This study evaluated the effects of replacing a concentrate supplement with fermented liquid feed (FLF) on gross feed efficiency (GFE), production performance, and blood parameters in lactating dairy cows. Twenty-four lactating Chinese Holstein cows with comparable body weight (657 ± 32.6 kg), milk yield (38.7 ± 4.7 kg/day), parity (2.8 ± 1.2), and days in milk (104 ± 10.8 d) were randomly assigned to either a control group (CON, total mixed ration without fermented products) or a treatment group (FLF, total mixed ration supplemented with FLF) in a randomized complete block design. Compared with the CON group, feeding FLF increased GFE, energy-corrected milk yield, milk fat and protein percentages, milk nitrogen efficiency, apparent total-tract digestibility of dry matter, crude protein, neutral detergent fiber, and acid detergent fiber, as well as fecal score. Additionally, compared with the CON group, ruminal total volatile fatty acid (VFA) concentration, microbial crude protein yield, and the molar proportion of propionate increased significantly in the FLF group, whereas ruminal pH, acetate proportion, and acetate-to-propionate ratio were decreased. Total VFA concentrations and the molar proportion of butyrate in the feces increased significantly, whereas pH was decreased. In addition, in the FLF group, the relative abundance of unclassified_Clostridia_UCG_014, Prevotella_7, and Lachnospiraceae_NK3A20_group in the rumen and the relative abundance of Bifidobacterium, Candidatus_Saccharimonas, unclassified_Clostridia_UCG_014, and unclassified_[Eubacterium]_coprostanoligenes_group in the feces increased significantly, whereas the relative abundance of NK4A214 group, Rikenellaceae_RC9_gut_group, Treponema, Succiniclasticum, and Isotricha in the rumen and UCG_005, Rikenellaceae_RC9_gut_group, and Bacteroides in the feces decreased. Compared with the CON group, supplement of FLF activated the secondary bile acid pathways, upregulating ursodeoxycholic acid, taurocholic acid, glycocholate, while downregulating deoxycholic acid, lithocholic acid, and chenodeoxycholate in the feces. Additionally, plasma glucose levels, total antioxidant capacity, and total superoxide dismutase activity significantly increased in the FLF group, while β-hydroxybutyric acid, nonesterified fatty acids, and malondialdehyde were significantly decreased. In conclusion, FLF effectively regulated the microbiota in the rumen and feces, improved lactation performance, GFE, and health condition of dairy cows, and provided a novel and promising nutritional regulation strategy for high-yielding dairy cows.

Dinosaur trackway fossils in the Otog Field Geological Heritage Museum, Inner Mongolia, China, have suffered severe weathering, with extensive surface powdering and granular disintegration. This study evaluated three TEOS-based hybrid materials modified with dodecyltrimethoxysilane (WD10), bis(triethoxysilyl)ethane (BEQ), and bis(triethoxysilyl)octane (BOQ), designated as 10%W5, 10%E5, and 10%O3. Performance was assessed in terms of penetration depth, hydrophobicity, mechanical strength, color difference, vapor permeability, salt resistance, and freeze–thaw durability. All three materials effectively improved the fossils’ properties, with 10%W5 showing the best performance: penetration depth of 28.4 mm within 5 min, water absorption reduced to 0.48%, contact angle of 125.73°, compressive strength improved by 67.73%, and excellent salt and freeze–thaw resistance. The exceptional performance of 10%W5 is attributed to its homogeneous and discontinuous siloxane gel network, which enhances intergranular bonding while preserving pore connectivity and vapor permeability.

This study evaluated the impact of fermented palm kernel meal (FPKM) on the lactation performance, blood biochemical indices, rumen microbiota, and metabolic functions of Holstein dairy cows, aiming to enhance the nutritional value of palm kernel meal (PKM) through solid-state fermentation. A 3 × 3 Latin square design was used, involving 12 multiparous Holstein cows (parity = 3; body weight = 625 ± 25.8 kg; days in milk = 103 ± 19.6 day(s); milk yield = 32.6 ± 1.58 kg/d) over three 28-day periods. Cows were randomly assigned to three isocaloric and isonitrogenous diets: a basal diet with wheat bran (WB group), a diet with wheat bran replaced by PKM (PKM group), and a diet with wheat bran replaced by FPKM (FPKM group). Solid-state fermentation improved PKM’s nutritional profile by reducing fiber and β-mannan content while increasing protein availability and ruminal degradability. Compared to the WB group, the PKM group showed lower dry matter intake, milk yield, and nutrient digestibility. In contrast, the FPKM group had higher DMI and milk yield than the PKM group, improved nutrient digestibility, and the highest energy-corrected milk yield due to increased milk protein and lactose production. The FPKM group also had higher concentrations of total volatile fatty acids, propionate, acetate, and microbial protein synthesis than the PKM group. Pro-inflammatory cytokines (tumor necrosis factor-α, interleukin-8) were elevated in the PKM group but were reduced to levels similar to the WB group in the FPKM group. Plasma immunoglobulin G levels were higher in both the FPKM and WB groups compared to the PKM group. The FPKM group also showed increased relative abundances of Prevotella, Fibrobacterota, and Verrucomicrobiota, while Bacillota and Ruminococcus were reduced compared to the WB group. Metabolomic profiling revealed that FPKM upregulated energy metabolism and inflammation-related pathways, increasing metabolites such as riboflavin and adenine and decreasing succinic acid and guanine compared to the PKM group. In conclusion, FPKM improved the feeding value of PKM-based material and showed more favorable responses than PKM, with generally comparable responses to WB, supporting its potential as an alternative feed ingredient for lactating dairy cows.

