Accurately identifying the potential functional genetic loci that significantly associated with animal economic traits holds both theoretical and practical significance for improving individual animal breeding outcomes and enhancing production performance. With the advancement of omics technologies, integrating multi-omics information has become an effective strategy to enhance the precision of genetic locus identification. Among these, expression quantitative trait locus (eQTL) analysis provides a new research perspective for analyzing the association mechanisms between genotypes and economic traits, as well as the genetic regulatory networks of gene expression, making it an important research direction in the post-genome-wide association study (Post-GWAS) era. This paper first introduces the basic principles and methods of eQTL analysis, then focuses on strategies and approaches for integrating eQTL and GWAS data to enhance the efficiency of functional genetic loci identification, as well as its applications in the genetic breeding of important livestock (cattle, pigs, chickens and sheep). Finally, it summarizes the problems that exist in practical implementation, which provides theoretical foundations for elucidating the genetic architecture of complex traits in animals and establishes a crucial basis for advancing innovative biobreeding technologies to revitalize the livestock breeding industry.

N6-methyladenosine (m⁶A) modification is a widespread RNA modification in eukaryotic cells and an important component of RNA epigenetic modification. m⁶A modification mainly regulates the transcription, processing, and degradation of mRNA and non-coding RNA (ncRNA), thus regulating a variety of physiological and metabolic activities. In livestock and poultry, m⁶A modification is involved in a variety of molecular mechanisms of life activities, such as muscle development, lipid metabolism, and reproduction. In this review, the regulatory role of m⁶A modification in livestock and poultry genetic breeding and reproduction is summarized. In addition, the application and future direction of m⁶A modification in livestock and poultry are also discussed.

With the rapid development of omics technology, high-throughput detection technologies of omics at all levels are emerging. As a new research field rising in the late 20th century, metabolomics can not only identify and detect all metabolites in organisms, but also use advanced analysis technology and data processing platform to analyze the collected metabolite information in depth and comprehensively. It can not only comprehensively and dynamically monitor the metabolic changes in pigs during growth and development, but also provide a new method for the mechanism analysis of economic traits by combining multi-omics data. This paper mainly summarizes the latest progress of metabolomics in the study of important economic traits such as pig growth, meat quality, reproduction and disease resistance, and discusses its challenges and application prospects, so as to provide reference for the application of metabolomics in the study of important economic traits of pigs in the future.

Post-sampling embryo cryopreservation technology is a critical component of preimplantation genetic testing (PGT), holding significant application value in embryo sex selection, genetic screening, genomic selection, and assisted reproduction. However, embryonic sampling procedures may lead to zona pellucida damage, reduced cell count, and epigenetic abnormalities, thereby limiting cryopreservation efficiency. Recent advancements, including optimized sampling methods (e.g., polar body biopsy, blastomere biopsy, and trophectoderm biopsy), improved culture medium formulations (e.g., supplementation with L-arginine and glutathione), and refined cryopreservation protocols (e.g., vitrification techniques), have markedly enhanced the cryopreservation efficiency of post-sampling embryos. Furthermore, non-invasive techniques (e.g., analysis of cell-free DNA in embryo culture medium) and epigenetic modifications (e.g., DNA methylation regulation) offer novel approaches to minimize embryonic damage. This review summarizes the mechanisms of cryopreservation-induced damage in post-sampling embryos and protective strategies, with a focus on cutting-edge advancements in zona pellucida repair, epigenetic regulation, and cryopreservation efficiency. By evaluating the strengths and limitations of existing technologies, this review aims to provide theoretical foundations and technical references for developing safe and efficient cryopreservation protocols, thereby promoting their application and adoption in livestock breeding and assisted reproductive technologies.

Early embryonic loss in cattle, as a major challenge in the cattle industry, has attracted extensive research attention in recent years and achieved significant progress. Studies have revealed that genetic factors, environmental factors (such as heat stress and nutritional imbalance), management negligence, and infectious pathogens are all factors contributing to early embryonic loss in cattle. To address this issue, researchers have proposed comprehensive prevention and control strategies including optimizing nutritional supply, reducing environmental stress, and strengthening disease prevention and control. Meanwhile, the innovation of modern biotechnology has opened up new avenues for in-depth exploration of the complex mechanisms of early embryonic loss in cattle. In the future, research in this field will focus on further revealing the mechanisms of embryonic loss, improving the precision and effectiveness of prevention and control technologies, and thus providing a solid guarantee for the sustainable development of the cattle industry.

Taurine is a sulfur-containing β-amino acid with multiple biological functions including antioxidant, anti-inflammation, anti-aging, hormone secretion and lipid metabolism regulation, immune function enhancement, and maintenance of intestinal health. This article systematicly reviews the sources, synthetic and metabolic pathways, biological functions of taurine, focuses on elucidating its dose-effect relationships and mechanisms of action in poultry production (broilers, laying hens, ducks, and quails).This study also systematically evaluates the optimal dietary taurine supplementation levels in poultry feed formulations, providing empirical evidence for precision nutrient supplementation and functional feed development in intensive poultry production systems. In broiler production, dietary inclusion of 0.05% to 0.25% taurine significantly enhances average daily gain (ADG) and feed conversion ratio (FCR), while effectively mitigating oxidative stress-induced muscular damage under chronic heat stress conditions. For laying hens, supplementation with 0.05% to 0.3% taurine demonstrates multifunctional benefits including improved egg production rate, enhanced eggshell integrity, and attenuation of hepatic lipid deposition. Notably, species-specific requirements were identified, with recommended taurine inclusion rates of 0.1% for ducks and 0.05%-0.1% for quail in formulated diets. These findings establish a scientific basis for optimizing taurine utilization efficiency and advancing practical applications in modern poultry nutrition strategies.

Zearalenone (ZEN) and its derivatives rank among the most hazardous mycotoxins worldwide, posing a significant threat to food safety and public health. Consequently, the development of efficient biodegradation strategies for ZEN has emerged as a critical research priority. Among these, enzymatic degradation approaches have garnered considerable attention due to their specificity and efficacy.ZEN-degrading enzymes primarily include laccases, manganese peroxidases, and lactonases. Compared to the first two enzyme classes, lactonases exhibit distinct advantages for the development of ZEN detoxification agents, including higher catalytic activity, well-defined degradation mechanisms, and non-toxic degradation byproducts. However, natural enzymes often suffer from limited stability under industrial processing conditions, hindering their applicability in large-scale production and practical settings. To address this challenge, enhancing enzyme activity and stability through rational or semi-rational design and optimization has become a key research focus.This review comprehensively examines microbial-derived ZEN lactonases, covering gene mining, biochemical characterization, molecular engineering, and practical applications. By evaluating the advantages and limitations of biological detoxification in industrial and livestock production, this analysis provides novel insights for the development of highly active, robust ZEN-detoxifying enzyme preparations.

The resistance gene mcr-1, as a key genetic element mediating bacterial resistance to "last-resort" antibiotic colistin, has become a research hotspot since its discovery and poses a potential threat to public health. This paper aims to investigate the fate of colistin resistance gene mcr-1 in the planting-breeding cycling system. First, we summarize the resistance mechanisms, horizontal gene transfer, fitness costs, and compensatory evolution of mcr-1. Subsequently, we examine its occurrence, fate, and horizontal transmission characteristics in the planting-breeding cycling system, along with the key factors influencing its dissemination. Finally, we propose corresponding control measures, providing insights for high-quality development of circular agriculture and public health security.

Brucellosis is a zoonotic bacterial disease caused by Brucella spp., which poses a serious threat to human health and the development of the livestock industry. Brucella is an intracellular pathogen, and its ability to evade the host's innate immune response and survive within host cells is a key aspect of its virulence. Lipopolysaccharide (LPS) is one of the major virulence factors encoded by Brucella. Compared to other Gram-negative bacteria, the main components, spatial structure, and linkage of Brucella LPS are distinct, which results in its unique low endotoxicity. Moreover, Brucella LPS plays a crucial role in helping the bacteria evade recognition by the host's immune system and in modulating immune responses. This review focuses on Brucella LPS, reviewing its unique structure, biosynthesis process, and immune evasion mechanisms, thereby laying the foundation for a comprehensive understanding of the biological functions of Brucella LPS.

Akabane virus (AKAV) is a vector-borne pathogen that primarily causes reproductive disorders in ruminants such as cattle and sheep. It is widely distributed in areas where blood- sucking insects such as mosquitoes and midges are concentrated in tropical and temperate regions. The infection situation is particularly severe in some Southeast Asian and East Asian countries. Regional epidemics have been found in China. With the increasing intensification of cattle and sheep farming, frequent trading and transportation, and the expansion of the breeding range of vector insects due to climate change, the disease has a further trend of spreading infection. In this paper, beginning with pathogenic characteristics and tissue tropism of AKAV, the interaction mechanisms between virus infection and the host′s responses were summarized emphatically. It could provide a reference for further exploration of the pathogenesis of AKAV and the development of corresponding therapeutic drugs and preventive preparations.

This study aimed to identify functional genes and key signaling pathways related to body weight at 16 weeks of age in Kangle yellow chickens based on omics data, in order to provide a basis for the selection and breeding of local chicken breeds for market weight traits. To detect SNPs significantly associated with body weight at 16-week in 434 Kangle yellow chickens, genome-wide association study (GWAS) was performed using the "Jingxin No. 1" 55K SNP chip. A total of 13 potential SNPs significantly associated with target traits were detected, located on chromosome 3 (81 641 565 bp), chromosome 4 (1 860 041 bp, 33 409 710 bp, 33 716 131 bp, 33 925 946 bp), chromosome 6 (23 563 123 bp, 35 324 210 bp), chromosome 11 (15 294 113 bp, 15 346 007 bp), chromosome 12 (2 503 065 bp, 2 504 749 bp) and chromosome 18 (1 994 686 bp, 2 307 607 bp). The 825 candidate genes near SNP sites were screened. Gene function annotation analysis showed the most significant enrichment of biological processes was motor activity. The significant enrichment of 17 KEGG pathways were found, including tight junction, glycosaminoglycan degradation and metabolic pathways (P < 0.05). Combined with previous transcriptomic sequencing data, and published literature, KCNQ5, P2RY10, ITM2A, RBPMS, PGAM1, CYP17A1, BNIP3, ATMIN, BCO1, TNNC1, GNAI2, MYH1F, MYH1A, MYH10, and PIK3R5 were preliminarily identified as important candidate genes for body weight traits at 16 weeks of age in Kangle yellow chickens. Neuroactive ligand-receptor interaction, metabolic pathway, adrenergic signaling in cardiomyocytes and tight junction were key pathways. In this study, 13 SNPs, 15 key candidate genes and 4 key pathways were preliminarily identified related to the body weight at 16-week of Kangle yellow chicken. These results provided theoretical basis and technical support for molecular marker-assisted breeding of growth traits in Kangle yellow chickens.

