Bulk RNA sequencing （bulk RNA-seq） is extensively used in livestock research to characterize transcriptomic variation at the tissue level. However， as it captures only the average gene expression across mixed cell populations， it fails to resolve cellular heterogeneity， thereby limiting the dissection of complex trait regulation at the cellular scale. In contrast， single-cell RNA sequencing （scRNA-seq）， with its high resolution and sensitivity， enables detailed profiling of cellular composition and uncovers regulatory mechanisms underlying cell-to-cell variability. Despite its advantages， large-scale adoption of scRNA-seq in livestock is hindered by high costs， complex biological systems， and technical barriers. Against this background， single-cell deconvolution algorithms have emerged. Depending on data characteristics， these methods leverage strategies such as reference expression profile construction， marker gene selection， or unsupervised learning to infer cellular composition proportions from bulk RNA-seq data， thereby overcoming its inherent limitation in resolving cellular heterogeneity. Moreover， advanced algorithms can further predict cell-type-specific expression patterns， enabling functional interpretation at single-cell resolution from population-level transcriptomes. In recent years， these methods have been applied in key domestic species such as pigs， cattle， and chickens， supporting studies in traits related to growth， reproduction， and immune function. This review systematically summarizes the principles， mainstream algorithms， and current applications of single-cell deconvolution in livestock， with a particular focus on challenges in reference matrix construction， cross-platform data integration， and model adaptation. Furthermore， we discuss the expanding potential of deconvolution in multi-omics integration and functional cellular analysis， aiming to provide a conceptual and methodological framework for advancing its use in livestock research.

Breed identification is fundamental to livestock conservation and plays a vital role in genetic resource management， breed purity assessment， breeding strategy implementation， product traceability， supply chain optimization， and ensuring food and biosafety. This review summarizes both traditional and modern methods for breed identification of common livestock and poultry （pig， cattle， sheep， dog， chicken）， and systematically reviews recent advances in machine learning-based breed identification research.Traditional methods， such as morphological assessment and physicochemical indicator analysis in animals，are increasingly inadequate for large-scale breed identification due to their limited accuracy，operational complexity，and time-consuming nature. With advancements in intelligent animal husbandry and high-throughput omics technologies， machine learning-based approaches have gained wide application. This paper reviews major achievements in breed identification using machine learning， analyzes the strengths and limitations of various algorithms， and highlights challenges in practical use. It offers insights for future development and supports molecular breeding and precision livestock management.

Yaks are well-adapted to the extreme environment of the Qinghai-Tibet Plateau， characterized by low temperatures， intense ultraviolet radiation， hypoxia， and nutritional stress， making them the dominant livestock species in high-altitude pastoral areas. Yak are multi-functional cattle breeds， possessing the performance of meat production， cashmere production， service use and milk production， however， their milk production performance has not undergone high-intensity breeding.Yaks have low milk yield but significantly higher milk fat， lactose and milk protein content.As the core organ for milk secretion， the development of yak mammary gland is regulated by complex network of genes， and the maintenance of its homeostasis and lactation is an intricate biological process of the organism， which relies on the coordinated functioning of multiple cells. With the development of omics technology and the deepening of functional genomics research，the core regulators of mammary gland development in human， mouse and common cow species have been analyzed， and the regulatory networks controlling key processes such as lactation and milk lipid secretion have been clarified. Although milk production is an economically important trait in yak， relatively few studies have been conducted on the regulatory network of mammary gland and lactation-regulating genes in yak.In this paper， the similarities and heterogeneity of yak mammary glands in terms of morphology， development， lactation mechanism， lactation performance and other aspects compared with mammary glands of other species were systematically summarized， and key scientific issues that need to be urgently addressed in yak-related aspects are presented with a view to providing important information for mammary gland biology research and identification of functional genes for milk production traits in yaks.

The intestinal tract is not only the central site for the digestion and absorption of nutrients， but also a natural barrier against pathogens， and its health is closely related to growth performance and immune capacity of livestock and poultry. Therefore， improving the intestinal health of livestock and poultry has become a research hotspot among scholars. Plant essential oils （PEO）， low molecular weight secondary plant metabolites extracted from plants， have been shown to have positive effects on improving intestinal health， such as enhancing intestinal barrier function and promoting the colonization of beneficial gut microbiota. In this paper， the effects of PEO on the intestinal health of livestock and poultry are systematically reviewed from the aspects of intestinal morphology， intestinal barrier function， and intestinal microbiota， and the potential mechanisms are discussed， in order to provide references for the development and application of PEO.

Rumen fermentation in ruminants results in a loss of 2% to 12% of feed energy as methane， which not only represents an energy loss but also has negative environmental impacts. Consequently， finding economical and effective methods to reduce methane emissions from ruminants has become the focus of current research. Proper feeding and grazing management strategies can reduce methane emissions by modulating the metabolic pathways of rumen microorganisms， inhibiting methanogenic activity and enhancing the host's nutrient utilization efficiency. This review summarizes current research on the effectiveness of different dietary strategies， including adjusting dietary composition， optimizing forage selection and utilization， and improving forage processing and preservation， as well as grazing management strategies such as stocking rate， grazing methods， and grazing intensity. It also discusses the applicability and potential of these strategies， providing theoretical insights and practical guidance for methane mitigation in ruminant production.

β-lactamase is an enzyme capable of hydrolyzing β-lactam antibiotics， and its residues in milk are increasingly concerning. The endogenous factors related to β-lactamase residues in milk are associated with the health status of the cows themselves， while exogenous factors mainly involve illegal human addition. The residues of β-lactamase may cause various health risks， including the accumulation of cytotoxic degradation products， the failure of antibiotics， and drug resistance development in humans. Currently， detection methods for β-lactamase residues in milk include microbial methods， iodometry， acidity meter method， instrumental detection method， and rapid detection method， etc.， but all these methods have limitations. This article reviews the biological characteristics， sources， and recent research progress on detection methods of β-lactamase in bovine milk. Also discusses the limitations of current detection methods and provides an outlook on future management measures and emerging detection technologies. The aim is to provide scientific references for detection personnel and researchers， ultimately enhancing dairy product safety.

Transport stress is a significant factor affecting cattle health and production performance， involving disruptions in multiple physiological systems such as metabolism， digestion， endocrinology， and immunity. During transportation， various stressors （such as fasting， dehydration， temperature fluctuations， overcrowding， and loading/unloading） can negatively impact cattle physiological functions， leading to the excessive production of stress hormones and changes in rumen microbial community abundance. This， in turn， reduces immune function and antioxidant capacity， causes tissue and organ damage， and ultimately affects overall health and production performance. This review summarizes the mechanisms underlying physiological imbalances in cattle caused by transportation stress， aiming to provide theoretical references for identifying target points for controlling transportation stress in cattle and improving animal welfare during transportation.

This study aimed to systematically evaluate the population genetic characteristics of Hetao Big-ear pigs using genomic approaches， providing scientific basis for the conservation of their germplasm resources. Based on 50K SNP chip data， Using software such as plink （v1.90）， GCTA （v1.93）， and MEGAX （v10.0） for analysis， PCA， genetic distance， kinship relationship， inbreeding coefficient， and phylogenetic tree information were obtained. Multi-dimensional analysis methods were used to detect 78 Hetao Big-ear pigs. Population structure analysis revealed that the population could be divided into 4 subgroups， with an average genetic distance of 0.793， and 6 sire lineages forming the foundation of its pedigree. Analysis of genomic runs of homozygosity （ROH） detected a total of 2 601 segments， with individuals carrying an average of 21 ROHs and an average length of 7.77 Mb. The population inbreeding coefficient （F_ROH ） was 0.13. Comprehensive analysis indicated that this population exhibits characteristics including a limited number of lineages， relatively few sire bloodlines， moderate kinship relationships， and low inbreeding levels. These findings at the genomic level provide important references for the population optimization and genetic improvement of Hetao Big-ear pigs.

This experiment aimed to investigate the effects of ZNF697on the proliferation and differentiation of porcine skeletal muscle satellite cells （MuSCs）. In this experiment， porcine MuSCs were used as the research object. Flow cytometry， Real-time quantitative PCR， Western blot and immunofluorescence staining were employed to detect the effects of ZNF697on the proliferation and differentiation of porcine MuSCs. The following results were obtained： 1） The expression of ZNF697mRNA dynamically changes with the proliferation of porcine MuSCs. Interfering with ZNF697inhibited the mRNA expression of Cyclin E （P<0.01） and Cyclin D （P<0.05） and the protein expression of Cyclin B （P<0.05）， promoted the mRNA expression of p21（P<0.01）. Overexpression of ZNF697showed the opposite results. 2） During the myogenic differentiation of porcine MuSCs， the expression of ZNF697gradually increased. Interfering with ZNF697significantly reduced the myogenic differentiation index （P<0.05） and inhibited the mRNA expression of MyoG （P<0.01）， the mRNA and protein expression of MyHC （P<0.05） and MyoD （P<0.01）. Overexpression of ZNF697showed the opposite results. The results show that ZNF697is a promoter of proliferation and myogenic differentiation in porcine MuSCs， which will provide a breeding target for high-lean-meat pig breeds.

The purpose of this study was to establish an efficient pig cell gene editing technology system and lay a foundation for the application of PE system in pig genetics and breeding. Multiple candidate sites were selected from the coding region of VIM gene， and sgRNA was designed strictly according to the target site design principle of CRISPR/Cas9 system. The candidate sgRNAs were scored by bioinformatics tools， and the two sgRNA sequences with the highest scores were screened out. Based on the above two sgRNAs， the corresponding pegRNAs were further designed. The cells were treated with drugs hydroxyurea and nocodazole to stay in the G1/S or G2/M phase， and the nCas9-RT fusion protein editor vector and pegRNA expression vector were electro-transfected into the cells. After 48 h， the genomic DNA of the mixed cells was extracted， the target sites were amplified by PCR， and the next-generation sequencing was performed. CRISPResso2 was used to analyze the editing results. The results showed that the editing efficiency of PE2 system in PK15 cells was between 0.16% and 14.03%. The editing efficiency in IPEC-J2 cells was between 0.08% and 0.58%. The editing efficiency of PEmax system in PK15 cells was between 0.63% and 23.25%. However， the editing efficiency of PE4 and PE4max systems in PK15 cells was poor， with the highest editing efficiency of 0.7% and 0.36%， respectively. Regulating the cell cycle could improve the editing efficiency of the PE2 system for the editing types of single base substitution and single base deletion of porcine VIM gene. The editing efficiency of the former had increased by 1.4 times， and the editing efficiency of the latter had increased by 2.6 times. According to the results， PE technology can perform a variety of different types of precise editing of pig VIM gene and improve its editing efficiency through cell cycle regulation. It provides a technical reference for the next step to combine somatic cell cloning technology to prepare pig populations with improved production performance.

