This study systematically analyzed the dynamic changes of milk fat globule membrane (MFGM) proteins in breast milk across different lactation stages. Using data-independent acquisition (DIA)-based quantitative proteomics, a total of 6445 MFGM proteins were identified. The total amount of MFGM proteins showed a significant decline over time. Bioinformatics analysis of differentially expressed proteins revealed that breast milk MFGM not only provides nutritional support but also plays multifunctional roles in regulating infant immunity, neurodevelopment, and gut health. Colostrum, rich in immuno-related proteins such as lactoferrin and IgA, facilitates gut microbiota colonization and establishes the newborn immune barrier. Transitional milk and mature milk progressively shift toward nutritional support and health maintenance, aligned with infant developmental stages. Through comprehensive analytical approaches, this study thoroughly elucidated the dynamic patterns of functional components in breast milk, providing a reference for optimizing the precise nutritional formulation of infant formula.

This study aims to explore the complex interactions between microbial communities and flavor characteristics in Mongolian cheese from five distinct geographical origins in Inner Mongolia. Through the integration of 16S rDNA/ITS high-throughput sequencing with SPME-GC-MS and UHPLC-MS technologies, comprehensive characterization was performed on bacterial and fungal compositions, as well as volatile organic compounds, free amino acids, and free fatty acids. Results indicated that Firmicutes was the dominant phylum and Lactococcus was the dominant genus. alpha- and beta-diversity analysis revealed that geographical location significantly influenced microbial structure, with samples from Xilingol League exhibiting the highest diversity. 91 flavor compounds were identified, including both non-volatile and volatile substances, among which organic acids were the primary contributors to the sour taste of Mongolian cheese. Orthogonal partial least squares (O2PLS) correlation analysis identified 8 bacterial genera and 14 fungal genera as the functional core microbiota, which were associated with the formation of key flavor compounds in Mongolian cheese. Acetobacter and Flavobacterium were positively associated with a variety of volatile compounds (P < 0.05), and Brochothrix and Kazachstania had strong correlations with several free amino acid catabolism (P < 0.05). This study has deeply explored the relationship between the microbial community and flavor compounds of Mongolian cheese, providing reference value for microbial regulation and the maintenance of traditional flavor characteristics.

Long-term maize storage causes oxidative deterioration, but its effects on intestinal redox status, systemic inflammation, and liver-related responses in ruminants remain unclear. Laminaria japonica polysaccharide (LJP) has antioxidant, immunomodulatory, and microbiota-regulating properties, but its efficacy during aged-maize feeding is unknown. This study evaluated whether LJP mitigates oxidative and inflammatory responses in Hu sheep fed aged maize and characterized cecal microbiome and metabolome alterations. Twenty-one Hu sheep (39.05 ± 3.55 kg) were assigned to three diets (n = 7) and fed for 10 weeks (a 14-day adaptation period followed by 8 weeks of treatment): normal maize (CK), aged maize (AM), or aged maize with 0.5% LJP (AML). Compared with CK, AM increased plasma lipopolysaccharide (0.428 vs. 0.379 EU/mL), TNF-α, and IL-1β, and raised ileal reactive oxygen species (248.79 vs. 166.23 fluorescence intensity/mg; p < 0.001) and malondialdehyde (1.87 vs. 1.63 nmol/L; p = 0.006), consistent with systemic inflammation and intestinal oxidative stress. AML lowered these inflammatory and oxidative indices and increased hepatic T-AOC (p = 0.009) and catalase activity (p = 0.013). Integrated 16S rRNA and untargeted metabolomic analysis revealed treatment-associated cecal microbe-metabolite associations. These findings indicate that aged-maize feeding was associated with intestinal and systemic redox-inflammatory changes in Hu sheep, whereas dietary LJP was associated with partial mitigation, potentially involving microbial and metabolic remodeling.