The aims of this study was to dissect the genetic architecture of body weight and age at the first egg. Phenotypic data were collected from an F₂ segregation population of Dongxiang blue-shelled chickens and White Leghorn. The 1 534 hens in F₂ generation were genotyped using 600K gene microarray. The SNPs data were quality cleaned. Missing data was imputed with BEAGLE software. SNPs associated with traits of onset of lay were obtained with single-step GWAS. Gene annotation and GO enrichment analysis were carried out on the significant SNPs to elucidate the biological roles of candidate genes. The body weight at first egg (BWFE) showed high levels of heritability. The heritability on BWFE ranged from 0.62 to 0.65. The age at first egg (AFE) showed moderate to high levels of heritability. The heritability on AFE ranged from 0.35 to 0.42. The genetic correlation coefficients between traits of onset of lay ranged from 0.24 to 0.37. There were one or more QTLs close to HTR2A or CAB39L on chromosome 1 that was significantly associated with the BWFE, which was supported by the additional 343 SNPs over the threshold of significant level. These QTL explained 11.74% and 9.34% of the phenotypic variance of BWFE, respectively. On chromosome 4, region close to NCAPGwas significantly associated with the BWFE, and reinforced by the additional 89 SNPs. The region explained 5.68% of the phenotypic variance. On chromosome 9, QTL was identified nearby the GPR149, and explained 1.33% of the phenotypic variance. Furthermore, QTL on AFE were identified near the STON2 gene on chromosome 5 and the PPM1L gene on chromosome 9, explaining 1.80% and 1.22% of the phenotypic variance of AFE, respectively. Single-step GWAS was used to detect the genetic structure of traits of onset of lay. Two novel QTL, mapped on 40 Mb of chromosome 5 and 22 Mb of chromosome 9, were identified with BWFE and AFE. GO enrichment analysis showed that the variations on BWFE were related to gonadal development, and the phenotypic variations on AFE was related to calcium ion transport.

This study aimed to investigate the targeting relationship between miR-376c-5p and PIK3CA to elucidate how miR-376c-5p influences 3T3-L1 preadipocyte differentiation, exploring its role and regulatory mechanisms in adipogenesis. In the early stage, the research group selected 6 healthy 3-year-old Bashbay sheep (large-tailed type) and 6 F2 generation sheep of wild Argali×Bashbay sheep (small-tailed type) under the same feeding conditions, their tail adipose tissues were collected, and screened out the differentially expressed miR-376c-5p through the sequencing data analysis results of the tail fat. Then, an miR-376c-5p mimic overexpression vector was constructed for transfection, and the expression levels of PIK3CA and adipogenic differentiation marker genes were detected by qPCR. The adipogenic capacity was detected by combining Oil Red O staining and Bodipy staining. The conservation of miR-376c-5p in different species was analyzed, and bioinformatics software was used to predict the target gene(PIK3CA) of miR-376c-5p and the binding sites between them. Additionally, dual-luciferase reporter assay was performed to verify the targeting relationship between miR-376c-5p and PIK3CA. Compared with the mimic NC group, qPCR results showed that overexpression of miR-376c-5p significantly downregulated the mRNA expression levels of key adipogenic differentiation marker genes (PPARγ, ADIPOQ, ACACA and FASN)(P < 0.001). Both Oil Red O and Bodipy staining results demonstrated a reduction in lipid droplet formation following miR-376c-5p overexpression, indicating that overexpression of miR-376c-5p inhibited adipogenic differentiation in 3T3-L1 preadipocytes. Bioinformatic prediction revealed a potential binding site between miR-376c-5p and the 3′UTR of PIK3CA. Dual-luciferase reporter assay further confirmed that overexpression of miR-376c-5p significantly suppressed the luciferase activity of a vector containing the PIK3CA 3′UTR fragment (P < 0.05). Moreover, overexpression of miR-376c-5p markedly reduced the mRNA expression level of the candidate target gene PIK3CA (P < 0.05). These findings suggest that miR-376c-5p may inhibit adipogenesis by targeting PIK3CA, providing a theoretical basis for further exploration of miRNA functions at the molecular level.

The aim of this study was to explore the molecular mechanism underlying the effect of methyltransferase like 14 (METTL14) on the myogenic differentiation of sheep skeletal muscle satellite cells(SMSCs) and to provide a theoretical basis for analyzing the molecular mechanism of METTL14 regulating the differentiation of SMSCs. SMSCs were isolated and identified. The overexpression plasmid of METTL14 and the control plasmid were transfected into SMSCs, and the overexpression efficiency and the function of METTL14 affecting SMSCs differentiation were detected by RT-qPCR and Western blot. RNA-seq was performed for METTL14-overexpressed SMSCs(OE-14) and control SMSCs(OE-NC). Differentially expressed genes (DEGs) were identified under the conditions of P < 0.05 and log₂|Fold Change| > 1, GO and KEGG enrichment analysis and protein-protein interaction (PPI) network interaction analysis for DEGs was conducted. Overexpression of METTL14 markedly elevated the expression of METTL14, as well as the mRNA and protein levels of myogenic differentiation-related genes (MyHC, MyoG) in ovine SMSCs, thereby demonstrating successful METTL14 overexpression and the enhancement of SMSCs differentiation mediated by METTL14. A total of 259 differentially expressed genes were identified between the two groups. Compared with the OE-NC group, the OE-14 group had 45 up-regulated genes and 214 down-regulated genes. Differential genes were mainly enriched in nuclear pathways, Rap1 signaling pathways, etc. PPI protein network interaction analysis identified 8 hub genes (Degree≥15), including MX1, RSAD2, IFIH1, DDX58, HERC5, ISG15, MX2, and IRF7. This study indicated that METTL14 can promote the differentiation of ovine SMSCs through influencing genes and signaling pathways related to cellular processes and muscle development.

This study aimed to reveal the adaptive changes in sex hormone secretion patterns and the metabolic mechanisms underlying the development of horn-supporting skin structures. Six adult buck, including 3 horned and 3 polled individuals, were selected. Blood samples were collected before morning feeding, and serum sex hormone levels were measured using enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA). Skin samples were taken from the horn base of horned bucks and the corresponding forehead region of polled bucks for transcriptome sequencing analysis. Hormone analysis showed that the serum concentrations of progesterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) were significantly lower in horned bucks than in polled bucks (P < 0.05). The testosterone-to-estradiol (T/E2) ratio was significantly higher in horned bucks (P < 0.05), but the levels of testosterone (T) and estradiol (E2) did not show significant differences. Transcriptome analysis identified 1 721 differentially expressed genes (DEGs), with 1 183 upregulated and 538 downregulated. These DEGs were significantly enriched in pathways related to metabolism (metabolic pathways), fatty acid elongation, fatty acid metabolism, extracellular matrix (ECM-receptor interaction), biosynthesis of unsaturated fatty acids, fatty acid degradation, and glycolipid metabolism. In conclusion, the findings suggest that horn-based skin requires more energy and keratin secretion for protein sheath growth, while maintaining higher structural stability by reducing cellular collagen and exoskeleton. In addition, horned bucks have reduced levels of P, FSH and LH, suggesting a shift in reproductive strategy towards horn-dependent morphological dominance and aggressive behaviour driven by high T/E2 ratios. The results of this study provide a theoretical basis for resolving the molecular regulatory mechanisms of horn growth and development in goats and the effects of horn traits on reproductive adaptations.

The objective of this study was to investigate the genetic diversity and population structure of Leizhou goats to provide the theoretical basis for conservation and sustainable utilization of their germplasm resources. In this study, whole-genome resequencing was performed on blood samples from 20 Leizhou goats, with an average sequencing depth of ~10×. The genome-wide data from 43 additional goats was downloaded, including Yunshang black goats, Jintang black goats, Jining grey goats, Tibetan goats, and Longlin goats, from the NCBI database. Using these whole-genome data, then linkage disequilibrium (LD) analysis was performed using PopLDdecay software. We calculated genetic distances using Plink software and constructed and visualized a genetic relationship matrix using GCTA software. In addition, we employed Plink for principal component analysis (PCA), Phylip to construct a neighbor-joining (NJ) phylogenetic tree, and ADMIXTURE for population structure analysis, collectively assessing the genetic diversity and population structure of goat populations. Finally, runs of homozygosity (ROH) were analyzed and fixation index (F_ST) values between populations were calculated. Subsequently, the identified genomic regions were annotated for candidate genes and subjected to functional enrichment analysis. We identified a total of 18 810 921 SNPs, with the majority (65.71%) located in intergenic regions. For Leizhou goats, we found the overall average values of H_O, H_E, MAF, Pi, PIC, Ne and F_IS were 0.298, 0.295, 0.211, 0.001 8, 0.808, 65.488 and 0.086, respectively. The higher H_O compared to H_E indicated normal levels of population genetic diversity. For LD analysis, we found the fastest LD decay in Leizhou goats, suggesting higher genetic diversity and weaker selection pressure on their genome. We calculated identity-by-state (IBS) distance matrices and genetic relationship matrices (G matrices), which demonstrated close genetic relatedness among Leizhou goats but significant genetic differentiation from other goat populations. Analysis of ROH revealed that the Leizhou goat population exhibited a reduced inbreeding level. Using PCA, we observed discrete clustering of Leizhou goats, clearly distinguishing them from other populations. We constructed a neighbor-joining (NJ) phylogenetic tree, which further supported the independent evolutionary branch of Leizhou goats. For population structure analysis, we determined that the optimal clustering occurred at K=3, revealing a unique genetic architecture in Leizhou goats. Genome-wide F_ST analysis identified significant selection signals in the ADGRL3, CXCR1, and HTR1F gene regions after screening the top 1% of highly differentiated loci, suggesting these genes may be associated with adaptation. In this study, we comprehensively analyzed the genetic diversity, kinship relationships, and population structure of Leizhou goats. The study results provide a scientific basis for understanding the genetic background of Leizhou goats and lay a theoretical foundation for developing effective conservation strategies and sustainable utilization plans.