Fresh-cut muskmelons have garnered significant attention due to the consumption trend of convenience and health, but they are prone to softening, browning and flavor deterioration, which limits their commercial value. L-Phenylalanine (L-Phe), the initial substrate for phenylpropane biosynthesis, is fundamentally important for plant stress resistance. This research investigated the impact of four consecutive (young fruit stage, early expansion stage, late expansion stage and mature stage) L-Phe spray throughout the melon fruit development phase on the postharvest quality of fresh-cut melons. The results indicated that L-Phe significantly increased the activities of NADPH oxidase (NOX), superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), enzymes related to the ascorbate-glutathione (AsA-GSH) cycle and the gene expression levels of aquaporins, it also promoted the accumulation of ascorbic acid (AsA), glutathione (GSH), total phenolics and flavonoids. This effectively eliminates excessive reactive oxygen species (ROS), reduces the O2 & sdot;- production rate and hydrogen peroxide (H2O2) levels. Meanwhile, L-Phe treatment enhanced the scavenging activity against DPPH, ABTS+ and FRAP free radicals, suppressed the accumulation of malondialdehyde (MDA), reduced cell membrane permeability, ultimately delayed the decline in hardness and total soluble solids (TSS) content, stabilized color and mitigated quality deterioration during storage. In conclusion, preharvest spraying of L-Phe can effectively improve the quality of fresh-cut melons by maintaining the ROS homeostasis of fruit and enhancing their antioxidant capacity. This study presents a safe, low-cost and feasible new strategy for preserving the quality of fresh-cut fruit and vegetables.

Ovum pick-up (OPU) technical variability limits the efficiency and consistency of bovine in vitro embryo production (IVP). Here, we established a sequential refinement framework for OPU by systematically evaluating key technical parameters across seven cumulative experiments in Holstein cows. An 18 G aspiration needle achieved significantly higher cumulus-oocyte complex (COCs) recovery than 20 G and 22 G needles (53.05% vs. 40.03-45.79%; p < 0.05). 90° improved recovery compared with non-vertical insertion (55.64% vs. 48.86%), while insertion to one-third to one-half follicle depth and bevel-down orientation further enhanced recovery efficiency (56.52% and 57.97%, respectively; p < 0.05). Polyvinyl chloride (PVC) tubing provided superior COCs recovery (63.29%) and morphological quality, with a tubing length of 1.2-1.5 m. Vacuum pressure of 50 mmHg resulted in higher COCs recovery (70.27%) and blastocyst rate (40.65%), significantly outperforming the previously used 70 mmHg setting. In contrast, excessive aspiration pressure (80 mmHg) was associated with mitochondrial dysfunction, disrupted cortical granule distribution, and increased apoptosis (p < 0.05), indicating impaired oocyte nuclear and cytoplasmic maturation under excessive mechanical stress. Notably, recovery efficiency progressively increased from 53.05% to 70.27% throughout the sequential optimization process throughout the sequential refinement process. Collectively, this study provides a field-based refinement approach for OPU procedures and demonstrates the association between aspiration parameters and bovine oocyte developmental competence under the tested conditions.

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.