This study aimed to investigate the effects of Escherichia coli （E. coli）， Clostridium butyricum （C. butyricum）， and their metabolites on mitochondrial function and related metabolic pathways in porcine intestinal epithelial cells （IPEC-J2）， providing a theoretical basis for regulating intestinal health. Initially， bacterial suspensions were screened to ensure logarithmic-phase growth （3 h） with optimal viability for subsequent experiments. IPEC-J2 cells were then treated with varying concentrations of Escherichia coli， Clostridium butyricum， lipopolysaccharide （LPS）， and short-chain fatty acids （SCFAs）， with 3 replicates per group. Cell viability and morphological integrity were assessed to determine the optimal treatment duration and concentration. Key metrics included ATP levels， reactive oxygen species （ROS）， mitochondrial membrane potential， and the expression of genes related to antioxidant defense and mitochondrial function. The results showed that： 1） The optimal concentrations for treating IPEC-J2 cells were determined as 10⁸ CFU·mL^-1 for Escherichia coli and 10⁶ CFU·mL^-1 for C. butyricum， with a treatment duration of 1 h. The treatment duration of SCFAs and LPS were also treated for 1 h. 2） Propionic acid and E. coli significantly increased ROS levels （P<0.05）， whereas C. butyricum and LPS reduced them （P<0.05）. 3） C. butyricum upregulated the expression of ACC1， NRF1， TFAM， HO-1， APAF-1， PIG3， and p53 （P<0.05）； E. coli downregulated the expression of TFAM and CtyC （P<0.05） but upregulated the expression of p53 （P<0.05）； Propionic acid and acetic acid had no significant effects on genes expression. The results indicated that SCFAs enhanced cellular metabolism， E. coli and LPS triggered oxidative stress and apoptotic pathways. C. butyricum activated mitochondrial lipid metabolism， antioxidant defenses， and apoptosis in IPEC-J2 cells. These findings offer novel insights into mitochondria-targeted strategies for gut microbiota regulation.

This study aimed to estimate the genetic parameters and genomic breeding values of “Min Gai” cashmere goat hair traits， compare the effects of GBLUP and Bayes methods on the accuracy of genomic breeding value prediction of hair traits of “Min Gai” cashmere goat， and provide a reference basis for optimising the genetic selection and breeding strategy of “Min Gai” cashmere goats. This study took the fleece traits of 847 “Min Gai” cashmere goats as the research object and used GBLUP， Bayes A， Bayes B， Bayes C， and Bayes LASSO to estimate the genetic parameters and genomic breeding values of fleece traits （fleece length， cashmere length， cashmere diameter， and cashmere yield）. Furthermore， the accuracy of the genomic breeding value estimates was evaluated using five-fold cross-validation. The results showed that the heritability of fleece length estimated by the 5 methods was 0.11-0.32， the heritability of cashmere length was 0.14-0.33， the heritability of cashmere diameter was 0.17-0.49， and the heritability of cashmere yield was 0.16-0.32. There was a strong positive genetic correlation between cashmere length and fleece length， with a correlation coefficient of 0.48-0.84. The genetic correlation between other traits was relatively small， ranging from -0.06 to 0.46. The genomic breeding value accuracy estimated by the GBLUP， Bayes A， Bayes B， Bayes C， and Bayes LASSO methods for fleece length， cashmere length， cashmere diameter， and cashmere yield were 0.623 4-0.950 9， 0.626 2-0.953 0， 0.830 8-0.953 1， and 0.741 8-0.961 4， respectively. The genomic prediction accuracy of each trait estimated by the Bayes method was significantly higher than that estimated by the GBLUP method， and the Bayes B method had the highest genomic prediction accuracy， reaching 0.950 9-0.961 4. In summary， the Bayes B method is recommended for the genomic selection of “Min Gai” cashmere goats， which may accelerate the genetic progress of the population.

This study aimed to develop a haplotype-segment matching-based maternal grandsire discovery approach applicable to different SNP array densities and segment window sizes in the absence of dam genotypes， thereby providing a reliable technical approach for pedigree reconstruction. The study utilized genomic data from 6 117 dairy cows to construct datasets of varying SNP densities， including low-density （5K， 10K） and medium-density （20K， 50K） SNP panels. For each dataset， the genome was partitioned into fixed-length haplotype segments of 5， 10， 35， 75， and 120 SNPs， followed by haplotype inference. Based on the known genotypes of the offspring's sire， paternal haplotypes were excluded to infer maternal haplotypes， which were then compared with those of candidate bulls on a segment-by-segment basis to calculate the haplotype matching rate （HMR）. The bull with the highest HMR was designated as the putative maternal grandsire （MGS）. A total of 345 individuals with complete and validated pedigree information were used to verify method accuracy. Furthermore， genomic relatedness coefficients and homozygosity levels were calculated to investigate the potential effects of kinship structure and inbreeding on identification outcomes. The haplotype-segment matching method achieved more than 90% assignment accuracy across most combinations of SNP-array density and segment window length. The 20K array with a 35-SNPs window yielded the highest accuracy at 98%， demonstrating robust performance. Nonetheless， the optimal decision thresholds for the haplotype match rate and the resulting accuracies varied substantially with parameter settings， indicating that SNP density and window length were key determinants of performance. Additional analyses showed that kinship structure and inbreeding levels significantly affected matching performance and assignment accuracy， suggesting that the population genetic background should be considered in practical applicatons. The haplotype-segment matching （HSM）-based method demonstrated high accuracy in maternal grandsire identification under conditions of missing maternal genotype data， making it a valuable tool for pedigree correction in large-scale commercial herds. Optimizing SNP density and haplotype window length based on the genetic background of the population is essential to enhance identification efficiency and robustness in practical applications.

This study aimed to systematically identify expression short tandem repeats （eSTRs） in dairy cattle that are significantly associated with gene expression， in order to explore their regulatory roles in gene expression and complex economic traits， thereby providing functional variant resources for molecular breeding. A total of 108 Chinese Holstein cows were sampled for tail vein blood collection. Genomic DNA and total RNA were extracted and subjected to 30×whole-genome resequencing and transcriptome sequencing， respectively. After quality control， 105 samples were retained for further analysis. STRs were annotated and genotyped at the population scale using TRF and HipSTR. Gene expression levels were quantified after quality filtering with Trimmomatic， alignment using STAR， and normalization with edgeR. Based on a ±1 Mb window around transcription start sites （TSS）， a linear model was used to assess the association between STR dosage and gene expression to identify cis-eSTRs. Weighted correlation network analysis （WGCNA） and Gene Ontology （GO） were further applied to explore the functional relevance of cis-eSTRs in complex traits. A total of 25 154 significant cis-eSTR-gene expression associations were identified. These cis-eSTRs were predominantly located in intronic regions and near TSSs. WGCNA revealed that cis-eSTR regulated genes clustered into several functional modules， some of which were significantly associated with traits related to milk production and immune. GO enrichment analysis revealed that these modules are broadly involved in biological processes such as immune response， lipid metabolism， cell cycle， DNA replication and stability. Several key genes （e.g.， IRF7， BoLA-DRB， CCT2） might play important functional roles in immune regulation and mammary gland function. cis-eSTRs play important roles in regulating gene expression and complex traits in dairy cattle. The cis-eSTR resource generated in this study provides a valuable foundation for the functional annotation of complex traits and offers potential molecular markers for breeding applications.

The aim of this study was to explore the impact of molecular networks constructed by DE-circRNAs related to milk fat synthesis on milk fat synthesis in dairy cows through in-depth analysis. In this study， we used the circRNA library and transcriptome sequencing data of Holstein dairy cows with high and low milk fat percentage constructed previously to screen and identify DE-circ RNAs（circPRKAA2） related to milk fat synthesis， and further predicted their ceRNA regulatory network using Targetscan and miRanda software. Real-time fluorescence quantitative PCR， RNA pull down assay and dual luciferase reporter assay were used to verify the targeting relationship. Oil red O and BODIPY staining were used to investigate the effect of target miRNA on lipid droplet secretion in BMECs. The results indicated that the regulatory relationship of the ceRNA （circPRKAA2/miR-22-3p/SLC27A1） network was successfully predicted and verified， and 9 miRNAs and 10 candidate mRNAs were predicted， and it was clarified that circPRKAA2 and miR-22-3p were mainly distributed in the cytoplasm of BMECs. circPRKAA2 negatively regulated miR-22-3p during the process of milk fat synthesis in BMECs. miR-22-3p inhibited the secretion of lipid droplets， the production of triglycerides and cholesterol in BMECs， and negatively regulated SLC27A1. In conclusion， this study analyzed and verified the targeting relationship of circPRKAA2/miR-22-3p/SLC27A1 and its regulatory effect on milk fat synthesis， with the hope of providing new insights for improving milk quality.

The aim of this study was to investigate the differential expression of genes in the blood of Yili horses before and after the 5 000 m race， and the correlation between cardiac parameters and the differentially expressed genes. Six elite Yili horses， which were among the top 8 horses in the 5 000 m speed race at the national level， were selected for the study. Echocardiograms were collected from the resting state of the horses before the 5 000 m speed race and the blood of the horses before and after the race by using the Myriad M6 Veterinary Portable Color Doppler Ultrasound System， which was used for transcriptome sequencing. The results showed that： 1） A total of 889 differentially expressed genes were screened， including 751 up-regulated genes and 138 down-regulated genes.GO and KEGG enrichment analyses revealed that the differentially expressed genes were mainly involved in the biological processes of cellular response to stimuli and immune regulation， as well as the MAPK and Jak-STAT signaling pathways involved in cellular immune response and inflammation. 2） CCL3， CSF1， CSF3， IFNG， IL4， IL10 were screened as key genes. In the correlation between the post-competition expression of key genes and cardiac structural indexes， CCL3 was extremely significantly positively correlated with PADs （P<0.01）， significantly positively correlated with LVIDd （P<0.05）， and significantly negatively correlated with RWTd （P<0.05）， CSF1 was extremely significantly positively correlated with PADd （P<0.01）， significantly negatively correlated with RWTd （P<0.05），CSF3 was significantly negatively correlated with IVSd and EDV （P<0.05）， IFNG was significantly positively correlated with PADd （P<0.05）， significantly negatively correlated with IVSd， MWTd， and RWTd （P<0.05）， IL10 was extremely significantly positively correlated with LVLD （P<0.01）， significantly positively correlated with MVD （P<0.05）， and significantly negatively correlated with EDV （P<0.05）. In the correlation between the post-race expression of key genes and cardiac function indexes， CSF3 was significantly negatively correlated with SV， SI， CI， and VTI （P<0.05）， IFNG was significantly negatively correlated with SI （P<0.05）， and IL10 was significantly negatively correlated with CO， CI， and VTI （P<0.05）. Moreover， CSF1 and IFNG post-race expression were significantly negatively correlated with the 5 000 m exercise performance of Yili horses （P<0.05）. Cardiac indexes and CCL3， CSF3， IL4， IL10 regulatory mechanisms play important roles in the 5 000 m race of Yili horses， and the expression levels of CSF1 and IFNG can be used as candidate genes to assess the athletic performance of Yili horses.