This study aimed to use the population-specific reference genome of Chinese indicine cattle and systematically evaluate its advantages in identifying SNPs. This study focused on whole-genome resequencing data from 60 Chinese indicine cattle samples to identify and compare SNPs via both the European taurine cattle reference genome (ARS-UCD1.2) and the population-specific reference genome of Chinese indicine cattle (Leiqiong breed: ASM3988116v1). For the multiallelic SNPs detected in ARS-UCD1.2, genomic coordinate mapping chain files between the genomes was established, converting these SNPs into biallelic SNPs on the basis of the coordinates of the population-specific reference genome, followed by functional annotation. Variants detected in Chinese indicine cattle populations via the ASM3988116v1 population-specific reference genome presented significant advantages over the ARS-UCD1.2 genome: 1) This approach could perform a more comprehensive identification of intronic and untranslated region variants, increasing the detection sensitivity for low-frequency and rare variants; 2) It could reduce false positives in variant identification caused by reference bias; 3) It facilitated the conversion of some multiallelic SNPs, previously filtered out owing to genomic reference bias, into biallelic SNPs. These SNPs were annotated to 8 352 genes, including important genes related to the growth, development, and environmental adaptability of indicine cattle, such as muscle development (CTNNA1), immunity (SIL1), blood circulation (VPS13A), muscle development and photoperiod regulation (EYA3), etc. The specific reference genome for Chinese cattle can increase variant detection sensitivity, reduce reference genome bias, and uncover more functionally significant loci. This provides a high-confidence data foundation for population genetics research and precision breeding, offering important theoretical and practical implications.

The study aimed to comprehensively evaluate the genetic diversity and genetic structure of the E. h. hemionus in Kalamaili Mountain Ungulate Wildlife Nature Reserve in Xinjiang (hereinafter referred to as "Kalamaili") and provide molecular genetic evidence for their conservation and management. This study employed non-invasive sampling methods, PCR amplification and sequencing techniques to analyze 161 freshly collected fecal samples with successfully extracted genomic DNA. Individual identification, genetic diversity indices, and genetic structure were evaluated. The results revealed that, based on 10 microsatellite loci, 159 genetically distinct individuals were identified, harboring 123 alleles in total. The average polymorphic information content (PIC) was 0.634, observed heterozygosity (Ho) was 0.533, expected heterozygosity (He) was 0.658, and the inbreeding coefficient (Fis) was 0.140 (P < 0.01). For mitochondrial markers, haplotype diversities (Hd) of CYTB, D-LOOP, and the concatenated gene (CYTB+D-LOOP) were 0.63, 0.82, and 0.84, respectively. The nucleotide diversities (Pi) were 0.005 24, 0.020 63, and 0.012 54, respectively. The genetic diversity of the E. h. hemionus population in Kalamaili was lower than that of population in Mongolia but higher than those in Iran, Indian, and Turkmenistan population, indicating an overall moderate-to-high level of genetic diversity. Genetic structure and phylogenetic analysis revealed that when K=2, the Kalamaili population had the optimal genetic structure division. Mitochondrial phylogenetic and haplotype network distribution analyses showed that the two major clades (Clade Ⅰ and Clade Ⅱ) within the Kalamaili population were closely related to the population in Mongolia, sharing haplotypes Hap 21, Hap 23 and Hap 24. Demographic analysis showed an "L"-shaped distribution of microsatellite allele frequencies, while mitochondrial neutrality tests (Tajima's D and Fu's Fs) yielded non-significant positive values (P>0.05). Mismatch distribution exhibits multimodal pattern, and Bayesian skyline plot indicated a relatively flat trend, collectively suggesting that the Kalamaili E. h. hemionus population has not undergone recent expansion or decline. In conclusion, the Kalamaili E. h. hemionus population exhibits high genetic diversity, with two ancestral lineages present and currently maintains a stable population size. These findings provide a molecular genetic basis for future conservation strategies and support evidence-based management decisions for the sustainable protection of this species.

The aim of this paper was to clone and analyse TIMP1, a member of the tissue inhibitor of matrix metalloproteinases (TIMPs) family. The regulatory role of TIMP1 on hair follicle growth-related genes in hair follicle papilla cells (DPCs) was explored by overexpression and knockdown of TIMP1. In this study, we cloned the coding sequence (CDS) of TIMP1 and analysed its bioinformatics function. After that, we constructed the overexpression vector pcDNA3.1-TIMP1 and designed and synthesized siRNA. We overexpressed and knocked down TIMP1 in DPCs to probe the expression of genes related to hair follicle growth and development, and detected the proliferation level of DPCs by EdU and CCK-8. The results showed that the CDS region of the rabbit TIMP1 gene was 624 bp in length, encoding a total of 207 amino acids. Bioinformatics analysis indicated the presence of a signal peptide in the TIMP1 protein, which did not contain a transmembrane structural domain and was homologous in different mammals. Overexpression of TIMP1 in DPCs was able to highly significantly up-regulate the mRNA expression levels of BMP2, SFRP2, and TGFβ1 genes (P < 0.01), and highly significantly down-regulate the mRNA expression level of WNT2 gene (P < 0.01), and knockdown of TIMP1 was able to significantly down-regulate the expression of mRNA of BMP2, SFRP2 genes (P < 0.05), and highly significantly down-regulated the mRNA expression of TGFβ1 gene (P < 0.01). In addition, EdU and CCK-8 results showed that overexpression of TIMP1 could inhibit the proliferation of DPCs, and knockdown of TIMP1 could promote the proliferation of DPCs. In this study, we successfully cloned the CDS sequence of rabbit TIMP1 gene and preliminarily predicted its bioinformatics function, analysed the regulatory effect of TIMP1 on hair follicle growth-related genes. We verified its effect on inhibiting DPCs proliferation, and provided a reference for elucidating the theoretical research of rabbit hair follicle growth and development.

This study aimed to investigate molecular structural changes in porcine mature oocytes before and after vitrification using Raman spectroscopy. Ovaries were collected from pigs at a slaughterhouse, and cumulus-oocyte complexes were isolated in the laboratory. After in vitro maturation, fresh oocytes and vitrified oocytes were collected, respectively. Laser confocal Raman spectrometer was used to obtain the Raman spectrum information of fresh oocytes and vitrified oocytes. The molecular structure changes of oocytes before and after vitrification were analyzed after quality control of the data. The results indicated that, compared to fresh oocytes, vitrified oocytes exhibited reduced carbohydrate components such as N-acetylgalactosamine within the carbohydrate signal range of 1 020-1 140 cm^-1. In the amide Ⅲ band signal range (1 230-1 300 cm^-1), β-sheet content increased markedly while α-helix structures decreased. Within the lipid signal range (2 800-3 100 cm^-1), the content of long-chain fatty acids decreased, while the content of branched-chain fatty acids increased. In summary, vitrification alters the secondary protein structure, fatty acid composition and other key components like N-acetylgalactosamine in porcine oocytes. These changes result in multiple cryodamage effects, including protein structural damage, altered lipid composition, and abnormal carbohydrate metabolism.

This study aimed to establish induced pluripotent stem cells (iPSCs) of wild boar with stable passage ability, and integrate exogenous genes into the X chromosome of iPSCs, which provides technical support for the conservation, development and utilization of wild boar resources. Eight pluripotency factors were introduced into wild boar fibroblasts by the PiggyBac transposon system to establish the iPSCs line and the pluripotency was identified. Plasmids expressing green fluorescent protein (GFP) were transferred into wild boar iPSCs by electroporation (3 procedures) and chemical transfection reagents lipofectamine 3000 (Lip 3000) and jetPRIME, respectively. And the optimal transfection protocol was screened with analysis of transfection efficiency by flow cytometry. Combined with CRISPR/Cas9 and site-directed integration technology independent of homologous recombination, the sgRNAs with high-efficiency were co-transfected with GFP expression elements into wild boar iPSCs. Then target fragments were amplified by PCR and the site-directed integration of GFP was identified by sequencing. In this study, human OCT4, SOX2, KLF4, C-MYC, NANOG, LIN28, NR5A2 and miR302/367 were introduced into wild boar fibroblasts. Twenty days post-induction, wild boar iPSCs were obtained with normal karyotype, positive alkaline phosphatase staining, stable expression of pluripotency factors OCT4, SOX2 and NANOG. Additionally, embryoid bodies, labeled with three germ layers marker (β Ⅲ tubulin, α-smooth muscle actin and GATA6), were observed in vitro. For different electroporation procedures, the cell transfection efficiency using CG104 ((9.73±0.23)%) was significantly higher than that of CA201 ((7.34±0.03)%) and CB150 ((6.69±0.10)%). The cell transfection efficiency with jetPRIME cationic polymer was (17.9±0.36)%, which was significantly higher than that of Lip 3000 ((14.07±0.85)%). After two plasmids with sgRNA or GFP expression elements were constructed successfully, they were co-transfected into wild boar iPSCs. PCR and sequencing results showed that GFP expression elements were inserted into the predetermined position of X chromosome and could be stably expressed. In conclusion, wild boar iPSCs were derived from fibroblasts. And GFP was knocked into the X chromosome of iPSCs by site-directed integration technology independent of homologous recombination. This study provided technical reference for gene editing in wild boar and breeding of domestic pig.

This experiment was conducted to investigate the effects of inulin on production performance, immune response, antioxidant status and hormone levels of lactating dairy cows under heat stress. Twenty-four healthy, high-yield Holstein cows with similar body conditions were grouped in 4 blocks of 6 cows based on days in milk (DIM), milk yield and parity (n=6 in each group) using a completely randomized grouping design. Within each block, cows were randomly assigned to 1 of the 4 treatment groups: a control group without inulin supplementation (CON), and three groups receiving inulin at doses of 200, 300 and 400 g·d^-1 (Inulin-2, Inulin-3 and Inulin-4). The trial period was 10 weeks, including 2 weeks of pre-feeding period and 8 weeks of formal trial period. The study was carried out during the hot summer, with an average temperature-humidity index (THI) greater than 68, indicating that the cows were experiencing heat stress. The results showed that feeding inulin had no significant effect on rectal temperature (RT) and respiratory rate (RR) of heat-stressed dairy cows (P>0.05). With the increase of inulin addition, the milk yield of dairy cows showed a quadratic increase trend, but the difference was not significant (0.05≤P < 0.1), while the milk yield of Inulin-3 and Inulin-4 groups were significantly higher than that of CON group (P < 0.05). With the increase of inulin feeding amount, serum creatinine concentration quadratic significantly decreased (P < 0.05), reached the lowest value in Inulin-3 group, and was significantly lower than that in CON group (P < 0.05). The concentration of serum interleukin (IL) - 1β quadratic significantly decreased (P < 0.05), and the lowest value was observed in Inulin-3 group, which was significantly lower than that in CON group (P < 0.05). The concentration of serum tumor necrosis factor (TNF) -α quadrtic decreased, but the difference was not significant (0.05≤P < 0.1), reaching the lowest value in Inulin-3 group, which was significantly lower than that in CON group (P < 0.05). The serum total antioxidant capacity (T-AOC) showed a quadratic upward trend, but the difference was not significant (0.05≤P < 0.1), reached the highest value in the Inulin-3 group, and was significantly higher than that in the CON group (P < 0.05). In summary, dietary inulin supplementation can alleviate the heat stress of high-yielding dairy cows by improving immune performance, antioxidant status and hormone secretion, and can also increase milk yield and improve milk composition. For lactating cows experiencing heat stress, the recommended dose of inulin is 300 g·d^-1.

