Search Results (45)

This study aimed to investigate the regulatory effects of dietary dandelion supplementation on the ruminal physiological status of lambs fed a high-concentrate diet. Twenty-two 6-month-old female crossbred lambs with an initial body weight of 32.05 ± 0.43 kg were randomly assigned to two groups (n = 11 per group): the HC group (fed a high-concentrate basal diet) and the DD group (basal diet supplemented with 2 g/kg dandelion powder), with a 14-day adaptation and 60-day experimental period. The results showed that compared with the HC group, dandelion supplementation significantly improved lamb growth performance and modulated ruminal fermentation parameters and tissue morphology. For inflammatory indices, the concentration of interleukin-10 (IL-10) was significantly increased, while the concentrations of interleukin-1β (IL-1β), lipopolysaccharide (LPS) and histamine (HIS) were significantly decreased (p < 0.05). Meanwhile, dandelion supplementation upregulated the mRNA expression of ruminal epithelial tight junction proteins and hexokinase II (HK-II), while downregulating the mRNA expression of myeloid differentiation factor 88 (MyD88), voltage-dependent anion channel 1 (VDAC1), inositol 1,4,5-trisphosphate receptor (IP3R), NLR family pyrin domain containing 3 (NLRP3), and caspase-1. Metagenomic analysis revealed that dandelion supplementation did not alter ruminal microbial α-diversity, but slightly modulated community composition and functional profiles. The abundances of dominant phyla Bacteroidota and Bacillota showed only slight fluctuations, while fiber-degrading genera (Xylanibacter, Quinella, Selenomonas) showed a trend toward enrichment, and proteolytic taxa were decreased. CAZyme analysis revealed an upward trend in families CE4, CE8, GH32, and GH13_46 (pectin/starch degradation) and a downward trend in GH73, GH27, and GT14 (oligosaccharide/peptidoglycan/polysaccharide metabolism) in the DD group. KEGG orthology annotation suggested that nominal KO differences were mainly associated with carbohydrate metabolism pathways, including starch and sucrose metabolism and peptidoglycan biosynthesis. None of these features, however, remained significant after FDR correction (raw p < 0.05; all q > 0.05). Collectively, dietary dandelion supplementation was associated with alterations in ruminal physiological status and microbial community in lambs fed a high-concentrate diet, providing foundational data for dandelion application in lamb diets under short-term feeding conditions. Full article

Genetic improvement of growth traits is a key objective in aquaculture breeding. In this study, we performed whole-genome resequencing of 282 Japanese flounder individuals from a single farmed cohort and obtained approximately 6.67 million high-quality SNPs. Single-trait and multi-trait GWAS were conducted for 11 growth traits: body weight (BW), total length (TL), body length (BL), body depth (BD), trunk length (TUL), head length (HL), snout length (SnL), caudal peduncle depth (CPD), eye diameter (ED), postorbital head length (PoL), and interorbital width (IW). Heritability estimates ranged from 0.172 (IW) to 0.487 (HL). Head length showed the highest heritability, followed by PoL (0.328), ED (0.323), BD (0.300), and CPD (0.291). Genetic correlation analysis revealed that BW was strongly positively correlated with most body size traits (r_g > 0.9), indicating that simultaneous improvement of these traits is feasible. Single-trait GWAS identified a total of 55 suggestive associated loci (top five SNPs per trait, combined across all 11 traits, at p < 1 × 10⁻⁵). Notably, we observed pronounced pleiotropic effects, where a single SNP or gene influences multiple growth traits. For example, multiple SNPs on chromosome 14 were associated with more than five traits (TL, BL, BD, TUL, BW), and the fras1 gene affected both head length and post-orbital head length, demonstrating a genetic basis of pleiotropy. Additionally, fras1 was associated with head length, and nkd1 with interorbital width. Multi-trait GWAS uncovered 12 novel loci not detected in single-trait analyses, with candidate gene annotation pointing to pathways such as ECM–receptor interaction (e.g., fras1, lamb4) and Wnt signaling (e.g., nkd1), suggesting that these biological processes may underlie the observed associations. These findings are preliminary and require validation in larger independent cohorts. KEGG enrichment analysis showed significant enrichment in ECM–receptor interaction and non-homologous end-joining pathways (corrected p < 0.05). Our results provide a genetic basis for growth trait improvement and candidate markers for breeding programs in Japanese flounder. Full article