This study aimed to investigate the anti-aging effects and underlying molecular mechanisms of reserve mesenchyme cells （RMCs） derived from the velvet antler of the sika deer on hydrogen peroxide-induced senescent human dermal fibroblasts （HDFs）. An HDF senescence model was established by treatment with 200 µmol·L^-1 H₂O₂ and confirmed via SA‑β‑galactosidase staining. Senescent HDFs were then co‑cultured with RMCs， and subsequent assays evaluated cell proliferation， migration， superoxide dismutase （SOD） activity， and malondialdehyde （MDA） content. The 4D-DIA proteomics technology was used to identify differential proteins between co-culture medium and senescent HDF medium， and bioinformatics enrichment analysis was performed. Functional analyses revealed that， compared with the model group， RMCs co‑culture significantly enhanced HDFs proliferation and migration （P<0.01）， increased SOD activity （P<0.05）， and decreased MDA levels （P<0.05）. Using 4D‑DIA proteomics， 289 differentially expressed proteins were identified in the conditioned media of the co‑culture versus senescent HDF alone， with key molecules such as ATP1A1， COL7A1， IGF2R， and MMP3 showing significant expression changes. KEGG enrichment analysis highlighted the proteasome， PPAR signaling pathway， extracellular matrix-receptor interaction， and focal adhesion pathways as principal regulatory axes. These findings suggest that velvet-antler-derived RMCs from the sika deer alleviate oxidative stress-induced dysfunction in senescent HDFs. It was speculated that RMCs may exert their effects by enhancing proteasome‑mediated clearance of damaged proteins， activating PPARγ to suppress inflammation and oxidative stress， promoting COL7A1‑dependent dermal-epidermal junction reconstruction， and inhibiting MMP3‑mediated extracellular matrix degradation， thus maintaining HDF homeostasis and function. This work provides experimental evidence and a theoretical foundation for developing deer antler-derived secretome-based strategies for skin anti‑aging.

This study aimed to investigate the impact of heat stress on the antioxidant capacity of porcine sertoli cells and to further elucidate the protective role and underlying mechanism of resveratrol （RES） in alleviating oxidative stress. Porcine sertoli cells were isolated and cultured primarily. The cells were divided into 3 groups： control group （NC， 34 ℃）， heat stress group （HS， 43 ℃ for 30 min）， and heat stress+resveratrol group （HS+200 nmol·L^-1 RES）. The levels of reactive oxygen species （ROS）， the content of malondialdehyde （MDA） and advanced glycation end products （AGEs）， as well as the activities of superoxide dismutase （SOD） and catalase （CAT） were measured. The expression levels of RAGE， NOX4， and NF-κB genes were determined using real-time fluorescence quantitative PCR. Compared with the control group， heat stress significantly increased ROS level （P<0.05）， MDA content （P<0.05）， and AGEs content （P<0.01）， whereas the activities of SOD and CAT were significantly decreased （P<0.01）， indicating that heat stress successfully induced oxidative stress in porcine sertoli cells. Pretreatment with 200 nmol·L^-1 RES significantly attenuated the heat stress-induced elevations in ROS， MDA， and AGEs levels （P<0.05）， and restored the activities of SOD and CAT （P<0.05）. Moreover， RES significantly downregulated the expression levels of RAGE， NOX4， and NF-κB genes induced by heat stress （P<0.05）. Heat stress disrupts redox homeostasis in porcine sertoli cells， leading to oxidative stress. Resveratrol alleviates this heat stress-induced oxidative stress by suppressing the AGEs-RAGE/NOX4/NF-κB signaling pathway. The findings provide a theoretical basis for utilizing resveratrol to improve boar semen quality during summer.

The objective of this study was to investigate the imprinting status and to search for potential DNA differentially methylated regions （DMRs） of the DLK1-DIO3 domain in bovine placentas. In this study， whole genome sequencing （WGS） was performed on 3 bovine placentas and their parents to mining the informative single nucleotide polymorphisms （SNPs）. The imprinting status of genes in the DLK1-DIO3 domain were determined by the combined analysis of WGS and transcriptome sequencing （RNA-seq） results. The DMRs were identified by combination of WGS and whole genome bisulfite sequencing （WGBS） data. WGS and RNA-seq results showed that MEG3， MEG8 and MEG9 genes were maternally expressed imprinted genes and DLK1， RTL1 and DIO3 genes were paternally expressed imprinted genes. The WGBS data analysis showed that 4 DMRs， Dlk1-DMR， IG-DMR， Gtl2-DMR and Meg8-DMR were conserved in human and mouse， and 5 new DMRs were identified， DLK1-DMR0 and DLK1-DMR1 located upstream of the DLK1 gene， MEG9-DMR in MEG9 promoter region， and DIO3-DMR1 and DIO3-DMR2 located upstream of DIO3 genes， respectively. The results indicate that the imprinted status and differentially methylated regions of bovine DLK1-DIO3 imprinted region are highly conserved to human and mouse. In addition， 5 potential DMRs were identified in bovine DLK1-DIO3 region， and their regulating mechanism on the DLK1-DIO3 imprinted region needs to be further investigated.

This study adopted an IF feeding method to investigate the effects and mechanisms of this dietary approach on ovarian function in mice， aiming to provide a theoretical basis for livestock reproduction management. In this study， 8-week-old female C57BL/6J mice with similar body weights were randomly divided into two groups： one group served as the normal diet control group （ND）， while the other group was subjected to a cyclic pattern of “1 day of restricted feeding （70% feed intake）-1 day of free feeding” to establish an intermittent mild fasting （IMF） mouse model. The growth performance of the mice was measured， and ovarian function as well as the expression of autophagy-related proteins were detected through Western blot （WB） and immunohistochemistry experiments. The results showed that IMF could significantly reduce the feed-to-gain ratio （82.85±2.68 & 93.02±7.40） and abdominal fat index （1.49±0.11 & 1.72±0.19）. At the same time， the autophagy markers LC3Ⅱ/LC3Ⅰ（1.06±0.12 & 0.89±0.02） and reproductive function proteins BMP-15 （1.05±0.08 & 0.75±0.10） and GDF-9 （0.53±0.02 & 0.44±0.02） were up-regulated. The mechanism study showed that IMF activated autophagy by inhibiting mTOR pathway （p-mTOR/mTOR 0.82±0.10 & 0.97±0.05）， and then improved ovarian function. This study is the first to elucidate the mechanism by which IMF enhances ovarian function through autophagy activation， offering a potential non-pharmaceutical strategy for developing reproductive regulation techniques in livestock.

The aim of this study was to investigate the effect of dietary rumen-protected fat （RPF） supplementation on fat deposition and related genes expression of fattening yaks. Twenty-four male yaks with similar body weight （275.63±9.84 kg） were randomly assigned to three treatment groups （8 in each group）， which were 1） Control group （CON）： fed a basal diet； 2） 1.5% RPF supplementation group （RPF 1.5）： fed the basal diet supplemented with 1.5% RPF； 3） 3.0% RPF supplementation group （RPF 3.0）： fed the basal diet supplemented with 3.0% RPF， respectively. The experimental period was 90 d. At the end of the trial period， the longissimus dorsi muscle and abdominal adipose tissues were collected for the analysis of nutritients， fatty acid composition， adipose tissue morphology and related genes expression. The results showed that there was no significant difference in the muscle energy and protein content， the intramuscular fat （IMF） content varied significantly and increased with RPF supplementation （P<0.05）. the cis-11，14-eicosenenoic acid in muscular fatty acid composition of RPF3.0 was significantly greater than that of CON group （P<0.05）， in RPF 1.5 group， the cis， cis-8，11，14-eicosatrienoic acid and cis-5，8，11，14，17-eicosapentaoic acid were significantly greater than those in the CON group （P<0.05）. RPF had no significant effect on the proportion of saturated and unsaturated fatty acids in muscle tissue. In terms of the adipose tissue morphology， the number of cells per unit area and mean cell area in IMF and mean cell area in abdominal fat increased gradually with the level of RPF addition. In longissimus dorsi muscle， the expression of transcriptional regulation factors PPARγ， SREBP-1， FABP4 were significantly higher in RPF 1.5 and RPF3.0 group （P<0.05）. Yaks in RPF 3.0 group had significantly lower ACC and FAS genes expression which are key enzymes in de novo fat synthesis compared with CON group. And DGAT-1 and CPT-1 （P<0.05） genes expression in RPF 1.5 and RPF 3.0 group were significantly higher than those in CON group （P<0.05）. The expression of transcriptional regulation factor C/EBPα gene in abdominal adipose tissue increased significantly with the supplemental dose of RPF （P<0.05）， genes expression of PPARγ and FABP4 were significantly higher in the RPF 1.5 and RPF3.0 groups than in the CON group （P<0.05）. The ACC gene expression which is the key enzyme for de novo fat synthesis in the RPF 1.5 and RPF3.0 groups were significantly lower than that in the CON group （P<0.05）. DGAT-1 gene expression increased significantly with the addition of RPF （P<0.05）. Yaks of the RPF 1.5 and RPF3.0 groups had significantly higher gene expression of LPL when compared to CON ones （P<0.05）. It can be seen that dietary RPF supplementation can effectively promote IMF and abdominal fat deposition in fattening yaks， by regulating the gene expression of transcriptional regulation factors and key enzymes involed in fat metabolism， reducing de novo fat synthesis， utilizing directly the fatty acids derived from RPF for fat synthesis. The feeding effect of 3.0% RPF supplementation is better than 1.5%.

This study aims to study the differences in rumen fermentation parameters and microbial communities between Holstein steer and Xiangxi cattle. It provided a reference for exploring the composition and functional characteristics of rumen microbial ecosystems in animals with different genetic backgrounds， and offered a scientific basis for the formulation of breed-specific feeding strategies. Twelve adult Holstein steer （241.3±9.1 kg） and twelve adult Xiangxi cattle （206.0±8.4 kg） with good body condition were selected as experimental subjects and all animals were fed their respective standard diets. The experimental procedure involved the collection of ruminal fluid for the determination of ruminal fermentation parameters. Additionally， 16S rRNA sequencing was applied to analyze the structure and function of the ruminal microbial community. The results showed that： 1） The dry matter digestibility （DMD）， organic matter digestibility （OMD）， neutral detergent fiber digestibility （NDFD）， and acid detergent fiber digestibility （ADFD） in Xiangxi cattle were significantly higher than those in Holstein steer （P<0.001）. 2） The ruminal pH in Xiangxi cattle was significantly higher than that in Holstein steer （P<0.001）. The concentrations of dissolved hydrogen （dH₂）， dissolved methane （dCH₄）， and ammonia nitrogen （NH₃-N） in Xiangxi cattle were significantly lower than those in Holstein steer （P<0.001）， while the dissolved hydrogen to dissolved methane ratio （dH₂/dCH₄） was significantly higher in Xiangxi cattle than Holstein steer （P<0.001）. In rumen fermentation products， the total volatile fatty acid （TVFA） concentration and molar ratios of propionate， butyrate， iso-butyrate， valerate， and iso-valerate were higher in Holstein steer， while the molar ratios of acetate and acetate to propionate ratio were higher in Xiangxi cattle （P<0.001）. 3） The results of α-diversity showed that Xiangxi cattle possessed richer and more diverse rumen microbial communities. In comparison， it was found that Holstein steer were primarily differentially enriched functional bacteria such as Agathobacter， Butyrivibrio and Lachnospira while functional bacteria differentially enriched in Xiangxi cattle included Fibrobacteres， Saccharofermentans， Lentimicrobium， Succiniclasticum and Acetatifactor （P<0.05）. 4） The pathways of carbohydrate metabolism and metabolism of other amino acids were significantly enriched in Holstein steer， and the nucleotide metabolism and biosynthesis of other secondary metabolites pathways were significantly enriched in Xiangxi cattle （P<0.05）.These results showed that there were significant differences in nutrient digestibility， rumen fermentation parameters and microbial communities between Holstein steer and Xiangxi cattle. Specifically， the rumen of Holstein steer was inclined to propionate-type fermentation mode and was more capable of digesting non-fiber plant polysaccharides and proteins， while the rumen of Xiangxi cattle was inclined to acetate-type fermentation mode and was more capable of digesting fiber.