The objective of this paper is to analyze the nutritional composition of fermented corn straw, determine its effective energy and standardized ileal digestibility (SID) of amino acids for growing-finishing pigs, and evaluate the feasibility of using fermented corn straw as an unconventional feedstuff in pig feeding. In Experiment 1, the substitution method was used to determine the digestible energy (DE) and metabolizable energy (ME) of fermented corn straw for fattening pigs. Twelve healthy crossbred barrows [Duroc×(Landrace×Large White)] (DLY) with an initial body weight of (65±3.08) kg were selected and randomly divided into two treatments using a 2×2 crossover design, with six replicates per treatment and one pig per replicate. The experimental diets included a corn-soybean meal-based diet and a fermented corn straw diet. In Experiment 2, a nitrogen-free diet method was employed to determine the apparent and standardized ileal amino acid digestibility of fermented corn straw for growing pigs. Six healthy DLY crossbred barrows with a T-cannula at the diatal ileum and an initial body weight of (42.7±3.3) kg were selected and randomly divided into two treatments using a 2×2 crossover design, with three replicates per treatment and one pig per replicate. The results showed that the DE and ME values of fermented corn straw for finishing pigs were 5.85 and 5.61 MJ·kg^-1, respectively. The SID of essential amino acids in fermented corn straw was 73.82%, the SID of non-essential amino acids was 65.09%, and the SID of total amino acids and crude protein was 70.33% and 61.60%, respectively. In conclusion, the results suggest that as a non-conventional feed ingredient, the fermented corn straw has a SID of amino acids similar to conventional feed ingredients such as wheat bran and corn DDGS in the Tables of Feed Composition and Nutritive Values in China (35^th Edition). Furthermore, its low cost and high feed value, which could provide theoretical support and technical assistance for addressing the shortage of feed ingredients in China.

This study aimed to investigate the effects of vitamin A supplementation during lactation in Tan lambs on production performance, fat deposition, muscle antioxidant capacity and meat quality. Forty-five healthy Tan lambs with body weight (4.38±0.23) kg were randomly divided into three groups of three replicates of five lambs each. The control group (CON group) was given normal milk intake, and the experimental groups were received 30 000 IU vitamin A (L group) and 60 000 IU vitamin A (H group) on the 1st day and the 1st month of age. All sheep were weaned on 70th day. After that, all animals were fed a basal diet for a fatten period of 120 days. The results indicated that: Compared with the control group, 1) the body weight of Tan lambs in groups L and H on the 1st day, 30th day and 60th day during the fattening period were significantly increased (P < 0.05), average daily gain was not significantly different (P>0.05). 2) Muscle catalase content of Tan lambs in L group was significantly increased (P < 0.05). Marbling score, meat color a_{45 min}^* and the pH at 24 h after slaughter were significantly increased (P < 0.05), intramuscular fat content tended to increase (0.05 < P < 0.1), and the muscle shear force was significantly decreased (P < 0.05). 3) The expressions of genes related to adipose deposition such as ZFP423, WNT10B, FABP4 and ACACA in the muscle of Tan lambs in groups L and H were significantly unregulated (P < 0.05). In conclusion, supplementing moderate amounts of vitamin A to lactating Tan lambs affects production performance during the fattening period, enhances content of intramuscular fat, improves the antioxidant capacity of the longest dorsal muscle, as well as enhances meat quality.

This experiment aimed to study the effects of replacing soybean meal by oil cake of flax seed under a high-precision feeding mode on nutrient apparent digestibility, rumen and fecal microbiota composition of fattening lambs. Sixty crossbred fattening lambs aged 2~3 months and weighing approximately 18 kg were randomly divided into three groups based on their body weight: Control group (CK group), Experimental Group Ⅰ, and Experimental Group Ⅱ. The proportions of oil cake of flax seed replacing soybean meal in the three groups were 0%, 50%, and 100%, respectively. The experimental period was 97 days, including a 7-day pretrial period and a 90-day formal trial period. The lambs were divided into early, middle, and late fattening stages based on their body weight. A digestion experiment was conducted from day 85~89 of the formal trial period, and the rumen fluid and feces were collected before the morning feed on the 90th day for microbial analysis. The results showed that: 1) There were no significant differences in the apparent digestibility of dry matter, neutral detergent fiber, acid detergent fiber, and crude protein among the groups (P>0.05). The ether extract digestibility increased with the increase of oil cake of flax seed replacing proportion (P=0.079). 2) At the phylum level, Bacteroidota and Firmicutes were the dominant microbial phyla in the rumen of all groups, the relative abundance of the top 10 phyla among the groups was not significantly different (P>0.05). At the genus level, Prevotella was the dominant bacterial genus in all groups, and there were no significant differences in the relative abundance of the top 10 bacterial genera among the groups (P>0.05). The relative abundance of Ruminococcus in Experimental Group Ⅱ was significantly higher than that in the CK group (P < 0.05). 3) At the phylum level of fecal microbiota, Bacteroidota and Firmicutes were the dominant bacterial phyla in all groups, accounting for 88.18%-91.88% of the total bacterial relative abundance. The relative abundance of Euryarchaeota in the CK group is significantly higher than that in the Experimental Group Ⅱ (P < 0.05), while the relative abundance of Actinobacteria in the Experimental Group Ⅱ is significantly higher than that in the CK group and Experimental Group Ⅰ (P < 0.05). There were no significant differences in the others relative abundance of the top 10 bacterial phyla among the groups (P>0.05). At the genus level, Muribaculaceae and Rikenellaceae_RC9_gut_group were the dominant bacterial genera in all groups, and there were no significant differences in the relative abundance of the top 10 bacterial genera among the groups (P>0.05). In summary, replacing soybean meal with oil cake of flax seed can improve the digestibility of ether extract and the relative abundance of Ruminococcus. Under the conditions of this experiment, oil cake of flax seed can completely replace soybean meal without negative effects on lambs.

The purpose of this study was to explore the anti-inflammatory activity of cinnamon essential oil (CEO) and cinnamaldehyde (CIN) and the protective effect on cellular inflammatory damage, and to provide theoretical reference for CEO and CIN to exert anti-inflammatory activity in livestock and poultry production. This study included 2 trials: 1) In vitro chemical anti-inflammatory study, the anti-inflammatory activity of CEO and CIN at four concentrations (6.25, 12.5, 25, 50 mg ·mL^-1) by measuring the inhibition of inflammation-related enzymes (cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX)) by ELISA method. 2) In the study of LPS induced inflammatory damage in RAW264.7 cells, the cytotoxicity tests were performed first to determine the CEO and CIN concentrations. Next, the RAW264.7 cells in the control group were cultured in a normal medium without any treatment. The cells in the LPS group were cultured in a normal medium and treated with LPS for 24 h. The cells in the CEO and CIN groups were cultured in a normal medium and treated with CEO or CIN of different concentrations (1, 3 and 5 μg·mL^-1) for 12 h, and subsequently treated with LPS for 24 h. The in vitroresults showed that both CEO and CIN exhibited dual inhibitory activity against COX-2/5-LOX, and showed dose-dependent reduction of COX-2 (P_L < 0.01) and 5-LOX (P_Q < 0.01) at 6.25-50 mg·mL^-1, and the inhibitory effect of CEO was significantly higher than (P < 0.01) CIN at the same concentration. The cell test results were showed as follows: 1) The results of cytotoxicity tests showed that CEO and CIN significantly increased (P < 0.05) cell survival rate within 0.781 25-6.25 μg·mL^-1, and 1, 3, and 5 μg·mL^-1 were selected for subsequent tests. 2) The observation of RAW 264.7 cell morphology showed that 1, 3 and 5 μg·mL^-1 CEO and CIN pretreatment could reduce the differentiation to varying degrees compared with the LPS group. 3) Compared with the LPS group, nitric oxide (NO) content was significantly decreased (P_Q < 0.05) in 3 and 5 μg·mL^-1 CEO and CIN groups, and the inhibitory effect of CEO was significantly higher than CIN (P < 0.01). 4) Compared with LPS group, the content of interleukin-1 β (IL-1 β), interleukin-6 (IL-6), and TNF-α (TNF-α) were significantly reduced (P < 0.05) in 1, 3 and 5 μg·mL^-1 CEO and CIN groups. In addition, the 3, 5 μg·mL^-1 CEO had significantly higher (P < 0.01) ability to reduce IL-1β and TNF-α content than CIN, but there was no significant difference on IL-6 (P>0.05). In conclusion, CEO and CIN had excellent anti-inflammatory activity, and the inhibitory effect of CEO on COX-2 and 5-LOX and the protective effect of LPS-induced inflammation in RAW264.7 cells were significantly stronger than that of CIN under the conditions of the current study.

This study aimed to evaluate the immune-enhancing effect of porcine soluble CD40L trimer protein on the Virus-like particles (VLPs) vaccine of Foot-and-mouth disease (FMD). In this study, the plasmid containing porcine CD40 ligand (CD40L) trimer (5B CD40L) gene was expressed in Escherichia coli BL21(DE3). The purified recombinant 5B CD40L protein was coupled to FITC and reacted with PBMCs. The biological activity of CD40L protein reacting with monocytes was studied by fluorescence microscopy. The guinea pigs were further immunized with 5B CD40L, VLPs and 5B CD40L-VLPs, and the effects of 5B CD40L on the immune effect of VLPs vaccine were evaluated by neutralizing antibodies, splenic lymphocyte proliferation and real-time RT-qPCR. SDS-PAGE and Western blot results showed that the size of recombinant 5B CD40L protein expressed was about 32 ku. The optimal expression conditions were 16℃, OD_{600 nm} value was 0.9, and concentration was 0.5 mmol L^-1 after IPTG induction for 12 h. Flow cytometry analysis showed that the recombinant CD40L protein could bind to CD40 protein molecules on the surface of porcine peripheral blood mononuclear cells (PBMCs). The combination of 5B CD40L and VLPs can accelerate the reaction of specific antibody and neutralizing antibody, and improve the proliferation level of spleen lymphocytes. In the 5B CD40L-VLPs group, the IL-2 and INF-γ mRNA of spleen cells are significantly higher than those in the VLPs group. In summary, 5B CD40L has the ability to enhance humoral and cellular immune responses of FMD VLPs vaccine, and has the potential as a vaccine adjuvant.