In this study, we investigated the protective effects of CPP-C3M4 against Salmonella-induced hepatic injury in lambs. Thirty 72-day-old Hu sheep were randomly divided into five groups: the control group (CON), the Salmonella-infected group (ST), and three CPP-C3M4 pretreatment groups at low, medium and high doses (CPP-C3M4-L, CPP-C3M4-M, and CPP-C3M4-H). The results showed that administration of CPP-C3M4 via duodenal fistula significantly alleviated liver enlargement and histopathological damage caused by Salmonella infection. CPP-C3M4 improved liver function by reducing alanine aminotransferase (ALT), aspartate aminotransferase (AST), and triglyceride (TG) levels. It enhanced the liver antioxidant capacity by increasing glutathione peroxidase (GSH-Px), catalase (CAT), and superoxide dismutase (SOD) activities. It also alleviated inflammatory responses by downregulating the mRNA expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2), toll-like receptor 2 (TLR2), and toll-like receptor 9 (TLR9). Transcriptomic Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis further confirmed that CPP-C3M4 exerts its protective effects by suppressing multiple inflammatory pathways, with the interleukin-17 (IL-17) signaling axis being a key target of regulation. These results indicate that CPP-C3M4 is a promising alternative antimicrobial substance for preventing and alleviating Salmonella-induced liver injury in lambs. Full article

Haemonchus contortus is one of the most prevalent gastrointestinal nematodes in all ecosystems in Mexico, where sheep are raised on pasture, and it poses a significant threat. This study aimed to examine the surgical isolation of a Mexican H. contortus strain, its morphological and molecular characterization, and the maintenance of this strain for future research. Biological behavior and some phenotypic aspects of the adults were considered. Fecal samples were obtained from naturally infected sheep, larval cultures were performed, and a nematode-free lamb was infected. Once the infection was established, the donor sheep were euthanized, the adults recovered, and H. contortus females and males were selected. A surgical transfer from H. contortus adult to the abomasum of a receptor lamb was performed, and the beginning of the egg excretion was confirmed three days post-transfer; fecal cultures from the receptor lamb were conducted to verify the purity of the strain. After three lambs were infected with 3000, 5000, and 10,000 L3, the pre-patent period, prolificacy, and measurements of the adult stages of the strain were studied. The molecular characteristics were evaluated by qPCR; primers were designed based on NCBI genomic DNA sequences of H. contortus to amplify a 176 bp fragment, and the amplicon was sequenced for taxonomic identification. The results of this study describe biological characteristics and some phenotypic aspects of the adults, as well as eggs and infective larvae, and molecular characteristics of the isolated strain, and establish a successful methodology for isolating and maintaining a pure strain of H. contortus (FESC-strain); it can be used as a reference in experimental infections or anthelmintic resistance studies. Full article

Junken meat sheep exhibit a characteristic postnatal coat-color transition, in which the initially black fleece gradually fades and develops into a white-trunk phenotype; however, the transcriptional basis of this developmental change in follicular pigment output remains unclear. In this study, three Junken meat sheep lambs showing a natural postnatal black-to-white coat-color transition were sampled longitudinally at the newborn black-fleece stage and the 179-day white-trunk stage, generating three matched biological pairs for RNA-seq analysis. Representative candidate genes were further validated by RT-qPCR. Differential expression analysis identified 1657, 400, and 1086 differentially expressed genes in the C11 vs. C1, C22 vs. C2, and C33 vs. C3 comparisons, respectively. Functional enrichment analysis indicated that these genes were mainly associated with tyrosine metabolism, ECM–receptor interaction, focal adhesion, Phosphoinositide 3-kinase-Akt signaling pathway (PI3K-Akt), arachidonic acid metabolism, estrogen signaling, and immune-related pathways. Integrated analysis of shared downregulated genes and expression patterns highlighted candidate genes related to pigmentation, the ECM/follicular microenvironment, and regulatory or metabolic processes. Pigmentation-related genes, including SOX10, TYR, TYRP1, PMEL, OCA2 and SLC45A2, were generally downregulated in 179-day white-trunk-stage skin, while changes in ECM- and metabolism-related genes suggested altered follicular microenvironmental regulation. These findings identify candidate transcriptional features associated with developmental coat-color fading in Junken meat sheep and support a follicular microenvironment–melanogenesis expression axis as a transcriptome-based framework for further investigation. Full article