Early-life nutrition is critical for the development and health of dairy calves, necessitating alternatives to in-feed antibiotics. This study investigated whether dietary glycerol monolaurate (GML) enhances growth performance, reduces diarrhea incidence, improves systemic antioxidant and immune status, and modulates hepatic immunometabolic function in calves. Twenty-four Holstein bull calves (7 ± 0.5 d of age) were randomly assigned by body weight and age to a control group or a GML-supplemented group, both fed milk replacer with starter feed provided throughout the 45-day trial. Calves in the GML group received GML at a dosage of 100 mg/kg of body weight, mixed into the milk replacer prior to feeding. Calves in the GML group had significantly greater final body weight, average daily gain, and starter intake during the latter period (d 23–45) compared with the control group. GML supplementation also significantly reduced the incidence of diarrhea and fever, alongside lower fecal scores and fewer antibiotic treatments. Plasma analysis revealed enhanced antioxidant capacity, as indicated by increased total antioxidant capacity and glutathione peroxidase, along with an improved immune profile characterized by elevated immunoglobulin G and reduced interleukin-2. Transcriptomic analysis of the liver showed that GML upregulated genes and pathways related to innate antiviral immunity, such as radical S-adenosyl methionine domain containing 2, interferon-stimulated gene 15, and MX dynamin like GTPase 1. Lipidomics further indicated that GML induced a targeted remodeling of hepatic lipids, including increased diacylglycerols and triacylglycerols and decreased specific phospholipids and sphingolipids, suggesting a metabolic shift supportive of immune activation and inflammatory control. In conclusion, dietary GML enhances growth and health in suckling calves, which is mediated through a coordinated immunometabolic reprogramming in the liver. GML represents a promising functional fat additive for sustainable calf rearing.

Microplastic pollution is increasingly serious worldwide, threatening human and animal health. The cow rumen is a key organ for nutrient digestion and absorption, and its fermentation is closely related to rumen microorganisms. Here, we investigated how polystyrene microplastics (PS-MPs) with varying particle sizes and concentrations affect rumen fermentation and the biodegradability of PS-MPs by rumen fermentation. The results reveal that exposure to PS-MPs lowered gas production and gas concentrations, as well as volatile fatty acid content, and these decreases were positively correlated with PS-MP concentration. However, higher PS-MP concentration and larger particle size increased the activity of carboxymethyl cellulose, β-glucosidase, and xylanase. Furthermore, PS-MP exposure reduced the abundance of certain rumen microorganisms and altered metabolic pathways and metabolites linked to PS-MP biodegradation. It was also found that PS-MP content decreased significantly after 24 h fermentation. Therefore, PS-MPs can inhibit rumen fermentation by affecting the rumen microbiome, and rumen microorganisms and their secreted enzymes can biodegrade PS-MPs to produce styrene and derivatives; such small molecules may further disrupt rumen homeostasis, thereby affecting lactation performance. In addition, rumen microbial degradation of PS-MPs provides a new idea to resolve future microplastic contamination challenges.

Long-term storage can impair the feeding value of corn. This study evaluated whether dietary Laminaria japonica polysaccharide (LJP) could alleviate the adverse effects of aged corn in Hu sheep. Twenty-one male Hu sheep (39.05 ± 3.55 kg) were assigned to a normal-corn diet (NC), an aged-corn diet (AC), or the AC diet supplemented with 0.5% LJP (AC+LJP; n = 7) for 70 d, including a 14-d adaptation period. Compared with NC, AC reduced average daily gain, gain-to-feed ratio, the apparent digestibility of crude protein, neutral detergent fiber and acid detergent fiber, ruminal total volatile fatty acids, acetate and butyrate, and muscle glutathione peroxidase activity. LJP supplementation restored average daily gain, gain-to-feed ratio, and crude protein and neutral detergent fiber digestibility to levels comparable with NC. It also increased ruminal ammonia nitrogen, microbial crude protein, total volatile fatty acids, acetate, butyrate, and isobutyrate relative to AC, while reducing muscle malondialdehyde concentration and increasing total antioxidant capacity. Dietary treatment had little effect on carcass traits, most meat-quality variables, or overall rumen bacterial diversity. In conclusion, supplementation with 0.5% LJP partially alleviated the decline in nutrient utilization and rumen fermentation associated with aged corn and improved muscle antioxidant status. These findings support the potential use of LJP as a functional feed additive to improve the utilization of aged-corn-based diets in sheep.

The milk fat globule membrane (MFGM) is rich in bioactive lipids essential for infants. Using Hebei as a reference region, this study investigated regional differences in MFGM phospholipid composition in human milk from Northeast China, where dietary patterns vary, including differences in fat sources and seafood intake. The results showed that although significant regional differences were observed in milk fat globule size and zeta potential, the overall microstructure remained largely consistent. Oleic acid (C18:1) was the predominant fatty acid across all regions. Hebei exhibited the highest saturated fatty acid content (48.85 +/- 0.07%), whereas Liaoning showed the lowest saturated fatty acids (37.97 +/- 0.11%). Phosphatidylcholine (PC) was dominant in Hebei and Liaoning, whereas sphingomyelin predominated in Heilongjiang. Orthogonal projections to latent structures-discriminant analysis identified 69-84 differential phospholipids among regions, with PC contributing most to regional discrimination. Network analysis revealed strong correlations between differential lipids and key hub molecules. These findings also provide a scientific basis for optimizing infant formula.

