The aim of this study was to investigate the differentially expressed genes associated with intramuscular fat deposition in the longissimus dorsi muscle of Xinjiang Brown Bulls. The longissimus dorsi muscles of 10 Xinjiang Brown Bulls were selected under the same feeding conditions. The intramuscular fat content of muscle samples was determined by the Soxhlet extraction method, for which 5 samples with high intramuscular fat content (HIMF group) and 5 samples with low intramuscular fat content (LIMF group) were selected. It was found that the intramuscular fat content of the HIMF group was 46.054% higher than that of the LIMF group. Muscle samples produced by paraffin sectioning were selected for morphological observation. It was found that the fat richness of the HIMF group was better than that of the LIMF group. Transcriptome sequencing technology was used to analyze the gene expression differences of longissimus dorsi muscle. Through in-depth analysis of the longissimus dorsi muscle by transcriptome sequencing technology, we screened a total of 165 differentially expressed genes. The results of Gene Ontology (GO) enrichment analysis showed that the differentially expressed genes in the two groups were mainly clustered in biological pathways related to carbohydrate metabolic processes, redox processes and oxidoreductase activities. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that the differentially expressed genes were significantly clustered in 15 metabolic pathways, which mainly covered fatty acid metabolism (related to lipid metabolism and glucose metabolism), the pentose phosphate pathway, the Peroxisome Proliferator-Activated Receptor (PPAR) signaling pathway and other important metabolic processes. The three genes that were predominantly enriched in the glycolipid metabolic pathway by analysis were SCD5, CPT1C and FBP2, all of which directly or indirectly affect intramuscular fat deposition. In summary, the present study investigated the differences in gene expression between high and low intramuscular fat content in the longissimus dorsi muscle of Xinjiang Brown Bulls by transcriptome sequencing technology and revealed the related signaling pathways. Therefore, we hypothesized that SCD5, CPT1C and FBP2 were the key genes responsible for the significant differences in intramuscular fat content of the longissimus dorsi muscles in a population of Xinjiang Brown Bulls. We expect that these findings will provide fundamental support for subsequent studies exploring key genes affecting fat deposition characteristics in Xinjiang Brown Bulls.

This study utilized MAC-T cells cultured in vitro as a model to investigate the effects of varying concentrations of valine on α-casein synthesis and its underlying regulatory mechanisms. In this experiment, MAC-T cells were subjected to a 12 h starvation period, followed by the addition of valine in a range of concentrations (a total of seven concentrations: 0.000, 1.596, 3.192, 6.384, 12.768, 25.536, and 51.072 mM, as well as in 10% Fetal Bovine Serum). The suitable range of valine concentrations was determined using enzyme-linked immunosorbent assays (ELISAs). Real-time fluorescent quantitative PCR (RT-qPCR) and Western blot analyses were employed to evaluate the expression levels and phosphorylation states of the casein alpha s1 gene (CSN1S1), casein alpha s2 gene (CSN1S2) and mTOR signaling pathway-related genes. The functionality of the mTOR signaling pathway was further validated through rapamycin (100.000 nM) inhibition experiments. Results indicated that 1× Val (6.384 mM), 2× Val (12.768 mM), 4× Val (25.536 mM), and 8× Val (51.072 mM) significantly enhanced α-casein synthesis (p < 0.01). Within this concentration range, valine significantly upregulated the expression of CSN1S1, CSN1S2, and mTOR signaling pathway-related genes including the RagA gene (RRAGA), RagB gene (RRAGB), RagC gene (RRAGC), RagD gene (RRAGD), mTOR, raptor gene (RPTOR), and 4EBP1 gene (EIF4EBP1), eukaryotic initiation factor 4E (EIF4E), and S6 Kinase 1 (S6K1) (p < 0.01). Notably, the expression of the eukaryotic elongation factor 2 (EEF2) gene peaked at 1× Val (6.384 mM), while the expression of other genes reached their maximum at 4× Val (25.536 mM). Additionally, valine significantly increased the phosphorylation levels of mTOR, S6K1, 4E-binding protein-1 (4EBP1), ribosomal protein S6 (RPS6), and eEF2 (p < 0.01), with the highest phosphorylation levels of mTOR, S6K1, and RPS6 observed at 4× Val (25.536 mM). Rapamycin treatment significantly inhibited mTOR phosphorylation and α-casein synthesis (p < 0.01); however, the addition of 4× Val (25.536 mM) partially mitigated this inhibitory effect. In conclusion, valine promotes α-casein synthesis by activating the mTOR signaling pathway, with an optimal concentration of 4× Val (25.536 mM).