In this experiment， the piglets weaned at 7 days of age were used， and the aim was to determine the effect which hydrolyzed wheat gluten （HWG） replacing different doses of whey protein concentrate （WPC） in artificial milks on growth performance， nutrient metabolism， immune function and antioxidant capacity of piglets. Forty-five healthy crossbred （Duroc×Landrace×Yorkshire） piglets at 7 days of age were randomly allotted into four treatments， including control group （n=15）， 33%HWG group （n=10）， 66%HWG group （n=10）， and 100%HWG group （n=10）， which were fed the artificial milks that HWG replaced 0%， 33%， 66% and 100% WPC for 13 days， respectively. The results showed that HWG replacing WPC in artificial milks could affect growth performance， nitrogen metabolism， lipid metabolism， kidney function， immune function and antioxidant capacity in piglets. Compared with the piglets of control group， HWG replacing 66% and 100% WPC significantly reduced average daily feed intake and average daily gain （P<0.05）， significantly enhanced feed to gain ratio and diarrhea index （P<0.05）， significantly decreased serum creatinine （P<0.05）， and significantly increased serum malondialdehyde concentrations （P<0.05） in piglets. In addition， serum immunoglobulin A and M levels were significantly reduced （P<0.05） when 66% of WPC was replaced by HWG， and serum high density lipoprotein cholesterol levels were significantly increased （P<0.05） when 100% of WPC was replaced by HWG. Compared with the piglets of control group， serum urea and high density lipoprotein cholesterol levels were significantly higher in the piglets fed the artificial milk that HWG replaced 33% WPC （P<0.05）. In summary， HWG replacing different doses of WPC in artificial milks might be not conducive to growth performance， nutrient metabolism， immune function and antioxidant capacity in the early-weaned piglets， but when HWG replacing 33% WPC in artificial milks had not significantly negative effect on piglets.

The study aimed to explore the effects of Pediococcus pentosaceus 368 on Archaea and metabolites in feces of Duroc×Saba pig. Twenty Duroc×Saba pigs which body weight were about 77 kg were randomly divided into 2 groups with 10 pigs per group. The pigs in experimental group drinked water added with Pediococcus pentosaceus 368（10¹⁰ CFU·L^-1）. The experiment lasted for 28 days. On the day at the end of the experiment， slaughterings were conducted. The effects of Pediococcus pentosaceus 368 on Duroc×Saba pig were analyzed through the Archaea in feces and the fecal metabolites. The results showed as follows： 1） PCoA （Principal Coordinates Analysis） analysis found that samples in the experimental group and control group could not be completely separated， and species similarity in the experimental group became consistent after the addition of Pediococcus pentosus 368.2） At the phylum level， the highest relative abundance of the two groups was the Candidatus_Thorarchaeota and at the genus level， the highest relative abundance of the two groups was Methanobrevibacter. When the Linear Discriminant Analysis value was highter than 3.0， the two groups obtained 14 biomarkers. Among them， 14 biomarkers were in the experimental group， and the largest LDA value in the experimental group was the Thermoplasmata_unclassified.3） KEEG metabolic pathway analysis found that when the LDA value was highter than 3.0， the functions of 4 genes were screened out： Folding_sorting_and_degradation， membrane transport，Cell motility and Signal transduction； Analysis of the level function of CAZy （Carbohydrate Active enzyme Database） showed that when the LDA value was highter than 3.0， a total of 21 biomarkers were obtained in the two groups， of which 13 were in the experimental group. The biomarker with the largest LDA value in the experimental group was CBM20. In the control group， the biomarker with the largest LDA value was GT2.4） Fecal short-chain fatty acid analysis showed that the contents of Butyric acid and Isovaleric acid in the experimental group were significantly decreased （P<0.05）. 5） KEGG pathway enrichment analysis of differential metabolites found that there were 12 significantly different terms， which were as follows： Degradation of flavonoids，Linoleic acid metabolism， Cutin， suberine and wax biosynthesis， Arginine biosynthesis， Valine， leucine and isoleucine biosynthesis， Retrograde endocannabinoid signaling， Biosynthesis of phenylpropanoids，Biosynthesis of unsaturated fatty acids， Autophagy-animal， Fatty acid biosynthesis，Kaposi sarcoma-associated herpesvirus infection； Gene Set Enrichment Analysis （GSEA） identified three significant genes： Glycerophosphates metabolism； Glycine， serine and threonine metabolism； and Biosynthesis of unsaturated fatty acids. （Conclusion） In summary， the addition of Pediococcus pentosaceus 368 can make the species similarity become consistent in the fecal Archaea of Duroc×Saba pigs. Improve the relative abundance of microbial functional genes related to Signal transduction， Cell motility， amino acid and fat metabolism in feces， and improve the probiotic function of bacteria and enzymes related to amino acid and fat metabolism.

The objective of this study was to investigate the effects of dietary Moutai dried distiller’s grains （MDDG） on growth performance， nutrient apparent total tract digestibility （ATTD）， meat quality， serum immunity and antioxidant function， and intestinal health in growing-finishing pigs. And find out the suitable addition ratio. A total of 389 healthy Duroc × Landrace × Large White growing-finishing pigs with body-weights about 97 kg were randomly assigned to 3 dietary treatments consisting of 4 replicates with 31 to 34 pigs per replicate. The control group was fed a corn-soybean meal basal diet， the other two groups were fed the diet with 5% and 10% MDDG supplementation which replacing a portion of corn， soybean meal and wheat bran， with equal energy and nitrogen to the basal diet. The experiment lasted for 44 d. The results showed as follows： 1） Compared with the control group， the average daily feed intake and average daily gain in 10% MDDG group increased by 5.37% and 4.61%， respectively. In addition， the mortality rate in 10% MDDG group was decreased by 0.78% compared with the control group. The inclusion of 10% MDDG decreased the meat production cost by 0.02 yuan per kg， but the difference is not significant （P>0.05）. 2） A reduction in ATTD of crude protein， crude ash and calcium was observed with 10% MDDG inclusion （P<0.05）. MDDG inclusion up to 10% showed a decreasing trend of ATTD of phosphorus （P<0.1）. MDDG inclusion up to 10% had no significant effect on ATTD of crude fat （P>0.05）. 3） Compared with control group， inclusion of 5% or 10% MDDG significantly decreased the serum levels of endotoxin， diamine oxidase， and interleukin-6 （P<0.05）. The serum content of catalase tended to be increased in the 10% MDDG group （P<0.1）. 4） Compared with the control group， inclusion of 10% MDDG had a tendency （P<0.1） to reduce drip loss， while inclusion MDDG in the diet at 5% significantly increased （P<0.05） the redness a^* value of longissimus dorsi muscle. 5） Dietary inclusion of 5% or 10% MDDG did not adversely affect the jejunum， liver and spleen histology. In conclusion， dietary inclusion of 10% MDDG increases the growth performance， fortifies intestinal barrier function， reduces inflammatory response， improves slaughter traits and meat quality of growing-finishing pigs， which enhances the economic benefits of swine feeding.

This study aimed to investigate the effects of gut microbiota on the production performance of aged laying hens during the late laying period by altering their intestinal microbiota structure through fecal microbiota transplantation （FMT）. Forty 28-week-old and 250 68-week-old hens were observed for 7 weeks to record their laying rate. The hens were divided into three groups based on the laying rate （LR） and age which recorded previously： 35-week-old high-yielding hens （YH， LR>90%， 37 hens）， 75-week-old high-yielding hens （OH， LR>90%， 76 hens）， and 75-week-old low-yielding hens （OL， 30%<LR<60%，104 hens）. FMT was then performed at the 8th week for 4 weeks to remodel the gut microbiota of aged hens： aged high-yielding hens received fecal microbiota suspension from young high-yielding hens （OH_FMT-YH group）， while aged low-yielding hens received fecal microbiota suspension from aged high-yielding hens （OL_FMT-OH group）. Control groups （OH and OL） were established， with each group comprising 5 replicates of 6 hens. The results were showed as follows： Compared to the OH group， aged high-yielding hens transplanted with microbiota from young high-yielding hens （OH_FMT-YH group） exhibited significant decreases （P<0.05） in laying rate， eggshell strength， eggshell thickness， serum sIgA and IL-10 levels， jejunal villus height， mucosal layer thickness， jejunal FOXP3 and ZO-1 mRNA expression， acetate content， and Bacteroidetes abundance. Conversely， yolk weight， serum IL-1β， IL-17A， and IL-6 levels， cecal mucosal layer and intestinal wall thickness， and Firmicutes abundance significantly increased （P<0.05）， the number of OTUs decreased， and the structure of the flora was obviously separated. In the OL_FMT-OH group， compared to the OL group， aged low-yielding hens transplanted with microbiota from aged high-yielding hens showed significant increases （P<0.05） in laying rate， eggshell strength， serum IL-10 levels， jejunal villus height and mucosal layer thickness， FOXP3 and ZO-1 mRNA expression， goblet cell count， cecal mucosal layer thickness， acetate content， and Bacteroidetes abundance. However， egg weight， egg longitudinal diameter， yolk weight， serum IL-1β levels， cecal intestinal wall thickness， jejunal STAT3 mRNA expression， and Firmicutes abundance significantly decreased （P<0.05）. β-diversity showed that the structure of the microbiota had changed. The LEfSe analysis revealed that Firmicutes showed the greatest contribution to the differences in both the OH_FMT-YH and OL groups， while Bacteroidetes were the most significant contributors to the disparities in the OH and OL_FMT-OH groups. In conclusion， FMT from young high-yielding hens to aged high-yielding hens increased Firmicutes abundance in the gut but reduced laying performance. Conversely， enhancing Bacteroidetes abundance in aged low-yielding hens through FMT from aged high-yielding hens improved gut microbiota structure， intestinal health， and laying performance. These findings suggest that modulating gut microbiota composition via FMT may serve as a potential strategy to enhance productivity in aged laying hens during the late laying period.