This study explored the generation characteristics, distribution of hydrolysis sites and potential pathways of action of antioxidant peptides derived from casein hydrolysates of different Lactobacillus casei. Compared with L. casei 1.0319, L. casei 1.0351 exhibited stronger casein hydrolytic activity, achieving a hydrolysis degree of (17.81 +/- 1.23)% within 48 h. Its casein hydrolysates exhibited strong antioxidant activity. Ultrafiltration separation revealed that peptides with molecular weights <3 kDa (F1 fraction) exhibited the strongest antioxidant activity. Further HPLC analysis identified 444 peptides from this fraction, primarily derived from alpha(S1)-casein and beta-casein. Bioinformatics screening identified 10 potentially high-activity antioxidant peptides (GPVRGPFP, MPIQAFLL, GPVRGPFPII, FPKYPVEPF, APFPEVFGK, HLPLPLL, NQFLPYPY, PHPHLSF, LHLPLPLL, PVVVPPFL). Network pharmacology analysis suggested their actions may involve key targets such as STAT3, CTSB, and ACE. KEGG pathway enrichment indicated these peptides may exert antioxidant effects by regulating pathways related to cardiovascular disease and apoptosis. Molecular docking results predicted that the peptide PVVVPPFL exhibits strong binding affinity with ACE (-9.3 kcal/mol), suggesting it may alleviate oxidative stress by inhibiting ACE activity. This study provides a theoretical basis for deepening the understanding of the generation mechanism of lactic acid bacteria-derived antioxidant peptides and their application in functional food development.

As a major contributor to methane production in agriculture, there is a need for a suitable methane inhibitor to reduce ruminant methane emissions and minimize the impact on the climate. This work aimed to explore the influence of cordycepin on rumen fermentation, gas production, microbiome and their metabolites. A total of 0.00, 0.08, 0.16, 0.32, and 0.64 g L-1 cordycepin were added into fermentation bottles containing 2 g total mixed ration for in vitro ruminal fermentation, and then the gas produced and fermentation parameters were measured for each bottle. Samples from the 0 and 0.64 g L-1 cordycepin addition were selected for 16S rRNA gene sequencing and metabolome analysis. The result of this experiment indicated that the addition of cordycepin could linearly increase the concentration of total volatile fatty acid, ammonia nitrogen, the proportion of propionate, valerate, and isovalerate, and linearly reduce ruminal pH and methane, carbon dioxide, hydrogen and total gas production, as well as the methane proportion, carbon dioxide proportion and proportion of butyrate. In addition, there was a quadratic relationship between hydrogen and cordycepin addition. At the same time, the relative abundance of Succiniclasticum, Prevotella, Rikenellaceae_RC9_gut_group, NK4A214_group, Christensenellaceae_R_7_ group, unclassified_F082, Veillonellaceae_UCG_001, Dasytricha, Ophryoscolex, Isotricha, unclassified_Eukaryota, Methanobrevibacter, and Piromyces decreased significantly after adding the maximum dose of cordycepin. In contrast, the relative abundance of Succinivibrio, unclassified_Succinivibrionaceae, Prevotellaceae_UCG_001, unclassified_Lachnospiraceae, Lachnospira, Succinivibrionaceae_UCG_002, Pseudobutyrivibrio, Entodinium, Polyplastron, unclassified_Methanomethylophilaceae, Methanosphaera, and Candidatus_Methanomethylophilus increased significantly. Metabolic pathways such as biosynthesis of unsaturated fatty acids and purine metabolism and metabolites such as arachidonic acid, adenine, and 2"-deoxyguanosine were also affected by the addition of cordycepin. Based on this, we conclude that cordycepin is an effective methane emission inhibitor that can change the rumen metabolites and fermentation parameters by influencing the rumen microbiome, thus regulating rumen methane production. This experiment may provide a potential theoretical reference for developing Cordyceps byproduct or additives containing cordycepin as methane inhibitors.