Dairy cow re-identification (ReID) aims to enable cross-view individual identification by matching the same individual across images captured by different cameras, which is crucial for intelligent farm management. However, the same individual can exhibit large intra-class appearance variations across viewpoints and body orientations, especially under opposite orientations. In addition, different individuals may also show high inter-class similarity, which jointly increases the difficulty of cross-view identification. To address these issues, this study proposes FOONet with three key components: Fine-grained Complementary-view Feature Aggregator (FCFA) is designed to fuse complementary-view features to alleviate cross-view visual bias and enhance the discriminability among visually similar individuals; Opposite Orientation Mask Mechanism (OOMM) is introduced to mitigate the mismatch between the generic ReID training paradigm and dairy cow ReID caused by opposite-orientation discrepancies of the same individual; and Orientation Weighting Strategy (OWS) is applied during retrieval to suppress false matches from visually similar negatives under opposite orientations. Furthermore, this study introduces an auxiliary evaluation metric, mAP-IOPS, to assess retrieval performance for positives with the same or adjacent orientations. To evaluate the proposed method, this study deployed eight cameras on a farm for data collection and constructed a cross-view dairy cow ReID dataset containing 16,330 images of 78 dairy cows. Experimental results demonstrate that FOONet achieves Rank-1, Rank-5, Rank-10, and mAP-IOPS scores of 96.9%, 98.5%, 100.0%, and 75.9%, respectively, verifying its effectiveness for cross-view dairy cow identification. The dataset is available at https://github.com/IPCLab-NEAU/Dairy-Cow-Re-Identification.

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.

This study explored the effects of preharvest L-Phenylalanine (L-Phe) sprays on cuticle development and associated lipid metabolism in muskmelon (Cucumis melo L.) fruit. Spraying 8 mM L-Phe during fruit growth increased cuticle thickness and the accumulation of hydroxy fatty acids, fatty alcohols, alkanes, and triterpenoids. L-Phe enhanced the activities of key enzymes (FAT, LACS, FAR, SQE, OSC) and upregulated genes involved in fatty acid elongation, hydroxylation, alkane and triterpenoid biosynthesis. It also increased ATP-binding cassette (ABCG) transporter and lipid transfer protein (LTP) expression, facilitating transmembrane transport and deposition of wax and cutin monomers. Consequently, treated fruits formed a denser epidermal barrier and improved peel integrity. These findings demonstrate that exogenous L-Phe regulates cutin and wax biosynthesis at both enzymatic and transcriptional levels, providing a theoretical basis for using amino acid treatments to strengthen fruit epidermis and enhance postharvest storability.

This study developed a Proofman-LMTIA assay for the rapid detection of Bifidobacterium longum subsp. infantis (BLI). Primers and Proofman probes were designed targeting specific sequences of the BLI genome. After optimizing reaction conditions, the specificity and sensitivity of the method were evaluated and compared with quantitative real-time PCR (qPCR). The results showed that the established Proofman-LMTIA method had no cross-reactivity with other common Bifidobacterium species at the optimal reaction temperature of 61 °C. The detection limit was 1 × 10⁻⁵ ng/µL, which was consistent with qPCR. This isothermal amplification method achieved detection within 20 min, representing a 3-fold improvement in amplification efficiency compared to qPCR. These findings demonstrate that the Proofman-LMTIA assay is suitable for ultra-fast detection of BLI.

Winter weaning presents significant physiological challenges for calves. The research investigated how replacing soybean meal (SBM) with fermented soybean meal (FSBM) in calf starter would affect its growth, plasma indices, fecal microbiota, and metabolites in pre- and post-weaning calves. Twenty newborn calves (3.9 ± 1.4 d, 43.2 ± 2.65 kg) were randomly assigned to two dietary treatments: starter containing SBM or FSBM (16% FSBM replacement on a dry matter basis. Growth indicators, blood, and feces were collected regularly, with fecal samples collected during the weaning period used for multi-omics analysis. Results indicated that in the pre- and post-weaning periods, body weight and feed efficiency increased in the FSBM treatment group. Furthermore, the FSBM treatment group exhibited reduced blood lipopolysaccharide (LPS), adrenocorticotropic hormone (ACTH), cortisol (COR), serum amyloid A (SAA), haptoglobin (HP), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and malondialdehyde (MDA) concentrations, while increasing immunoglobulin-A (IgA), immunoglobulin-G (IgG), and total antioxidant capacity (T-AOC) levels (P < 0.05). Concurrently, during the weaning period, the replacement of FSBM facilitated changes in microbial communities of calves, including unclassified_Muribaculaceae and Christensenellaceae_R_7_group, and activated amino acid-related metabolic pathways (P < 0.05). In conclusion, replacing SBM with a part of FSBM in the starter improved growth performance, related plasma parameters, hindgut bacterial communities, and metabolites in weaning calves, offering a nutritional strategy to mitigate weaning stress during winter.

