This study investigated the effects of chemically protected sodium butyrate (CSB) on growth performance, serum indicators, rumen fermentation parameters, and microbial communities in Gangba sheep. Twenty-four healthy 5-month-old male Gangba sheep with an initial body weight of 19.54±1.04 kg were randomly assigned to 4 groups and fed diets containing 0, 1.0, 5.0, and 10.0 g/kg CSB for 74 days. After a 14-day adaptation period, the daily feed intake of Gangba sheep was recorded. Fecal samples were collected during the last 7 days of the trial, and serum and rumen fluid were sampled on the final day. CSB improved final body weight, average daily dry matter intake, and average daily gain while reducing the feed conversion ratio of Gangba sheep (p < 0.05). In addition, CSB improved the apparent digestibility of crude protein and neutral detergent fiber (p < 0.05). Moreover, CSB increased serum immunoglobulin G levels while decreasing tumor necrosis factor-α levels (p < 0.05). Meanwhile, CSB increased serum growth hormone-releasing hormone levels (p < 0.05). Moreover, CSB increased rumen fluid pH and enhanced concentrations of ammonia nitrogen, acetic acid, iso-butyric acid, butyric acid, and total volatile fatty acids, as well as activities of lipase and butyrate kinase (p < 0.05). Besides, CSB enhanced the rumen microbiota structure by increasing the relative abundance of Fibrobacter, unclassified _Oscillospiraceae, and Ruminococcus. CSB improved serum immune and hormone indicators, enhanced the rumen ecological environment, and increased feed digestibility and utilization, thereby promoting the growth of Gangba sheep. It is recommended to use a dosage of 5.0 g/kg dry matter for optimal growth performance and health benefits.

Intramuscular fat (IMF) deposition is a key factor for meat quality traits like juiciness, tenderness, and flavor. Although fibro/adipogenic progenitor cells (FAPs) participate in adipogenesis, the roles of other stromal cells in the muscle microenvironment are unclear. This study aimed to characterize cellular heterogeneity in cattle IMF deposition and identify key cell types and molecular mechanisms regulating intramuscular adipogenesis. We performed single-nucleus RNA sequencing (snRNA-seq) on Longissimus dorsi muscle samples from three cattle breeds-Angus, Hereford, and Simmental-which exhibit divergent IMF content. Cell populations were annotated using multiple reference-based methods. We applied Scissor to identify phenotype-associated cells, CellChat to infer intercellular communication, hdWGCNA to detect co-expression modules, and scFEA to evaluate metabolic activity. Virtual gene perturbation was conducted using scTenifoldKnk, and drug-responsive cell subpopulations were assessed with scRank. We identified ten distinct cell types in bovine skeletal muscle. Angus cattle, known for high marbling, had elevated proportions of adipocytes, FAPs, and vascular endothelial cells (VEndoCs), while Hereford cattle were enriched in type II fibers. VEndoCs were consistently linked to high IMF and acted as central signaling hubs, interacting extensively with adipocytes and FAPs. These cells also showed high glucose metabolic activity. Co-expression analysis within VEndoCs identified 40 candidate regulators, including nine BMP signaling pathway genes. Virtual knockdown highlighted BMPR1A as the top regulator, altering expression of 13 lipid metabolism-related genes. Neurocytes were most sensitive to resveratrol treatment, largely via ALDH2, whereas FAPs were least responsive. Our findings establish VEndoCs as active regulators of intramuscular adipogenesis via a BMPR1A-mediated signaling pathway. These results provide new insights into the cellular and molecular basis of marbling in cattle and highlight BMPR1A as a potential target for genetic or nutritional strategies aimed at improving meat quality.

Microtubule dynamics regulator protein 2 (RMDN2) plays a crucial role in cell division, cytoskeleton maintenance, and various other cellular processes, establishing it as a candidate gene influencing chicken follicle development in our previous studies. This research aims to explore single-nucleotide polymorphisms (SNPs), perform phylogenetic analysis, and assess sequence characteristics of RMDN2, offering valuable insights for molecular marker-assisted breeding and enhancing the understanding of its regulatory mechanisms. SNPs within the RMDN2 coding sequence (CDS) region were identified in an F2 resource population. Bioinformatics tools were employed to investigate the effects of SNP mutations on the structure and function of RMDN2 protein, and a phylogenetic tree was constructed to elucidate the potential mechanisms underlying the role of RMDN2 in chicken laying traits. Four novel exonic SNPs were discovered: SNP1 (c.250G>A, p.Val84Ile), SNP2 (c.270G>C, p.Lys90Asn), SNP3 (c.533G>T, p.Gly178Val), and SNP4 (c.606G>A). The heterozygous genotypes of SNP1, SNP3, and SNP4 were significantly associated with increased egg number at 66 weeks (EN66) (p < 0.05). In contrast, the heterozygous genotype of SNP2 is associated with higher body weight at first egg (BWFE) (p < 0.05). Notably, the H1H1 haplotype combination was associated with lower BWFE, body weight at 105 days (BW105), and first egg weight (FEW) (p <0.05). Missense mutations in SNP1, SNP2, and SNP3 were speculated to potentially influence the hydrophilic/hydrophobic properties, transmembrane regions, functional domains, and secondary structure of the RMDN2 protein, potentially reducing its stability. Phylogenetic analysis demonstrated 100% sequence homology between chicken and quail, with substantial conservation within species of the same order, but a marked decrease across different taxonomic orders. These findings enrich the candidate gene pool for regulating chicken laying traits. However, further validation through in vivo and in vitro experiments remains necessary to strengthen the theoretical foundation for molecular breeding strategies.