This study examined branched-chain fatty acids (BCFAs)’ effects on oxidative stress, energy metabolism, inflammation, tight junction disruption, apoptosis, and Toll-like receptor 4/nuclear factor kappa-B (TLR4/NF-κB) signaling in lipopolysaccharide (LPS)-induced calf small intestinal epithelial cells (CSIECs). Eight groups were used: a control group, an LPS-induced group, and six BCFA treatment groups (12-methyltridecanoic acid (iso-C14:0), 13-methyltetradecanoic acid (iso-C15:0), 14-methylpentadecanoic acid (iso-C16:0), 15-methylhexadecanoic acid (iso-C17:0), 12-methyltetradecanoic acid (anteiso-C15:0), and 14-methylhexadecanoic acid (anteiso-C17:0)) with LPS. The BCFA pretreatments significantly increased CSIEC activity compared to the LPS-induced group, with iso-C14:0 showing the highest activity (89.73%). BCFA reduced Reactive Oxygen Species (ROS) generation and malondialdehyde (MDA) levels and improved the superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) activities and glutathione (GSH) levels. Iso-C16:0 optimized total antioxidant capacity (T-AOC). BCFA enhanced the mitochondrial membrane potential, Adenosine Triphosphate (ATP) enzyme activity, and ATP content, with iso-C14:0 increasing ATP by 27.01%. BCFA downregulated interleukin (IL)-1β, IL-8, tumor necrosis factor (TNF)-α, and interferon (INF)-γ gene expression, reduced IL-6 levels, and increased IL-10 expression. Myeloid differentiation factor 88 (MyD88) mRNA levels were reduced. BCFA alleviated Zonula Occludin (ZO-1), Claudin-1, and Claudin-4 decrease and increased Occludin levels. BCFA mitigated LPS-induced increases in Caspase-3 and BCL2-Associated X (BAX) mRNA levels, reduced Caspase-8 and Caspase-9 expression, and increased B-Cell Lymphoma-2 (BCL-2) mRNA levels. The Entropy Weight-TOPSIS method was adopted, and it was discovered that iso-C15:0 has the best effect. In summary, BCFA supplementation mitigated oxidative stress and enhanced mitochondrial function. BCFA inhibited TLR4/NF-κB signaling pathway overactivation, regulated inflammatory cytokine gene expression, reduced cellular apoptosis, preserved tight junction integrity, and supported barrier function.

Objective The study aimed to assess effects of supplemented co-fermented edible plants and probiotics (AEPP) on growth performance, disease resistance, plasma and rumen metabolites, and bacterial communities in the rumen and feces of pre-weaned calves. Methods Twenty female Holstein calves (7±0.50 d, 41.65±6.20 kg) were randomly assigned to one of two treatments: the control group or the treatment group (30 g/head/day AEPP supplementation). Growth performance, blood, and fecal samples were measured on regular basis. On day 30 of the trial, rumen fluid and fecal samples were collected for multi-omics analysis. Results Dietary supplementation with AEPP enhanced calf growth and improved disease resistance, as evidenced by a reduced incidence of respiratory disease and diarrhea and a decreased frequency of antibiotic therapy (p<0.05). The treatment group exhibited enrichment of rumen microorganisms Prevotella, Ruminococcus, and Xylanibacter (linear discriminant analysis>2, p<0.05), along with increased activity in beneficial metabolites such as indoleacetic acid, which activated starch and sucrose metabolism and tryptophan metabolism pathway. This intervetion significantly improved average daily gain, feed efficiency, immunoglobulin G, total superoxide dismutase, and glutathione peroxidase activities, as well as significantly reduced levels of tumor necrosis factor-alpha and interleukin-6 (p<0.05), promoting calf growth and health. The elevated abundance of fecal microorganisms, Subdoligranulum and Bifidobacterium, in the treatment group altered fecal pH, short-chain fatty acids, and butyrate proportions (p<0.05). Conclusion Feeding AEPP improved growth performance, disease resistance, and antioxidant function. It altered the bacterial communities and metabolic profiles in the rumen and feces of preweaning dairy calves, providing a data reference for the use of AEPP in young ruminant production.

