Single-cell RNA sequencing (scRNA-seq) is revolutionizing poultry biology research by resolving cellular heterogeneity and dynamic gene expression at single-cell resolution. This review systematically summarizes the advances in scRNA-seq applications for poultry: the technology enables cell-by-cell analysis, offering novel strategies to enhance egg production efficiency, improve meat quality, and boost disease resistance. However, current research faces bottlenecks including incomplete cellular atlas coverage, scarce functional validation systems, and high agricultural conversion costs. Future efforts should prioritize developing low-cost sequencing, integrating multi-omics data (e.g., scATAC-seq and metabolomics), and implementing targeted validation via CRISPR/Cas9-mediated avian models. These approaches will drive intelligent design of precision breeding and reproductive regulation. This review provides theoretical foundations and technical prospects for deciphering cell fate determination mechanisms in poultry and their practical applications.

Gene modification refers to the precise editing of an organism's DNA sequence through human intervention in order to modify its genomic structure or function in a targeted manner. In recent years, with the development of molecular biology and genetic engineering technology, gene modification technology (e.g., transgenic technology and gene editing technology) combined with assisted reproduction technology has shown great potential in the field of disease-resistant breeding in cattle and small ruminants. This technology can not only precisely improve disease resistance traits and enhance the health and production performance of livestock but also make up for the shortcomings of traditional selection and breeding methods in terms of low efficiency and long cycle time. However, compared with disease resistance breeding in pigs and chickens, cattle and small ruminants still have more room for progress in this field. Currently, optimizing the genome of cattle and Small ruminants through gene modification technology is becoming a cutting-edge direction to break through the bottleneck of traditional prevention and control. The review describe the current development of gene modification technology from the aspects of animal transgenics and gene editing technology and briefly summarize the status of the application of this technology in disease resistance breeding in cattle and small ruminants, which will provide some references for future research on disease resistance breeding in cattle and small ruminants.

Fault bars are structural deformities formed during feather growth due to disrupted keratin synthesis, leading to weakened or fractured barbules, loss of vane integrity, and consequently impaired flight capacity and survival adaptability in birds. Studies on wild birds have shown that the occurrence of feather fault bars may be associated with multiple factors, such as nutritional deficiencies, environmental stressors, disease infections, and genetic predispositions. Although the exact etiology remains unclear, it has been widely accepted that physiological or psychological stressors during feather growth can induce fault bars formation. In modern farming systems, poultry are frequently exposed to multiple stressors in the high-density environment, compromising their welfare status. However, evaluating stress level in poultry remains a persistent challenge in practical welfare assessments. Recently, fault bars have gained attention as a non-invasive, practical, and promising biomarker for poultry welfare evaluation. This review synthesizes the influencing factors and formation mechanisms of fault bars, as well as their current applications in poultry welfare assessment, aiming to provide insights for improving poultry welfare evaluation systems.

Extracellular vesicles (EVs) belong to the membrane structure derived from cell secretion, which are the carriers of information material transfer between cells, playing crucial roles in mammalian reproduction process. Mammalian reproduction is a rather complex process, regulated by multiple factors. With the continuous deepening of the research on EVs in animal reproduction, their application in pig reproduction has gradually demonstrated great potential. EVs mainly achieve balance in the reproductive process by participating in a variety of physiological functions, ensuring the smooth progress of gametogenesis, fertilization, blastocyst implantation, embryo implantation, and delivery in the process of pig reproduction. This article reviews the biological characteristics of EVs and their roles in various porcine reproduction stages, aiming to provide some theoretical reference for deep research and application of EVs in porcine reproduction.

Second litter syndrome (SLS) in sows is widespread worldwide. Sows with SLS have a lower litter size of the second parity than the first parity, the sows may have a lower estrus rate and a high culling rate, which waste the reproductive performance of low-parity sows, and the lifelong reproductive performance of sows with SLS is relatively reduced, while the breeding cost increases accordingly. In this paper, the causes and mechanisms of the formation of SLS are systematically reviewed, and targeted prevention and treatment measures are also put forward, so as to provide theoretical basis and technical support for the stable development of sow reproductive performance, as well as the stable and healthy development of pig industry. Exploring the occurrence mechanism of SLS in different pig farms supplies scientific research approach to deal with SLS, and to avoid the negative effect of SLS on reproductive performance in the subsequent parity. Precise feeding, nutritional regulation, and drugs research and development in reproductive regulation and their reasonable utilization will be the key points to solve the second litter syndrome.

Flavonoids are a class of active ingredients derived from natural plants, which has excellent pharmacological effects such as antiox idant, anti-inflammatory, reducing oxidative stress, improving the structure of intestinal flora, protecting intestinal mucosa, regulating immunity and promoting growth. This paper provides a systematic review of the role of flavonoids in animal intestinal health and barrier protection, briefly describes their unique physicochemical properties, summarises their digestive, absorptive and metabolic pathways in living organisms, focuses on the interactions between flavonoids and gut microorganisms, and provides insight into how flavonoids can influence the physiological function of the intestinal barrier by mediating the intestinal microbiota and generating key metabolites, such as short-chain fatty acids, bile acids and their receptors and tryptophan and other key metabolites to influence the physiological function of the intestinal barrier. The aim is to lay a solid theoretical foundation for the development and utilization of plant-derived flavonoids in animal feeding and production practices, especially in the study of intestinal health in young animals.

Catechins are phenolic active substances extracted from plants. Because of their significant antioxidant, anti-inflammatory, antibacterial and anti-tumor biological properties, they perform excellent in protecting intestinal health and homeostasis. A stable and healthy intestinal environment is crucial for the growth, development, and overall health of animals. In recent years, a large number of clinical studies at home and abroad have reported the mechanism of catechins in alleviating intestinal injury, including regulating intercellular tight junctions, stimulating inflammation and oxidative stress-related pathways to regulate immune function, and regulating the balance of intestinal flora to stabilize intestinal homeostasis. However, there are few reports on the application of catechins in animal production. This paper reviews the biological characteristics of catechins, the effect and mechanism of repairing intestinal barrier, and looks forward to the research progress of the application of catechins in animal production, so as to provide reference for the rational use of catechins in livestock farming.

