Oxidative stress is a risk factor for mammary health, resulting in decreased milk yield and milk quality. Application of exogenous bioactive compounds has been a research focus of antioxidation of animals in the mammary gland. Quercetin is a flavonoid extracted from vegetables, fruits and tea and has been shown to have a variety of biological activities, but the effect of quercetin on redox imbalance in mammary epithelial cells is unclear. In this study, cells of HC11, a mouse mammary epithelial cell line, were treated with quercetin, and the effects and molecular mechanisms of quercetin protection on hydrogen peroxide-induced oxidative stress were studied. Results showed that 20 μΜ quercetin attenuated hydrogen peroxide-induced lactate dehydrogenase release and reactive oxygen species (ROS) accumulation and alleviated the reduction of cell viability and antioxidant capacity. Quercetin significantly restored the activation of mitogen-activated protein kinase (MAPK) and nuclear factor E2-related factor 2 (Nrf2) pathways induced by hydrogen peroxide. Importantly, the inhibitors of p38 MAPK and extracellular regulated protein pathways affected the activation of Nrf2 pathway. All inhibitors of MAPK and Nrf2 pathways reduced the protective effects of quercetin on cell proliferation, the activity of catalase and the expression of glutamate-cysteine ligase modifier subunit. Meanwhile, the effects of quercetin on the production of ROS and expression of glutamate/cystine reverse transporter light chain were mainly dependent on Nrf2 pathway. In summary, the protective effect of quercetin in mammary epithelial cells was mediated via MAPK and Nrf2 pathways.

Mastitis in dairy cows is an important factor restricting the healthy development of dairy industry. Natural extracts have become a research hotspot to alleviate and prevent diseases because of their unique properties. The purpose of this study was to investigate the effects of resveratrol (RES) on the mitochondrial biosynthesis, antioxidation, and anti-inflammatory in bovine mammary epithelial cells (BMECs) and its mechanism involved. Blood samples were collected from six healthy cows and six mastitis affected cows, respectively, and lipopolysaccharide (LPS) was used to treat BMECs to construct inflammation models, gene interference is achieved by transfection. The results showed that messenger RNA (mRNA) expression of peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) was down-regulated and mitochondrial biogenesis-related gene expression was disrupted in the blood of mastitis cows and LPS-induced BMECs. RES is the best active substance to activate PGC-1α. The addition of RES can effectively alleviate the production of BMECs reactive oxygen species (ROS) and mitochondrial damage induced by LPS, and improve the antioxidation and anti-inflammatory ability, while the alleviation effect of RES is inhibited after interfering with protein kinase AMP-activated catalytic subunit α 1 (PRKAA1). In summary, our study emphasizes that PRKAA1 is a key gene mediating the activation of PGC-1α by RES, which regulates mitochondrial biosynthesis, inhibits ROS release, attenuates mitochondrial damage, and improves mitochondrial antioxidant capacity through the activation of PGC-1α by PRKAA1, thus attenuating the inflammatory response in BMECs.

This study evaluated the effects of dietary yeast culture (YC) supplementation on rumen microbiota and lactation performance in dairy goats. Twenty mid-lactation dairy goats were selected and divided into two groups: the control (CON) group was fed a basal diet; the YC group was supplemented with 10 g of YC in 1 kg of basal diet. The administration of YC was associated with a significant increase in dry matter intake, milk yield, milk protein yield, and milk lactose yield in dairy goats (P < 0.05). Additionally, serum total protein, albumin, creatinine, glucose, superoxide dismutase, and catalase levels were increased (P < 0.05). Furthermore, there was an increase in rumen pH and NH3-N levels (P < 0.05), while volatile fatty acid levels were observed to decrease (P < 0.05). The study found no significant difference in the α-diversity of bacteria and fungi between the YC and CON groups (P > 0.05). However, 15 bacterial genera and 13 fungal genera were upregulated (P < 0.05), while 4 bacterial genera and 11 fungal genera were downregulated (P < 0.05) in the YC group. The relative abundance of pathogenic fungi Dipodascus and Gibberella decreased (P < 0.05). Correlation analysis revealed that the bacterial genera were not significantly correlated with lactation performance (P > 0.05), whereas fungal genera Dipodascus and Gibberella were significantly (P < 0.05) correlated with lactation performance. In conclusion, the study demonstrated that YC can influence the rumen microbial composition, reduce the abundance of harmful fungi in the rumen, and improve lactation performance in dairy goats, suggesting that the addition of YC to dairy goat diets has good application prospects.