Hepatic oxidative stress is a key driver in liver injury pathogenesis, with D-galactose (D-gal) modeling serving as an established inducer of accelerated oxidative damage. Silibinin (SLB), a flavonolignan from milk thistle, shows therapeutic promise through potent antioxidant activity and gut-liver axis modulation. This study investigated whether the hepatoprotective effect of SLB against oxidative stress depends on gut microbiota regulation. Using mouse models with gut microbiota ablation by oral antibiotics or direct oxidative stress induction by D-gal (150 mg/kg), SLB treatment (200 mg/kg) was administered. The protective mechanisms were evaluated through the Nrf2/ARE pathway, target gene expression, gut microbiota profiling, and cecal metabolomics. Results demonstrated that SLB significantly alleviated D-gal-induced hepatic oxidative stress (e.g., reduced MDA by 33.3%), but this protection was markedly weakened after antibiotic-induced microbiota depletion (e.g., a loss of efficacy exceeding 50%). Integrated omics revealed that antibiotics caused a severe reduction in unclassified_Muribaculaceae (a butyrate producer, decreased by 80%), impairing butyrate-mediated Nrf2/Keap1 activation. Simultaneously, the absence of Parabacteroides led to accumulated primary bile acids and inhibited secondary bile acid production (e.g., taurochenodeoxycholate reduced by 75%), further disrupting redox homeostasis. Conclusion: Silibinin's mitigation of hepatic oxidative stress is gut microbiota-dependent, highlighting the therapeutic potential of microbiota-targeted antioxidant strategies for oxidative stress-related pathologies.

Background The objective of this study was to evaluate the effects of dietary fatty acids（FA） saturation and lysophospholipids supplementation on growth, meat quality, oxidative stability, FA profiles, and lipid metabolism of finishing beef bulls. Thirty-two Angus bulls（initial body weight: 623 ± 22.6 kg; 21 ± 0.5 months of age） were used. The experiment was a completely randomized block design with a 2 × 2 factorial arrangement of treatments: 2 diets with FA of different degree of unsaturation [high saturated FA diet（HSFA） vs. high unsaturated FA diet（HUFA）] combined with（0.075%, dry matter basis） and without lysophospholipids supplementation. The bulls were fed a high-concentrate diet（forage to concentrate, 15:85） for 104 d including a 14-d adaptation period and a 90-d data and sample collection period.Results No interactions were observed between dietary FA and lysophospholipids supplementation for growth and meat quality parameters. A greater dietary ratio of unsaturated FA（UFA） to saturated FA（SFA） from 1:2 to 1:1 led to lower DM intake and backfat thickness, but did not affect growth performance and other carcass traits. Compared with HSFA, bulls fed HUFA had greater shear force in Longissimus thoracis（LT） muscle, but had lower intramuscular fat（IMF） content and SOD content in LT muscle. Compared with HUFA, feeding the HSFA diet up-regulated expression of ACC, FAS, PPARγ, and SCD1, but down-regulated expression of CPT1B. Compared with feeding HSFA, the HUFA diet led to greater concentrations of c9-C18:1 and other monounsaturated FA in LT muscle. Feeding HUFA also led to lower plasma concentrations of cholesterol, but there were no interactions between FA and lysophospholipids detected. Feeding lysophospholipids improved growth and feed conversion ratio and altered meat quality by increasing muscle pH 24h , redness values（24 h）, IMF content, and concentrations of C18:3, C20:5 and total polyunsaturated fatty acids. Furthermore, lysophospholipids supplementation led to lower malondialdehyde content and up-regulated the expression of ACC, FAS, and LPL in LT muscle.Conclusions Results indicated that supplementing a high-concentrate diet with lysophospholipids to beef bulls can enhance growth rate, feed efficiency, meat quality, and beneficial FA. Increasing the dietary ratio of UFA to SFA reduced DM intake and backfat thickness without compromising growth, suggesting potential improvements in feed efficiency.

Thermal treatment ensures food safety, but high temperatures affect the structure and activity of protein-polyphenol complexes. This study used whey protein isolate (WPI) to examine the impact of thermal treatments (pasteurization and sterilization) on the interactions between WPI with varying degrees of enzymatic hydrolysis (WPIHs) and epigallocatechin gallate (EGCG), and the functional properties of the formed complexes. The results indicated that thermal treatment significantly altered the protein structure, promoting aggregation and unfolding. The binding of WPI/WPIHs to EGCG follows a static quenching mechanism, with hydrophobic interactions playing a dominant role. Sterilization (121 °C, 15 min) shifted the primary interaction force of partially hydrolyzed WPI (WPIH1) to electrostatic interactions, while transforming highly hydrolyzed WPI (WPIH2) to van der Waals forces and hydrogen bonds. The binding capacity of WPIH1 with EGCG can reach 3.798 × 105 ± 0.084 × 105 M-1. Under thermal treatment, the number of binding sites was consistently one. In terms of functional properties, WPIH1 combined with EGCG exhibited better antioxidant activity, enzyme inhibitory capacity and emulsifying properties. Specifically, the WPIH1-EGCG complex prepared at 65 °C showed the highest ABTS (94.16 % ± 0.33 %) and DPPH (84.55 % ± 0.45 %) scavenging rates, with α-glucosidase (72.17 % ± 1.00 %) and α-amylase (69.70 % ± 0.46 %) inhibition, and its emulsifying properties increased by 47.49 %. In summary, appropriate thermal treatment and enzymatic hydrolysis render the functional properties of the complexes more stable. This study contributes to understanding the binding mechanism between WPI (varying hydrolysis degrees) and EGCG. It provides insights for optimizing thermal processing parameters of functional foods containing EGCG and whey protein derivatives.