Leucine, an essential branched-chain amino acid, serves not only as a substrate for protein synthesis but also as a key regulator of placental function and fetal development. This study investigated the effects of dietary supplementation with RP-Leu during late gestation on placental development and offspring performance in Hu sheep. Sixty twin-pregnant ewes at day 80 of pregnancy were randomly assigned to either a control group (fed a basal diet) or an RP-Leu group (fed a basal diet supplemented with 19 g/day RP-Leu). The feeding trial lasted for 60 d. The ewes were slaughtered at day 140 of gestation. Maternal slaughter traits and fetal organ weights were recorded. Blood and milk samples were collected for milk composition analysis and targeted metabolomic profiling. Leucine supplementation significantly increased the percentage of milk fat content, total solid content, and the birth weight of lambs (p < 0.05). Improvements in placental morphology and antioxidant capacity were observed, including a significant increase in cotyledon density and a significant enhancement of catalase (CAT) activity (p < 0.05). Gene expression analysis indicated that the NOS3, SLC38A1 and FABP4 genes in the placental cotyledons (p < 0.05), and the VEGFA, NOS3, SLC27A1 and FABP4 genes were significantly upregulated in the maternal caruncles (p < 0.05). Plasma metabolomic profiling revealed increased L-glutamic acid levels and alterations in several amino acids, with pathway enrichment indicating involvement in amino acid metabolism and membrane transport processes. Transcriptomic analysis identified 739 differentially expressed genes, which were mainly enriched in the PI3K/Akt signaling pathway, ECM–receptor interaction pathway, and cytokine–cytokine receptor interaction pathway. Collectively, these findings suggest that RP-Leu supplementation during late gestation may enhance offspring growth by modulating amino acid metabolism, promoting placental development, and improving placental nutrient transport capacity, thereby supporting fetal growth and development. Full article

Early weaning in intensive lamb production improves reproductive efficiency but predisposes lambs to diarrhea, oxidative stress, and intestinal barrier dysfunction, highlighting the need for non-antibiotic strategies to protect gut health. This study evaluated whether a sheep-derived mixed probiotic could alleviate early weaning–induced intestinal injury and clarified its potential molecular mechanisms. Early weaning reduced body weight, average daily gain and feed efficiency, increased diarrhea, decreased plasma and colonic catalase (CAT), glutathione peroxidase (GSH-PX), and superoxide dismutase (SOD) activities, increased malondialdehyde (MDA), elevated interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α), reduced interleukin-10 (IL-10) and transforming growth factor-β (TGF-β), increased plasma and mucosal immunoglobulin A, M, and G (IgA, IgM, IgG), and increased colonic lipopolysaccharide (LPS) with reduced diamine oxidase (DAO). Intestinally, EW induced villus atrophy, deeper crypts, lower villus height-to-crypt depth ratios, goblet cell loss, higher histopathological scores, and decreased colonic mucin 2, zonula occludens-1, claudin-1, and occludin. Probiotic supplementation partially reversed these alterations, restoring antioxidant enzyme activities, improving villus architecture and barrier protein expression, and rebalancing cytokine and immunoglobulin profiles. Transcriptomic and network analyses showed that early weaning activated Cytokine–cytokine receptor, NF-κB, TNF and Th17 pathways, whereas probiotics suppressed a weaning-responsive inflammatory gene module, downregulated key hub genes, and enhanced peroxisome proliferator-activated receptor (PPAR) signaling. These results show that supplementing early-weaned lambs with a mixed probiotic generated from sheep is an efficient nutritional strategy to reduce intestinal oxidative and inflammatory damage associated with weaning and to enhance their health and performance. Full article