Feed intake, a critical factor for dairy cows during the postpartum period, is intricately linked to the rumen microbiome. However, the specific roles of rumen metagenome and metabolome in modulating feed intake in postpartum dairy cows remain unclear. In the current study, 20 postpartum dairy cows were divided into low feed intake (n = 5) and high feed intake (HFI, n = 5) groups to investigate the role of ruminal microbial composition, function, and metabolism on feed intake using a combined approach of metagenomics and metabolomics. Our analysis revealed a significant enrichment of Bacteroides and Fibrobacter in HFI cows (p < 0.05), contributing to enhanced protein and energy metabolism. Metabolomic analysis disclosed that HFI cows exhibited a higher relative concentration of rumen metabolites, such as alpha-tocopheryl acetate (fold change = 9.2, p = 0.008), linoleic acid (fold change = 5.96, p = 0.007), and leucine (fold change = 4.14, p = 0.004). Spearman correlation analysis pinpointed a positive correlation between specific microbiota (Succinivibrionaceae and Prevotellaceae) and metabolites involved in amino acid and peptide metabolism, fatty acid metabolism, and conjugates. Furthermore, co-occurrence network analysis showed that the unclassified_f_Succinivibrionaceae, Succinatimonas, and Ruminobacte were significantly associated with dry matter intake-associated metabotypes, including rumen metabolites involved in fatty acids and conjugates, favonoids, and gycerophosphocholines. The feed intake variation explained by the rumen microbiome, functions, and metabolites were 29.63%, 27.30%, and 33.50%, respectively. These findings provide comprehensive insights into rumen metagenomics at different feed intake levels in postpartum dairy cows, potentially guiding strategies to manipulate the rumen microbiome for feed intake and production improvement.

Gut microbiome changed dramatically during pregnancy and played important roles in metabolic status and reproductive endocrinology in mammals. However, investigating the functional microbiota and metabolites to improve the reproductive performance and understanding the host–microbiota interaction are still arduous tasks. This study aims to reveal the dominant strains and metabolites that improve the reproductive performance. We analyzed the fecal microbiota composition and metabolic status of higher yield Chinese pig breed Meishan (MS) sows and lower yield but widespread raised hybrid pig breed Landrace × Yorkshire (L × Y) sows on days 28 and 100 of gestation. Results showed that MS sows had higher litter sizes and steroid hormone level but lower short-chain fatty acid level in feces. Fecal metabolomic analysis revealed that MS sows showed a different metabolic status compared with L × Y sows both at early and late pregnancy, which enriched with phenylpropanoid biosynthesis, bile secretion, steroid hormone biosynthesis, and plant secondary metabolite biosynthesis. In addition, 16S rDNA and internal transcribed spacer sequencing indicated that MS sows showed different structures of microbiota community and exhibited an increased bacterial α-diversity but non-differential fungal α-diversity than L × Y sows. Moreover, we found that the litter sizes and bacteria including Sphaerochaeta, Solibacillus, Oscillospira, Escherichia–Shigella, Prevotellaceae_UCG-001, dgA-11_gut_group, and Bacteroides, as well as fungi including Penicillium, Fusarium, Microascus, Elutherascus, and Heydenia both have positive association to the significant metabolites at the early pregnancy. Our findings revealed significant correlation between reproductive performance and gut microbiome and provided microbial and metabolic perspective to improve litter sizes and steroid hormones of sows.

The rise of sequencing technologies has greatly contributed to our knowledge of the microbiota and its role in animal health and production. However, many members of the microbiota have historically been considered “unculturable.” Culturomics can be utilized to bring these fastidious microbes into cultivation and can be used in conjunction with culture-independent methods to study the microbiota in a more comprehensive manner. This review paper details culturomics’ role in revolutionizing human, swine, and bovine microbiota research and how its use has greatly increased the bacterial repertoire. Additionally, it describes how culturomics can be applied to develop microbiota-derived therapeutics, such as next-generation probiotics, and to study the role of the microbiota. Finally, this review provides potential methods and considerations for designing future culturomics studies.

The sophisticated relationship between the intake of dietary carbohydrates and host immunity is mediated in large part by the gut microbiota. This comprehensive review explores the multifaceted connections linking human gut microbiota with nondigestible carbohydrate metabolism and immune responses, highlighting the critical importance of this symbiotic relationship in maintaining overall host health. Understanding the mechanisms by which gut microbiota act as a bridge between carbohydrate intake and host immunity has significant implications for precision nutrition strategies and the development of therapeutic interventions.

Ruminant-derived foods, predominantly milk and meat, are globally recognized as staples of a high-quality diet. Despite their widespread popularity, there is a notable deficiency in comprehensive standards addressing the nutritional values and safety of these products. This gap significantly limits both the supply of and demand for premium quality milk and meat. This review endeavors to highlight the benefits of utilizing metabolomics for the evaluation of quality and safety in milk and meat products from ruminants. It identifies critical metabolites, genetic signals, and metabolic pathways related to the synthesis of ruminant-derived milk and meat, proposing their potential as nutritional or regulatory targets and biomarkers. These biomarkers are instrumental in predicting and assessing the quality and safety of dairy and meat products, offering guidance for quality-based pricing and food safety inspections in the market. This review offers a critical overview of current metabolomics-based platforms and tools for interpreting the quality and safety of ruminant foods. The core metabolic biomarkers and biological biosynthetic processes of milk and meat enhance our understanding of the interplay between conventional food production from animals and new synthetic biological technologies.