The huge milk thistle meal (MTM) production requires proper processing. The study utilized response surface methodology to optimize fermentation conditions and assessed complex probiotics and cellulase impacts on nutrients, fermentation, digestibility, and in vitro ruminal features of fermented MTM (FMTM) for advancing clean bioresource technologies and feed utilization. Optimal solid-state fermentation conditions for maximum neutral detergent fiber reduction (14.25 %) were: 1.0 g/kg complex probiotics, 7.1 g/kg enzyme, 38 degrees C, 456.9 g/kg moisture, and 96 h. Under fermentation, the contents of crude protein (254.0 vs. 268.8 g/kg DM; P = 0.002), total amino acid (201.7 vs. 249.2 g/kg DM; P < 0.001), ammonia-N (9.92 vs. 19.2 g/kg TN; P < 0.001), and in situ degradation rates of DM (371.0 vs. 410.9 g/kg; P = 0.006) and crude protein (576.7 vs. 614.2 g/kg; P < 0.001) of FMTM were elevated. In vitro gas production experiments have shown that compared with TMR including MTM, TMR involved FMTM had lower methane concentration (200.40 vs. 182.54 g/kg; P = 0.095), higher ammonia-N concentration (40.06 vs. 44.18 mg/dL; P < 0.001), total volatile fatty acid production (79.72 vs. 83.63 mmol/L; P = 0.0031), and molar ratios of propionate (24.41 vs. 24.96 %; P = 0.0002) in the rumen. Moreover, TMR containing FMTM increased microbial diversity, and enrichment of norank_f__F082, Succiniclasticum, Christensenellaceae_R_7_group, Ruminococcus, NK4A214_group, and unclassified_f__Lachnospiraceae. In conclusion, solid-state fermentation using complex probiotics and cellulose enables sustainable MTM utilization, making it a high-quality protein source for ruminants and providing a reference for other Chinese herb residue applications.

This study investigated the effects of Clostridium butyricum (C. butyricum) supplementation on the bacterial community composition and fermentation characteristics of the digestive system, assessed blood antioxidant capacity, and evaluated growth performance and digestibility in pre-weaning Angus calves. Sixteen male Angus calves were randomly divided into two groups (n = 8). The control group was fed a basal diet, while the other group was supplemented with 2 × 107 colony-forming unit/d C. butyricum in the basal diet. After 8 weeks of feeding, body growth and nutrient intake were measured. Rumen fluid samples, blood samples, and fecal samples were collected and analyzed. Fecal scores were recorded daily, and the frequency of diarrhea treatments was documented. The results showed C. butyricum supplementation improved the digestibility of neutral detergent fiber, crude protein, and ether extract in calves, as well as the growth hormone content in the blood. Furthermore, supplementation elevated isovaleric acid concentrations in the rumen, modulated the microbiome, and increased pre-weaning body weight and average daily gain of calves. Meanwhile, C. butyricum supplementation reduced myeloperoxidase and superoxide dismutase levels while increasing total antioxidant capacity in the blood, alleviating oxidative stress, and enhancing the antioxidant capacity of the body. Intestinal health was also improved, as evidenced by an increase in butyrate content, a reduction in enterotoxigenic Escherichia coli, lower fecal scores, and a decreased need for diarrhea treatments. In summary, dietary C. butyricum supplementation may support digestive system development, enhance antioxidant capacity, and improve growth performance and health status in pre-weaning Angus calves.

