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  • Rui Zhou, Li Zhe, Yves Mercier, Liang Hu, Ran Li, Hong Chen, Xiaoling Zhang, Lingjie Huang, Lun Hua, Yong Zhuo, Jian Li, Shengyu Xu, Yan Lin, Bin Feng, Lianqiang Che, De Wu, Zhengfeng Fang
    Animal Nutrition. 2025, 20(1): 145-157.

    Methionine (Met) metabolism is vital for one carbon metabolism, redox status and fetal development. Hence, this study investigated the effects of different levels and sources of Met on maternal metabolism, anti-oxidative capacity and fetal survival in pregnant sows. Forty primiparous sows were assigned to the following four groups: control group (basal diet, CON), 1.5S-OHMet group (supplemented methionine hydroxy analogue [OHMet] at 1.5 g/kg diet), 3.0S-OHMet group (supplemented OHMet at 3.0 g/kg diet), and 3.0S-Met group (supplemented L-Met at 3.0 g/kg diet) (n = 10). The trial lasted from day 60 of gestation to the farrowing day. Maternal 1.5S-OHMet consumption had the lowest stillborn ratio and the highest serum glucose levels during farrowing. Further analysis revealed that dietary 1.5S-OHMet consumption elevated the serum contents of glucose-6-phosphate, citric acid, butyric acid, malic acid, 3-methyladenine, 1-methyladenosine, ferulic acid and salicylic acid, but reduced the serum contents of succinic acid, oxoglutaric acid, 9(S)-hydroperoxylinoleic acid, 13(S)-hydroperoxy-octadecatrienoic acid, uric acid and urea nitrogen when compared to contents observed in the 3.0S-OHMet and 3.0S-Met groups (P < 0.05). Serum metabolomics analysis was conducted to determine the enriched differential metabolites and an enrichment analysis was performed using Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. The results showed that the enriched metabolites were mainly associated with central carbon metabolism, amino acid metabolism, lipid metabolism, and nucleotide metabolism. Moreover, maternal 3.0S-OHMet or 3.0S-Met consumption upregulated the trans-methylation pathway by elevating the S-adenosyl-methionine (SAM) level and the ratio of SAM to S-adenosyl-homocysteine (P < 0.05) at day 114 of gestation, while increasing homocysteine concentration (P < 0.001). However, compared to the 3.0S-Met group, maternal 3.0S-OHMet consumption elevated fetal survival and glutathione peroxidase (P < 0.05). Thus, this study provided new insights into the mechanisms through which sows fed with a 1.5S-OHMet diet during mid-to late-gestation period had high fetal survival, such as improvements in maternal amino acid, nucleotide and glycolipid metabolism.

  • Zhantao Yang, Yuhui Zheng, Kai Ren, Wei Wang, Shengli Li
    Animal Nutrition. 2025, 20(1): 171-181.

    Heat stress can lead to decreased feed intake, apoptosis of mammary epithelial cells, and decreased milk yield and quality. Selenium is an important element in the composition of at least 25 selenoproteins. Hydroxy-selenomethionine (HMSeBA) is a novel organic selenium that has been shown to have a better deposition effect. However, whether HMSeBA alleviates damage to the mammary gland blood-milk barrier caused by heat stress and how this affects the performance of dairy cows remain largely unexplored. Therefore, 32 healthy Holstein cows with similar gestation days (150.41 ± 20.07 d), milk yield (36.15 ± 3.02 kg) and parity (3.25 ± 0.51) were selected and randomly divided into two total mixed rations with different selenium (Se) sources: sodium selenite (SSe) and HMSeBA. This study evaluated the outcomes of HMSeBA on antioxidant capacity, immunity, and blood-milk barrier damage in dairy cows during heat stress by collecting the samples of blood, rumen fluid and mammary gland biopsy. The experiment was conducted over 35 d, including a 5-day pre-feeding period and a 30-day experimental period. The temperature and humidity index (THI) were all above 80 throughout the experiment period. The results showed that HMSeBA decreased the respiratory rate (P < 0.001) and the content of inflammatory cytokines in the serum and increased the content of immune factors and antioxidant capacity (P < 0.05). In addition, HMSeBA reduced the expression of inflammatory cytokines and heat shock proteins in mammary gland (P < 0.05). Hematoxylin-eosin-stained pathological sections showed massive thickening of acinar walls and severe destruction of glandular structures in the SSe group, but the structure of the acinar mammary gland in the HMSeBA group was intact. Furthermore, HMSeBA promoted the expression of the phosphatidylinositol 3-kinase (PI3K, P < 0.001)/protein kinase B (AKT, P = 0.011)/mammalian target of rapamycin (mTOR, P = 0.008) pathway and improved the expression of zonula occludens-1 (ZO-1, P = 0.014) and occluding (OCLN, P = 0.012) in the mammary gland, suggesting less damage caused by heat stress to the blood-milk barrier. Our results demonstrated that HMSeBA can improve the antioxidant capacity and immunity of dairy cows and the expression of tight junction proteins in mammary gland to help alleviate the blood-milk barrier damage by heat stress, which could reduce the damage of heat stress on milk yield.