The methane (CH₄) emission prediction method, using predicted CO₂ emissions and the CH₄:CO₂ concentration ratio, faces challenges in evaluating the efficacy of CH₄-reducing feed additives due to CO₂ prediction bias associated with energy utilization efficiency. We hypothesized that incorporating dry matter intake (DMI), along with metabolic body weight (MBW) and energy-corrected milk (ECM) as explanatory variables, would eliminate this bias. The primary objective was to compare the performance of CO₂ emission models with and without including DMI. The secondary objective was to assess the CO₂-based method's applicability for quantifying CH₄-reducing effects, through a case study of 3-nitrooxypropanol (3-NOP). Prediction models for CO₂ emissions were developed including DMI, MBW, and ECM as explanatory variables, based on 219 records obtained from previous experiments with Holstein cows using respiration chambers or headboxes. Model performance was evaluated using cross-validation. Bias associated with energy utilization efficiency was assessed. The applicability of the CO₂-based method to quantify the CH₄-reducing effect of 3-NOP was assessed using data obtained from the literature, including 10 studies with 22 treatment and control mean comparisons. The agreement between the observed and predicted CH₄ reductions was assessed. Incorporating DMI along with MBW and ECM improved the predictive performance of CO₂ emissions. While the models without DMI showed bias associated with energy utilization efficiency, the bias was eliminated when DMI was incorporated. Applicability assessment demonstrated that the models without DMI systematically underestimated the CH₄-reducing effect of 3-NOP. In contrast, the models that included DMI showed smaller discrepancies between observed and predicted CH₄ reductions. This study highlights the importance of incorporating DMI as an explanatory variable to achieve accurate and unbiased predictions of CO₂ emissions. These findings would contribute to the appropriate application of the CO₂-based method for evaluating the CH₄-reducing effects of feed additives.

Negative energy balance in transition cows elevates the circulating concentrations of non-esterified fatty acids (NEFA), which can trigger mastitis and pose a severe threat to the dairy industry. Resveratrol is a polyphenolic compound with anti-inflammatory properties, yet its role in NEFA-induced inflammation in bovine mammary epithelial cells (BMECs) remains unclear. This study aimed to investigate the protective effects of resveratrol on mastitis and elucidate its underlying mechanisms. BMECs were pre-treated 100 µM RES for 24 h and then treated 0.9 mM NEFAs for 4 h, and a PINK1 inhibitor was used to assess the underlying mechanisms. Furthermore, a mouse model of mastitis was utilized to further evaluate the hepatoprotective effects of resveratrol against mastitis in vivo. Resveratrol significantly attenuated the NEFA-induced inflammatory response, as evidenced by reduced levels of NLRP3 inflammasome components (NLRP3, caspase1, IL-1β) and pro-inflammatory cytokines (IL-6, IL-1β and TNF-α). Mechanistically, resveratrol promoted mitophagy by upregulating levels of LC3-II, PINK1, and Parkin, and downregulating P62 expression. Crucially, the anti-inflammatory effect of resveratrol was reversed upon inhibition of PINK1. The in vivo experiments confirmed that resveratrol alleviated mammary gland inflammation and enhanced PINK1-mediated mitophagy. This study demonstrates that resveratrol mitigates NLRP3-mediated inflammatory responses by activating PINK1-mediated mitophagy, suggesting its potential as a promising therapeutic candidate for mastitis in perinatal dairy cows experiencing negative energy balance.