The gastrointestinal tract is the largest microecosystem in animals, where complex microbial communities constitute the animal’s intestinal microbial barrier. 5-Hydroxytryptamine (5-HT), commonly known as serotonin, is a neurotransmitter in animals, with more than 90% produced by the intestine, which exhibits various biological functions including the regulation of intestinal peristalsis, secretion processes, immune responses, and maintenance of intestinal barrier integrity.As a pivotal signaling molecule in the gut-brain axis, 5-HT can transmit information through the nervous, endocrine, and immune systems; thus playing a vital role in maintaining neural and intestinal homeostasis while modulating disease states. Currently, research on 5-HT mainly focuses on fields such as biology, medicine, ethology, and psychology, however, studies pertaining to its implications in livestock and poultry remain scarce. This paper systematically reviews the regulatory mechanisms of 5-HT in neurological abnormal behaviors and intestinal diseases of rodents, livestock, and poultry. It also discusses the regulation of intestinal 5-HT synthesis and microbial balance by adding tryptophan precursors such as rumen-protected 5-hydroxytryptophan (5-HTP), probiotics, or plant extracts (such as Astragalus polysaccharide, natural capsaicin) to alleviate intestinal disorders such as post-weaning diarrhea in piglets and necrotic enteritis in broilers. Under stress scenarios such as heat stress or transport stress, enhancing 5-HT levels via probiotic supplementation or regulating intestinal 5-HT receptor gene expression through colostrum feeding can mitigate stress-induced behavioral abnormalities and immune dysfunctions. These findings aim to provide theoretical support for practical applications in animal husbandry.

Rumen is a unique digestive organ of ruminants，housing a diverse variety of microorganisms that together form a complex and highly efficient micro-ecosystem. The normal functioning of the ruminal microbial fermentation is a crucial prerequisite for ensuring the health and production performance of ruminants. The contamination of mycotoxins in the feeds and forages for ruminants is relatively common，and these toxins primarily affect the rumen and its microbiota once ingested by ruminants，consequently impairing the digestive function and health of the animal. This review summarizes the research on the impact of various mycotoxins on the rumen fermentation and ruminal microbiome of ruminants，and discusses the potential related mechanisms based on relevant studies and evidences. The aim is to provide new insights for the prevention and control of mycotoxin-induced harms to ruminants.

The UL7 gene is highly conserved among members of the Herpesviridae family, and its encoded pUL7 protein plays a critical biological role in the pathogenic regulation of viral infection. This review systematically outlines the structural characteristics of the UL7-encoded protein and comprehensively summarizes the functions of pUL7 in key stages of the viral life cycle, including review systematically summarizes the structural characteristics of UL7-encoded proteins and the replication, transcription, protein expression, virion assembly, and release. Additionally, it explores the interaction mechanisms between pUL7 and other viral or host proteins, aiming to provide valuable insights for further research on the UL7 gene in herpesviruses.

Coronaviruses, infecting diverse species with broad infection spectra, continue to devastate human and animal health. Porcine intestinal coronaviruses, represented by porcine epidemic diarrhea virus (PEDV), pose serious challenges to the global swine industry. The newly identified porcine deltacoronavirus (PDCoV) has been shown to possess zoonotic potential in experimental settings, highlighting the risk of interspecies transmission. As obligate intracellular parasites, viral infection involves complex processes, including attachment, entry, replication, assembly, and release. Among these stages, the interaction between viral envelope spike proteins and specific receptors on host cell membranes not only determines tissue tropism and host range but also constitutes the fundamental molecular basis affecting cross-species transmission. This review primarily focuses on the methodological advances, current research landscapes, and unresolved challenges regarding the functional receptors and host entry factors of these porcine enteric coronaviruses. Through this systematic analysis, we aim to provide a reference for elucidating the infection and pathogenic mechanisms of coronaviruses, particularly for designing receptor-blocking therapeutics and genome-editing-based breeding programs.

DNA/RNA methylation and lysine acetylation modifications are both important epigenetic modifications that play a crucial role in regulating gene expression and protein activity in life processes. DNA/RNA methylation and lysine acetylation modification are also highly promising drug targets. This article will mainly review the research progress on DNA/RNA methylation modification and histone acetylation modification in regulating the development and transformation of Apicomplexan parasites, in order to provide a basis for the screening of new anti-coccidial drug targets.

Avian pathogenic Pasteurella multocida, a widespread Gram-negative short bacillus, causes avian cholera and leads to significant economic losses on the global poultry industry. The long-term reliance on antibiotic treatment for avian cholera has led to an increase in antibiotic-resistant bacteria in poultry. This article compiles data on the prevalence and antibiotic resistance of avian P. multocida from 17 regions domestically and internationally over the past two decades. Statistics indicate that avian P. multocida is prevalent across all continents, particularly in areas with common poultry farming and consumption, where the prevalence rate exceeds 43.50%. Domestically, it is highly prevalent in Jiangsu Province (53.73%) and Guangdong Province (48.97%), with rates over 48%. Genomic analysis of avian Pasteurella multocida in the database revealed a total of 31 sequence types (STs), among which ST159 (115/733,15.69%) and ST128 (82/733,11.19%) exhibited host preferences for chickens (n=109) and ducks (n=65), respectively. Both domestic and international studies have found high resistance rates to commonly used antibiotics, especially kanamycin (99/165, 60%), chloramphenicol (124/230,53.91%), and tetracycline (258/556,46.40%). Genome analysis has detected 29 resistance genes covering eight classes of antibiotics, with sul2 (13.64%) and tet(B) (10.64%) being the most prevalent. In conclusion, there are differences in the prevalence of avian P. multocida between domestic and international regions, and resistance rates to multiple antibiotics are high. Prevention and control measures such as vaccines and bacteriophages can be implemented based on local prevalence characteristics, and the use of antibiotics with high resistance should be reduced to achieve effective treatment and establish green, sustainable farming practices.