  • Milan Kandel, Shemil P. Macelline, Mehdi Toghyani, Peter V. Chrystal, Mingan Choct, Aaron J. Cowieson, Sonia Yun Liu, Peter H. Selle
    Animal Nutrition. 2025, 20(1): 342-354.

    Feedstuffs derived from canola, predominantly canola meals plus whole, "full-fat" canola seed, and even canola protein isolates and/or concentrates, have the potential to decrease soybean meal inclusions in diets for broiler chickens. The protein content of soybean meal exceeds that of canola meal; however, canola meal contains more methionine and cysteine in absolute and relative terms. The purpose of this review is to explore this potential as Australian chicken-meat production is uniquely positioned to take advantage of this opportunity to the extent that it can be realised. Australia harvests ample quantities of canola, the bulk of which is exported as seed; alternatively, soybean production is very limited; therefore, large quantities of soybean meal are imported as the principal source of dietary protein for broiler chickens. This importation of soybean meal is not sustainable; however, canola meal inclusions in broiler diets do not usually exceed 100 g/kg. Regression equations derived from 15 recent studies indicate that dietary inclusions of 150 g/kg solvent-extracted canola meal would compromise weight gain by 4.04% and feed conversion ratio (FCR) by 4.72%. The foremost factors driving these depressions in canola meal are probably (1) high fibre contents coupled with low energy densities and (2) the presence of glucosinolates, which may be converted into toxic metabolites including thiocyanates. Moreover, regression equations from nine studies suggest that calculated dietary glucosinolate concentrations of 2.00 μmol/g would compromise weight gain by 5.72% and FCR by 6.56%. The nutritive value of canola meal could be enhanced by improvements in canola breeding programs, processing methods in canola meal production, and dietary formulations including judicious application of exogenous enzymes. Consideration is given to these aspects in this review as any improvements would increase the extent to which canola meal can feasibly replace soybean meal in broiler diets. An additional pathway to decrease the reliance on soybean meal could be the adoption of reduced-crude protein (CP) diets containing canola meal. The combined strategy of canola meal replacing soybean meal in reduced-CP diets, if successful, would tangibly decrease soybean meal requirements in global chicken-meat production.

  • Hao Cheng, Hao Li, Yujie Zhao, Kai Yang, Jing Wang, Bie Tan, Xiaokang Ma
    Animal Nutrition. 2025, 20(1): 226-238.