This study aimed to investigate, using in vitro technique and high-throughput sequencing, the regulatory effect of fermented total mixed ration (FTMR) prepared using synergistic fermentation of lactic acid bacteria (LABs) and cellulase on rumen fermentation, methane emission, and rumen microbiome. The concentrate-to-forage ratio of the TMR was 6: 4, net energy was 1.66 Mcal (for lactating cows), and metabolizable energy and protein allowable milk yield predictions were 36.3kg or 37.6kg from Cornell-Penn-Miner dairy model (CPM dairy). The LABs (106 colony-forming units (cfu)/g, Lactobacillus buchneri and Lactobacillus plantarum, at a ratio 1:1) and 1g fibrolytic enzyme (10,000 U/g activity) per fresh matter were used for ensiling the TMR. The results show that the synergistic fermentation of cellulase and lactic acid bacteria reduces the proportions of structural carbohydrate, neutral detergent insoluble protein, and acid detergent insoluble protein in high concentrate diet while increasing the proportions of crude protein, soluble protein, and non-protein nitrogen. The FTMR with LABs and cellulase addition diet had higher ammonia nitrogen, acetic acid, lactic acid contents, and lower pH than TMR. Also, the in situ effective degradation rate of neutral detergent fiber, dry matter, and crude protein was elevated. At the same time, the abundance of lactic acid bacteria was improved when LABs and cellulase were added to FTMR. The in vitro experiments showed that synergistic fermentation increased the proportion of propionic acid (26.47 vs 22.39, P = 0.0166) and ammonia nitrogen concentration (12.25 vs 10.24, P = 0.0144) and reduced ruminal pH (6.56 vs 6.68, P = 0.0249). In addition, the proportion of acetic acid (60.34 vs 66.38, P = 0.0059) and the ratio of acetic acid to propionic acid concentration (2.29 vs 2.97, P = 0.0077) was lowered when FTMR was prepared by LABs and cellulase addition. Fermented total mixed ration with LABs and cellulase changed the rumen fermentation pattern and reduced methane production (184.33ml vs 219.38ml, P=0.0003) by regulating in vitro ruminal bacteria's abundance and the diversity of protozoa and methanogens. Therefore, for a high-concentrate diet, adopting a fermented total mixed ration technology with LABs and cellulase addition is a clean option that could help reduce greenhouse gas production and protect the environment.

Little information exists on the variation in morphological characteristics, nutritional value, ruminal degradability, and molecular structural makeup of diverse whole-plant silage corn (WPSC) cultivars among different growing regions. This study investigated the between-regions (Beijing, Urumchi, Cangzhou, Liaoyuan, Tianjin) discrepancies in five widely used WPSC cultivars in China (FKBN, YQ889, YQ23, DK301 and ZD958), in terms of 1) morphological characteristics; 2) crude protein (CP) chemical profile; 3) Cornell Net Carbohydrate and Protein System (CNCPS) CP subfractions; 4) in situ CP degradation kinetics; and 5) CP molecular structures. Our results revealed significant growing region and WPSC cultivar interaction for all estimated morphological characteristics (P < 0.001), CP chemical profile (P < 0.001), CNCPS subfractions (P < 0.001) and CP molecular structural features (P < 0.05). Except ear weight (P = 0.18), all measured morphological characteristics varied among different growing regions (P < 0.001). Besides, WPSC cultivars planted in different areas had remarkably different CP chemical profiles and CNCPS subfractions (P < 0.001). All spectral parameters of protein primary structure of WPSC differed (P < 0.05) due to the growing regions, except amide II area (P = 0.28). Finally, the area ratio of amide I to II was negatively correlated with the contents of soluble CP (δ = -0.66; P = 0.002), CP (δ = -0.61; P = 0.006), non-protein nitrogen (δ = -0.56; P = 0.004) and acid detergent insoluble CP (δ = -0.43; P = 0.008), in conjunction with a positive correlation with moderately degradable CP (PB1; δ = 0.58; P = 0.01). In conclusion, the cultivar of DK301 exhibited high and stable CP content. The WPSC planted in Beijing showed high CP, SCP and NPN. The low rumen degradable protein of WPSC was observed in Urumchi. Meanwhile, above changes in protein profiles and digestibility were strongly connected with the ratio of amide I and amide II.