This study aimed to analyze the genetic mechanism of key growth traits in Large White pigs through genome-wide association study (GWAS) and provide a theoretical basis for molecular marker-assisted breeding. A total of 5 701 Large White pigs were used in this study. Phenotypic data on age at 100 kg body weight (AGE), backfat thickness at 100 kg body weight (BF), and loin muscle area at 100 kg body weight (LMA) were collected. Combined with genotyping data from the Illumina 50K SNP chip, GWAS was performed using a mixed linear model to identify SNPs and candidate genes associated with pig growth traits. The heritability values of AGE, BF, and LMA were 0.50, 0.45, and 0.43， respectively, all of which were medium to high heritability traits. The genetic correlation coefficient between AGE and BF was -0.13, and the phenotypic correlation coefficient was -0.20; between AGE and LMA, the genetic correlation coefficient was -0.09, and the phenotypic correlation coefficient was -0.31; between BF and LMA, the genetic correlation coefficient was 0.22, and the phenotypic correlation coefficient was 0.06. A total of 13 significant SNPs were identified through GWAS, including 8 significant SNPs for AGE, which were associated with 11 candidate genes including PIK3CG; 3 significant SNPs for BF, which were associated with 6 candidate genes including IFN-ALPHA-15; and 2 significant SNPs for LMA, which were associated with ALDH6A1. This study identified 20 candidate genes related to growth traits in Large White pigs, including PIK3CG, ELOVL2, IFN-ALPHA-15, and ALDH6A1, through GWAS. The results provide an important reference for molecular marker-assisted selection in pig breeding.

Using primary ovine cells as materials, this study aimed to verify the gene editing efficiency of precise repair through homology-directed repair (HDR) with single-stranded oligodeoxynucleotide (ssODN) as the donor mediated by the CRISPR/Cas12i system, and to explore the effects of small molecule drugs on the efficiency of insertions and deletions (Indels) and the knock-in efficiency. In this study, two types of primary ovine cells were used as experimental subjects to verify the gene editing efficiency of the CRISPR/Cas12i system at 25 target gene loci. Then, 3 target sites (SOCS2-gRNA1, SOCS2-gRNA3, and TBXT-gRNA5) in ovine fetal fibroblasts and 5 target sites (MSTN-gRNA2, MSTN-gRNA5, Myf6-gRNA4, Myf6-gRNA5 and MyoG-gRNA4) in ovine hindlimb muscle cells were selected to detect the effects of 5 small molecule drugs (SCR7, Nocodazole, RS-1, Vorinostat, and Entinostat) on the Indel efficiency at each target site. Finally, the effect of the 5 small molecule drugs on the gene knock-in efficiency of precise repair through HDR with ssODN as the donor mediated by the CRISPR/Cas12i system was detected at the MSTN-gRNA2 target site. This study verified that the CRISPR/Cas12i system had high gene editing activity at 25 different target sites in the two types of primary ovine cells. After treatment with small molecule drugs, SCR7 decreased the Indels efficiency at each target site in primary ovine cells, while treatment with RS-1, Entinostat, and Vorinostat increased the Indels efficiency at each site. Nocodazole had inconsistent effects on the two types of cells. After treatment with the 5 small molecule drugs, the knock-in efficiency at the MSTN-gRNA2 site was increased to varying degrees. Among them, the treatment groups with 2 μmol·L^-1 Vorinostat and 4 μmol·L^-1 Entinostat had the highest enhancement effects, with the knock-in efficiencies being 7.67% and 7.73%, respectively. This experiment provides a reference for the application and promotion of the CRISPR/Cas12i gene editing system, offers new ideas for the creation of gene-edited sheep with excellent traits, and provides technical support for accelerating the work of precise molecular breeding.

The study aimed to identify genes associated with superior traits holding significant value for genetic improvement and practical breeding programmes. This study utilized 93 Qira black sheep, 33 Pishan red sheep, and 13 Waghgir sheep. Blood samples were collected from the jugular vein, followed by DNA extraction and genotyping. Genotype data quality control was performed using PLINK software (quality control standards: excluding individuals with detection rates below 90%, SNP detection rates below 95%, minimum allele frequencies below 5%, and SNPs with P<1×10^-6 for Hardy-Weinberg equilibrium). Principal component analysis (PCA) was conducted to construct evolutionary trees, analyze population ancestral components, and assess linkage disequilibrium (LD). Simultaneously, based on genome-wide runs of homozygosity (ROH) analysis and population genetic differentiation index (F_ST) analysis, the top 10% ROH segments were selected as high-frequency regions, and the top 5% loci by value were designated as selected regions. Reference was made to annotated genes in the sheep genome Oar_v4.0 for Gene Ontology (GO) and KEGG pathway analysis. The F_ST and F_ROH results indicated a low level of genetic differentiation among the 3 groups, with a clear genetic background differentiation emerging at K=4. Concurrently, genes including ACVR1, ACVR1C, UPP2, CRY1, and NR4A2 were identified as candidate genes for genetic adaptation in local sheep breeds of southern Xinjiang within arid desert environments. This study revealed the genetic variation characteristics of local sheep breeds in southern Xinjiang through analysis of population genetic structure diversity and selection signals. It identified relevant superior genes from a multidimensional perspective of genetic variation, providing important reference for sheep germplasm resource conservation, new variety development, and enhancement of resource diversity.

This study aimed to reveal the regulating role of CTCF in the differentiation of ovine preadipocytes and its regulatory mechanism. In this study, adipose tissues from different parts of 4 8-month-old Guangling Large-tailed sheep were collected, and the expression level of CTCF was detected by quantitative real-time fluorescent quantitative PCR (qRT-PCR). Primary preadipocytes were isolated from 1 healthy 3-day-old Guangling Large-tailed sheep. CTCF gene was overexpressed or interfered in preadipocytes, with 3 replicates set for each treatment group. The regulatory effect of CTCF gene on the differentiation of sheep preadipocytes was detected using qRT-PCR, Western blotting and Oil Red O staining techniques. On this basis, transcriptomic sequencing analysis on sheep preadipocytes with CTCF overexpression was performed to explore the molecular mechanisms by which CTCF regulates lipid metabolism-related signaling pathways. The results showed that the mRNA expression of CTCF gene in the tail adipose of Guangling Large-tailed sheep was significantly higher than that in the perirenal and subcutaneous adipose tissues (P<0.05). After overexpression of CTCF, the expression level of adipogenic marker genes was significantly increased (P<0.05), and the amount of intracellular lipid droplets was increased. The opposite was true when the CTCF is interfered with. Transcriptome results showed that there were 1 079 differentially expressed genes between two groups. These differentially expressed genes were enriched in several pathways related to fat metabolism, including the sphingolipid signaling pathway, the FoxO signaling pathway, and the AMPK signaling pathway. Candidate genes affecting fat metabolism, such as CCND2, ITGA7, BRCA1, LAMA5, and CCNE2, were screened out. In summary, this study results shows that CTCF can promote the differentiation of sheep preadipocytes, and reveals the related pathways and candidate genes affecting CTCF-mediated fat metabolism.