Compound yeast culture (CYC) is known to enhance animal health, but its effects on hepatic immune function are unclear. This study systematically examined CYC’s regulatory effects on the liver of weaned lambs using transcriptomics and integrative bioinformatics. Ten lambs were randomly assigned to a control diet or a basal diet supplemented with 30 g/d per head of Saccharomyces cerevisiae and Kluyveromyces marxianus co-culture (CYC group) for 42 days. Histological analysis showed that CYC improved hepatocyte arrangement and sinusoidal integrity, suggesting enhanced hepatic tissue stability. Cytokine analysis revealed CYC significantly increased IL-6 and IL-1β while reducing IL-10, TGF-β1, TNF-α, and CXCL9, indicating a bidirectional modulation of the immune response. Additionally, CYC enhanced antioxidant defenses by increasing T-SOD, GSH-Px, and T-AOC activities and decreasing MDA content. Transcriptomic sequencing indicated that CYC reshaped hepatic gene expression. Upregulated genes were enriched in immune-regulatory and structural pathways, including PI3K-AKT signaling, ECM–receptor interactions, Toll-like receptor pathways, and cell adhesion molecules. Protein-level validation further confirmed activation of PI3K and AKTAKT phosphorylation with limited engagement of NF-κB signaling. Conversely, downregulated genes were mainly associated with oxidative stress and energy metabolism, such as ROS-related pathways and MAPK signaling. WGCNA identified key hub genes (PTPRC, CD86, and ITGAV), which correlate with pro-inflammatory factors and participate in immune recognition, T-cell activation, and cell adhesion. These data suggest that CYC promotes hepatic immune homeostasis by enhancing immune signaling, stabilizing tissue architecture, and modulating oxidative stress/metabolic processes. This study provides mechanistic insights into CYC’s regulation of liver immune function and supports its targeted application as a functional feed additive for ruminants. Full article

The Slit2/Robo signaling pathway acts as a tumor suppressor in various cancers. This study identified an 8-amino acid peptide, LT1-3, derived from the Slit2 LamG domain, and demonstrated its ability to inhibit lung cancer cell proliferation and invasion independently of Robo receptors. Notably, LT1-3 was non-toxic to normal cells (Beas-2B, MRC5, and HUVECs). Combination treatment of LT1-3 and cisplatin synergistically inhibited the proliferation of lung cancer cells (CL1-5, A549, H1355, H460, H23, H661), but had no inhibitory effect on H1299 and H1975. Furthermore, combination therapy prolonged the median survival of tumor-bearing immunodeficient nude mice from 27.5 days (control) to 37.5 days (LT1-3 or cisplatin) and further to 47.5 days (LT1-3/cisplatin combination). The tumor suppressor TP53 positively influences LT1-3-mediated proliferation inhibition, while MAPK8 (JNK1) and PRKACA (PKA) have been identified as negative regulators. With the exception of the p53R273 variants, most TP53 mutants retained their function in this context. The p53 reactivator APR-246 restores sensitivity of p53R273H-expressing cells to LT1-3. JNK inhibition sensitizes p53-deficient or p53R273H-expressing cells to LT1-3-mediated proliferation inhibition. LT1-3, alone or in combination with a JNK inhibitor, enhances cisplatin efficacy, even in the presence of p53 mutations. Therefore, LT1-3 possesses multifunctional antitumor properties, directly inhibiting tumor cells and enhancing the efficacy of cisplatin, without causing toxicity to normal cells. Combining LT1-3 with cisplatin holds promise as a first-line therapy for lung cancer, while LT1-3 alone may be suitable for maintenance therapy. Full article