Agriculture is at the pivot point between anthroposphere, biosphere, and atmosphere. Innovative solutions are needed to reduce agricultural emissions and improve sustainability. Microalgae animal feed could be such a solution. This study aimed to evaluate the effects of 10 freshwater microalgae: Auxenochlorella protothecoides, Chlamydomonas pulvinate, Chlorella luteoviridis, Chlorella variabilis, Euglena mutabilis, Parachlorella kessleri, Stichococcus bacillaris, Tetradesmus acuminatus, Tetradesmus obliquus, and Tetraselmis gracilis, on ruminal methane (CH4) production, nutrient digestibility, and rumen fermentation using the in vitro Hohenheim gas test. The microalgae were cultured in a carbon dioxide (CO2) incubator at 2% CO2, at the optimal conditions for each strain. The highest producers were P. kessleri and T. obliquus, with a biomass concentration of 0.69 and 0.73 g/L·d, respectively. Their PUFA contents ranged from 33.2% to 69.1% of total fatty acids. Microalgae were tested at a 15% replacement in a control basal diet of 40.0% DM grass silage, 40.0% maize silage, 15% hay, and 5% concentrate. Data were analyzed using a mixed model in R. Ruminal CH4 production was reduced by 15.4%, 17.4%, and 16.4% in diets containing A. protothecoides, C. luteoviridis, and P. kessleri, respectively, compared with the control diet. Similarly, these diets reduced in vitro organic matter digestibility by 3.5%, 5.2%, and 5.4%, respectively. However, only A. protothecoides reduced CH4/CO2 ratio by 3.5% compared with the control diet. Propionate molar proportion was decreased by 2.4, 3.0, 2.5, and 2.5 percentage points for diets containing Ch. pulvinate, E. mutabilis, P. kessleri, and T. obliquus, respectively. Marginal effects of dietary variables were analyzed using the generalized additive model framework, revealing a negative relationship between dietary PUFA, sulfur content, and CH4 production, and a negative relationship between dietary PUFA and CH4/CO2 ratio. Incorporating high-PUFA microalgae in ruminant diets shows potential for reducing enteric CH4 emissions, warranting further investigation.

In this study, muskmelon plant and fruit were sequentially sprayed with 8 mM phenylalanine (Phe) four times during fruit development. The effect of preharvest Phe spraying on chilling injury (CI) of harvested muskmelons was assessed and the mechanism involved was investigated. We found that Phe spray activated NADPH oxidase (NOX), superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX) and glutathione reductase (GR), and increased glutathione (GSH) and ascorbic acid (AsA) levels during fruit chilling. The spray increased endogenous Phe, total phenolic and flavonoid content, and DPPH and ABTS(+) scavenging capacity. In addition, the spray decreased O-2(center dot-) production rate, H2O2 levels, cell membrane permeability and malondialdehyde (MDA) content, and significantly reduced CI index in fruit, which was 16.5 %, 16.6 %, 13.5 %, 20.2 % and 26.5 % lower than the control after 28 d, respectively. In conclusion, Phe spraying alleviates CI in harvested muskmelons by maintaining ROS homeostasis.

Enteric methane (CH4), the major contributor to on-farm greenhouse gas emissions, is a key mitigation target due to its high short-term global warming potential. The objectives of this study were to investigate the combined effects of 3-nitrooxypropanol (3-NOP) and Acacia mearnsii tannin extract (TAN), and their interactions with dairy cattle breed [Brown Swiss (BS) vs Holstein Friesian (HF)] on lactational performance and CH4 emissions. Sixteen multiparous mid-lactation cows, including 8 BS and 8 HF cows, were used in a split-plot design, with breed as the main plot. Cows within each subplot were arranged in a replicated 4 × 4 Latin Square design with a 2 × 2 factorial arrangement of treatments across four 24-d periods, including 3-d of sampling. The experimental diets were: (1) CON (basal total mixed ration), (2) 3-NOP (60 mg/kg DM), (3) TAN (3% of DM), and (4) 3-NOP + TAN. Spot samples of urine, faeces, and gas emissions (via GreenFeed) were collected at the end of each period 8 times over 3 days. No 3-NOP × TAN × Breed interactions were observed for DM intake (DMI), milk production, or enteric gas emissions, except for CH4 yield (g/kg DMI) and CO2 production. Breed influenced DMI, milk production, and component yields, with HF cows consuming 3.7 kg/d more DMI, producing 9.3 kg/d more milk, and achieving greater feed efficiency and higher milk component yields than BS cows. Milk yield and energy-corrected milk (ECM) tended to increase in HF but tended to decrease in BS cows by 3-NOP. Cows fed TAN had 1 kg/d lower DMI with the tendency for 3-NOP × TAN that showed greater reduction when TAN was fed alone, but milk yield, ECM, and feed efficiency remained unchanged. Cows fed TAN exhibited 18% lower milk urea nitrogen (N) concentration and 23.0% lower urinary N but 36.7% greater faecal N excretions as a percentage of daily N intake. A 3-NOP × Breed interaction was observed in CH4 production (g/d), with a 21.7% reduction in HF, and a 13.0% reduction in BS. Similarly, there were 3-NOP × Breed tendencies in CH4 yield and intensity (g/kg ECM), with reductions in HF cows of 21.8 and 23.4%, respectively, compared to 11.0 and 10.8% in BS cows. In conclusion, there were no synergistic or additive effects between 3-NOP and TAN on enteric CH4 mitigation. The enteric CH4 emission mitigating effect of 3-NOP was more pronounced in HF cows than in BS cows. Further research is needed to understand breed-specific responses and to optimise CH4 mitigation strategies for inclusion in national greenhouse gas inventories.