The purpose of this study was to investigate the growth and development of down feather, feather follicle characteristics and the expression of genes related to feather sac development in growing Sanhua geese and Wanxi White geese. Taking Wanxi White geese and Sanhua Geese as the research objects, the changes of velvet length, thousand velvet weight and velvet diameter of geese at 90 days of age were analyzed. Hematoxylin-eosin staining (HE staining) and trichrome staining (Masson staining) were performed on geese skin and the growth changes of feather follicles at each stage were observed. The expression of Wnt5a, IGF-1, BMP2, and FGF5 genes in white geese skin tissues at different growth stages was studied by fluorescence quantitative PCR. The results showed that: 1) By observing the morphology of down feathers in different parts, it was found that the down feathers of Wanxi White geese were more fluffy and larger than those of Sanhua geese. 2) Observing the microstructure of down feathers, it was found that the nodes of down twigs of Sanhua geese down feathers at 60 days of age were forked nodes, and the nodes of down twigs at 90 days of age were triangular nodes. At 60 days of age, the down twig nodes of Wanxi White geese down feather showed two kinds of triangle and fork nodes; at 90 days of age, the down twig nodes showed triangle nodes and fork nodes at the end. 3) It was found that the down feather traits of different parts of 90-day-old Sanhua geese and Wanxi White geese were significantly better than those of 60-day-old Sanhua geese and Wanxi White geese. 4) By measuring the diameter of different feather follicle, it was found that the secondary feather follicle in different parts of Sanhua geese and Wanxi White geese at 90 days of age were significantly higher than those in the same part of the same breed at 60 days of age.5) The results of fluorescence quantitative PCR showed that the expression of Wnt5a gene was the highest in the skin of Wanxi White geese at 60 days of age, but there was no significant difference in the expression of Wnt5a gene in the skin of 90 day-old Sanhua geese and Wanxi White geese at 90 days of age (P>0.05). The expression levels of BMP2 and IGF-1 genes in the skin of 60-day-old Wanxi White geese were significantly higher than those of 60-day-old Sanhua geese and 90-day-old Wanxi White geese (P<0.01). Among them, the expression levels of BMP2 and IGF-1 genes in the skin of Sanhua geese were the lowest at 90 days of age. There was no significant difference in the expression level of the FGF5 gene in the skin of Sanhua geese and Wanxi White geese at different ages (P>0.05). From the measurement indicators of down feathers, it is found that the older the age, the better the quality of the down feathers. Whether it is the Sanhua geese or the Wanxi White geese, the quality of the down floss on the back is the best. It can be seen from the genetic level that the expression levels of Wnt5a, BMP2 and IGF-1 genes in the dorsal skin of 60- and 90-day-old Wanxi White geese were significantly higher than those of Sanhua geese. In addition, the quality of geese down feathers is closely related to the development of feather follicle, and the development of feather follicle is regulated by multiple genes.

This study aimed to investigate the genetic diversity and inbreeding status of the Huoyan geese (Wulong geese) conservation population using whole-genome resequencing, evaluate the effectiveness of current germplasm conservation strategies, and provide actionable insights for future breeding programs. A total of 40 male Huoyan geese (Wulong geese) were randomly selected from the Shandong Provincial Conservation Farm for whole-genome resequencing. Subsequent analyses included assessments of genetic diversity, population genetic structure, and runs of homozygosity (ROH). The results showed that: a total of 40 slit-eyed geese (Wulong geese) were sequenced with an average depth of 13×, and 13 251 291 SNP loci were identified after quality control. The average polymorphism information content (PIC) of this conservation population was 0.264, the average observed heterozygosity (Ho) was 0.323, and the average expected heterozygosity (He) was 0.327, with an inbreeding coefficient (F) of 0.012. LD decay analysis showed that the maximum R² value was 0.47, with a decay distance of 162 kb. A total of 309 runs of homozygosity (ROH) were detected in the Huoyan Goose (Wulong Goose) conservation population. The inbreeding coefficient (F_ROH) calculated based on ROH was 0.009, indicating a low level of inbreeding within the population. The results of the IBS distance matrix and G matrix showed simi-lar trends, with only a small proportion of individuals exhibiting close kinship. Principal component analysis (PCA) and neighbor-joining (NJ) clustering analysis further confirmed that the conservation population was generally in a dispersed state. Overall, the genetic diversity of the Huoyan goose (Wulong goose) conservation population surveyed in Laiyang was favorable, with a low level of inbreeding, indicating effective conservation efforts.

This study aimed to comparatively analyze the genetic diversity and the genetic basis of small stature between Ningqiang ponies and Jeju ponies. This study compared and analyzed the genetic structure, genetic diversity, and genetic differentiation of Ningqiang ponies and Jeju ponies based on the whole-genome resequencing data from 199 modern domestic horses, focusing on comparing the genetic basis of small stature. Population structure analysis indicated that Ningqiang ponies belong to pony breeds of Southwestern China, while Jeju ponies had ancestral components that are distinct from those of the Southwestern ponies. Nucleotide diversity, inbreeding coefficient, and ROH analysis indicated that Jeju ponies had undergone strong artificial selection and breeding recently, with low genetic diversity, while Ningqiang ponies had high genetic diversity. Furthermore, the genetic distance between Ningqiang ponies and Debao ponies is the closest (F_ST=0.02), while Jeju ponies had a moderate degree of genetic differentiation from 11 domestic horse breeds (F_ST>0.05). Whole-genome selection scanning detected multiple differential candidate regions in the genomes of Ningqiang ponies and Jeju ponies, and identified genes related to growth and development (TBX5, TBX3, MEIS1, CECR2) and immunity (ADA2, AP3B1). In conclusion, Ningqiang ponies and Jeju ponies have different ancestral components and degrees of selection and breeding, with obvious genetic differentiation, and the genetic basis for small stature is also different. These results are of great significance for an in-depth understanding of the genetic basis of small stature traits in ponies and for the protection and utilization of horse breeds in China.