This experiment aimed to study the effects of anthocyanins from red elephant grass （Pennisetum purpureum Schumab cv. Red） on the growth performance， antioxidant capacity， and cecal microbial microbial structure of white-feathered broilers. A total of 210 healthy and similar-sized Shengze “901” 1-day-old male chicks were randomly divided into 5 groups， with 6 replicates in each group and 7 chicks in each replicate. Each group was fed the same basal diet， and the control group had no anthocyanin added to the drinking water. The drinking water of the treatment groups 1， 2， and 3 contained 50， 100， and 200 mg·kg^-1 of anthocyanin extracted from red elephant grass， respectively， while the drinking water of the treatment group 4 contained 100 mg·kg^-1 of anthocyanin from purple sweet potato （Ipomoea batatas L.）. The treatment period was 42 days. The results showed that： 1） The addition of red elephant grass anthocyanin to the drinking water had no significant effect on the growth performance of white-feathered broilers （P>0.05）. 2） At 21 days of age， the serum and liver malondialdehyde （MDA） content of the treatment group 2 was significantly lower than those of the control group （P<0.05）， and the serum glutathione peroxidase （GSH-Px） content and liver catalase （CAT）， glutathione （GSH）， and superoxide dismutase （SOD） contents were significantly higher than those of the control group （P<0.05）； at 42 days of age， the serum SOD content and liver GSH， GSH-Px content of the treatment group 2 were significantly higher than those of the control group （P<0.05）. 3） Compared with the control group， the relative expressions of SOD1， SOD2， GPX1， and HO-1 mRNA in the treatment groups at 21 days of age and the relative expressions of NQO1， GPX1， SOD1， and HO-1 mRNA in the treatment groups at 42 days of age were significantly increased （P<0.05）. 4） Compared with the control group， the Firmicutes in the treatment group 3 at 21 days of age were significantly reduced （P<0.05）， while the Bacteroidetes and Bacteroidetes were significantly increased （P<0.05）. At 42 days of age， Colidextribacter and Lactobacillus in treatment group 1， as well as Colidextribacter in treatment group 3， were significantly increased （P<0.05）. Conversely， Eisenbergiella in treatment groups 1， 2， and 4 were significantly decreased （P<0.05）. In conclusion， under the condition of the current study， supplementation of red elephant grass anthocyanins can enhance the antioxidant capacity in the serum and liver of white-feathered broilers， modulate the intestinal microbial community structure， and improve their resistance to oxidative stress.

The purpose of this experiment is to investigate the metabolic energy requirements of 1-21-day-old rapid yellow feathered broiler chickens. A total of 900 healthy 1-day-old Xiangjia Yellow Chicken chicks （450 male and 450 female chicks） with similar body weights were randomly allocated into 5 groups for each gender， each consisting of 6 replicates with 15 birds per replicate. A single-factor completely randomized design was employed to feed different levels of metabolic energy diets over an experimental period of 21 days（Male：11.51、11.92、12.34、12.76、13.18 MJ·kg^-1；Female：11.30、11.72、12.13、12.55、12.97 MJ·kg^-1）. The results demonstrated that： 1） Growth performance analysis revealed quadratic patterns （P<0.01） in body weight （BW） and average daily gain （ADG） of male chicks with increasing dietary metabolizable energy （ME） levels during 1-21 days， while females exhibited significant positive linear relationships （P<0.01）. Both gender demonstrated linearly decreasing trends （P<0.01） in average daily feed intake （ADFI） and feed conversion ratio （FCR） as ME levels elevated. 2） Regarding carcass characteristics， dressing percentage， eviscerated yield， and abdominal fat rate in males increased linearly with ME supplementation （P<0.05）. Notably， females demonstrated greater sensitivity to ME stimulation in abdominal fat deposition，the abdominal fat percentage significantly increased （P<0.01）. 3） Serum biochemical profiling identified a unique quadratic fluctuation （P<0.05） in uric acid concentration of male chicks. Based on the relationship between metabolic energy intake and average daily weight gain， the metabolic energy prediction equation for rapid yellow feathered broiler chickens at this developmental stage can be established， which is： Male： Metabolic Energy Requirements （MEr）=1.653 8BW^0.75（g）+12.489ADG（g）； Female： MEr=3.360 5BW^0.75（g）+7.736 4ADG（g）. The corresponding metabolic energy requirement for male chicks is 379.32 kJ·d^-1， while for female chicks， it is 351.66 kJ·d^-1， corresponding to feed metabolic energy levels of 12.50 MJ·kg^-1 for male chicks and 12.47 MJ·kg^-1 for female chicks.

Porcine reproductive and respiratory syndrome virus （PRRSV） is a major pathogen that poses a serious threat to the global swine industry. Its non-structural protein NSP4， as a 3C-like serine protease （3CLSP）， cleaves viral multiproteins at the junctions between other conserved domains， playing a critical role in viral transcription and replication. This study aims to elucidate the structural features and enzymatic activity of the NSP4 protein， providing a theoretical basis for the development of antiviral small-molecule drugs targeting NSP4. This study used the PRRSV NADC30-like strain as the research object. The PRRSV nsp4 gene was cloned into the pGEX-6p vector， induced for expression in Escherichia coli BL21（DE3）， and purified to obtain high-purity protein. Crystallization conditions were screened using the drop-plate method， and the 1.47 Å high-resolution crystal structure of NSP4 was determined using X-ray crystallography. Its enzymatic activity was measured in vitro using fluorescence resonance energy transfer （FRET）. The results showed that PRRSV NSP4 consists of three domains： the N-terminal β-barrel domain （I， residues 12-69）， the middle β-barrel domain （II， residues 89-153）， and the C-terminal α/β domain （III， residues 159-190）. Further active site prediction identified the substrate-binding pocket of NSP4 as composed of Ser-19， Met-20， and His-39， providing a target for structure-based inhibitor design. In vitro enzyme activity experiments showed that the catalytic efficiency of PRRSV NADC30-like strain NSP4 toward the substrate， Kcat/Km， was 411.38 L·（mol·s）^-1， and the substrate affinity， Km， was 23.28 μmol·L^-1. This study reports the high-resolution structure and enzyme kinetic characteristics of the PRRSV NADC30-like strain NSP4， laying an important foundation for structure-based antiviral drug design and screening.

Porcine reproductive and respiratory syndrome virus （PRRSV） remains a pathogen causing substantial economic losses to China’s swine industry. Porcine reproductive and respiratory syndrome virus type 2 （PRRSV-2） currently predominates in China， exhibiting a complex epidemiological pattern characterized by co-circulating multiple lineages， which significantly complicates disease control measures. This study aimed to evaluate the diagnostic potential of metatranscriptomic sequencing in discriminating PRRSV-2 mixed infections involving multiple lineages. A PRRSV-2-positive serum sample collected from a swine farm in Sichuan Province was subjected to metatranscriptomic sequencing. This approach successfully recovered complete viral genome sequences. Subsequent comprehensive analysis， including phylogenetic reconstruction （IQ-TREE）， similarity comparison （Megalign）， and recombination analysis （RDP4/SimPlot）， was employed to elucidate the evolutionary and recombination characteristics of the viruses. The results demonstrate the successful identification of three distinct PRRSV-2 strains from this single serum sample： SC01-2025 （15 023 bp）， SC02-2025 （14 954 bp）， and SC03-2025 （15 293 bp）—the phylogenetic classification assigned these strains to lineages 1.8， 3， and 8.7， respectively.SC01-2025 was identified as a recombinant between lineage 1.8 （NADC30） and lineage 5 （VR2332）. While its full genome shared 88.09% nucleotide similarity with the NADC30 strain， regions encompassing NSP3-7， NSP9-12， and ORF3 exhibited higher similarity to the VR2332 strain. SC02-2025 was found to be a triple-recombinant virus （lineages 1.8/3/8.7）. This strain displayed the highest similarity to the NADC30 strain within the NSP2 and NSP3 regions， to the JXA1 strain within NSP1 and NSP7-10 regions， and the QYYZ strain in the remaining genomic segments. SC03-2025 showed 99% full-genome nucleotide similarity to the JXA1-P80 vaccine strain， with no evidence of recombination detected. This study， leveraging metatranscriptomic sequencing， precisely identified a mixed infection involving multiple PRRSV-2 lineages within a single pig farm in Sichuan. It revealed the complex epidemiological scenario involving the concurrent presence of a vaccine strain （JXA1-P80） and field strains （NADC30-like and QYYZ-like viruses）. These findings demonstrate that metatranscriptomic sequencing provides high-resolution diagnostic support for PRRSV disease surveillance and monitoring.

Porcine epidemic diarrhea virus （PEDV） is a coronavirus that causes acute diarrhea， vomiting， and dehydration in piglets， among other clinical signs， and has caused severe economic losses to the global swine farming industry. More and more studies have reported that long non-coding RNAs （lncRNAs） play important regulatory roles in porcine coronaviruses， however， the role and mechanism of lncRNA 663 in PEDV infection remain unclear. Based on prior transcriptome sequencing， we identified a co-expression pattern between lncRNA 663 and the tight junction protein CLDN10. This study aimed to explore their roles and regulatory relationship during PEDV infection. The qRT-PCR and Western blot results showed that the expression levels of lncRNA 663 and CLDN10 were up-regulated after PEDV infection of IPEC-J2 cells （P<0.05）， DSS treatment significantly reduced the expression levels of lncRNA 663 and CLDN10 （P<0.05）. Subcellular localization prediction revealed that lncRNA 663 was mainly localized in the cytoplasm， and RPISeq prediction revealed that lncRNA663 and CLDN10 were highly interoperable （SVM： 0.98， RF： 0.8）. Functional experiments demonstrated that siRNA-mediated knockdown of lncRNA 663 led to a significant decrease in both CLDN10 expression and PEDV replication （P<0.05）. Furthermore， structure prediction using AlphaFold3 and ChimeraX revealed potential hydrogen bond interactions between CLDN10 and the PEDV spike （S） protein， suggesting a role in viral entry or infection. Collectively， these results suggest that lncRNA 663 may promote PEDV replication by regulating CLDN10 expression. These findings expand our understanding of PEDV-host interactions and provide a theoretical basis for future antiviral strategies targeting host non-coding RNAs.