Reproductive and growth traits are key economic traits in sheep. This study aims to identify key single nucleotide polymorphisms (SNPs) and candidate genes associated with reproductive and growth traits in Tianmu polytocous sheep through a genome-wide association study (GWAS). The findings are expected to provide both a theoretical foundation for molecular breeding in this breed and novel insights into the genetic basis of ovine reproductive and growth performance. This study took 483 adult Tianmu polytocous ewes as the research subjects, collected their lambing records, measured their phenotypic values of growth traits (3 weight and 11 body size traits), and collected their blood samples for whole-genome resequencing to identify SNPs in the Tianmu polytocous sheep genome. The results identified a total of 9,499,019 (3× coverage) and 27,413,216 (30× coverage) high-quality SNPs in the Tianmu polytocous sheep genome. Subsequently, the association analysis between SNPs and reproductive and growth traits was conducted using a mixed linear model. A total of 92, 66, 18, 28, 6, 42, 3, 3, 6, 1, 12, 3, 22, 8, 6, and 3 SNPs were found associated with litter size at first parity, litter size at second parity, litter size at third parity, litter size at fourth parity, birth weight, weaning weight, body height, withers height, body length, head length, head width, cannon bone circumference, forelimb height, chest girth, chest depth, and withers width, respectively. Further, based on SNP annotation, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, candidate genes associated with the reproductive and growth traits were identified. Among these genes, 11 LOC, DEPTOR, GNG12, GRM7, PTH, PTH2R, WWOX, INHA, and NRG3 are candidate genes associated with litter size at first parity or litter size at third parity. These genes are involved in the G protein-coupled receptor signaling pathway, G protein-coupled receptor activity, ovarian tissue development, and hormone secretion. Additionally, TFRC and NTN1 are candidate genes associated with birth weight, while five UGT1A and CASR are candidate genes associated with weaning weight. These candidate genes are primarily involved in lipid metabolism. Finally, the following genes were identified as candidates associated with specific traits: DLG2, TMEM126A, and TMEM126B with body height; DSCAM and SCN8A with body length; BARX1 with cannon bone circumference; four LOC genes with forelimb height; EPHA4 with chest depth; and MRS2 with withers width. Full article

To investigate the gene regulatory mechanisms underlying muscle development in Oula sheep at different growth stages, under supplementary feeding, particularly the shift in core regulatory mechanisms governing muscle development from the fetal stage to the postnatal period, we conducted transcriptomic sequencing and comparative analysis of the longissimus dorsi muscle collected during the embryonic, lamb, and adult stages. Differentially expressed genes (DEGs) potentially associated with muscle growth and development were identified across various age phases. Furthermore, Short Time-series Expression Miner (STEM) analysis was employed to decipher the temporal expression patterns of these DEGs. The results indicated that metabolic processes related to carbohydrates, energy, and amino acids were enhanced with increasing age in Oula sheep muscle. Comparative analysis between different growth stages revealed that the functional enrichment of DEGs was directly associated with changes in skeletal muscle development, with significant enrichment in biological pathways such as ECM–receptor interaction, PI3K-AKT signaling pathway, and protein digestion and absorption. Additionally, we observed that PTPRC, IL10, NDUFAB1, BUB1, BUB1B, CDK1, ITGB3, and ITGB2 may play pivotal roles in the regulation of muscle growth and development across different stages in Oula sheep. These findings provide theoretical support for the understanding of the genetic regulatory mechanisms underlying muscle development in Oula sheep. Full article

Feeding HC diets has been found to induce metabolic dysregulation in the colon. However, the mechanisms by which changes in colonic flora and metabolites damage the colonic epithelium are poorly studied. Therefore, the present experiment used a multi-omics technique to investigate the mechanism of colonic injury induced by high-concentrate diets in lambs. Twelve male Dumont lambs were randomly split into two groups: a low-concentrate diet (LC = concentrate/forage = 30:70) group and a high-concentrate diet (HC = concentrate/forage = 70:30) group. The results showed that the HC group presented significantly increased lipopolysaccharide (LPS) concentrations in the colonic epithelium and significantly decreased serum total cholesterol (TC), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) levels (p < 0.05), which led to cavities and inflammatory cell infiltration in the colonic epithelium. The HC group had significantly lower pH and less VFAs in colon contents, as well as a significantly increased abundance of bacteria of the genera [Eubacterium]_coprostanoligenes_group, Rikenellaceae_RC9_gut_group, Treponema, Clostridia_UCG-014, Alistipes, Ruminococcus, Christensenellaceae_R-7_group, UCG-002, Bacteroidales_RF16_group and Lachnospiraceae_AC2044_group compared to the LC diet group. These microorganisms significantly increased the level of metabolites of cholic acid, chenodeoxycholic acid, LysoPA (P-16:0/0:0), methapyrilene, and fusaric acid. A transcriptome analysis showed that cytokine–cytokine receptor interaction, glutathione metabolism, and the peroxisome signaling pathway were downregulated in the colon epithelium of the lambs fed the HC diet. Therefore, the HC diet caused epithelial inflammation and oxidative damage by affecting the interaction between the microbial flora of the colon and metabolites and the host epithelium, which eventually disrupted colon homeostasis and had a negative impact on sheep health. Full article