This study aimed to optimize the in vivo embryo cryopreservation method for Rongchang pigs to improve the hatching rate of blastocysts after embryo freezing. The experiment used 50 Rongchang pigs to obtain in vivo embryos through synchronized estrus, artificial insemination, and surgical flushing. The effects of different cryo-carriers (SOPS and Cryotop), different vitrification cryoprotectants (Medium-199, NCSU23 and TL-PVA), lipid centrifugal polarization in pre-cryopreserved morula and zona pellucida (ZP) digestion of vitrified-warmed (V-W) embryos were compared, and the survival rate, continued development ability and blastocyst cell number of V-W embryos were determined to screen the optimal embryo V-W method in Rongchang pig. An average of 9.22 blastocysts per donator were obtained after embryo flushing, that raised to 12.16 blastocysts per donator combining with in vitro culture. The survival and development rate of V-W embryos using Cryotop were significantly higher than those of SOPS (P<0.05). V-W embryos of NCSU23 group gained the highest survival rate (85.91%±3.27%) and development rate (80.61%±10.06%) comparing to Medium-199 and TL-PVA groups. The abilities of embryonic cell division were NCSU23>Medium-199>TL-PVA. This study proofed that NCSU23 was the most suitable cryoprotectant for Rongchang pig embryos. Lipids in morula were polarized by centrifugation before vitrification, while post-V-W embryonic development was not observed. The time cost of ZP digestion was prolonged after V-W, ZP hardening affected the expansion and hatching of V-W embryo, while ZP digestion offset this disadvantage and promoted the expansion of V-W embryo. In vivo embryo production combining with in vitro culture raised the efficiency to 12.16 blastocysts per donator. V-W embryos using Cryotop and NCSU23 reached the highest survival rate (85.91%±3.27%) and expansion and hatching blastocyst rate (80.61%±10.06%) and the highest level of embryonic cell division. The ZP digestion promoted V-W embryo expansion after warming.

The study aimed to evaluate the application effect of somatic cell cloning technology (SCNT) in the rapid expansion of Topigs E line super boars. Using 12 super boars as donors, 1 100 embryos were isolated, cultured, frozen, amplified and subcultured, and transferred to 6 sows. Finally, 4 sows delivered 29 piglets (cloning efficiency 2.63%). The growth and develo-pment, semen quality and reproductive performance of cloned and non-cloned pigs were compared and analyzed. The results showed that there was no significant difference in growth and development between cloned pigs and donor normal offspring. There was no significant difference in semen volume ((188.17±49.27) mL) and effective sperm number ((486.35±159.56) million) between cloned pigs and donor siblings and donor half-sibs. There was no significant difference in reproductive performance of sows after mating. SNP detection showed that the inconsistency rate between cloned pigs and donor genes was below 0.14%, indicating that genetic information was derived from donors. In summary, there was no significant difference in reproductive performance and growth performance between cloned pigs and donor offspring. SCNT technology can be effectively used for super boar expansion production.

This investigation aimed to optimize the in vitro culture system for small cohorts of ovine COCs(5 per group), and enhance in vitro maturation quality and subsequent developmental potential. Experimental variables included culture volume (50 COCs/500 μL,5 COCs/500 μL, 5 COCs/50 μL,) supplemented with exogenous BMP15 and GDF9 proteins (200 ng·mL^-1), and maturation durations tested (18, 21, 24 h). Comprehensive evaluations encompassed cumulus expansion index, HAS2 gene expression profiles, apoptosis-related gene expression (BAX, BCL2, C-Myc) in cumulus-oocyte complexes, oocyte redox homeostasis parameters (GSH, ROS, GPX-1, GSR), and embryonic developmental indexes (cleavage rate, blastocyst rate, mean cell numbers/blastocyst). Addition of the combination of BMP15 and GDF9 into maturation medium significantly enhanced cumulus expansion indexes (P<0.01), upregulating HAS2 expression and improving redox balance through elevated GSH levels and reduced ROS accumulation. The optimized protocol employing 500 μL culture medium with 21 h maturation achieved superior outcomes, yielding cleavage rate of 70.23%±2.78% and blastocyst rate of 35.33%±1.53% at 48 h after in vitro fertilization of oocytes cultured in small group. Notably, the 21-hour maturation group exhibited significantly lower BAX expression and BAX/BCL2 ratios compared to the 24-hour group. Culturing 5 COCs in 500 μL microdrops yielded better results than culturing 5 COCs in 50 μL microdrops. Co-supplementation with BMP15 and GDF9 proteins reduced apoptosis and enhanced intracellular redox capacity. Shortening the in vitro maturation (IVM) duration from 24 h to 21 h reduced apoptosis, promoted OCT4 and CCNB1 gene expression in blastocysts, and increased the average blastocyst cell count.

This study aimed to compare the effects of ellagic acid (EA) and its metabolites urolithin A (UA), urolithin B (UB), and urolithin C (UC) on promoting proliferation, enhancing antioxidant activity, inhibiting apoptosis in Guizhou black goat ovarian granulosa cells (OGCs), in order to identify a superior alternative to EA.Ovarian tissues from healthy, sexually mature goats were collected to isolate and culture OGCs, which were validated for subsequent experiments. At the same time, based on previous research findings of the research group, 1 μmol·L^-1 concentration of EA had the optimal biological effect on OGCs. Therefore,cells were divided into 5 groups: control (CON), EA, UA, UB, and UC, with treatment groups supplemented with 1 μmol·L^-1 of the corresponding compounds. Proliferation was assessed via CCK-8 assay at 0, 24, 36, and 48 h. Cell migration was evaluated using a scratch assay. Intracellular ROS levels were measured using ROS detection kits. RT-qPCR was employed to analyze mRNA expression of anti-apoptotic (Bcl-2), pro-apoptotic (Bax, Caspase-3, Caspase-9), inflammatory factors (IL-1β, IL-6, TNF-α), and reproduction-related genes (GDF9, BMPR-1B, CYP19A1, FSHβ). UA exhibited the strongest pro-proliferative effect on OGCs, followed by EA and UB, with UC showing the weakest activity. The scratch assay revealed that UA significantly enhanced cell migration, whereas UB inhibited migration; no significant difference was observed between EA and UA. ROS levels were markedly reduced in UA and EA groups. qRT-PCR results demonstrated that UA, UB, UC, and EA significantly downregulated pro-apoptotic genes (CASP3, CASP9, BAX) compared to the CON group. UA upregulated BCL-2 expression most prominently. Among inflammatory factors, UA, UC, and EA significantly suppressed IL-1β expression (with UA being the lowest), there was no significant differences between CON group and other groups, while TNF-α levels decreased across all groups. UA and EA significantly enhanced the expression of reproduction-related genes (GDF9, BMPR-1B, CYP19A1, FSHβ), with UA showing the highest efficacy. In this study, compared to EA and other metabolites, UA demonstrated superior effects in promoting OGCs proliferation and migration, reducing oxidative stress and apoptosis, and modulating reproductive-inflammatory gene expression. These findings highlight UA as a promising candidate for further research as a reproductive regulatory additive in mammals.