    The current study aims to investigate the potential interaction between glycosylation profiles of the Ningxiang breed (NX) and Western Duroc × Landrace × Yorkshire breed (DLY) weaned piglets, and their characteristic microbes, employing integrated analyses of transcriptomics and metagenomics. Twenty-four (12 NX and 12 DLY) at 28 days of age were transported into an experimental house and fed the same weaned piglet diet. The trail period was 7 days. Results revealed that the NX piglets had a higher growth-to-feed ratio, body weight gain scale, and lower pathological score of intestinal injury compared with the DLY piglets (P < 0.01). DLY piglets displayed elevated mRNA expression levels of MUC2 and MUC5AC in colonic mucosal tissue than NX piglets (P < 0.05). Within the O-linked glycosylated differentially expressed genes (DEGs), FNTA, GALNT18, POMGNT1, POMGNT2, and POMT1 were significantly upregulated in DLY piglets relative to NX piglets (P < 0.05). Conversely, C1GALT2, GALNT1, KMT2C, and OGT were significantly downregulated in DLY piglets compared to NX piglets (P < 0.05). The KMT2C gene was hardly expressed in the transcriptome of DLY piglets. At the phylum taxonomic level, NX piglets had a higher abundance of Firmicutes, while DLY piglets had a higher abundance of Proteobacteria. At the genus taxonomic level, NX piglets had a higher abundance of Lactobacillus, whereas DLY piglets had a higher abundance of Collinsella, Enterococcus and Escherichia. The results of the correlation between intestinal differential bacteria and O-chain glycosylated DEG showed that C1GALT2, GALNT1 and KMT2 were associated with Lactobacillus_pontis showed a positive correlation (R = 0.67). Through comparative analysis of differentially glycosylated genes and their associated functions, this study highlights the potential role of reduced expression of GALNT1 and KMT2C genes, involved in O-linked protein and glycan reactions, in impairing the intestinal barrier function of DLY piglets. Furthermore, members of the Lactobacillus and Prevotella genera may actively contribute to the regulation of piglet colon glycosylation profiles.

  • Shaobo Zhang, Manhua You, Youming Shen, Xinghua Zhao, Xin He, Juxiang Liu, Ning Ma
    Animal Nutrition. 2025, 20(1): 182-199.

    Bacteroides fragilis (B. fragilis), a crucial commensal bacterium within the gut, has shown connections with hepatic lipid metabolism and inflammation regulation. Nonetheless, the role of B. fragilis in the progression of fatty liver hemorrhagic syndrome (FLHS) remains unknown. This study aims to explore the ameliorative effects of B. fragilis on FLHS in laying hens, as well as its underlying mechanisms. This is the first study to employ a chicken FLHS model, combining microbiomics and oxylipin metabolomics to investigate the mechanism of action of intestinal symbiotic bacteria. Exp. 1: 40 laying hens at 25 weeks old were randomly divided into five treatment groups (eight replicates per group and one hen per replicate), including the control group (basal diet), the high-energy and low-protein (HELP) group, and the HELP group with three different levels (108, 109, and 1010 CFU) of B. fragilis. Exp. 2: 18 chickens at 25 weeks old were randomly divided into three treatment groups (six replicates per group and one hen per replicate) including the control group (basal diet), the model group (HELP diet), and the arachidonic acid (AA) group (HELP diet with 0.3% AA). The experiment period of Exp. 1 and Exp. 2 were 8 weeks. B. fragilis significantly improved body weight of seventh week (P = 0.006), liver lipid degeneration, blood lipid levels (triglycerides, cholesterol, and low-density lipoprotein cholesterol; P < 0.05), and liver function (alanine aminotransferase and aminotransferase; P < 0.05) in laying hens. B. fragilis downregulated the expression of lipid synthesis-related genes fatty acid synthase, acetyl-CoA carboxylase, and liver X receptor α, and inflammation-related genes tumor necrosis factor α, interleukin (IL)-1β, IL-6, and IL-8 in the liver of FLHS-affected hens (P < 0.05), while upregulating the expression of lipid oxidation-related genes carnitine palmitoyl transferase-1, peroxisome proliferator activated receptor (PPAR) α, and PPARγ (P < 0.05). The in-depth analysis indicated alterations in oxylipin pathways triggered by B. fragilis, as evidenced by changes in the expression of pivotal genes arachidonate 15-lipoxygenase, arachidonate 5-lipoxygenase (P < 0.05), subsequently causing modifications in relevant metabolites. This included a decrease in pro-inflammatory substances such as 15-oxoETE (P = 0.004), accompanied by an increase in AA (P = 0.008). B. fragilis regulated the homeostasis of intestinal flora by increasing the abundance of Bacteroides and decreasing the abundance of Succinatimonas and Faecalicoccus (P < 0.05). The integrated analysis revealed a robust positive correlation between Bacteroides abundance and AA levels (P = 0.007). This relationship was corroborated through in vitro experiments. Subsequently, the beneficial effect of AA in mitigating FLHS was confirmed in laying hens with FLHS, further supported by reverse transcription-polymerase chain reaction analysis demonstrating gene expression patterns akin to B. fragilis intervention. This study demonstrated that B. fragilis exerts an anti-FLHS effect through modulation of oxylipin metabolism and gut microbiota stability, with a pivotal role played by AA.

