytic and cytotoxic activity. We selected three lead AMPs from this list (named TeRu4, TeBi1, and PeNi4), and evaluated their in vitro and in vivo efficacy, safety, and immunomodulatory potential for use in poultry farming. In animal experiments, AMPs were administered via in ovo injection on d 18 of embryonic development. In APEC challenge trials, yolk sacs were inoculated with APEC post-hatch to assess early chick mortality, while in pen trials, birds were raised in a commercial production setting for 35 d. For challenged birds, TeBi1 (10 μg/egg) significantly reduced culture-positive rates for APEC in the air sac and pericardium, increased body weight by 50% and reduced cytokine transcript levels by 10%-30% on d 7 post hatch. In HD11 chicken macrophage-like cultured cells, TeRu4 (16 μg/mL) suppressed lipopolysaccharide (LPS)-induced pro-inflammatory cytokine transcript levels. In pen trials, TeRu4 (20 μg/egg) increased the survival probability of female birds by 4.9%, while TeBi1 (20 μg/egg) increased the survival probability of all birds by 4.4%, by d 35. Gene expression analysis revealed AMP- and sex-specific cytokine responses. In pen trials, no significant differences were observed in mean weights, feed conversion ratio (FCR), and flock uniformity on d 35. By integrating high-throughput in ovo automation with large-scale commercial pen trials, this study provides a systematic translational bridge from in silico AI discovery to field-relevant poultry production interventions.

These findings demonstrate that TeBi1 and TeRu4 are promising antibiotic alternatives that improve survival, modulate immune responses, and maintain normal growth performance in broiler chickens in this experimental setting.

Weaning piglets are highly susceptible to enterotoxigenic Escherichia coli (ETEC) infections, which can cause intestinal barrier function dysfunction and death. However, there is still a lack of efficient, economical, and safe nutritional interventions. This study aimed to investigate the effects of combining butyrate with niacin on intestinal barrier function repair and resistance to ETEC infection in weaned piglets. In this study, two 14-d animal experiments were designed to observe the optimal butyrate-to-niacin ratio and assess their responses to the ETEC challenge.

Supplementation with butyrate and niacin at a ratio of 100:2 (2,000 mg/kg butyrate and 40 mg/kg niacin, BN2) increased the average daily gain (ADG) and reduced the diarrhea incidence. We also observed an increase in the levels of nicotinamide adenine dinucleotide (NAD) in the colon of weaned piglets. Notably, BN2 promoted amino acid anabolism in the colon and enhanced glycolysis and the tricarboxylic acid (TCA) cycle by increasing the acetylation of key enzymes in the TCA. Furthermore, BN2 enhanced the expression of indispensable genes for the colonic mucosal barrier, including antimicrobial peptides such as porcine β defensin 1 (pBD1), porcine β defensin 2 (pBD2), and proline-arginine rich 39-amino acid peptide (PR39), tight junction proteins, and improved colonic microbiome composition. Based on these findings, we found that BN2 alleviated growth restriction and diarrhea, and modulated the expression of antimicrobial peptides, tight junction proteins, and cytokines to reduce colonic barrier function dysfunction in weaned piglets challenged with ETEC. Mechanistically, we confirmed that BN2 elevated the protein expression of acetylation of histone 3 lysin 27 (H3K27ac) and enhanced the binding of acH3K27 to the promoter regions of pBD1 and PR39.

Supplementation with BN2 improved growth performance, supported colonic barrier function repair, and enhanced disease resistance in weaned piglets challenged with ETEC. This offers new insights into nutritional strategies for intestinal barrier function repair of piglets infected with ETEC.

Baicalin is a bioactive flavonoid from Scutellaria baicalensis Georgi with antioxidant, anti-inflammatory and antibacterial properties. However, its bitter taste and susceptibility to ruminal degradation limit its practical use in mammals. Enteric coating technology might overcome these limitations by enabling targeted intestinal release. This study investigated effects of dietary supplementation with baicalin and coated baicalin on rumen fermentation, gastrointestinal microbiota, immune function and growth performance in Hu sheep.