The aim of this study was to investigate the replication characteristics and pathogenicity of a cell-adapted strain derived from a prevalent PEDV isolate. The viral strain isolated from a large-scale pig farm was serially passaged in Vero cells to different generations. The biological characteristics of the virus were evaluated in vitro， and the 25th （F25） and 141st （F141） passages were inoculated into 3-day-old piglets to compare their pathogenic differences. The results showed that typical syncytial cytopathic effects （CPE） caused by PEDV were observed in Vero cells at the 4th blind passage of the clinical inoculum. By the 25th passage， both the CPE morphology and viral titers had stabilized. Upon reaching the 141st passage， the viral titer increased significantly and induced a more rapid cytopathic effect. Indirect immunofluorescence assay （IFA） specifically detected the expression of the PEDV N protein， and genomic sequence analysis identified the strain as genotype G2b. Pig challenge experiments demonstrated that， compared with the low-passage parental strain （YJH/2017/F25）， the YJH/2017/F141-inoculated group showed no significant clinical symptoms or histopathological lesions， along with a significant reduction in viral shedding via anal swabs. All assessed parameters showed no significant difference from those of the DMEM control group， indicating reduced pathogenicity and favorable safety profile of the strain. These findings indicate that after serial in vitro passaging， the prevalent PEDV strain acquired enhanced cell adaptability while exhibiting decreased pathogenicity in neonates. This study provides a promising candidate strain for the development of an attenuated PEDV vaccine， and its complete genomic sequence and biological characteristics establish a foundation for further research on recombinant viruses.

The non-structure protein （NSP） 12 of feline infectious peritonitis virus （FIPV） serves as the catalytic core component of the RNA-dependent RNA polymerase （RdRp）. However， the molecular mechanisms regulating its interactions with NSP7 and NSP8， as well as the enzymatic activity and function of the RdRp complex， remain to be fully elucidated. This study aims to investigate the molecular characteristics of NSP12， explore its interaction with NSP7 and NSP8， and verify the enzymatic activity of the RdRp complex. Firstly， the physicochemical properties and structural features of NSP12 were predicted by bioinformatics analysis. To investigate the interaction between NSP7， NSP8 and NSP12 in the FIPV RdRp complex， pcDNA3.1-Flag-NSP7， pcDNA3.1-HA-NSP8 and pcDNA3.1-Myc-NSP12 plasmids were constructed and expressed in 293T cells. The protein interactions and co-localization were detected using co-immunoprecipitation （Co-IP）， pull-down， and indirect immunofluorescence assay （IFA）. Secondly， the NSP7， NSP8 and NSP12 genes of FIPV were amplified by PCR and homologously recombined into the pET-30α vector to construct prokaryotic expression recombinant plasmids. The induced expression conditions were further optimized， and the proteins were purified by nickel column affinity chromatography and concentrated using an ultrafiltration tube to obtain the RdRp complex. Finally， an in vitro RNA elongation system for FIPV RdRp was established， and the enzymatic activity of the RdRp complexes obtained above was tested using the gel method. Bioinformatics analysis showed that NSP12 was a stable hydrophilic protein， containing 43.51% α helices， 45.02% random coils and 90 potential phosphorylation sites. The Co-IP， pull-down， and IFA assays confirmed the interaction and co-localization of NSP7， NSP8 and NSP12 in 293T cells. NSP7 and NSP8 were expressed as inclusion bodies， with molecular masses of 10 and 23 ku， respectively. In contrast， NSP12 was expressed as a soluble protein， with a molecular mass of 105 ku. The results of the enzyme activity assay demonstrated that the RdRp complex exhibited the capacity to extend the RNA template strand. Conversely， NSP12 alone exhibited no such activity. In this study， the FIPV RdRp complex was purified by prokaryotic expression and an enzyme activity detection system was established in vitro. It was confirmed for the first time that NSP12 from the FIPV-DF2 strain requires the formation of a complex with NSP7 and NSP8 to exert its RdRp function. This study lays the foundation for further exploring the biological function of FIPV RdRp and developing antiviral drugs targeting RdRp.

This study aimed to achieve soluble prokaryotic expression of the N-terminal extracellular domain of the pseudorabies virus （PRV） receptor Nectin1 protein， and to screen high-affinity， high-specificity DNA aptamers targeting it， thereby providing a novel molecular tool for blocking PRV adsorption. The porcine Nectin1 gene was cloned into the prokaryotic expression vector pET28a containing a maltose-binding protein （MBP） solubility-enhancing tag to construct the recombinant plasmid pET28a-MBP-Nectin1. The correctly digested and sequenced recombinant plasmid was transformed into Escherichia coli BL21 （DE3） for induced expression. The recombinant protein was validated by Western blot and pull-down assays. A DNA aptamer specifically targeting rMBP-Nectin1 was selected using magnetic bead-coupled Systematic Evolution of Ligands by EXponential enrichment （SELEX）. The affinity and specificity of the selected aptamer Apt-Nectin1-6 were evaluated by magnetic bead-based ELASA. Nectin1 knockdown cell lines （sh-Nectin1-1 and sh-Nectin1-2） were used as controls to verify the binding ability of Apt-Nectin1-6 to cell surface Nectin1 by flow cytometry. Additionally， the inhibitory effect of the aptamer on on adsorption of pseudorabies virus to host cells was assessed by flow cytometry and viral genome copy number analysis. Western blot and pull-down assays confirmed the correct expression and biological activity of the recombinant MBP-Nectin1 protein. Through SELEX， the DNA aptamer Apt-Nectin1-6， exhibiting high affinity to rMBP-Nectin1 with an equilibrium dissociation constant （Kd） of （0.11±0.02） nmol and no cross-reactivity with rMBP-gD， rMBP-S1， rMBP-CD163， or rMBP alone， was successfully screened. Molecular docking analysis indicated that Apt-Nectin1-6 binds to Nectin1 via hydrogen bonds. Further flow cytometry， viral genome copy number quantification， and virus titration collectively demonstrated that Apt-Nectin1-6 inhibits PRV adsorption to host cells in a dose-dependent manner. （P<0.05）. In this study， a DNA aptamer Apt-Nectin1-6 with high specificity and affinity for the Nectin1 protein was successfully screened and verified to effectively block PRV adsorption， providing a foundation for developing novel anti-PRV strategies.

This study aimed to analyze the genetic relatedness of key antigenic and virulence genes in the attenuated FJ19 strain of infectious laryngotracheitis virus （ILTV）， and to evaluate its safety and immunogenic efficacy in specific-pathogen-free （SPF） chickens， thereby providing valuable groundwork for the development of an attenuated live vaccine derived from the ILTV FJ19 strain. The ILTV FJ19 strain was adapted through allantoic cavity inoculation in chicken embryos， with sequencing confirming immunogenic and virulence genes. Safety tests were assessed in 14-day-old SPF chickens. Protective efficacy was demonstrated by challenging vaccinated chickens （ocular/oral routes at 14 days） with virulent ILTV strain at 21 days post-vaccination （dpv）. Vaccine durability was also determined. After serial passages in 10-day-old SPF chicken embryos via allantoic cavity inoculation， the EID₅₀ values of harvested P0， P1， P2， and P3 viral suspensions were 10^5.0， 10^6.5， 10^6.9， and 10^6.5·mL^-1， respectively. Genetic and phylogenetic analysis revealed： key immunogenic genes （gB， gC， gD， UL32） and virulence-associated genes （gE， gI， TK， ICP4， gG， gH， gK， gL， gM， gN） of FJ19 strain clustered closely with attenuated strains K317， Nobilis Laringovac^®， LT Blen， Poulvac ILT^®， Laryngo Vac and Chinese virulent isolates LJS09 （2009）， ck/CH/LHLJ/120305 （2012）， WF03 （2019）. Safety assessment demonstrated： Ocular inoculation （10^4.0 EID₅₀ per bird） caused mild conjunctivitis in 40% of 14-day SPF chickens， while oral administration showed no symptoms. Intratracheal challenge （10^4.0 EID₅₀ per bird） of FJ19 strain in 21-day-old SPF chickens caused transient respiratory reactions in a minority （30%）， with no severe ILTV-specific symptoms. Efficacy evaluation confirmed： Immunization （0.2 or 1 dose via ocular/oral routes） provided at least 90% protection against intratracheal challenge with virulent WF03 （10^4.0 EID₅₀ per bird） at 21 dpv， with no clinical signs in immunized groups. However， at least 90% of unimmunized virus-challenged chickens developed characteristic severe ILTV infection symptoms. Vaccine-induced immunity persisted for ≥6 months. The results demonstrated that： The ILTV strain FJ19 achieves high-titer cultivation via allantoic cavity inoculation. Its major antigenic and virulence genes show no significant variation compared to early Chinese isolates and vaccine strains. The FJ19 strain demonstrated excellent safety and immunogenicity in 14-day-old SPF chickens， immunity duration reached 6 months， with oral administration maintaining safety and efficacy. Thus， adaptation to allantoic cavity inoculation of FJ19 strain yielded a virus suspension with optimal safety and immunogenicity， the attenuated ILTV FJ19 strain represents an ideal candidate for oral attenuated live vaccine development.

Infectious bursal disease （IBD） severely restricts the economic development of the poultry industry. Therefore， the development of safe and efficient IBD vaccines has become a key topic of considerable interest. To explore the feasibility of using Eimeria tenella as a live vector for the expression and delivery of infectious bursal disease virus （IBDV） antigens， this study constructed a genetically modified strain of E. tenella （Et-VP2） expressing the VP2 protein of the IBDV. An expression plasmid was constructed， digested with enzymes， and then transfected into E. tenella using nuclear transfection. Flow cytometry was utilized to sort and select positive strains expressing the red fluorescent reporter gene （mCherry）， and the expression of the VP2 gene was identified at both the genetic and protein levels. Results showed that after three passages in vivo in chicks， the transgenic E. tenella expressing VP2 protein had a positive rate of fluorescence exceeding 95%， confirming successful integration and stable expression of the foreign gene at both the genetic and protein levels （with fluorescence rates reaching 95% in the offspring population）. Indirect immunofluorescence results indicated that the fusion protein （Mic2-linker-VP2） was localized at the apical end of the sporozoites of E. tenella. This study successfully established a stable transfection system for E. tenella， laying the foundation for subsequent research on the immune protection of genetically modified strains against infectious bursal disease.

Rabbit coccidiosis is a worldwide-distributed parasitic protozoal disease， which has caused huge economic losses to rabbit industry. Traditional methods of anticoccidial treatment result in drug resistance and drug residue， vaccination is a promising alternative. The rEiAMA1， rEiIMP1， EmGAM56， and EmROP17 were respectively displayed on the surface of B. subtilis spores by using a coat protein （CotB） as an anchor. Then the recombinant spores were used to oral immunize rabbits， the protective effects of B. subtilis-rEiAMA1， -rEiIMP1， and -rEiAMA1+rEiIMP1 against Eimeria intestinalis were evaluated； and the protective effects of B. subtilis-EmGAM56， -EmROP17， and -EmGAM56+ EmROP17 against Eimeria magna were evaluated. The results showed that B. subtilis-rEiAMA1 immunized rabbits displayed better anti-E. intestinalis protective effect， the oocyst output and lesion score were significantly decreased （P<0.05）， the oocyst output decrease ratio was 59.91%. B. subtilis-EmGAM56 immunized rabbits displayed better anti-E. magna protective effect， the oocyst output was significantly decreased （P<0.05）， the oocyst output decrease ratio was 68.54%. Moreover， the immunized rabbits showed significantly increased specific serum IgG levels and specific IgA levels （P<0.05）. The four recombinant B. subtilis spores can stimulate the body to produce specific intestinal and systemic humoral immune responses. B. subtilis-rEiAMA1 and B. subtilis-EmGAM56 showed better immunoprotection against E. intestinalis and E. magna， respectively. Therefore， B. subtilis is a potential live vector for rabbit coccidiosis vaccines.