The purpose of this study is to investigate the effects of silage inoculants (FJ) and natural fermentation (CK) on the quality and microbial community of whole-plant corn silage under different fertilization treatments, including conventional fertilization (CK), liquid microbial inoculant and conventional fertilization (JJ), and microbial organic fertilizer and conventional fertilization (YJ). After 30 days of room-temperature fermentation, parameters including pH, LA, CP, starch, ADF, NDF, and the microbial community were determined. The results showed that after 30 days of ensiling, silage inoculants significantly affected the nutritional components and fermentation parameters of whole-plant corn silage under different fertilization treatments. Furthermore, the two factors (silage inoculants and different fertilization treatments) exhibited a significant interaction effect. Simple effects analysis revealed that the significant interaction was mainly driven by a more pronounced differential effect of fertilization treatments on the nutritional indicators (starch, CP, ADF, and NDF) under silage inoculant (FJ) addition than under natural fermentation (CK) (p < 0.05). Among all silage treatments, the silage inoculants + microbial solution drip irrigation and conventional fertilization (FJJJ) group exhibited relatively superior silage quality. Specifically, the FJJJ group had the lowest contents of pH, ADF, and NDF, along with the highest contents of lactic acid (LA) and ether extract (EE). The addition of silage inoculants under different fertilization treatments consistently increased the abundance and reinforced the dominance of Lactobacillus in the microbial community. This effect was most pronounced in the FJJJ group, which showed the highest relative abundance. In contrast, the relative abundance of genera such as Pantoea, Acinetobacter, Klebsiella, and Pseudomonas decreased significantly. In summary, appropriate fertilization treatments combined with the addition of silage inoculants contribute to enhancing the quality of whole-plant corn silage and improve the fermentation microbial community of the silage. These findings provide a theoretical basis for producing high-quality corn silage.

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.

The high prevalence of diarrhea in calves raises serious welfare concerns and imposes substantial economic losses on dairy farms. Probiotic strains that produce butyric acid may lower diarrhea incidence and improve gut health. This study evaluated the effects of direct-fed Clostridium beijerinckii R8 on growth performance, plasma biochemistry, and fecal microbiota in neonatal calves. Sixty newborn female calves were blocked by birth weight and randomly allocated to 4 treatments: (1) control (0 cfu/d), (2) low dose (1 × 109 cfu/d), (3) medium dose (1 × 1010 cfu/d), and (4) high dose (1 × 1011 cfu/d). Diarrhea frequency and duration exhibited a significant quadratic response to supplementation, with the lowest values observed at 1 × 1010 cfu/d. Quadratic treatment contrasts were significant for growth performance. Calves receiving 1 × 1010 cfu/d achieved the highest BW, ADG, DMI, body diagonal length, and hip width, together with the lowest feed conversion ratio. Plasma total cholesterol, high-density lipoprotein, BUN, and insulin increased linearly with dose. At 28 d of age, IgG, IgM, total antioxidant capacity, superoxide dismutase, and catalase levels showed linear increases, whereas malondialdehyde declined linearly as dose increased. Fecal microbiota showed both dose-dependent and temporal shifts. On d 28, Lachnoclostridium and Collinsella decreased linearly with dose, whereas Eubacterium_coprostanoligenes_group and Escherichia-Shigella displayed quadratic declines, reaching their lowest relative abundance at 1 × 1010 cfu/d. By d 56, Lachnospiraceae_NK4A136_group and Intestinimonas increased linearly with dose, whereas Bifidobacterium and Lactobacillus showed quadratic responses, with their lowest abundance at 1 × 1011 cfu/d. In conclusion, supplementation of 1 × 1010 cfu/d Clostridium beijerinckii R8 optimally reduced diarrhea, enhanced growth performance, and improved intestinal health by modulating the gut microbiota.

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.