Thirty-six lambs with similar body weight (33.01 ± 2.68 kg) were randomly assigned to three groups (4 replicates per group, 3 sheep per replicate). The control group was fed basal diet (CON) while treatment I (BAI) and treatment II (C-BAI) groups were fed a basal diet supplemented with 0.1% baicalin and coated baicalin, respectively. After a 60-d feeding trial, baicalin and coated baicalin supplementation improved total weight gain and average daily gain compared with CON group (P < 0.05). In addition, BAI and C-BAI groups exhibited higher total antioxidant capacity (P < 0.05) and catalase activity (P < 0.05) with associated lower malondialdehyde levels (P < 0.05). Immunoglobulin G and anti-inflammatory cytokines interleukin-4 (IL-4) were also increased (P < 0.001). Notably, IgM, IL-10 and IL-4 in the C-BAI group exceeded those of the BAI group (P < 0.001). Microbiome analysis revealed that baicalin supplementation enriched abundance of beneficial bacterial taxa including Firmicutes and Lachnoclostridium (P < 0.05) and reduced potential pathogen abundance, e.g., Treponema and Ralstonia (P < 0.05). The C-BAI group also showed increased abundance of the beneficial Bradyrhizobium compared with CON (P < 0.05). Metabolomic analysis revealed that baicalin altered propionate and tyrosine metabolic pathways (P < 0.05), while coated baicalin modulated penicillin metabolism and glyceride metabolism in jejunum (P < 0.05) increasing ATP production. Overall, these results indicated enhanced nutrient metabolism and gut health in the presence of dietary baicalin.

Dietary supplementation with baicalin and coated baicalin improved growth performance, antioxidant status, immunity and beneficially modulated the microbiome-metabolome crosstalk in Hu sheep. Notably, uncoated baicalin exerted more pronounced effects on growth performance and supported a role for baicalin as a potential and functional feed additive.

The regulatory effects of glucose absorption at different sites (rumen vs. small intestine) on lipid metabolism exhibit significant variation in beef nutrition. This study aimed to investigate the regulatory pathways of rumenprotected or unprotected glucose on lipid metabolism through the rumen-jejunum axis in Xinjiang Brown cattle.

Thirty Xinjiang Brown cattle (females) with similar initial weight (410 ± 22.4 kg) were randomly assigned to 3 treatment groups (n = 10 animals per group). The experimental groups were fed a basal diet with the following daily supplements per head: 150 g palmitate coating (CON group), 150 g palmitate coating plus 150 g rumen-unprotected glucose (RUG group), and 300 g rumen-protected glucose (containing 50% glucose; RPG group). The experiment lasted for 70 d. Supplementation with both rumen-unprotected glucose and rumen-protected glucose increased chest width (P = 0.001), chest girth (P = 0.013), abdominal girth (P = 0.002), backfat thickness (P = 0.041), omental fat weight (P = 0.047), as well as serum concentrations of insulin (P < 0.001), glucagon-like peptide-1 (P < 0.001), and 5-hydroxytryptamine (P < 0.001), while decreasing the content of deoxycholic acid (P < 0.001) in the jejunum. The inclusion of rumen-unprotected glucose resulted in a trend toward higher intramuscular fat (IMF) (P = 0.064) in the longissimus thoracis, along with significant increases in C14∶0 (P = 0.042), C15∶0 (P = 0.014), and marbling score (P = 0.048), as well as significant reductions in drip loss (P = 0.022) and shear force (P = 0.041). These changes were accompanied by significant increases in dry matter intake (P = 0.001), ruminal concentrations of acetate (P = 0.022) and propionate (P = 0.011). The rumen-protected glucose supplementation elevated serum glucose (P < 0.001) level, while enhancing digestibility of ether extract (P = 0.027) and neutral detergent fiber (P = 0.027). Neither rumen-unprotected glucose nor rumen-protected glucose affected the alpha diversity and beta diversity of ruminal and jejunal microbiota (P > 0.05), but the differential bacterial biomarkers were either positively or negatively correlated with chest girth, abdominal girth, marbling score, backfat thickness, and deoxycholic acid (P < 0.05).

Rumen-unprotected glucose supplementation enhances IMF deposition and meat quality attributes through microbial volatile fatty acid-driven metabolic reprogramming.