  • Dongdong Lu, Cuiping Feng, Yu Pi, Hao Ye, Yujun Wu, Bingxu Huang, Jinbiao Zhao, Dandan Han, Nicoline Soede, Junjun Wang
    Animal Nutrition. 2025, 20(1): 318-331.

    Maternal inulin intake has been shown to alleviate oxidative stress in piglets, but the role of bile acids (BAs) in this process remains unknown. This study aimed to investigate the roles of gut microbiota and BAs metabolism in the amelioration of intestinal oxidative stress in piglets through a maternal inulin diet. A total of 40 sows were allocated into two dietary treatments from day 85 of gestation until the end of lactation: CON (control diet) and INU (diet with 2% wheat bran replaced by inulin). An oxidative model was further established on the intestinal porcine epithelial cell-jejunum 2 cell line (IPEC-J2) to examine the effect of bacterial BAs on intestinal oxidative stress. Results showed that the maternal inulin diet promoted the average daily gain of piglets during suckling and reduced diarrhea rate during weaning (P = 0.026 and P = 0.005, respectively). Piglets from the INU group had lower serum levels of reactive oxygen species (P = 0.021), malondialdehyde (P = 0.045), along with higher serum levels of glutathione peroxidase (P = 0.027), catalase (P = 0.043), and total superoxide dismutase (P = 0.097). Compared to the CON group, maternal inulin intake increased fecal ursodeoxycholic acid (UDCA) by 10.84%, hyodeoxycholic acid (HDCA) by 250.64% (P = 0.026), and lithocholic acid (LCA) by 16.41% (P = 0.048) in piglets. Moreover, the fecal abundance of Ruminococcus and Christensenellaceae_R-7_group increased by 167.08% and 75.47% in INU piglets (P = 0.046 and P = 0.037, respectively). Furthermore, the in vitro study using IPEC-J2 cells demonstrated that UDCA, LCA, and HDCA attenuated intestinal oxidative stress by mediating kelch-1ike ECH-associated protein 1/nuclear factor E2-related factor 2 signaling. In conclusion, our results suggested that maternal dietary inulin intake during late gestation and lactation alleviates intestinal oxidative stress of piglets by regulating gut microbiota and BA metabolism.

  • Xin Yi, Yueming Li, Yue Liu, Minzhe Zhang, Zhenming Zhou, Qingxiang Meng, Hao Wu
    Animal Nutrition. 2025, 20(1): 42-53.

    The present study investigated whether replacing dietary rice straw with peanut vine (PEV) and Broussonetia papyrifera silage (BPS) reduces the use of soybean meal and explored its effects on the growth performance, blood biochemical indicators, serum metabolomics, and meat quality of fattening bulls. Forty-five Simmental crossbred bulls (initial body weight = 484.29 ± 8.49 kg) were randomly allotted into three dietary treatment groups (n = 15): (1) CON, 5% rice straw (DM basis); (2) PEV, 5% peanut vine (DM basis); and (3) BPS, 5% B. papyrifera silage (DM basis). The remaining roughage for all three treatment groups was supplemented with 25% corn silage (DM basis). The experiment lasted for 123 d, with the first 14 d serving as an adaptive period. Throughout the experiment, dietary BPS decreased the average daily dry matter intake (P < 0.001) and feed cost (P < 0.001). Serum metabolomics analysis showed that PEV affected the phenylalanine, tyrosine, and tryptophan biosynthesis pathways (P = 0.021) and lysine degradation pathway (P = 0.042), whereas BPS affected the phenylalanine, tyrosine and tryptophan biosynthesis pathways (P = 0.004), lysine degradation pathway (P = 0.012), and serotonergic synapse pathway (P < 0.001). Regarding meat quality, the redness (P = 0.025) and hue angle values (P < 0.001) of the longissimus dorsi muscle were lower in the BPS group than in the CON and PEV groups. The yellowness of the longissimus dorsi muscle was lower in the BPS group than in the PEV group (P = 0.024), and the shear force was lower in the PEV group than in the BPS group (P = 0.014). However, lysine content in beef was higher in the BPS group than in the CON group (P = 0.005). In conclusion, replacing rice straw with PEV and BPS reduced the use of soybean meal but had no adverse effects on growth performance. BPS affected the amino acid metabolism of bulls, thus decreasing feed intake and increasing the lysine content in meat. The PEV group showed better meat quality than the BPS group.