Introduction: This study examines the effects of steam-flaked corn starter on pre-weaned Simmental calves' growth, immunity, and metabolism. Despite benefits shown in adult cattle, research on calves is limited. The goal is to optimize calf feeding for better growth, health, and nutrient use.Methods: Thirty-two Simmental bull calves (avg. wt: 50.50 ± 4.50 kg, avg. age: 21 ± 7 days) were divided into four groups of eight. The 127-day study included a 7-day pretest and a 120-day trial. Calves had unlimited access to starter feed, alfalfa hay, and water. Groups received starter diets with 0% (CK, control), 33% (SFC33, low), 66% (SFC66, medium), or 100% (SFC100, high) steam-flaked corn replacing regular corn. Other conditions were consistent.Results: Feeding 33% steam-flaked corn to pre-weaned Simmental calves led to highest daily weight gain (0.80 kg/d), significantly boosting serum globulin, cholesterol, urea nitrogen, glucose, immunoglobulins, GH, INS, and IGF-1 (P < 0.05). Compared to control, 31 metabolites differed in SFC33 group, mostly up-regulated, including glycerophospholipids, linoleic/arachidonic acid metabolism, cholesterol pathway molecules, L-glutamine in multiple pathways, and mannose in fructose/mannose metabolism.Discussion: In summary, feeding 33% steam-flaked corn-based starter can improve the growth performance, enhance immunity, and improve sugars, lipids, and proteins metabolism of pre-weaned Simmental calves.

This study aimed to investigate the effects of feeding high-moisture corn (HMC) on weight performance, serum immune and antioxidant indices, rumen fermentation, microbial community, and metabolomics in Kazakh rams. A total of 32 healthy Kazakh rams were randomly divided into a control group (CT, diet with only ordinary crushed corn) and an experimental group (GS, diet with 50% ordinary crushed corn + 50% HMC), following a 7-day adaptation period and a 120-day trial period. Results showed that the F/G was significantly lower in the GS group than in the CT group (p < 0.05). FBW, net weight gain and ADG increased by 4.58%, 8.69%, and 8.70%, respectively, while ADFI decreased by 7.04% (p > 0.05). Regarding serum immune indices, IgA in the GS group was significantly higher at 40 d (p < 0.01), and IgM was significantly higher at 40, 80, and 120 d (p < 0.05). For antioxidant indices, the SOD activity in the GS group was significantly higher than that in the CT group at 120 d (p < 0.01). The CAT activity in the GS group was significantly higher at 40, 80, and 120 d (p < 0.01). Among rumen fermentation parameters, the concentration of butyric acid in the GS group was significantly lower than in the CT group (p < 0.01). Microbial diversity analysis indicated no significant differences in Alpha- and Beta-diversity of rumen microorganisms between the two groups. However, the relative abundance of Firmicutes_A at the phylum level was significantly higher in the GS group (p < 0.05), and the abundance of Cryptobacteroides was significantly higher than in the CT group (p < 0.01). Rumen metabolomic analysis identified a total of 1357 differential metabolites, among which 1130 showed significant differences, with 459 upregulated and 671 downregulated. These were mainly enriched in pathways such as Glutathione metabolism, Beta-alanine metabolism, Sphingolipid metabolism, and lysine degradation. In conclusion, feeding HMC can improve feed conversion efficiency and weight performance in Kazakh rams, regulate the structure of dominant rumen microorganisms, and enhance immune and antioxidant capacities.