This study aimed to prepare monoclonal antibody against bovine kobuvirus (BKV) and establish a double antibody sandwich ELISA method for detection. The pET-30a (+) vector was utilized to express the BKV structural proteins VP0, VP3, and VP1 in a prokaryotic expression system. Three structural proteins were purified and used for immunization in BALB/c mice respectively. After three immunizations, the spleen cells of the mice were isolated and fused with SP2/0 cells to prepare hybridoma cells. Then the hybridoma cells were screened by the indirect ELISA method, the prepared monoclonal antibodies were used as capture antibody and detection antibody respectively, the conditions such as antibody concentration and incubation time were optimized, and a double antibody sandwich ELISA method for specifically detecting BKV was established. Results were as follows: Two hybridoma cell lines stably secreting monoclonal antibodies were obtained, designated 2A12 and 5E11, both of which were anti-VP1 protein antibodies; The results of the superimposed ELISA and antibody variable region sequence analysis indicated that they recognized different antigen sites. The results of monoclonal antibody specificity detection showed that the two monoclonal antibodies were specifically bound to BKV and were not bound to other bovine diarrhea viruses. One hundred and seven clinical samples were detected by RT-PCR method and the double antibody sandwich ELISA method established in this study. The results demonstrated that the positive coincidence rate between the two methods was 92.2%, the negative coincidence rate was 95.3%, and the overall coincidence rate was 93.5%. In conclusion, two monoclonal antibodies against BKV were prepared in this study, and a double antibody sandwich ELISA method for BKV was established. This method has good specificity, repeatability and sensitivity, which lays a foundation for the rapid diagnosis, prevention and control of BKV.

Mycoplasma gallisepticum(MG) infection causes chronic respiratory disease (CRD) in chickens, adversely affecting feed conversion efficiency, egg production rate, hatchability, and chick activity, leading to significant economic losses in the global poultry industry. This study aims to analyze the genomic characteristics and biological features of the LC strain and explore the epidemiological patterns of MG in China. Pipped failure embryos were collected in hatchery for MG isolation. The isolate underwent whole-genome sequencing (WGS) and analysis of adhesion-related protein variations. Additionally, multilocus sequence typing (MLST) analysis was conducted to determine the prevalence of MG strains in China. Genomic analysis revealed a compact MG genome of 953.4 kb with a GC content of 31.5%. Annotation identified 721 coding regions with an encoding density of 89.3%, 33 tRNAs, and 2 CRISPR sequences. Functional enrichment in translation, metabolism, and signal transduction indicated efficient protein synthesis and environmental adaptability, with potential genes related to virulence regulation. Moreover, 11 amino acid mutations were identified in the adhesion-related proteins P1, P30, and hwm3. MLST analysis classified the isolate as ST-81, along with ST-78 and ST-36, which constitute the prevalent MG genotypes circulating in China. MG has become as a significant contaminant in hatcheries, along with their genetic diversity notably increasing control challenges. This study provides insights into the genomic and biological characteristics of the LC strain and elucidates the dominant MG sequence types in China, offering valuable scientific evidence for pathogen biology research and vaccine strain selection.

In order to explore the mechanism of abortion caused by Brucellasecreted protein, a BrucellaBPE005 deletion strain was constructed to preliminarily explore the effect of secreted protein BPE005 on GPR126, a key protein in embryonic development. In this experiment, the BPE005 deletion strain S2308ΔBPE005 was constructed by homologous recombination method of replacing the target gene with Kan gene, and its genetic stability was tested by continuous culture for 15 generations. At the same time, the pBBR1MCS-4-BPE005 plasmid was constructed and electrotransposed into the BPE005 deletion strain S2308ΔBPE005, and the BPE005 backfill strain S2308▲BPE005 was successfully constructed. Every 4 hours, measure the OD_{600 nm} of strains S2308, S2308△BPE005, and S2308▲BPE005, and plot the growth curves of each strain based on time and OD_{600 nm}.The multiplicity of infection (MOI) was 100, and the strains S2308, S2308ΔBPE005 and S2308▲BPE005 were used to infect trophoblast cells HTR-8, and the intracellular survival of the three strains was detected by plate counting. Protein and RNA samples were collected from trophoblast cells at 0, 4, 8, and 12 h after infection, respectively, and the protein expression of GPR126 was detected by real-time PCR and western blot. In this experiment, the deletion strain and the supplemental strain of Brucella secreted protein BPE005 were successfully constructed, and the gene could be stably inherited for 15 generations. The results of the growth curve showed that compared with S2308, the growth viability of the BPE005 strain lacking Brucellasecretory protein was reduced from 12 h to 44 h, and the growth activity was significantly decreased from 24 h to 44 h plateau stage (P < 0.001), but the resupplementation strain reversed this result. The results of intracellular survival assay showed that the survival of S2308ΔBPE005 in HTR-8 cells decreased compared with S2308, and decreased significantly at 12 h and 24 h after infection (P < 0.001). The results of real-time fluorescence quantification and western blotting showed that compared with the control group, the expression of GPR126 protein in S2308 strain increased significantly at 4 h after infection (P < 0.001), and then began to decrease. Compared with the S2308 strain, the expression of GPR126 protein in the S2308ΔBPE005 strain was significantly decreased at 4 h after infection (P < 0.001), while the expression of GPR126 protein was significantly increased at 8 h (P < 0.001). No significant differences are noted between the S2308▲BPE005 and S2308 strains.. In this study, the stable genetic Brucellasecreted protein BPE005 deletion strain and the supplementary strain were successfully constructed, the Brucellasecreted protein BPE005 could promote the reproduction of Brucella and promote the intracellular proliferation of Brucella, and the BPE005 deletion strain inhibited the expression of GPR126 protein in the early stage of infection, but could promote the expression of GPR126 in the middle stage. The results of this study lay a foundation and provide ideas for the effect of Brucellasecreted protein on the interaction of GPR126 protein and the pathogenic mechanism of Brucellain trophoblast cells.

The purpose of this study is to establish efficient and rapid detection systems for brucellosis that does not rely on professional equipment and personnel, and finished rapid detection of clinical samples within 40 minutes. First, the genomes of different Brucellaspecies were compared, and the core consensus sequence of Brucella was found. The positive plasmid was constructed using the BrucellaS2 strain gene SEQ NO.6 (CN 105018489A) as a template for subsequent experiments; CRISPR/AsCas12a and LwaCas13a proteins were expressed by prokaryotic expression methods; the recombinase-mediated isothermal nucleic acid amplification technology (RAA) and in vitro expression technology combined with the AsCas12a/LwaCas13a DETECTOR system were used to optimize the efficient detection system and the detection was determined by the infinite dilution method. The designed RAA primers can effectively amplify DNA fragments in samples. Optimal DETECTOR systems obtained 200 nmol·L^-1 AsCas12a protein, 200 nmol·L^-1 gRNA; also needs 300 nmol·L^-1 LwaCas13a and 400nM crRNA. The optimal reaction time of the detection system of RAA-AsCas12a and RAA-LwaCas13a was 35 min, the optimal detection time of the lateral flow strip was 20 min, the minimum detection concentration of bacteria was 10 Copies ·μL^-1, and it had good detection specificity. The detection test of clinical samples showed both methods had good repeatability and detection accuracy, but LwaCas13a had higher precision, but there was no significant difference between the two methods. This study established two RAA-CRISPR/Cas Brucellanucleic acid rapid detection systems, which can be well applied in clinical, and the RAA-LwaCas13a detection method has higher accuracy.

In recent years, bacterial drug resistance has become an increasingly significant issue threatening animal husbandry and human health. However, the regulatory mechanisms of bacterial drug resistance are extremely complicated. This study aims to analyze the correlations between virulence genes, drug resistance and biofilm formation ability of diarrheagenic Escherichia coli(DEC). Virulence genes, phylogenetic grouping, drug sensitivity testing, biofilm formation ability assay and hemolysis tests were carried out on 76 strains of E. coli from young livestock with diarrhea in Xinjiang. The results showed that 76 strains of DEC included ETEC (n=28), STEC (n=6), EPEC (n=5), ETEC/STEC (n=23), ETEC/EPEC (n=1), ETEC/EPEC/STEC (n=13), mainly belonging to group B1 (40.8%) and group A (28.9%). The resistance rates to amoxicillin, ampicillin, tetracycline and cefotaxime were 53.9%-80.3%, to streptomycin, florfenicol, sulfamethoxazole, ceftriaxone, ceftazidime, gentamicin and ciprofloxacin were 21.1%-47.4%, and to levofloxacin, ampicillin sulbactam, amikacin, fosfomycin and polymyxin B were 1.3%-15.8%. About 73.7% of the isolates had strong biofilm formation ability, 17.1% had moderate biofilm formation ability, 7.9% had weak biofilm formation ability, and 1.3% had no biofilm formation ability. Fourteen strains showed β-hemolysis. Strong biofilm-forming strains were more common among DEC strains carrying STb, stx1, hly and eae (P＜0.001). The biofilm formation ability was related to hemolysis (P＜0.05), not related to multidrug resistance (P>0.05), significantly related to ceftazidime resistance (P＜0.001), and related to ampicillin-sulbactam (P=0.027 9, P＜0.05). The research results indicated that the types of DEC in young livestock with diarrhea in Xinjiang were complex, and most strains had the biofilm formation ability. The biofilm formation ability was related to virulence genes and hemolysis, and the related to resistance to cefotaxime and ampicillin sulbactam. The biofilm formation ability might be associated with the failure of antibiotic treatment and persistent infection. This study provides a reference for the prevention and treatment of diarrhea in young livestock.