This study aims to evaluate the biological characteristics of Trichinella spiralis nuclear hormone receptor-23 （NHR-23） and its potential for vaccine development. The recombinant TsNHR-23 protein （rTsNHR-23） was obtained via cloning and expression， followed by purification. Its immunoreactivity was assessed via Western blot， while transcriptional levels of the TsNHR-23 gene across different developmental stages of T. spiralis were analyzed using qPCR. In vitro experiments were conducted to analyze the inhibitory effect of anti-rTsNHR-23 polyclonal antibody on the molting of muscle larvae and to explore the effect of this protein on the function of mouse macrophages. The immune-protective efficacy of PLGA nanoparticle-encapsulated rTsNHR-23 was tested by immunizing ICR mice and analyzing parasite burden. The TsNHR-23 gene was transcribed across all developmental stages of T. spiralis， with the highest expression observed in muscle larvae （ML） stage. Anti-rTsNHR-23 antibodies significantly reduced larval molting rates and inhibited molting progression. The rTsNHR-23 protein significantly enhanced the transcription levels of IL-1β， IL-6， and IL-10 in mouse macrophages. Immune protection experiments demonstrated that mice immunized with the rTs-NHR-23 protein exhibited a highly significant increase in serum IgG antibody levels compared to the control group. After the first immunization， IGF-1 levels markedly decreased， while IL-6 and TGF-β levels increased. Following the second immunization， IGF-1 partially rebounded， whereas TGF-β showed significant downregulation. Post-challenge infection， compared to the control group， the immunized mice displayed significant reductions in the number of T.spiralis adult worms， the average number of newborn larvae produced per female worm， and the muscle larval burden. rTsNHR-23 exhibits potent inhibitory effects on ML molting， modulates macrophage functions， and significantly reduces parasitic load in immunized hosts， demonstrating promising potential as a candidate vaccine against T. spiralis.

This study aimed to identify the suspected pathogens causing rabbit mortality in a large-scale meat rabbit farm in Ningxia and analyze their pathogenicity. The causative pathogens and their pathogenicity were determined through necropsy of deceased rabbits， bacterial isolation and culture， 16S rRNA sequencing analysis， PCR identification， antimicrobial susceptibility testing， and challenge experiments in mice and rabbits. Results showed that 3 Gram-negative bacterial strains were isolated and identified as capsular type A Pasteurella multocida， Proteus mirabilis， and Escherichia coli through 16S rRNA sequencing and PCR identification. These strains were designated as QF24-Pm01， QF24-Ps02， and QF24-Ec03， respectively. Antimicrobial susceptibility testing by the Kirby-Bauer （K-B） method revealed that QF24-Pm01， QF24-Ps02， and QF24-Ec03 exhibited resistance to commonly used clinical drugs such as penicillin， kanamycin， and enrofloxacin， but were sensitive to ceftriaxone. Detection of resistance genes showed that all three isolates carried resistance genes aadB， strA， strB， bla_TEM， and sul1. Virulence gene analysis indicated that Pasteurella multocida carried 16 virulence genes， including exbB， exbD， and hgbA； Proteus mirabilis carried ureC， zapA， atfC， atfA， and pmfA； and Escherichia coli carried eaeA and fimC. Mouse pathogenicity experiments revealed LD₅₀ values of 9.27×10⁵ CFU， 1.43×10⁶ CFU， and 1.88×10⁷ CFU for QF24-Pm01， QF24-Ps02， and QF24-Ec03， respectively. The results of the mixed-infection pathogenicity assay in mice showed that mortality occurred at 9 h， 18 h， and 16 h post-inoculation in the QF24-Pm01， QF24-Ps02， and QF24-Ec03 groups， respectively. In contrast， the QF24-Pm01+QF24-Ec03 and QF24-Ps02+QF24-Ec03 co-infection groups exhibited mortality at 6 h and 13 h post-inoculation， respectively. Histopathological analysis revealed that both single and mixed infections resulted in tissue damage in the lungs， liver， and intestines. The primary pathological changes included pulmonary interstitial edema， capillary congestion， and neutrophil infiltration； hepatic steatosis and disordered hepatocyte cord arrangement； as well as intestinal epithelial shedding and villous rupture. In the rabbit pathogenicity assay， all rabbits in the QF24-Pm01 + QF24-Ec03 co-infection group and the QF24-Pm01 group succumbed at 24 and 36 h post-inoculation， respectively. The mortality rates at 7 days post-inoculation for the QF24-Ps02+QF24-Ec03， QF24-Ec03， and QF24-Ps02 groups were 60%， 20%， and 40%， respectively. Necropsy findings demonstrated that rabbits in the QF24-Pm01+QF24-Ps02+QF24-Ec03 co-infection group exhibited pulmonary congestion and edema with a ground-glass appearance， as well as splenic atrophy and dark brown discoloration. These pathological changes were consistent with the lesions observed in clinical necropsy examinations of affected rabbits. This study demonstrates that the pathogenicity of Pasteurella multocida is enhanced when co-infected with Escherichia coli， and that Escherichia coli and Proteus mirabilis exhibit increased virulence when co-infected. Therefore， co-infection with Pasteurella multocida，Proteus mirabilis， and Escherichia coli may be the primary cause of high mortality rates in meat rabbits at the studied facility. Consequently， in addition to preventing Pasteurella multocida infection， control measures against Escherichia coli and Proteus mirabilis should be reinforced to reduce the risk of mixed infections and associated mortality.

This study aims to evaluate the immunogenicity of membrane surface protein P4 from Mycoplasmacapricolum subsp. capripneumoniae （Mccp） and its potential as a candidate antigen for vaccine. Preliminary laboratory work identified the membrane surface protein P4. In this study， recombinant protein was prepared using a prokaryotic expression system， after purification， the recombinant protein was emulsified with an adjuvant to prepare vaccine. Immunization and challenge experiments were conducted on goats， and the immune efficacy was evaluated based on clinical symptoms， pathological changes， serum antibodies， bacterial load in lung， and cytokines. The results showed that the rP4 protein was successfully expressed and purified， and could specifically react with the His-tag antibody， rP4 polyclonal antibody and Mccp infection antibody. Immunization with the rP4 stimulated high levels of antibody production in goat but did not alleviate clinical symptoms， with a survival rate of only 40%， the same as the PBS control group. Pathological observations indicated that both acute and chronic cases in the P4 group were more severe than those in the control group. The average bacterial load in the lungs was higher in the P4 group than that in the inactivated vaccine group and the PBS control group. Serum cytokines analysis revealed that the P4 group had upregulated IL-2 level and downregulated IL-4 level post-immunization. In conclusion， this study successfully expressed and purified rP4 and the prepared P4 protein induced antibody production in goats but failed to protect them from Mccp challenge， even exacerbating the pathogenic effects of Mccp on the goats. The findings provide a reference for the design and development of future new type vaccines for contagious caprine pleuropneumonia.

The aim of this study was to identify the Lactobacillus strains that are resistant to Salmonella and provide antibiotic alternatives for the chicken industry. Lactobacillus were isolated from chicken jejunal contents using MRS medium， candidate strains were screened by in vitro bacterial inhibition， acid resistance， enzyme production and adhesion， selected strains were orally administered to one-day-old AA chickens for 28 days to evaluate their effects on growth performance. At 42 days， a Salmonella challenge test was conducted to validate the inhibitory effect against salmonellosis， finally the Lactobacillus strain resistant to Salmonella was identified. Results A total of 27 Lactobacillus strains （including 13 strains of Ligilactobacillus salivarius， 11 strains of Limosilactobacillus reuteri， 2 strains of Lactobacillus johnsonii， and 1 strain of Lactobacillus gasseri） were isolated and identified， among which L. salivarius S31 could significantly inhibit Salmonella Typhimurium and Salmonella Gallinarum， with survival rate of more than 50% at pH 3.0， and produce protease and lipase. This suggests that S31 strain not only has a strong ability to inhibit Salmonella， but also has gastrointestinal adaptability and potential probiotic function. The results of animal experiments showed that the transplantation of L. salivarius S31 strain for 28 d significantly increased the body weight （P<0.000 1）， leg muscle weight （P<0.001）， breast muscle weight （P<0.01）， jejunum length （P<0.01）， intestinal villi length （P<0.05）， goblet cell counts （P<0.05）， and abundance of Lactobacillus in jejunum of the chickens （P<0.05）， which suggests that S31 strain can promote the growth of chickens. The Salmonella attacking was carried out at 42 days， and bacterial load of the spleen （P<0.05）， the transcription levels of TLR-4， MyD88 and AP-1 signaling pathways in the ileum （P<0.01）， and the transcription levels of pro-inflammatory factors IL-1β， IL-6 and IL-12 （P<0.05） in L. salivarius S31 transplantation group were significantly lower than those in the control group， and the jejunum and ileum structures were intact. L. salivarius S31 strain exhibits robust Salmonella-inhibitory capacity， favorable gastrointestinal adaptability， and superior growth-promoting properties， demonstrating significant potential for practical applications.

This work aimed to prepare nanosilver and evaluate its therapeutic effect on methicillin-resistant Staphylococcus aureus infections in canine skin wounds. The nanosilver was prepared by biosynthesis method using Ganoderma lucidum extracts （40% polysaccharide content） as reducing agent and polyvinylpyrrolidone as stabilizing agent， followed by characterization using ultraviolet visible absorption spectroscopy， transmission electron microscopy and dynamic light scattering. Subsequently， the invitro antimicrobial activity of nanosilver was evaluated by determining the minimum inhibitory concentration. Finally， 12 Beagles weighing about 10 kg were randomly divided into 3 groups of 4 dogs each， namely， saline control group， enrofloxacin positive control group and nanosilver group. After establishing the model of wound infection with methicillin-resistant Staphylococcus aureus， wound healing， bacterial load， histopathological morphology， collagen fibers and angiogenesis， and macrophage polarization levels were tested and analyzed after 12 consecutive days of treatment. The results showed that the prepared nanosilver had a maximum absorption wavelength at 433 nm， and were spherical and uniformly distributed， with a particle size of （20.01±6.99） nm and zeta potential of （-2.56±0.32） mV. Nanosilver exhibited good antibacterial activity against common skin infection causative organisms containing methicillin-resistant Staphylococcus aureus， with a minimum inhibitory concentration of 16-32 μg·mL^-1. The results of in vivo experiment showed that nanosilver could effectively reduce the bacterial load of wound tissues， promote collagen deposition and angiogenesis， and improve the immune microenvironment of wounds， which in turn promoted the healing of infected wounds. The results suggested that the prepared nanosilver can effectively treat methicillin-resistant Staphylococcus aureus induced canine skin wounds infections， laying a material and practical foundation for its translation and application to the clinical treatment of pet skin infections.