Compare cold adaptation mechanisms between cold-tolerant Hezuo and cold-sensitive Bama pigs. Lung histology, W/D ratio, oxidative and inflammatory markers, apoptosis, and transcriptomics were analyzed. The results showed that the lung of Hezuo pigs displayed less severe alveolar septal thickening, inflammatory infiltration, and fine bronchial fold extension during cold exposure compared to Bama pigs. The W/D ratio dramatically decreased in Hezuo pigs and increased in Bama pigs. Hezuo pigs exhibited significantly higher AQP-1(Aquaporin-1) and AQP-5(Aquaporin-5) expressions than Bama pigs in the middle and late phases. Bama pigs displayed increased ROS(Reactive Oxygen Species), MDA(Malondialdehyde), TNF-α(Tumor Necrosis Factor-alpha), IL-1β(Interleukin-1 beta) and decreased GSH(Glutathione). However, Hezuo pigs maintained stable GSH and showed no significant late-phase inflammatory marker changes. Bama pigs had a higher apoptosis density and number of TUNEL-positive cells than Hezuo pigs, which was related to the down-regulation of Bcl-2(B-Cell Lymphoma 2) and the up-regulation of Bax(BCL2-Associated X Protein) and Caspase-3. Furthermore, Transcriptomic analysis revealed that, in Bama pigs, the distinctive genes-MUC5B(Mucin 5B), MMP9(Matrix Metallopeptidase 9), AMCF-II(Alveolar Macrophage Chemotactic Factor-II), IL22RA1(Interleukin 22 Receptor Subunit Alpha 1), CCL16(C-C Motif Chemokine Ligand 16), SOX9(SRY-Box Transcription Factor 9), KRT5(Keratin 5) function to drive mucus hypersecretion, extracellular matrix degradation, and sustained inflammatory chemotaxis, thereby exacerbating tissue damage. In contrast, the distinctive genes of Hezuo pigs-ALDH1A2(Aldehyde Dehydrogenase 1 Family Member A2), ACSL6(Acyl-CoA Synthetase Long Chain Family Member 6), ACSM5(Acyl-CoA Synthetase Medium Chain Family Member 5), AKR1C1(Aldo-Keto Reductase Family 1 Member C1), NR4A3/2(Nuclear Receptor Subfamily 4 Group A Member 3/2), GNG4(G Protein Subunit Gamma 4), GYS2(Glycogen Synthase 2) primarily enhance lipid metabolism, facilitate aldehyde detoxification, and mitigate oxidative stress, collectively orchestrating a cellular protection mechanism. Hezuo pigs exhibit protective molecular mechanisms, suggesting candidate targets for cold-resistance breeding.

Acetyl-CoA synthetase 2 (ACSS2) is the obligatory gatekeeper for converting rumen-derived acetate into acetyl-CoA in ruminants. However, whether ACSS2 actively regulates the transcriptional networks governing lactation, beyond its catalytic role, remains unclear. This study aimed to elucidate the molecular characteristics of buffalo ACSS2 and investigate its function as a central node in the metabolic-transcriptional circuitry of buffalo mammary epithelial cells (BuMECs). The complete coding sequence of buffalo ACSS2 was characterized, and its expression was analyzed across lactation stages. Subcellular localization was determined via high-resolution confocal microscopy. We utilized siRNA-mediated knockdown in BuMECs to assess cell viability, triglyceride (TAG) content, and the expression of core metabolic and regulatory genes to dissect the underlying molecular mechanisms. ACSS2 expression was highly enriched in lactating mammary tissue, and the protein exhibited a dual nucleocytoplasmic distribution. ACSS2 knockdown induced a "dual collapse" of cellular function: it severely impaired lipogenesis (significantly reducing intracellular TAG and downregulating FASN, ACACA, SCD, CD36, LPL, FABP3, DGAT1, DGAT2 and AGPAT6) and arrested cell proliferation (downregulating the G1/S phase regulators CCND1, CCNE1, CDK2 and CDK4). Mechanistically, ACSS2 depletion dismantled the transcriptional machinery itself, suppressing the mRNA levels of master regulators SREBF1 and PPARG. Crucially, this collapse was accompanied by the paradoxical upregulation of the SREBP1-inhibitor INSIG1, suggesting that metabolic stress triggers an INSIG1-mediated blockade of the feedback loop. This study establishes ACSS2 as a critical metabolic checkpoint in the buffalo mammary gland, rather than a passive enzyme. We propose a model where ACSS2 maintains a reciprocal positive feedback loop with SREBP1 and PPARG. By ensuring sufficient acetyl-CoA to suppress INSIG1 and support histone acetylation (implied by nuclear localization), ACSS2 couples substrate availability to the stability of the lipogenic program and cell cycle progression. These findings reveal an evolutionarily conserved metabolic-epigenetic axis essential for high-efficiency lactation in ruminants.