This study aims to explore the prevalence and critical control points of Salmonella within the pork production chain, encompassing pig farms, slaughterhouses, and pork markets, through Salmonella isolation and molecular typing. Samples from pig farm, slaughterhouse and pork markets with interconnected were collected for Salmonella isolating and serotyping. Pulsed field gel electrophoresis (PFGE) methods were used for molecular typing of Salmonella isolates to analyze critical control points of contamination. The effect of increase washing times on Salmonella depletion was evaluated by comparing the changes of Salmonella isolation. The results showed that 417 Salmonella strains were isolated from 950 different types of samples with a total isolation rate of 43.9%. Among of them, 80 fecal samples from farm, 470 pig carcasses and environmental wipe samples from slaughterhouse and 400 pork samples from supermarkets showed isolation rate of 28.8%, 34.5% and 56.8%, respectively. In different slaughtering stages, samples after splitting showed the highest separation rate of 51.0% while the separation rate of the samples after chilling was significantly reduced to 23.0% (P < 0.05). Nine different serotypes were identified from the Salmonella isolates, and the predominant serovars were Salmonella Derby (39.6%), Salmonella Typhimurium (38.1%), Salmonella London (10.3%) and Salmonella Rissen (8.4%). There were 32 pulsotypes were identified in 67 Salmonella Derby isolates by PFGE analysis, of which the isolates from slaughterhouse samples contain the most widely pulsotypes distribution. The isolates from the same sampling visiting appeared in the same pulsotypes, and there were pulsotypes contains isolates from farms, slaughterhouse, and markets with similarity more than 95%, indicating the characteristic of spreading along the production chain. Two times more washing operations for 1 min were added after splitting, which significantly reduced the average isolation rate of Salmonella from 41.7% to 20.0% (P < 0.01) in the subsequent slaughtering process. These results indicate that Salmonella can be spread along the pork production chain via cross-contamination at critical control points such as splitting, offering vital insights and guidance for the prevention and control of Salmonella in pork.

This study aims to obtain soluble and highly immunogenic mutants of Clostridium septicum alpha toxin. Site mutations in the alpha toxin protein were predicted using the MPEPE prediction tool. Expression vectors were constructed and expressed in E. coli for validation. The solubility, toxicity, and immunogenicity in mice of various rCsa mutants were compared. The results showed that the rCsa mutants were correctly expressed in E. coli, with all bands appearing at approximately 47 ku. The soluble expression levels of rCsaS398D, rCsaK405M, and rCsaN397D accounted for 9.5%, 9.4%, and 10.5% of total expressed proteins in the bacterial lysate supernatant, respectively. The ratios of protein amounts in the supernatant to those in the pellet were 2.5, 2.6, and 2.9, respectively. rCsaN397D showed the highest soluble expression level among the three rCsa mutants, and it was also superior to the 40% solubility of rCsa observed in previous experiments. Mouse toxicity tests indicated that none of the three rCsa mutants lost their toxicity. Vero cell toxicity tests showed that concentrations as low as 0.0016 μg·mL^-1 of each rCsa mutant could induce significant cytopathic effects (CPE). ELISA detection of IgG antibody levels in mouse serum 21 days after the second immunization revealed that the S/N values of the rCsaN397D group serum were significantly higher than those of the rCsaS398D and rCsaK405M groups (P < 0.01). Neutralizing titers of mouse serum from the second immunization tested on Vero cells showed that the neutralizing titer of rCsaN397D was 6400 MLD·mL^-1, which was significantly higher than those of the rCsaS398D and rCsaK405M groups (P < 0.0001), and also higher than the neutralizing titer of unmutated rCsa from previous experiments. In summary, rCsaN397D outperformed the other two rCsa mutants in terms of solubility and immunogenicity, and its immunogenicity was better than that of the unmutated rCsa. Therefore, rCsaN397D can be considered a candidate antigen for a genetically engineered subunit vaccine against Clostridium septicum.

This study aimed to investigate the types of Clostridium perfringens isolated from bovine sources in 10 dairy farms in Hebei and Henan, China, and to investigate the information of five isolates by whole gene sequencing. The isolates were identified using PCR and sequencing techniques from 89 bovine disease cases suspected of having C. perfringens disease collected from large-scale dairy farms nationwide. Combined with the genomes of 17 strains of bovine C. perfringens obtained from NCBI public databases, bioinformatics techniques were used to analyze the isolates for pangenomic, phylogenetic, virulence, and drug-resistance genes. The whole genome sequencing analysis showed that the genome size of the isolates ranged from 2.98 to 3.19 MB, with an average of 60 tRNAs and 2 873 coding regions of genes (CDS) per genome. MLST (multilocus sequence typing) showed that the isolates in this study belonged to a different type from the 17 C. perfringens strains of bovine origin downloaded from NCBI, with the exception of PAN2309B1CP belonged to ST777 type, and the remaining 4 strains belonged to ST72 type. A total of 13 drug resistance genes and 30 virulence factors were detected in 22 strains of C. perfringens of bovine origin, among which the tetracycline resistance gene, tetA(P), had the highest carrier rate of 86.3%. Analysis of the pan-genomic results showed that the 5 isolates in this study contained a total of 7 927 genes, of which the number of core genes was 1647 (21.0%). The analysis of the core genome and plc gene evolution tree showed that PAN2305I2CP, PAN2304V2CP, PAN2404F3CP, and PAN2404F4CP were all in the same branch, and PAN2309B1CP was in another branch. This study is the first case of isolation, identification and bioinformatics analysis of Clostridium perfringens from dairy cows, which further enriches the domestic database of epidemic strains of Clostridium perfringens from dairy cows, as well as its genomic structure, composition and evolutionary information. It also provides reference value for the prevention and treatment of C. perfringens disease and further genomic research.

To investigate the impact of porcine pseudorabies virus (PRV) infection on gene expression in the brain tissue of Jiangkou radish pigs, 12 two-month-old piglets were randomly divided into an infected group and a control group. The infected group was intramuscularly inoculated with 1 mL of the PRV-HLJ strain (with a titer of 1×10⁴ TCID₅₀ ·100 μL^-1), while the control group received an equal volume of PBS. Seven days post-infection, brain tissues were collected for histopathological, immunohistochemical, and transcriptomic sequencing analyses. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were conducted to explore the functions and pathways of differentially expressed genes, and quantitative real-time PCR (qRT-PCR) was used to validate the expression trends of key genes. The results showed a significant increase in the number of oligodendrocytes in the brain tissue of the infected group, along with clear PRV-positive expression. Transcriptomic sequencing identified a total of 269 differentially expressed genes, including 149 upregulated genes and 120 downregulated genes. GO analysis revealed that these genes were primarily involved in biological processes such as signal transduction, transmembrane transport, apoptosis, and neuroactive ligand-receptor interactions. KEGG analysis indicated that the differentially expressed genes were significantly enriched in pathways related to neuroactive ligand-receptor interactions, ferroptosis, and the IL-17 signaling pathway. qRT-PCR results demonstrated that in the neuroactive ligand-receptor interaction pathway, the genes LPAR3, GZMA, P2RY2, and NMB were downregulated, while CNR1, CALCR, and NTSR1 were upregulated. In the ferroptosis pathway, SLC7A11 and SLC40A1 were upregulated, while TF was downregulated. In the IL-17 signaling pathway, FOS, FOSB, and MAPK15 were all downregulated. These findings of this study provide new molecular evidence for further research into the pathogenic mechanisms of PRV and offer important data references for the prevention, control, and treatment strategies of PRV.

Previous genome-wide CRISPR screening in human hepatocellular carcinoma cells identified human interferon-induced transmembrane protein 3 (IFITM3) as a critical host factor regulating porcine epidemic diarrhea virus (PEDV) infection. However, its functional role in porcine-derived cells remains unexplored. Here, we generated a porcine IPEC-J2 cell line stablely overexpressing hIFITM3 (IPEC-J2^hIFITM3) and systematically evaluated its impact on viral proliferation. The recombinant lentiviral vector encoding hIFITM3 was co-transfected with packaging plasmids pMD2.G/psPAX2 into HEK293T cells to produce replication-incompetent lentiviral particles. Following transduction of IPEC-J2 cells and puromycin selection, monoclonal cells were established via limiting dilution. Robust hIFITM3 overexpression was confirmed by Western blot, indirect immunofluorescence assay (IFA), and RT-qPCR. CCK-8 assays verified no significant cytotoxicity associated with hIFITM3 expression. Functional validation demonstrated that IPEC-J2^hIFITM3 cells potently suppressed vesicular stomatitis virus (VSV-GFP) replication, consistent with IFITM3's canonical antiviral function. Strikingly, challenge with PEDV DR13-GFP revealed a paradoxical pro-viral effect: IPEC-J2^hIFITM3 cells exhibited a 1 000-fold increase in viral titer (TCID₅₀ ·mL^-1) compared to wild-type cells at 24 hpi. This study establishes a porcine intestinal epithelial model stably expressing hIFITM3, which unexpectedly potentiates PEDV replication. These findings not only highlight the species-specific functional divergence of IFITM3 but also provide a unique platform for investigating PEDV-host interactions, optimizing viral cultivation, and developing targeted antiviral therapeutics.

The aim of this study was to explore the effect of porcine epidemic diarrhea virus (PEDV) infected piglets on the immune organs, eight healthy 5-day-old piglets were randomly divided into control group (n=4) and PEDV-infected group (n=4). The clinical symptoms of piglets infected with PEDV were observed, and the thymus, spleen, mesenteric lymph nodes, inferior iliac lymph nodes, inguinal lymph nodes, anterior shoulder lymph nodes, mandibular lymph nodes and hilar lymph nodes were collected after 2 days of viral infection. Hematoxylin-eosin (HE) staining was used to examine the pathological changes of the immune tissues and organs. Immunohistochemistry (IHC), Western blot and RT-qPCR were used to detect the distribution pattern of PEDV, PEDV-N mRNA and protein levels, and transcription levels of tumor necrosis factor α (TNF-α), interferon β (IFN-β), interleukin 1 (IL-1) and interleukin 6 (IL-6) in the immune system. (Result) Compared with the control group, the infected piglets exhibited symptoms of watery diarrhea beginning at 1 day postinfection (dpi), and the lymph nodes were congested with different degrees at 2 dpi. The increased red pulp, the rupture of cells in splenic sinus, and the indistinct boundary between splenic cord and splenic sinus were observed in the infected spleen via HE staining. Trabecular hyperplasia was detected in mesenteric lymph nodes. Furthermore, the lymph nodes became unclear in structure and decreased in number. IHC and RT-qPCR results showed that PEDV was distributed in multiple immune organs, and the viral load was the highest in mesenteric lymph nodes. Unexpectedly, PEDV-N protein was not detected in all immune tissues using Western blot. In addition, it was found that the transcription levels of TNF-α, IFN-β, and IL-1 increased in the infected spleen and mesenteric lymph nodes. This study demonstrated that PEDV infection destroyed the structure of immune organs and had a significant effect on the mRNA levels of TNF-α, IFN-β, and IL-1, and provided important information in understanding the pathogenic mechanism of PEDV.