  • Jian Chen, Kanwar K. Malhi, Xiaowei Li, Xiangwen Xu, Jianxun Kang, Bichen Zhao, Yaru Xu, Xuenan Li, Jinlong Li
    Animal Nutrition. 2025, 20(1): 95-109.

    Weaning stress causes substantial economic loss in the swine industry. Moreover, weaning-induced intestinal barrier damage and dysfunction of the gut-liver axis are associated with reduced growth performance in piglets. Metasilicate-based alkaline mineral water (AMW) has shown potential therapeutic effects on gastrointestinal disorders; however, the mechanisms involved and their overall effects on the gut-liver axis have not been explored. Here, sodium metasilicate (SMS) was used to prepare metasilicate-based AMW (basal water + 500 mg/L SMS). A total of 240 newly weaned piglets were allocated to the Control and SMS groups (6 replicate pens per group and 20 piglets per pen) for a 15-day trial period. Histopathological evaluations were conducted using hematoxylin and eosin staining. To analyze the composition of the gut microbiota, 16S rRNA PacBio SMRT Gene Full-Length Sequencing was performed. Western blotting and immunofluorescence were employed to assess protein expression levels. Our results indicated that metasilicate-based AMW effectively alleviated weaning-induced colonic or liver morphological injury and inflammatory response, as well as liver cholesterol metabolism disorders. Further analysis showed that metasilicate-based AMW promoted deoxycholic acid (DCA) biosynthesis by increasing the abundance of Lactobacillus_delbrueckii in the colon (P < 0.001). This consequently improved weaning-induced colon and liver injury and dysfunction through the DCA-secondary bile acid (SBA) receptors (SBAR)-nuclear factor-kappaB (NF-κB)/NOD-like receptor family pyrin domain-containing 3 (NLRP3) pathways. Growth performance parameters, including final body weight (P = 0.034) and average daily gain (P < 0.001), in the SMS group were significantly higher than those in the Control group. Therefore, metasilicate-based AMW maintains gut-liver axis homeostasis by regulating the microbiota-mediated SBA-SBAR pathway in piglets under weaning stress. Our research provides a new strategy for mitigating stress-induced gut-liver axis dysfunction in weaned piglets.

  • Weibin Wu, Muhammad Faheem Akhtar, Jiahong Geng, Huizhen Lu, Muhammad Ajwad Rahim, Jianbo Cheng, Xiaoling Ding, Shengyong Mao, Yanfeng Xue
    Animal Nutrition. 2025, 20(1): 263-276.