This study aimed to assess the interaction between chicken myofibrillar protein and cranberry bean powder (CBP) processed through different drying methods oven-drying (OD) and freeze-drying (FD) along with cranberry bean protein isolates (CBPI), on the rheological properties of chicken myofibrillar protein gels (MPGs). Two experiments were done in this study. The extracted myofibrillar protein gel (MPG) treated with different levels of CBP in different drying methods was prepared for the first experiment. The MPG with CBP and CBPI with different drying methods was prepared and compared to SPI for the second experiment. The cooking yield (CY), gel strength (GS), viscosity, SDS-PAGE and microstructure were measured in both experiments. The results showed that FDI exhibited the best gel strength, cooking yield, and viscosity, which were comparable to those of soy protein isolate (SPI). FDI demonstrated superior protein-protein interactions, with SDS-PAGE and scanning electron microscopy analyses revealing well-formed structures. FD performed better than OD in terms of gel strength and viscosity but still lower than FDI. OD samples, which contained higher levels of carbohydrates and starch, displayed lower gel strength and less stable gel networks, indicating that starch may interfere with protein matrix formation. The addition of FDI or SPI was shown to be the most effective in enhancing the rheological properties of MPGs, and the higher levels of powder addition also improved the results, including viscosity, gel strength, and cooking yield.

To determine how dietary vitamin A (VA) level and duration affect intramuscular fat (IMF), meat quality, storage stability, and antioxidant gene expression in Yanbian yellow cattle. Twenty 15-month-old Yanbian yellow cattle (314.13 ± 13.30 kg) were assigned to five treatments: CON (supplemental VA 2200 IU/kg DM), NVA1 (0 IU/kg DM supplemental VA for 180 d), NVA2 (0 IU/kg DM supplemental VA for 240 d), LVA1 (supplemental VA 1100 IU/kg DM for 180 d) and LVA2 (supplemental VA 1100 IU/kg DM for 240 d). Growth performance, carcass traits, physicochemical characteristics, and storage stability were measured. Serum biochemical and muscle antioxidant indexes were analyzed, and the mRNA expression of antioxidant-related genes (FOXO1, GSTA1, SOD) was quantified by qPCR. Statistical significance was set at p < 0.05. All VA-restricted groups showed higher IMF, marbling score, and lower muscle fiber diameter, drip loss, and shear force than the CON group (p < 0.05). Serum SOD and glutathione peroxidase (GSH-Px) levels in the NVA1/NVA2 group were lower than those in the LVA groups and CON (p < 0.05). During storage, NVA2/LVA1/LVA2 had lower drip loss and shear force on days 1, 3, and 5 (p < 0.05); on day 7, b* was higher in NVA1/NVA2 than CON, and drip loss, cooking loss, and shear force were lower in NVA2, LVA1, and LVA2 than CON (p < 0.05). At the transcriptional level, antioxidant-related genes were upregulated across all VA-restricted groups, with FOXO1 and GSTA1 peaking in LVA1 and SOD elevated in all restricted groups (p < 0.05). Restricting dietary VA to 50% of the recommended level for 180 days significantly improved marbling, tenderness, and oxidative stability without compromising growth performance. These findings highlight a feasible nutritional strategy to enhance beef quality and extend shelf life in Yanbian yellow cattle.

This study investigated the role of ferroptosis, an iron-dependent form of regulated cell death, in meat quality by comparing the effects of pre-slaughter administration of teriflunomide, an inhibitor of dihydroorotate dehydrogenase (DHODH), which can induce ferroptosis, and dietary vitamin E, an antioxidant that can suppress ferroptosis, on the post-mortem biochemical properties of broiler chicken muscle. Broiler chickens were randomly assigned to three groups: control, teriflunomide-treated, and vitamin E-supplemented. Teriflunomide was administered subcutaneously before slaughter, and vitamin E was provided in the diet. Post-mortem thigh muscles were analyzed for oxidative status such as lipid peroxide levels and the ratio of reduced to oxidized glutathione (GSH/GSSG); the expression of ferroptosis-related genes such as glutathione peroxidase 4 (GPX4) and acyl-CoA synthetase long-chain family member 4 (ACSL4); and meat quality traits such as pH, color, and water-holding capacity. Teriflunomide significantly decreased the GSH/GSSG ratio (p<0.001) and increased ACSL4 expression compared with the control and vitamin E groups (p<0.05), indicating elevated oxidative stress. Vitamin E significantly increased GPX4 expression compared with teriflunomide (p<0.05). Lipid peroxidation was numerically lower in the vitamin E group than in the other groups, although the differences were not statistically significant (p=0.062). Teriflunomide slowed post-mortem pH decline (p<0.001) and decreased L* and b* values, whereas a* values were higher (p<0.05). Vitamin E maintained higher L* and b* values and slightly lower a* values than the control (p<0.05). No statistically significant differences were observed in water-holding capacity among the groups. These findings indicate that ferroptosis-related processes play a critical role in regulating post-mortem meat quality in broiler thigh muscles. DHODH inhibition by teriflunomide and antioxidant supplementation with vitamin E produced opposing effects.