This study aims to evaluate the effects of different levels and timings of Broussonetia Papyrifera (BP) leaf supplementation on the production performance and meat quality of yellow-feathered meat chickens, in order to promote the application of BP leaves in yellow-feathered meat chickens. A total of 360 one-day-old yellow-feathered meat chickens were selected and randomly divided into four treatment groups, with six replicates each group. The four treatment groups were fed with a corn-soybean meal-based diet (control group) throughout the entire period (1-63 d), a diet containing 5% BP leaves throughout the entire period (full 5% group), a diet containing 15% BP leaves throughout the entire period (full 15% group), and a diet containing 15% BP leaves in the later period (36-63 d, later 15% group). The results showed that compared with the control group, the body weight and average daily gain of yellow chickens in the full 5% and full 15% groups were significantly reduced (P<0.05), and their feed-to-gain (F/G) ratio was significantly increased (P<0.05), while the production performance of the late 15% group was not significantly different from the control group (P>0.05). Compared with the control group, the yellowness value (b^*) of the pectoral muscle and leg skin of birds in the three BP leave groups was significantly increased (P<0.05), but there was no significant difference in pH, drip loss and shearing force of the pectoral muscle (P>0.05). Compared to the control group, the proportion of monounsaturated fatty acids in the breast muscle of yellow chickens in the BP leaf treatment groups was significantly reduced (P<0.05), and the proportion of polyunsaturated fatty acids was significantly increased (P<0.05). There were no significant differences in the protein or amino acid content of the breast muscle (P>0.05). Additionally, feeding 15% BP leaves in both the full period and the later period significantly increased the content of inosine monophosphate (IMP) in the breast muscle of yellow chickens (P<0.05). In summary, feeding 15% BP leaves during the later stage had no significant impact on the production performance of yellow-feathered meat chickens, but it can increase the yellowness value of the breast muscle and skin, enhance the proportion of polyunsaturated fatty acids in the breast muscle, and increase the content of IMP, thus having an improving effect on the quality of the muscle.

The objective of the present study was to determine the chemical compositions and standardized ileal amino acid digestibility (SIAAD) of 10 soybean meals from different sources for broilers at 13 d and 28 d of age, and establish the prediction equations. A completely randomized design was used in this study. A total of 990 1-d-old male Arbor Acres broilers in good health were randomly allotted to 1 of 11 treatment groups (1 nitrogen-free diet treatment group and 10 treatment groups for soybean meal). The experiment was divided into two experimental periods of 10-13 d and 25-28 d, in which there were 6 replicates of 10 chickens per treatment group during the 10-13 d, and 6 replicates of 5 chickens per treatment group during the 25-28 d. On 13 and 28 days of age, the ileal contents were collected to determine the SIAAD. The results showed that: The chemical compositions of soybean meal from different sources differed. There was significant interaction (P<0.05) between the source of soybean meal and broiler age on the SIAAD except for the standardized ileal digestibility (SID) of His in soybean meal. The differences in SIAAD among different sources of soybean meals were significant (P<0.05). Except for the SID Lys and SID Met, broiler age had an effect (P<0.05) on the SIAAD in soybean meal, and the SIAAD of soybean meal were higher on d 28 than on d 13 (P<0.05). The 12 and 16 prediction models of SIAAD in soybean meal were successfully developed for broilers at 13 and 28 d of age, respectively. The prediction models with highest fitting degree at 13 d and 28 d are as follows: SID Arg=254.056-1.179CP + 1.212EE-0.409ADF-0.586Ash-0.08TI + 0.885PA-1.509NFE-3.265GE (13 d, R²=0.999，P<0.000 1)；SID Trp=- 107.513 + 11.121GE (28 d, R²=0.467, P=0.029 3). In summary, these results indicate that the difference in the chemical compositions among different sources of soybean meal was great. The fitting degree of the prediction model at 13 d of age was higher than that at 28 d of age, and they have good reference value for predicting SIAAD of soybean meal in broilers.

This experiment was conducted to prepare a multi-epitope nanofusion protein of foot-and-mouth disease virus serotype A (FMDV-A) and to establish a magnetic particle CLIA antibody detection method, which was to provide technical support for clinical immunization monitoring, prevention and control of FMDV. In this study, a magnetic particle chemiluminescent immunoassay (CLIA) for detecting antibodies against foot-and-mouth disease virus serotype A (FMDV-A) was established based on the expressed multi-epitope fusion protein of FMDV-A and its polyclonal antibody. Reaction conditions were optimized, and clinical samples were simultaneously tested using this method and commercial kits to compare the coincidence rate between the two methods. Finally, a highly soluble FMDV-A multi-epitope nanofusion protein and specific polyclonal antibody with a titer of 1∶5 120 were successfully prepared. A magnetic particle CLIA for FMDV-A antibodies detection was established based on the principle of competitive immunoassay. The optimal concentration of streptavidin magnetic beads (SA-Beads) was 0.5 mg·mL^-1, the optimal concentration of biotin-labeled recombinant FMDV-A protein antigen was 5 μg·L^-1, and the optimal concentration of acridinium ester (AE)-labeled polyclonal antibody was 2 μg·L^-1. The optimal sample volume was 10 μL, and the optimal reaction time was 15 minutes. The sensitivity test results show that the sensitivity of this method is 97.47%, and the sensitivity is good. No cross-reactivity was observed with other clinically similar pathogens, demonstrating strong specificity. The results of repeatability test demonstrated that coefficient of variation (CV) values were less than 5%, which had good stability and repeatability The results of the clinical trials indicated that the overall coincidence rate was 96.76% when compared with the LPB-ELISA method developed by Lanzhou Veterinary Research Institute. This research has established a technical foundation for the research and rapid diagnosis on type A foot-and-mouth disease in China. This study successfully prepared a VP1 multi epitope fusion nanoparticle protein of FMDV-A strain and obtained its high titer polyclonal antibody. The protein and polyclonal antibody were used to establish a detection method for FMDV-A magnetic particle CLIA antibody.