The rumen microbiome has attracted tremendous interest among microbiologists and ruminant nutritionists because of its crucial role in mediating feed digestion and fermentation and supplying most of the energy, nutrients, and precursors for producing ruminant products. The application of various omics technologies, including metataxonomics, metagenomics, metatranscriptomics, metaproteomics, and metabolomics, have enabled unprecedented investigations into this ecosystem, shedding new light on its interactions with diet and animals and its relationships with key production traits. Despite the valuable insights these omics technologies provide, each has its unique utility and inherent limitations. Achieving a holistic characterization of the rumen microbiome and deciphering its causal relationship with diet and key animal production traits remain an ongoing endeavor. In this perspective review paper, we highlight the limitations of individual technologies and advocate for an integrated multi-omics approach and data analyses in studying the intricate relationships between diet, rumen microbes, and ruminant nutrition. This approach, termed “rumen microbiome nutriomics,” aims to comprehensively understand the rumen microbiome in the context of diets and animal productivity. Our emphasis lies in recognizing the necessity of integrated analysis across multiple data layers, encompassing data of diet, rumen microbiome features, animal genotypes, and production traits and identifying the causal relationship among them. We also call for collaborative efforts to develop a comprehensive rumen microbiome genome database, including prokaryotes, protozoa, fungi, and viruses. Furthermore, standardization of processes and analyses is crucial to address the variability observed in the literature, facilitating comparison of results among future studies and enabling robust data reanalysis through advanced data analytics.

The gastrointestinal microbiota plays a crucial role in host nutrition and health. While culture-independent techniques have advanced microbial research, they often overlook low-abundance bacteria in the microbial community. Furthermore, empirical studies and mechanistic research rely on bacterial isolates obtained through culturing. The introduction of culturomics in 2012 significantly advanced culture-dependent techniques, contributing to microbial research. However, these methods remain labor-intensive, time-consuming, and costly, similar to traditional culture methods. This review provides an overview of the contributions of culturomics, summarizes procedures for optimizing sample and culture processes, and offers guidance on utilizing metagenomic data to enhance culturomics workflows. By collating and synthesizing these developments, this review aims to provide valuable insights into improving the practicality and productivity of culturomics for capturing gastrointestinal bacterial communities.

Milk fat is a high-value component of the U.S. dairy market. It is the major energy component of milk and is responsible for many organoleptic and technological characteristics of milk and dairy products. In addition, milk fat is unquestionably distinctive among all dietary fats that humans consume, as it is not only comprised of several hundred different fatty acids (FAs) but also contains a wide and unique array of bioactive lipids. Milk fat is dispersed in milk primarily in the form of fat globules. These cytoplasmic lipid droplets originate from mammary epithelial cells (MECs) and are secreted into the alveolar lumen surrounded by a membrane. Many advances in our knowledge of specific enzymes involved in milk lipid synthesis, the selectivity of the triacylglyceride (TAG) synthesis enzymes for specific FAs, the molecular mechanisms behind the uptake of long-chain FAs into the cells and the milk lipid secretion process have led to an improved understanding of the biology of milk fat synthesis. However, research to provide deeper insights into the mechanism of lipid synthesis in MECs is warranted and might lead to novel strategies to alter milk fat content and quality to benefit the dairy industry and meet dietary recommendations and consumer demands for foods that positively impact health. In this review, we aimed to provide a general overview of our current knowledge of the molecular aspects of milk lipid synthesis in MECs, from the uptake of blood-derived precursors to the intracellular formation of TAG-rich fat droplets secreted into milk as milk fat globules. We also highlight some current gaps in the knowledge that warrant further exploration. Given the importance of dairy food in the human diet, a better understanding of these processes could help develop novel strategies to alter milk fat composition in ways that benefit both human health and dairy producers.