Background C16:0 and cis-9 C18:1 may have different effects on animal growth and health due to unique metabolism in vivo. This study was investigated to explore the different effects of altering the ratio of C16:0 and cis-9 C18:1 in fat supplements on growth performance, lipid metabolism, intestinal barrier, cecal microbiota, and inflammation in fattening bulls. Thirty finishing Angus bulls(626 ± 69 kg, 21 ± 0.5 months) were divided into 3 treatments according to the randomized block design:(1) control diet without additional fat(CON),(2) CON + 2.5% palmitic acid calcium salt(PA, 90% C16:0), and(3) CON + 2.5% mixed fatty acid calcium salt(MA, 60% C16:0 + 30% cis-9 C18:1). The experiment lasted for 104 d, after which all the bulls were slaughtered and sampled for analysis.Results MA tended to reduce 0–52 d dry matter intake compared to PA(DMI, P = 0.052). Compared with CON and MA, PA significantly increased 0–52 d average daily gain(ADG, P = 0.027). PA tended to improve the 0–52 d feed conversion rate compared with CON(FCR, P = 0.088). Both PA and MA had no significant effect on 52–104 days of DMI, ADG and FCR(P > 0.05). PA tended to improve plasma triglycerides compared with MA(P = 0.077), significantly increased plasma cholesterol(P = 0.002) and tended to improve subcutaneous adipose weight(P = 0.066) when compared with CON and MA. Both PA and MA increased visceral adipose weight compared with CON(P = 0.021). Only PA increased the colonization of Rikenellaceae, Ruminococcus and Proteobacteria in the cecum, and MA increased Akkermansia abundance(P < 0.05). Compared with CON, both PA and MA down-regulated the m RNA expression of Claudin-1 in the jejunum(P < 0.001), increased plasma diamine oxidase(DAO, P < 0.001) and lipopolysaccharide(LPS, P = 0.045). Compared with CON and MA, PA down-regulated the ZO-1 in the jejunum(P < 0.001) and increased plasma LPS-binding protein(LBP, P < 0.001). Compared with CON, only PA down-regulated the Occludin in the jejunum(P = 0.013). Compared with CON, PA and MA significantly up-regulated the expression of TLR-4 and NF-κB in the visceral adipose(P < 0.001) and increased plasma IL-6(P < 0.001). Compared with CON, only PA up-regulated the TNF-α in the visceral adipose(P = 0.01). Compared with CON and MA, PA up-regulated IL-6 in the visceral adipose(P < 0.001), increased plasma TNF-α(P < 0.001), and reduced the Ig G content in plasma(P = 0.035). Compared with CON, PA and MA increased C16:0 in subcutaneous fat and longissimus dorsi muscle(P < 0.05), while more C16:0 was also deposited by extension and desaturation into C18:0 and cis-9 C18:1. However, neither PA nor MA affected the content of cis-9 C18:1 in longissimus dorsi muscle compared with CON(P > 0.05).Conclusions MA containing 30% cis-9 C18:1 reduced the risk of high C16:0 dietary fat induced subcutaneous fat obesity, adipose tissue and systemic low-grade inflammation by accelerating fatty acid oxidative utilization, improving colonization of Akkermansia, reducing intestinal barrier damage, and down-regulating NF-κB activation.

This study evaluated the effects of different proportions of palmitic (C16:0) and oleic (cis-9 C18:1) acids in fat supplements on rumen fermentation, glucose (GLU) and lipid metabolism, antioxidant function, and visceral fat fatty acid (FA) composition in Angus bulls. The design of the experiment was a randomized block design with 3 treatments of 10 animals each. A total of 30 finishing Angus bulls (21 ± 0.5 months) with an initial body weight of 626 ± 69 kg were blocked by weight into 10 blocks, with 3 bulls per block. The bulls in each block were randomly assigned to one of three experimental diets: (1) control diet without additional fat (CON), (2) CON + 2.5% palmitic calcium salt (PA, 90% C16:0), (3) CON + 2.5% mixed FA calcium salts (MA; 60% C16:0 + 30% cis-9 C18:1). Both fat supplements significantly increased C18:0 and cis-9 C18:1 in visceral fat (P < 0.05) and up-regulated the expression of liver FA transport protein 5 (FATP5; P < 0.001). PA increased the concentrations of insulin (P < 0.001) and aspartate aminotransferase (AST; P = 0.030) in bull’s blood while reducing GLU (P = 0.009) concentrations. PA increased the content of triglycerides (TG; P = 0.014) in the liver, the content of the C16:0 in visceral fat (P = 0.004), weight gain (P = 0.032), and up-regulated the expression of liver diacylglycerol acyltransferase 2 (DGAT2), stearoyl-CoA desaturase 1 (SCD1; P < 0.05). MA increased blood superoxide dismutase activity (SOD; P = 0.011), reduced the concentration of acetate and total volatile FA (VFA) in rumen fluid (P < 0.05), and tended to increase non-esterified FA (NEFA; P = 0.069) concentrations in blood. Generally, high C16:0 fat supplementation increased weight gain in Angus bulls and triggered the risk of fatty liver, insulin resistance, and reduced antioxidant function. These adverse effects were alleviated by partially replacing C16:0 with cis-9 C18:1.