To investigate the effects of new wild-type PEDV infection on the intestinal mucosal injury and intestinal cell methuosis of piglets, 8 healthy piglets aged 5 days were randomly divided into Mock and PEDV-infected groups. The clinical symptoms were monitored. After 2 days of virus infection, the piglets were euthanized and the duodenum, jejunum, ileum, cecum, colon, and rectum were collected. Hematoxylin-eosin (HE) staining, immunohistochemistry (IHC) staining, RT-qPCR and Western blot were used to detect the pathological changes of the intestinal tissues, the viral location in the intestines, and its effect on methuosis. HE staining revealed that PEDV-infected group had different degree of damage to the intestinal mucosa, and the jejunum showed more obvious pathological damage. Compared with other intestinal segments, the viral load was the highest in jejunum after PEDV infection using RT-qPCR. IHC staining showed that PEDV mainly distributed in the upper cortex of small intestinal mucosa, and the most positive signals were observed in jejunum segment, which was consistent with the result of RT-qPCR. Further study revealed that the expression levels of megavesicular death-related genes Rac1(Ras-related C3 sarcoma virus oncogene 1), Rab7(Ras-related protein 7) and LAMP1(lysosomal-related membrane protein 1) in PEDV infection group increased significantly in jejunum, which was further confirmed by Western blot. The above results indicate that the new wild-type PEDV mainly located in the small intestine, caused the most serious pathological damage to jejunum and induced methuosis of the jejunal cells. This study could provide a reliable basis for further understanding of the pathogenesis of PEDV infection.

Porcine reproductive and respiratory syndrome virus (PRRSV), comprising two distinct species, PRRSV-1 (European genotype) and PRRSV-2 (North American genotype), exhibits significant genetic variability and poses a substantial threat to global swine production. In this study, a PRRSV-1 strain, designated GZ0308, was isolated from the lung tissue of infected piglets on a farm in Guizhou Province using porcine alveolar macrophages (PAMs). The complete genome of GZ0308 spans 14 970 nucleotides, displaying 86.99% nucleotide identity with the PRRSV-1 reference strain LV and 66.49% with the PRRSV-2 strain VR2332. Phylogenetic analysis positioned GZ0308 within a novel branch of PRRSV-1 subtype 1. Comparative genomic analysis revealed that GZ0308 harbors 43 discontinuous amino acid deletions in the NSP2 coding region and a 5-amino-acid deletion in the overlapping region of ORF3 and ORF4 relative to the LV strain. To assess pathogenicity, five-week-old piglets were inoculated intramuscularly and intranasally with 2 mL of viral suspension (10^5.5TCID₅₀·mL^-1 per route). Compared to the control group, GZ0308-infected piglets exhibited transient pyrexia, lethargy, and a significant reduction in average daily weight gain. Postmortem examination identified characteristic diffuse interstitial pneumonia lesions, confirming the strain’s pathogenicity in piglets. These findings contribute novel data to the molecular epidemiology of PRRSV in China and underscore the need for continued surveillance of emerging PRRSV variants.

To study the variation of porcine pseudorabies virus (PRV) local isolates, a PRV virus strain named JH2403 was successfully isolated from diseased materials sent to a pig farm in Yantai City. Perform genetic evolution analysis on its gB, gC, gD, TK genes and study of the pathogenicity of the isolates in mice and piglets. Phylogenetic analysis showed that the gB, gC, gD, TK genes of the strain had 99.9% to 100% homology with the vaccine strain Bartha-K61, with a total of 3 amino acid mutations. The pathogenicity test of mice showed that 1×10³ TCID₅₀·mL^-1 virus solution with a concentration of 0.2 mL per mouse was infected, and the mortality rate on the 6th day was 100%. HE staining of brain tissue showed lymphocyte infiltration. Pathogenicity test of piglets proved that there were no deaths although there were obvious symptoms. A strain of porcine pseudorabies virus was successfully isolated, which had the highest homology with the vaccine strain Bartha-K61, which showed enhanced pathogenicity in mice and lower pathogenicity in piglets. This study provides a reference for the genetic evolution analysis and epidemiological study of PRV in Shandong Province.

Infectious bronchitis is an acute and highly contagious respiratory disease caused by infectious bronchitis virus (IBV). In order to establish a suitable method for the field detection of IBV, we designed primers and probes targeting M, a conserved gene of IBV, and combined RPA and colloidal gold technology to create an RPA-test strip method for IBV detection within 1 hour. The specificity test demonstrated that the RPA-test strip method only detected IBV, but did not react with the avian influenza virus (AIV), chicken bursal disease virus (IBDV), Newcastle disease virus (NDV), avian leukosis virus (ALV), and goose astrovirus (GAstV). The sensitivity test proved that the lowest detection line of this RPA-test strip method was 8.26 TCID₅₀·mL^-1. Comparison of 31 clinical samples showed that the positive detection rates of this RPA-strip method, qPCR and conventional PCR were 74.2%, 64.5% and 54.8%, respectively. The above data showed that the RPA-strip method for the rapid detection of IBV established in this study is highly specific, sensitive and easy to operate, which provides a new method and technology for the efficient diagnosis of IBV and its immunological prevention and control.

Avian influenza (AI) is a significant threat to the healthy development of the poultry industry in China. It spreads rapidly and widely, with the potential for cross-species transmission, posing a severe threat to the health of both animals and humans. In particular, zoonotic influenza virus infections are highly hazardous and can cause substantial economic losses and social impacts. Therefore, the rapid and sensitive detection of common avian influenza viruses is of great significance for controlling the spread of the disease. This study aims to develop an efficient and convenient detection method for avian influenza, addressing the actual needs of prevention and control. Based on the conserved sequences of the H5N1 HA and H9N2 HA genes of the avian influenza virus (AIV), primers and probes were designed. A detection method for AIV using reverse transcription recombinase polymerase amplification (RT-RAA) combined with lateral flow dipstick (LFD) was established. This method can complete the detection in just 15 minutes at 37℃. There was no cross-reactivity between the AIV H5 subtype and the AIV H9 subtype, as well as between AIV and other viruses such as Newcastle disease virus (NDV), infectious bronchitis virus (IBV), and infectious bursal disease virus (IBDV), indicating good specificity. The sensitivity of the RT-RAA-LFD method can reach 100 copies per reaction, which is approximately 100 times higher than that of conventional RT-PCR. Furthermore, after testing 120 clinical samples, a 99% concordance rate was achieved between RT-RAA and commercialised RT-qPCR as well as conventional RT-PCR. The results demonstrate that this method can rapidly, accurately, conveniently, and intuitively detect AIV H5 and H9 subtypes, providing an effective and intuitive approach to the detection of avian influenza.