This study aimed to establish a CRISPR/Cas9 gene-editing system for Larix kaempferi (Lamb.) Carr. (Japanese larch). We screened L. kaempferi U6 promoters and used them to drive sgRNA expression in the CRISPR/Cas9 gene-editing system. The L. kaempferi embryogenic callus was used as the receptor material for genetic transformation, and the frequency and types of gene editing were then analyzed. The results showed various mutations in the transgenic materials, including base substitutions and deletions, and the editing frequency ranged from 5% to 14.29%. In summary, we established a CRISPR/Cas9 gene-editing system for L. kaempferi. Our results demonstrate that the CRISPR/Cas9 system can efficiently edit genes in L. kaempferi, with significantly higher editing frequencies observed when sgRNA expression is driven by endogenous LaU6 promoters compared to the exogenous promoter ProAtU6-26. This work provides technical support for the study of L. kaempferi gene functions and genetic improvement. Full article

The production and release of cortisol during stress responses are key regulators of growth in teleosts. Understanding the molecular responses to cortisol is crucial for the sustainable farming of rainbow trout (Oncorhynchus mykiss) and other salmonid species. While several studies have explored the genomic and non-genomic impacts of cortisol on fish growth and skeletal muscle development, the long-term effects driven by epigenetic mechanisms, such as cortisol-induced DNA methylation, remain unexplored. In this study, we analyzed the transcriptome and genome-wide DNA methylation in the skeletal muscle of rainbow trout seven days after cortisol administration. We identified 550 differentially expressed genes (DEGs) by RNA-seq and 9059 differentially methylated genes (DMGs) via whole-genome bisulfite sequencing (WGBS) analysis. KEGG enrichment analysis showed that cortisol modulates the differential expression of genes associated with nucleotide metabolism, ECM-receptor interaction, and the regulation of actin cytoskeleton pathways. Similarly, cortisol induced the differential methylation of genes associated with focal adhesion, adrenergic signaling in cardiomyocytes, and Wnt signaling. Through integrative analyses, we determined that 126 genes showed a negative correlation between up-regulated expression and down-regulated methylation. KEGG enrichment analysis of these genes indicated participation in ECM-receptor interaction, regulation of actin cytoskeleton, and focal adhesion. Using RT-qPCR, we confirmed the differential expression of lamb3, itga6, limk2, itgb4, capn2, and thbs1. This study revealed for the first time the molecular responses of skeletal muscle to cortisol at the transcriptomic and whole-genome DNA methylation levels in rainbow trout. Full article

Clostridium perfringens alpha toxin (CPA), which causes yellow lamb disease in sheep and gas gangrene and food poisoning in humans, is produced by all types of C. perfringens and is the major virulence determinant of C. perfringens type A. CPA induces hemolysis in many species, including humans, murines, sheep and rabbits, through its enzymatic activity, which dissolves the cell membrane. Recent studies have shown that some pore-forming toxins cause hemolysis, which is achieved by the activation of purinergic receptors (P2). However, the relationship between P2 receptors and non-pore-forming toxin hemolysis has not been investigated. In the present study, we examined the function of P2 receptors in CPA toxin hemolysis and found that CPA-induced hemolysis was dependent on P2 receptor activation, and this was also true for Staphylococcus aureus β-Hemolysin, another non-pore-forming toxin. Furthermore, we use selective P2 receptor antagonists to demonstrate that P2X1 and P2X7 play important roles in the hemolysis of human and murine erythrocytes. In addition, we found that redox metabolism was mainly involved in CPA-induced hemolysis using metabolomic analysis. We further demonstrate that CPA activates P2 receptors and then activates NADPH oxidase through the PI3K/Akt and MEK1/ERK1 pathways, followed by the production of active oxygen to induce hemolysis. These findings contribute to our understanding of the pathological effects of CPA, clarify the relationship between P2 activation and non-pore-forming toxin-induced hemolysis, and provide new insights into CPA-induced hemolysis. Full article