This study aimed to understand the drug susceptibility, the carriage of resistance genes, and the correlation between resistance phenotypes and genotypes of Escherichia coli (E. coli) from geese in Guangdong. We conducted antimicrobial susceptibility testing on 387 strains of E. coli isolated and identified from 2015 to 2023 using the K-B disk diffusion method for 25 drugs and detected 19 resistance genes using PCR. The correlation between resistance phenotypes and genotypes of the isolated strains was statistically analyzed. The results showed that the sensitivity rate of the isolated strains to polymyxin B and meropenem was the highest, with a resistance rate of only 1.29% and 1.03%, respectively; the resistance rate to someβ-lactams (penicillin, amoxicillin, ampicillin, cefradine, cefazolin) and tetracyclines (tetracycline, doxycycline) reached over 90%; the resistance rate to most other drugs was between 40% and 70%. The results of multiple drug resistance statistics showed that the resistance spectrum of the isolated strains reached 314 types, with 99.22% of the isolated strains exhibiting resistance to two or more drugs. The detection results of resistance genes showed that among the 19 resistance genes, the detection rates of bla_CTX-M, aadA1, aph(3′)-1, qnrS, floR, tetA, sul2 were relatively high, at 60.72%, 70.03%, 86.30%, 70.54%, 73.13%, 89.66%, and 73.64%, respectively, while the detection rate of mcr-1 was the lowest, at 5.68%. The results of the correlation analysis indicate that the presence of resistance genes bla_CTX-M, bla_NDM, bla_TEM and bla_OXA has an extremely significant or significant correlation with the production of amoxicillin/clavulanic acid (P < 0.01); the carriage of the resistance gene aadA1 has an extremely significant correlation with the production of neomycin, streptomycin, spectinomycin, and apramycin-resistant strains (P < 0.01); the carriage of the resistance gene aph(3')-1 has an extremely significant correlation with the production of neomycin, streptomycin, spectinomycin, and apramycin-resistant strains (P < 0.01). This study provides a reference for the clinical medication program for the prevention and treatment of E. colidisease in geese in Guangdong.

The aim of this study was to investigate the antibiotic resistance of Escherichia coli isolated from Tibetan pigs in Xizang. A total of 180 fecal samples were collected from Tibetan pigs in Shannan, Changdu, and Linzhi in Xizang. Bacterial strains were isolated and identified using bacterial culture, Gram staining, and 16S rRNA gene amplification. Antibiotic resistance was assessed using the K-B disk diffusion method, while virulence genes and resistance genes carried by the strain were detected through PCR amplification. The results showed that, a total of 42 diarrheagenic E. coli strains were isolated from 180 fecal samples, with the overall isolation rates of Enteroaggregative Escherichia coli (EAEC) and atypical Enteropathogenic Escherichia coli (EPEC) being 25.36% and 5.07%, respectively. Antibiotic resistance analysis showed that EAEC and atypical EPEC exhibited imipenem resistance rates of 97.14% and 100%, respectively. The most common resistance phenotypes for EAEC and atypical EPEC were TE+DX+MI+AM+IPM+SIZ and TE+DX+AM+IPM+SIZ, respectively. All isolates carried the β-lactamase bla_TEM gene, with a detection rate of 100.00%. In summary, diarrheagenic E. coli isolated from Tibetan pigs in Xizang exhibited high prevalence and multidrug resistance, with diverse resistance phenotypes and varying levels of resistance gene detection. EAEC was the predominant strain. This study provides valuable data for the prevention and control of diarrheagenic E. coli in Tibetan pigs in Xizang, contributing to the improvement of public health and the optimization of clinical practice.

This study aimed to establish a density stress model with a stocking density of 7 goslings per square meter. The effects of Astragalus, Epimedium, and Fructus Ligustri Lucidi (AEF) extract and Lactobacillus plantarum (LP) on body weight, immune function, antioxidant capacity, meat quality, intestinal morphology, and intestinal barrier integrity of goslings were investigated. The same batch of 42 one-month-old Ma′gang goslings were randomly divided into six groups: control group (c group), AEF treatment group (aef group), AEF and LP treatment group (aef+lp group), density stress group (s group), AEF and density stress group (s+aef group), and AEF, LP, density stress group (s+aef+lp group). The experiment lasted for 30 days. Goslings in the aef and s+aef groups were fed with 0.1 g ·mL^-1 AEF water solvent, goslings in the aef+lp and s+aef+lp groups were fed with 0.1 g ·mL^-1 AEF and 0.01 g ·mL^-1 LP water solvents once daily. The results indicated that supplementation with AEF only and supplemented with AEF and LP together alleviated stress-induced weight loss and reduced serum level of CORT and decreased the expression of inflammatory factors. Furthermore, addition of AEF and LP significantly enhanced relative mRNA expression of the tight junctional proteins Occludin in the jejunum. Intestinal morphology results showed that villi in the duodenum and jejunum of goslings were shorter and sparser under stress, and the villi height to crypt depth ratio (Vh/Cd) decreased significantly as well. Treatment with AEF and AEF+LP increased villus height and improved Vh/Cd in the small intestine. In summary, AEF and AEF+LP adding to the water enhanced gosling weight, immune performance, antioxidant capacity, and improved intestinal health under stress. These findings provide new insights for enhancing productivity and preventing stress in goslings.

This study aimed to analyze the pyroptosis of endometrial epithelial cells induced by lipopolysaccharide (LPS) in dairy cows. Endometrial epithelial cells of dairy cows were treated with 0.1 μmol·L^-1 LPS. The mRNA transcription levels of related inflammatory factors (IL-1β, Nod-1, TNF-α, IL-18 and IL-6) were detected by real-time fluorescence quantitative PCR (qPCR). The protein levels of related inflammatory factors (IL-1β, IL-6 and TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). The morphological changes of endometrial epithelial cells of dairy cows were further observed by scanning electron microscopy. The expression levels of pyroptosis-related proteins (NLRP3, IL-18, IL-1β, GSDMD, Cleaved caspase-1 and Cleaved GSDMD) were detected by Western blot. The results showed that the mRNA transcription levels of inflammatory factors (IL-1β, Nod-1, TNF-α, IL-18 and IL-6) in endometrial epithelial cells of dairy cows increased after LPS stimulation for 12 h. Cell swelling and rupture; the expression levels of inflammatory factors IL-1β, IL-6 and TNF-α were increased. The expression of pyroptosis and inflammatory-related proteins (NLRP3, IL-18, IL-1β, GSDMD, Cleaved caspase-1 and Cleaved GSDMD) was significantly enhanced (P < 0.05). LPS stimulation can induce pyroptosis in bovine endometrial epithelial cells.

This study aimed to evaluate the feasibility of placenta-derived phospholipids as carriers for antibacterial drugs and explore their application in the treatment of endometritis. An in vitroinflammatory model of bovine endometrial epithelial cells and an in vivomodel of endometritis in mice were established using Escherichia coli (E. coli), and the therapeutic effect of gentamicin-placenta lipid complex (L-GEN) was evaluated. Results were as follows: Gentamicin liposomes (L-GEN) were prepared by the film dispersion method; the L-GEN suspension was milky white with a bluish tinge and good fluidity. The particle size of L-GEN was measured by a nanoparticle size and zeta potential analyzer to be 293.7 nm±2.4 nm, with a polydispersity index of 0.21±0.02. No significant change in the particle size of L-GEN was observed within 30 days, indicating the stability of the liposomes. Transcriptome sequencing analysis showed that L-GEN had a smaller impact on bovine endometrial cells compared to the control group. KEGG pathway enrichment analysis revealed that L-GEN mainly enriched in the PPAR signaling pathway, affecting cellular lipid metabolism, reducing cell damage and inflammation, and enhancing antibacterial activity. In vivo experiments found that L-GEN could alleviate the congestion and swelling of the mouse uterus tissue and inflammatory cell infiltration after E. coli perfusion, and the expression levels of inflammatory-related genes IL-2, IL-6, IL-1β, TNF-α, and TLR2 decreased (P≤ 0.05), thereby alleviating endometrial damage and inflammatory response in mice. Through the endometritis cell model and the in vivomodel of endometritis in mice, it was found that the placenta-derived lipid drug delivery complex can effectively alleviate endometrial inflammatory responses and enhance therapeutic effects, providing a reference for the subsequent use of tissue-derived liposomes to carry antibiotics for the treatment of endometritis.

This study aimed to explore the effects of light disturbance and biological clock on osteoarthritis (OA) based on the relationship between the length of sunshine duration (SD) and osteoarthritis (OA) in dairy farms in different regions. By investigating the incidence of bone and joint diseases of lactation cows and local monthly SD in 11 regions across the country from July 2018 to June 2021, the correlation between its incidence and SD was analyzed through Pearson. At the same time, 64 SD male rats were randomly divided into N group (n=32) and OA group (n=32). The OA group used the method of anterior cruciate ligament cleavage (ACLT) to establish an early OA model, and N group performed sham surgery. Each group is divided into two subgroups, and two light-dark period regulation modes (normal light LD and disordered light illumination Dis-LD) are established, namely N+LD, N+Dis-LD, OA+LD and OA+Dis-LD groups, with 16 individual groups. The test cycle was 6 weeks, and the weight changes in rats were recorded, the levels of inflammatory factors and cartilage degradation markers were detected in rat serum, and the articular cartilage was stained with frairo O-solid green and OARSI scores were used to detect the levels of proteins related to cartilage metabolism in rats, and the effects of key biological clock proteins in the heart, liver, spleen, lung and kidney of OA rats were detected. Among the 11 regions surveyed, except for Yinchuan, the other 10 regions were positively correlated with SD, with 6 regions moderately positive correlation (0.4 < R² < 0.6) and 4 regions weakly correlated (0.2 < R² < 0.4). The results of rat experimental research showed that light disorders can significantly slow down the weight gain in rats, increase the content of proinflammatory factors (IL-1β, TNF-α, IL-6, iNOS) and cartilage degradation markers (COMP and CTX-Ⅱ) in rats, promote the catabolism of articular cartilage matrix (MMP-3, MMP-13, ADAMTS-4) in rats in OA group and reduce the synthesis of cartilage collagen fibers (COL2A1). The key biological clock proteins (BMAL1, NR1D1, CRY1 and PER3) in the heart, liver, spleen, lung and kidney of OA rats showed varying degrees. Epidemiological investigations revealed that the incidence of bone and joint diseases in dairy cows was significantly positively correlated with SD. Rat experiments showed that light disorders lead to abnormal biological clocks, promote pathological damage to the cartilage and accelerate the development of OA. This experiment revealed the significant impact of light changes on bone and joint diseases in mammals, and provided a reference for dairy farms to develop appropriate artificial light cycle plans to reduce the incidence of bone and joint diseases in dairy cows in the future.