    Nutrition consistently affects microbe-host interactions in the gastrointestinal tract. This study aimed to unravel how undernutrition reshapes the microbial composition and the homeostasis of epithelium in the jejunum and ileum. Sixteen late-gestation Hu-sheep were randomly assigned to the control group (n = 8, 100% ad libitum feeding levels) or the undernutrition group (n = 8, which received 30% ad libitum feeding levels). After 15-d treatment, all ewes were slaughtered, and jejunal and ileal digesta and epithelium samples were collected for 16S rRNA gene sequencing and transcriptome sequencing, respectively. Results indicated that undernutrition decreased the jejunal and ileal tissue weights (P = 0.005 and P = 0.022) and the levels of volatile fatty acids (P = 0.019 and P = 0.007) and microbial protein levels (P = 0.019 and P = 0.031) in jejunal and ileal digesta. The relative abundance of acetate producing microbiota, including Clostridia UCG-014 norank, Ruminococcus, [Ruminococcus] gauvreauii, and Lachnospiraceae_Blautia, were significantly reduced (P < 0.05) in the jejunum and ileum. Undernutrition up-regulated (P < 0.05) the expression of genes involved in amino acid synthesis and fatty acid oxidation, but down-regulated (P < 0.05) the expression of genes associated with amino acid degradation, fatty acid synthesis, and extracellular structures in jejunal and ileal epithelium. In the jejunal epithelium, genes associated with extracellular matrix–receptor interactions, cell growth, and immune response were down-regulated (P < 0.05) upon undernutrition. Taken together, undernutrition changed the microbial community in the jejunum and ileum, which altered the fermentation mode and the production of volatile fatty acids and microbial protein. These affected the energy and protein system in the epithelium and reprogrammed substance metabolism and extracellular structures, which probably further influenced cell growth and immune response. These insights provide a foundation for completely clarifying the crosstalk between small intestinal microbiota and the host.

  • Xi Sun, Chaohui Wang, Sijing Li, Xiaoying Liu, Yun Li, Yumeng Wang, Yuxin Niu, Zhouzheng Ren, Xin Yang, Xiaojun Yang, Yanli Liu
    Animal Nutrition. 2025, 20(1): 54-65.

    Multiple stressors are believed to deteriorate production performance and cause substantial economic losses in commercial poultry farming. Folic acid (FA) is an antioxidant compound that can improve oocyte function and regulate gut microbiota composition. The current study was conducted to investigate the role of FA in alleviating stress and improving production performance. Sixty Hyline Brown laying hens at 21 weeks of age were randomly divided into three groups, with 10 replicates in each group and each replicate containing two chickens. Each group received basic diet and saline injection (Con group), basic diet with dexamethasone (DXM) injection (DXM group), or basic diet supplemented with FA (13 mg/kg in the premix) with DXM injection (FA group). The feeding trial lasted five weeks. Birds in the DXM and FA groups receiving subcutaneous DXM injections at a dosage of 4.50 mg/kg per day during the first seven days of the trial. Results showed that the levels of corticosterone, triglyceride, total cholesterol, and malondialdehyde in serum were significantly increased in the DXM group (P < 0.05), while the concentrations of FA and 5-methyltetrahydrofolate were decreased in the DXM group (P < 0.05). Laying hens in the DXM group had lower laying rates and egg quality, including egg weight, eggshell thickness, eggshell strength, albumen height, and Haugh units (P < 0.05). Conversely, FA alleviated these negative impacts. Through transcriptome analysis, a total of 247 and 151 differentially expressed genes were identified among the three groups, and 32 overlapped genes were further identified. Moreover, 44 and 59 differential metabolites were influenced by DXM and FA, respectively. Kyoto Encyclopedia of Genes and Genomes enrichment from the transcriptome and metabolomics showed that the reduced production performance may be due to the disturbance of oocyte production, calcium metabolism, and oxidative stress. Analysis of 16S rRNA gene amplicon sequences revealed the differential microbial composition and potential functional changes among the different groups. LEfSe analysis showed that Mucispirillum and Nautella were the predominant bacteria in the DXM group, while Clostridium was the predominant bacteria in the FA group. Functional prediction demonstrated that stressors enhanced fatty acid biosynthesis, while betaine biosynthesis and retinol metabolism were elevated in the FA group. Dietary FA reversed the elevated levels of bile acids (BA), including cholic acid, taurodeoxycholic acid, and taurochenodeoxycholic acid (P < 0.05). The DXM group showed an overall decrease in short-chain fatty acids (SCFA), but FA restored the concentrations of acetic acid, propionic acid, and isobutyric acid (P < 0.05). In conclusion, this study reveals that dietary FA can alleviate the degradation of production performance caused by stress through improving circulating antioxidant capacity, maintaining intestinal microbiota homeostasis, and regulating SCFA and BA biosynthesis. Thus, highlighting the prominent role of gut microbe-host interactions in alleviating multi-stresses.