This study investigated the impact of diallyl disulfide (DADS) on oxidative stress induced by hydrogen peroxide (H2O2) in ovine rumen epithelial cells (RECs). Initially, the effects of DADS were evaluated on cellular reactive oxygen species (ROS) levels, antioxidant capacity in RECs were estimated. Then, RNA-seq analysis was conducted in DADS-treated and untreated cells to analyze the differential gene expression, as well as Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Finally, the effects of DADS on Kelch-like ECH associated protein 1/the nuclear factor erythroid 2-related factor 2 (Keap1/Nrf2) signaling pathway in RECs were evaluated. Results showed that DADS remarkably enhanced superoxide dismutase (SOD) activity and total antioxidant capacity (T-AOC) (P < 0.05) while reducing ROS and malonaldehyde production (P < 0.05) in H2O2-treated RECs. Transcriptomic analysis revealed that DADS might influence glutathione synthesis through cysteine and methionine metabolism, thereby affecting the transcription of genes involved in immunity and oxidative stress. The DADS treatment resulted in increased nuclear translocation of Nrf2 and upregulation of mRNA and protein levels of quinone oxidoreductase 1, heme oxygenase 1, and Nrf2. The Nrf2-specific inhibitor nullified the protective effects of DADS on malonaldehyde formation induced by H2O2 and decreased T-AOC and SOD activities. In conclusion, DADS demonstrated the ability to alleviate oxidative stress in RECs by promoting antioxidative capacity through the Keap1/Nrf2 signaling pathway.

Rumen microbial biohydrogenation (RBH) is the major factor responsible for the bovine milk rich in saturated fatty acids (FAs). Here, we evaluated the effects of nutritional manipulation of ruminal propionogenesis and methanogenesis, two primary hydrogen sinks, on the RBH and milk FA profiles in vivo and in vitro using three propionogenesis enhancers (fumarate [FUM], biotin and monensin) and one methanogenesis inhibitor (N-[2-(nitrooxy)ethyl]-3-pyridinecarboxamide [NPD]). The in vivo results showed that inclusion of FUM in lactating dairy goat diet could protect dietary unsaturated FAs against RBH with increased proportions of C18:2n − 6 (by 33.5%), C18:3n − 3 (by 38.1%) and RBH intermediates (e.g. trans-10 C18:1 and trans-11 C18:1) in rumen contents. Additionally, FUM supplementation increased the milk Δ9 desaturase index (by 15.5%) with higher cis-9 monounsaturated FAs in the milk. As a result, FUM increased the proportions of polyunsaturated and monounsaturated FAs in the milk with lower atherogenicity index (by −15.3%) and thrombogenicity index (by −19.5%). Conversely, supplementing NPD increased RBH completeness (by 7.4%) with higher milk atherogenicity index (by 10.5%) and thrombogenicity index (by 8.7%). The adverse effects of NPD on the milk FA profiles can be eliminated when supplemented in combination with FUM. The metagenomic analyses showed that neither FUM nor NPD affect the rumen microbial α- or β-diversity at the strain or gene level. The in vitro study showed that the conversion rate of FUM to propionate was increased from 54.7% to 80.6% when FUM supplemented in combination with biotin and monensin, resulting a higher anti-RBH potential. Accordingly, manipulation of ruminal methanogenesis and propionogenesis can redirect hydrogen toward or away from RBH and thereby influence the milk FA profiles. FUM is a promising feed additive in ruminant not only to reduce the methane emissions as previously proved but also to improve the nutritional desirability of the milk FA profiles for human health.

Somatic cells (SCs) in milk are a heterogeneous population composed of several subsets of cells. However, a complete understanding of this heterogeneity in cow’s milk remains elusive. This study aimed to characterize heterogeneity within mammary epithelial (MEC) and immune cell subpopulations from healthy Holstein cows. An initial cell characterization of SC populations was completed using a single milk collection (3.8 L) from a base population of 25 multiparous Holstein cows to identify MEC and immune cells using flow cytometry with Butyrophilin 1A1 (BTN) and CD45 as cell surface markers. From the base population, 5 multiparous cows (≥300 days in milk (DIM), ≤162 × 103 SC/mL, and milk yield (MY) ≥ 25 kg/d) were selected for fluorescence activated cell sorting and single-cell RNA sequencing (scRNA-seq) analysis. A single-cell-suspension of approximately 1,000 sorted cells was prepared from each cow for characterization using scRNA-seq. Gel beads and barcodes were generated, cDNA amplified, cDNA sequencing libraries constructed and sequenced. After data normalization, scaling, and filtering control, two CD45+ databases were generated. The CD45+ databases contained 923 and 851 single cells, each comprising 17,771 and 12,156 features, respectively. Principal component analysis revealed seven and eight distinguishing clusters. Based on marker expression, most immune cells present in the samples were T cells (CD3E and PTPRC). Three different T cell subpopulations were revealed: helpers (CD4), cytotoxic (CD8A and CD8B), and regulatory T cells (IL2RA). The remaining four clusters were composed of granulocytes (neutrophils, eosinophils, and basophils; TLR4 and CXCL8), macrophages (PTPRC, CD14, CD68, TL2, IL1B), and a small population of B cells (CD19, CD22, and MS4A1). The study and characterization of immune cell subpopulations present in milk provide a basis for developing greater insights into mammary gland immune function, offering potential avenues for enhancing animal health and milk production in the future.