Amino acids play a dual role in milk protein synthesis, functioning as both metabolic precursors and signaling molecules. This study aimed to elucidate the molecular mechanism by which glycine regulates α-casein production in bovine mammary epithelial cells (MAC-T). Under serum-free conditions, MAC-T cells were exposed to graded concentrations of glycine (1.105, 2.209, 4.418, 8.836, 17.673, and 35.345 mM) for 24 h. α-Casein levels in cell lysates and culture supernatants were quantified by ELISA. Transcriptional activity of casein-encoding genes (CSN1S1, CSN1S2) and PI3K-AKT-mTOR pathway components was assessed by RT-qPCR. Phosphorylation status of pathway proteins was analyzed by Western blot. The functional involvement of the PI3K-AKT-mTOR pathway was validated using the specific inhibitor LY294002. Glycine stimulated α-casein synthesis and secretion in a concentration-dependent manner, with maximal efficacy at 4.418 mM. At this concentration, glycine upregulated CSN1S1, CSN1S2, and PI3K-AKT-mTOR pathway gene expression, and enhanced phosphorylation of the corresponding proteins. Inhibition of PI3K-AKT-mTOR signaling by LY294002 abolished glycine-induced α-casein synthesis, and this effect was reversed by glycine co-treatment. These findings demonstrate that glycine enhances α-casein synthesis through activation of the PI3K-AKT-mTOR pathway.

Grapevine branch and leaf silage (GBLS), a polyphenol-rich unconventional forage, exhibits antimicrobial and antioxidant properties that can benefit animal health and productivity. A total of 60 healthy six-month-old Kazakh rams (43.29 ± 4.55 kg, p > 0.05 for initial body weight among groups) were randomly assigned to three dietary groups, each consisting of four replicates with five rams per replicate. The control group (CK) was fed a basal diet based on whole-plant corn silage, whereas the experimental groups received diets in which 50% (GBLS50%) or 100% (GBLS100%) of the corn silage was replaced with GBLS. A 10-day adaptation period preceded the 90-day formal feeding trial. Results showed a significant quadratic response for average daily gain (ADG) and average daily feed intake (ADFI) across GBLS substitution rates (p < 0.05), with the 50% level yielding the highest values. Specifically, ADFI at the 50% replacement level was significantly higher than that of the control (p < 0.05), confirming an inverted U-shaped response with 50% as the optimal substitution rate. However, in-depth analysis of serum biochemical parameters revealed that GBLS supplementation significantly reduced serum concentrations of total cholesterol, triglycerides, urea nitrogen, interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and malondialdehyde (MDA), while significantly increasing levels of immunoglobulins (IgA, IgM, IgG), superoxide dismutase (SOD), and catalase (CAT) (p < 0.05). Rumen fermentation analysis showed that the GBLS50% group had significantly lower concentrations of acetate, butyrate, and total volatile fatty acids (VFA) (p < 0.05). In the rumen microbiota study, no significant differences were observed in alpha or beta diversity or at the phylum level between groups (p > 0.05); however, the abundance of Lactobacillus gasseri was significantly reduced in the GBLS50% group (p < 0.05). Metabolomic profiling identified 43 significantly altered metabolites—27 upregulated (e.g., PE (18:1(9Z)/0:0) and 12,14-pentacosadiynoic acid) and 16 downregulated (e.g., deoxyadenosine). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis highlighted purine metabolism as a significantly altered pathway (p < 0.05), providing insight into the potential metabolic mechanisms underlying the physiological effects of GBLS in rams. In conclusion, replacing 50% of whole-plant corn silage with grapevine branch and leaf silage improves growth performance trends and significantly enhances immunity and antioxidant capacity in Kazakh rams.