This study aimed to investigate the inhibitory effects of Forsythiaside A (FTA) on the replication of H9N2 avian influenza virus (H9N2 AIV) in pulmonary microvascular endothelial cells (PMVECs) and explore its underlying mechanisms. PMVECs infected with H9N2 AIV were treated with varying concentrations of FTA. Cytotoxicity was assessed using the MTT assay, and viral replication was analyzed via RT-qPCR. Protein expression levels of TLR3, TRIF, and NF-κB were evaluated using Western blot, while cytokine secretion (TNF-α, IL-6) and type Ⅰ interferons (IFN-α, IFN-β) were measured through ELISA. The results showed that FTA significantly inhibited the replication of H9N2 AIV (P < 0.05) in a dose-dependent manner. It downregulated the overexpression of TLR3, TRIF, and NF-κB proteins (P < 0.05), suppressed the secretion of pro-inflammatory cytokines (P < 0.05), and enhanced the production of type Ⅰ interferons (P < 0.05). FTA exhibits significant antiviral activity against H9N2 AIV by modulating the TLR3-NF-κB signaling pathway and regulating cytokine expression, highlighting its potential as a novel host-targeted antiviral agent.

This study aims to investigate the occurrence of heat stress and the therapeutic effects and mechanisms of Sheng Mai San (SMS) in treating heat stress-induced lung injury, providing theoretical support for the application of SMS in this context. Forty-eight SD rats were selected to establish heat stress models based on the recovery time of heat stress. Another 60 SD male rats were randomly divided into 6 groups (n=10 per group). Each group was given intragastradized administration 2 hours before heat stress every day, and the Control group and heat stress group (HS) were given normal saline. SMS group was administered at SMS-H (5.04 g·kg^-1), SMS-M (2.52 g·kg^-1) and SMS-L (1.26 g·kg^-1) doses, and N-acetylcysteine (NAC) (150 mg·kg^-1) was set up as a positive control group. Sampling was performed after the end of heat stress on day 7. Biological techniques including HE staining, Masson staining, Western blot, and real-time quantitative PCR were employed to evaluate the effects of SMS on heat stress-induced lung injury and its potential mechanisms. Heat stress caused significant lung injury between 6-12 h post-exposure, with the most severe damage observed at 6 h post-stress (P<0.05). Real-time quantitative PCR, Western blot, and biochemical analyses revealed that SMS significantly reduced the expression levels of inflammatory cytokines TNF-α, IL-1β, IL-10, IL-6, and HSP70, alleviating the abnormal accumulation of ATP (P<0.05). Moreover, SMS notably decreased p-mTOR expression and p62 accumulation, activated LKB1 and AMPK phosphorylation, and increased Beclin1 and LC3 expression levels (P<0.05). Heat stress-induced lung injury was most severe at 6 h post-exposure. SMS demonstrated therapeutic effects by activating the AMPK-mTOR pathway, promoting autophagy, and clearing damaged lung cells, thereby mitigating heat stress-induced lung injury.

This study aims to assess the effectiveness of a Urinary prescription canned food as an adjunctive treatment for feline lower urinary tract disease (FLUTD). A total of twelve healthy cats were randomly assigned to two groups: the Urinary prescription canned food group (referred to as the urinary group) and a control group, with each group consisting of six cats. Throughout the feeding period, daily water intake and emotional states of the cats were monitored. Urinalysis, urine sediment examination, and measurements of magnesium (Mg²⁺), oxalate (C₂O₄^2-), phosphate (PO₄^3-), and calcium (Ca²⁺) concentrations in urine were conducted both prior to and following the feeding regimen. Additionally, metabolomics analysis was performed on urine samples collected post-feeding. The urinary group demonstrated a higher cumulative water intake compared to the control group. Furthermore, there was a 5% increase in the proportion of positive emotional states, particularly "content", in the urinary group relative to the control group. Post-feeding analysis revealed a significant reduction in the urinary sediment of the urinary group, including crystalline substances, casts, and urethral epithelial cell counts (P<0.05), with significant differences when compared to the control group (P<0.05). The concentrations of Mg²⁺, C₂O₄^2-, PO₄^3-, and Ca²⁺ in the urine of the urinary group significantly decreased after feeding (P<0.05), again showing significant differences from the control group (P<0.05). Metabolomics analysis identified a total of 5 767 metabolites, with 381 differentially expressed metabolites that were significantly enriched in the tryptophan metabolism pathway. The findings of this study indicate that the Urinary prescription canned food effectively inhibits the formation of struvite and calcium oxalate uroliths, reduces the presence of urethral epithelial cells and casts in urine, and positively influences the emotional well-being of felines. Moreover, it significantly affects the tryptophan-serotonin metabolic pathway, thereby playing a beneficial role in the regulation of stress and anxiety in cats. This dietary intervention holds potential clinical significance in the adjunctive management of FLUTD.

This study aimed to analyze the serum metabolome changes following laparoscopic hepatic lobectomy in Beagle dogs and explore the underlying mechanisms of these metabolic alterations. Eight healthy Beagle dogs of similar weight were selected to establish a liver lobectomy model. Serum samples were collected preoperatively, and at 7 and 14 days post-surgery. Ultra high performance liquid tandem chromatography quadrupole time of flight mass spectrometry(UHPLC-QTOFMS) was employed to detect changes in serum metabolites during the perioperative period. The results demonstrated that: Principal component analysis (PCA) revealed a clear separation between the 7-day post-surgery group and the preoperative group, indicating significant differences in metabolites between these time points. In contrast, the overlap between the 14-day post-surgery group and the preoperative group suggested a gradual return to preoperative status by day 14. The orthogonal partial least squares discriminant analysis (OPLS-DA) score plot and model validation indicated that the constructed model possessed good explanatory and predictive capabilities without overfitting. Differential metabolites were identified using the variable importance in projection (VIP) score>1 for the first principal component of the OPLS-DA model, with a significance level of P<0.05 by t test. Compared to the preoperative period, 35 differential metabolites were detected at 7 days post-surgery, with 13 down-regulated and 22 up-regulated. Notably, L-phenylalanine was involved in phenylalanine, tyrosine, and tryptophan biosynthesis, as well as phenylalanine metabolism and aminoacyl-tRNA biosynthesis, while 3α, 7α-dihydroxy-5b-cholestane was implicated in primary bile acid biosynthesis. At 14 days post-surgery, 42 differential metabolites were identified, with 19 down-regulated and 23 up-regulated, including 1-methylnicotinamide and nicotinamide, which were involved in niacin and nicotinamide metabolism. Fewer metabolites showed changes at 14 days compared to 7 days after surgery. Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway enrichment results showed that the differential metabolites were significantly enriched in the metabolic pathways of phenylalanine, tyrosine, and tryptophan biosynthesis, and primary bile acid biosynthesis at 7 d postoperatively as compared to preoperatively; Laparoscopic hepatic lobectomy resulted in significant alterations in several metabolites. The biosynthesis of phenylalanine, tyrosine, tryptophan, and primary bile acids played important roles in the metabolic mechanisms underlying liver injury, while the metabolic pathway of nicotinate and nicotinamide contributed to the liver recovery process.

The aim of this study was to investigate the effects of maternal supplementation with Lactiplantibacillus plantarumX86 during pregnancy and lactation on early offspring development and gut microbiota. Sixteen gregnant SD rats were selected and divided into control group (n=8) and L. plantarumX86 group (n=8). Rats in the L. plantarumX86 group were given Lactiplantibacillus plantarumX86 gavage from late pregnancy until early lactation. The impact on offspring growth, intestinal barrier function, and gut bacterial community structure were evaluated by various indexes. Results showed that L. plantarumX86 supplementation increased body weight at 7 and 14 days of age, and elevated heart and liver coefficients of the offspring; However, it reduced the white blood cell counts and hemoglobin content. Additionally, L. plantarum X86 supplementation downregulated IL-6 mRNA expression and upregulated Occludin mRNA expression in the intestines of rat offspring. 16S rRNA sequencing analysis revealed that the maternal supplementation with L. plantarumX86 during late pregnancy and lactation significantly altered the gut microbiota in rat offspring, increasing the relative abundance of Lactobacillus and Romboutsia while reducing that of Escherichia-Shigella, Staphylococcus, and Akkermansia. A broad reduction in KEGG metabolic pathway abundance was also observed. In conclusion, consumption of L. plantarumX86 during late pregnant and early lactation promotes offspring growth and development, and exerts beneficial effects on the gut barrier function and bacterial microbiota composition.

Bovine viral diarrhea virus (BVDV) is one of the major pathogens responsible for significant economic losses in cattle herds, particularly affecting the health and development of calves. This study aimed to establish a universal SYBR Green I-based real-time quantitative PCR (qPCR) assay for BVDV detection and to investigate the epidemiology, genotypes, and subtypes of BVDV in samples collected from free-range cattle herds in selected regions of China. A universal BVDV SYBR Green I qPCR assay was developed based on the 5′ untranslated region (5′UTR) sequences of various BVDV genotypes obtained from GenBank. Using this method, a total of 712 serum samples from free-range cattle, as well as tissue samples from calves exhibiting symptoms such as coughing, fever, and diarrhea collected between June 2022 and December 2024, were tested and subjected to phylogenetic analysis. Additionally, virus isolation, indirect immunofluorescence assay (IFA), and hematoxylin-eosin (HE) staining were performed on calf samples. The established BVDV SYBR Green I qPCR assay demonstrated high sensitivity, strong specificity, good linearity, and excellent reproducibility. The positive detection rate in serum samples was 19.10%, and the BVDV-24GS strain was successfully isolated. Phylogenetic analysis of the 5′UTR gene sequences revealed that the BVDV strains belonged to subtypes 1c, 1m, 1o, 1i, 1q, 1b, and 2. These findings indicate the successful development and application of a universal BVDV SYBR Green I qPCR assay, and the presence of varying degrees of BVDV infection among free-range cattle in parts of China. The identification of viral genotypes enriches the molecular epidemiological data of BVDV and provides a scientific basis for effective disease control, vaccine development, and optimization of immunization strategies.