This study aimed to investigate preventive effects of aspirin eugenol ester （AEE） on oxidative stress and inflammatory response in the ovaries of hens with fatty liver hemorrhagic syndrome （FLHS）. A total of sixty-five 160-day-old Hy-Line Brown laying hens were randomly assigned to five groups， each containing 13 hens. The control group （Control） was fed a basal diet， while the model group （high-energy low-protein diet， HELP） and low-， medium-， and high-dose AEE treatment groups （AEE-L， AEE-M， AEE-H） were all fed a high-energy low-protein diet supplemented with 0， 25， 50， and 100 mg·kg^-1 aspirin eugenol ester， respectively. The results showed that， compared with the control group， the ovaries of laying hens in the HELP group exhibited distinct lipid vacuoles of varying sizes. In contrast， the number of lipid vacuoles in the ovarian tissue was significantly reduced in the AEE-treated groups compared with the HELP group. Compared with the control group， the activities of SOD， GSH， and CAT in the ovaries of hens in the HELP group were significantly decreased， while the level of MDA was significantly increased （P<0.05）. In contrast， AEE treatment groups demonstrated significantly enhanced SOD， CAT， and GSH activities and reduced MDA levels and significantly upregulated SOD and CAT mRNA transcription compared with the HELP group （P<0.05）. Compared with the Control group， the expression levels of antioxidant-related genes and proteins Nrf2， NQO1， and HO-1 were significantly decreased in the ovaries of hens in the HELP group， while the expression of Keap1 was significantly increased （P<0.05）. In contrast， the AEE-treated groups showed significantly increased expression of Nrf2， NQO1， and HO-1 and decreased expression of Keap1 compared with the HELP group （P<0.05）. Compared with the control group， the mRNA transcription levels of inflammatory cytokines IL-1β， IL-6， and TNF-α were significantly increased in the ovaries of hens in the HELP group. In addition， the expression levels of inflammation-related genes and proteins-including TLR4， NF-κB， and p-p65 were also significantly elevated （P<0.05）. In contrast， AEE treatment groups exhibited significantly reduced expression of IL-1β， IL-6， TNF-α， TLR4 and NF-κB genes， along with significantly decreased p-p65 protein expression compared to the HELP group （P<0.05）. In summary， fatty liver hemorrhagic syndrome in laying hens can induce ovarian oxidative stress and inflammatory responses. Supplementation with varying doses of AEE demonstrated preventive effects against ovarian damage in laying hens with fatty liver hemorrhagic syndrome.

Cyclophosphamide （CTX） is a widely used chemotherapeutic agent for treating canine tumors； however， its potent immunosuppressive effects can impair immune function and disrupt gut microbiota homeostasis， increasing the risk of secondary complications. Lactobacillus murinus （L. murinus） has been reported to modulate immune cell activity and protect the gastrointestinal tract， highlighting its potential as a functional food additive. In this study， a probiotic strain with optimal characteristics， L. murinus ZNL-13， was isolated from the fresh feces of healthy dogs. The aim was to evaluate the effects of this canine-derived strain on CTX-induced immunosuppression and gut microbiota dysbiosis in dogs. Serum immune parameters and fecal 16S rRNA sequencing were used for analysis. Results showed that prophylactic administration of L. murinus ZNL-13 significantly improved CTX-induced reductions in white blood cells and lymphocytes （P<0.05）， alleviated liver and kidney injury， reduced oxidative stress （P<0.05）， and enhanced intestinal barrier function by lowering serum levels of DAO and D-lactic acid （P<0.01）. In addition， L. murinus ZNL-13 mitigated the CTX-induced decline in serum IgG and key immune cytokines （P<0.05）， increased the abundance of beneficial gut bacteria， and restored microbial homeostasis. Meanwhile， the levels of beneficial bacteria such as Blautia_A， Holdemanella， and Peptacetobacter were increased， contributing to the restoration of gut microbiota homeostasis. In conclusion， L. murinus ZNL-13 effectively counteracts CTX-induced immunosuppression， oxidative stress， and gut barrier dysfunction， offering a promising strategy for the development of novel functional food additives to support immune health in dogs.

This study aimed to analyze the medication law of the core medication for the treatment of dairy cow mastitis prescription. This study used Excel and GraphPad Prism software to analyze the basic information of the prescription for the treatment of dairy cow mastitis. IBM Spss Statistics 26.0， IBM Spss Modeler， and Cytoscape software were used for analyzing the performance and association of Chinese herbal medicines in the prescription. Data mining was conducted using association rules with IBM SPSS Statistics 26， IBM SPSS Modeler， and Cytoscape software to identify the core drugs. The clinical efficacy of the core drug dandelion was also validated. The results showed that a total of 239 prescriptions were included in the analysis， involving 185 different traditional Chinese medicines. The total frequency of drug use was 2 317. The core drugs identified were dandelion， angelica， licorice， honeysuckle and forsythia， with a frequency of ≥100 times. These drugs have different properties， including cold， warm， bitter， sweet， and pungent， and mainly belong to the liver， stomach， and lung meridians. The combination of dandelion-Zingiber officinale， honeysuckle drug pair had the highest confidence. Cluster analysis categorized the core drug into 7 groups， with dandelion being the most central drug in its use.Clinical efficacy tests demonstrated that the core drug， dandelion， significantly reduced the number of somatic cells in the milk of cows with subclinical mastitis and showed a good therapeutic effect on subclinical mastitis in dairy cows. In this study， dandelion was identified as the core drug for the treatment of dairy cow mastitis. This provides a reference for scientifically guiding the rational use of medication in the clinical treatment of dairy cow mastitis， as well as offering data support for the development of Chinese veterinary medicine for the prevention and treatment of dairy cow mastitis.

The aim was to study the effects of the ethanol extract of chicory （CIL）， a medicinal and edible homology herb， on the hepatic lipid metabolites of non-alcoholic fatty liver disease （NAFLD） induced by high-fat diet （HFD）， and to analyze the mechanism of chicory action in ameliorating the disorders of lipid metabolism. HFD was induced to establish a NAFLD model in C57BL/6J mice， and the therapeutic effects of CIL （800 mg·kg^-1 and 2 g·kg^-1） and simvastatin （SVT， 20 mg·kg^-1） were evaluated in NAFLD mice. Measurement of liver aspartate aminotransferase （AST） and alanine aminotransferase （ALT） enzyme activity， catalase （CAT） activity， malondialdehyde （MDA） content， and tumor necrosis factor-α （TNF-α） and interleukin-10 （IL-10） content. The composition of hepatic lipid metabolites was detected using UPLC-MS/MS lipidomics； qRT-PCR was used to detect the expression levels of mRNA genes for lipid synthesis-related genes （PPARγ， ACCα， Srebp1c and CD36） and lipid oxidation-related genes （PPARα and CPT1α）. The results of in vivo experiments showed that high-dose chicory alcohol extract group （HCIL） suppressed HFD-induced abnormal elevation of AST and ALT highly significantly （P<0.01）， and there was no significant difference compared with the SVT group enzyme activity （P>0.05）. The CIL treatment highly significantly reduced blood and liver triglyceride （TG） levels （P<0.01）， and there was no significant difference in TG levels with the SVT group （P>0.05）. The HCIL treatment obviously alleviated hepatic steatosis and fat cell volume； remodeled the lipid composition of liver， lowered hepatic TG and DG levels， and elevated the levels of PG， PE， LNAPE， LPE， and HexCer. Both the HCIL group and the SVT group highly significantly lowered the hepatic expression levels of PPARγ， ACCα， Srebp1c， and CD36 mRNA genes （P<0.01）， targeted inhibition of lipid uptake by CD36 protein， and molecular docking with high binding energy. Therefore， CIL regulates lipid metabolism disorders by targeted inhibition of CD36 lipid uptake， improving liver lipid composition. The pharmacological effects of CIL treatment for NAFLD are not significantly different from those of SVT drug treatment， which will promote the development and application of chicory products in the livestock and poultry farming industry.

This study aimed to systematically evaluate the effects of compound Chinese herbal medicine on growth performance and intestinal flora of chickens. Based on the PRISMA guidelines， this study conducted a Meta-analysis of 21 Chinese literatures （including randomized controlled trials of chickens） published in CNKI， VIP， Wanfang， PubMed and Web of Science databases from the establishment of the database to September 2025. The RevMan 5.4 software was used to calculate the weighted average and 95% confidence interval by random effect model， and to evaluate the heterogeneity and publication bias. The results showed that compound Chinese herbal medicine could significantly increase ADFI （MD=2.42 g·d^-1， 95%CI［1.99， 2.85］） （P<0.01）， ADG （MD=3.23 g·d^-1， 95%CI［2.48， 3.98］） （P<0.01）， and VH/CD （MD=0.87， 95%CI［0.40， 1.35］） （P<0.01） compared with the blank group. Significantly reduced F/G （MD=-0.19， 95%CI［-0.27， -0.11］） （P<0.01） and the number of E.coli （MD=-1.48， 95%CI［-1.87， -1.09］） （P<0.01）， but there was no significant difference between the Chinese herbal medicine group and the blank group in improving Shannon index， Chao1 index， Simpson index and the number of lactic acid bacteria （P>0.05）. Compared with the antibiotic group， there was no significant difference in ADFI， ADG and F/G （P>0.05）. In this study， the effects of compound Chinese herbal medicine on production performance and intestinal health of chickens were evaluated by Meta-analysis system. The results showed that the compound Chinese herbal medicine had a significant effect on ADFI and ADG of broiler and broiler-egg dual-purpose chickens， and the effect was prominent in the brooding period and the growing period. At the same time， it could significantly reduce the F/G of broilers， but had no significant effect on the related indexes of laying hens. Compound Chinese herbal medicine could selectively inhibit the proliferation of Escherichia coli and significantly increase the ratio of intestinal villus height to crypt depth （VH/CD）， but had no significant effect on the diversity index of flora and the number of lactic acid bacteria. The results of this study provide a systematic reference for the application of compound Chinese herbal medicine in chicken production， which is of great significance for the application and in-depth study of Chinese herbal medicine in chicken breeding.

From 2023 to 2024， 10-20-day-old ducks in a meat duck farm in Jiangsu Province， were suspected of being infected with duck plague virus （DPV）. This study aims to isolate and identify the virus and analyze genomic characteristics. Viral isolation and genome sequencing analysis were performed on the disease material in this study. Results were as follows： Duck plague virus was detected in visceral tissue， and a duck plague strain was successfully isolated， named QL2312. It has the same pathogenicity as the virulent strain. Sequence analysis of the isolate genome revealed that the UL2 gene of QL2312 strain was the same as that of the vaccine strain C-KCE strain， while the UL56/LORF5 gene was the same as the classical strain CHv and CV strains. This study successfully isolated a duck plague mutant strain， providing a basis for further research on related diseases.