This study aimed to shorten the detection cycle for subgroup J/K avian leukosis virus (ALV-J/K) purification and accelerate the avian leukosis (AL) purification process, this study established a rapid blood nucleic acid screening technique (ALV-J/K blood quantitative polymerase chain reaction, ALV-J/K-B-qPCR) suitable for low-prevalence scenarios of ALV-J/K. By selecting conserved regions of ALV-J and K to design specific primers and TaqMan probes, and optimizing the reaction conditions, a TaqMan qPCR detection method for ALV-J/K was established. Using blood samples identified as ALV-J/K positive through cell culture virus isolation as experimental materials, anticoagulated whole blood DNA, blood cell DNA, and plasma cDNA were prepared and subjected to ALV-J/K TaqMan qPCR detection to identify the optimal detection template. Nucleic acids were extracted using a blood room-temperature lysis kit and a universal column-based genomic DNA extraction kit to screen for the optimal nucleic acid extraction method. The effect of varying dilution factors of positive samples on detection results was investigated to assess the feasibility of mixed sample detection. A simulated blood nucleic acid screening test with an expected prevalence rate of 1% to 2% was conducted using ALV-J/K-B-qPCR, and the results were compared with those obtained by the virus isolation method. The specificity, sensitivity, and repeatability tests of ALV-J/K TaqMan qPCR demonstrated that this method specifically amplifies only ALV-J and ALV-K, with detection limits of 8.95×10¹ copies·μL^-1, 8.16×10¹ copies·μL^-1 for positive standards, respectively. The sensitivity was 100 times higher than that of conventional PCR, and both intra- and inter-assay coefficients of variation <2%. The detection results of 96 clinical samples revealed that the ALV-J/K TaqMan qPCR had a detection rate of 21.9%, which was higher than that of conventional PCR (15.6%) and ALV p27 ELISA (18.8%). In the detection template screening test, anticoagulated whole blood DNA was identified as the optimal detection template. In the nucleic acid extraction method screening test, the universal column-based genomic DNA extraction kit yielded the best nucleic acid detection results. In the feasibility assessment test for mixed sample detection, the method established in this study was capable of detecting samples with low viral loads when the number of mixed samples <10. The simulated nucleic acid screening test for blood with an expected prevalence rate of 1% to 2% showed that the ALV-J/K-B-qPCR had a detection rate of 4%, which was 2.5% higher than that of the virus isolation method. The ALV-J/K-B-qPCR technology established in this study offers the advantages of simultaneous detection and differentiation of ALV-J and ALV-K, high sensitivity, strong specificity, high detection efficiency, and cost-effectiveness, aiming to provide a novel technique for AL purification in breeding poultry farms.

To investigate the evolutionary trends and molecular characteristics of Avian leukosis virus (ALV), this study collected 1 600 blood samples from a poultry farm in Shanghai. Viruses were isolated by inoculating chicken fibroblast cells (DF-1), followed by subgroup identification through PCR amplification of the gp85 gene. Biological characterization was performed using indirect immunofluorescence assay (IFA) and multi-step growth curves. Coding and non-coding regions of the isolated strains were compared with representative ALV subgroups for sequence alignment and molecular conformation analysis. The results showed that six ALV-J strains were successfully isolated and identified via PCR and IFA. Notably, strains ALV-J-345 and ALV-J-480 exhibited significant sequence divergence, with strain ALV-J-480 demonstrating stronger replication efficiency in DF-1 cells than strain ALV-J-345. Further comparison with other ALV-J strains revealed 46 distinct amino acid variations in the gp85 and gp37 protein between the two isolates, and strain ALV-J-480 displayed lower homology to other ALV-J strains. Additionally, a premature termination codon at position 866 in the pol gene of strain ALV-J-480 truncated the protein by eight amino acids. In non-coding regions, strain ALV-J-345 harbored a 7-nucleotide deletion in the E(XSR) element region of the 3'UTR, whereas strain ALV-J-480 carried a 19 nt nucleotide insertion at positions 358-376 in the 5'UTR. These sequence variations likely contributed to the enhanced replication capacity of strain ALV-J-480, endowing it with a competitive advantage to emerge as a potential epidemic strain in poultry populations. Therefore, this study provides updated molecular insights into ALV-J, which will aid in elucidating its evolutionary dynamics and inform the development of targeted control strategies.

This study aims to investigate the interaction between FCV NS2 protein and host cells. In this study, a eukaryotic expression plasmid of FCV NS2 was transfected into CRFK cells, with an empty vector as the control. Three samples from each group were collected 24 hours post-transfection for transcriptomic sequencing. Results were as follows: Comparison of the sequencing results revealed 218 differentially expressed genes (DEGs) in the experimental group transfected with NS2 protein, including 200 upregulated genes and 18 downregulated genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses showed that the significantly differentially expressed genes were mainly involved in intracellular viral invasion and immune evasion pathways. This study reveals that the NS2 protein significantly regulates multiple signaling pathways at the transcriptional level, including endosomal transport, TGF-β, and RIG-I, in CRFK cells, systematically reveals the global transcriptional regulation patterns of host cells by feline calicivirus (FCV) NS2 protein. These findings provide a new perspective for future research on the interactions between FCV and host cells.

This experiment mainly carried on isolation and identification Bacillus subtilis (B.S) from healthy egg chicken fecal samples, and explored the protective effect on E.coli O78 induced diarrhea in chicks. The experiment used morphological identification and 16S rDNA sequencing to isolate, screen and identify B.S. The tolerance to pH, bile salts, and artificial gastrointestinal fluids and hemolysis was assessed. Ninty-six healthy 1-day-old Dawu Jinfeng laying hens were selected for animal experiments and randomly divided into eight groups, 12 chickens per group: The control group (fed with basal diet, CON), low dose group (fed with basal diet and orally administeres 1 mL×10⁹ CFU of B.S JZH 9, LBS), medium dose group (fed with basal diet and orally administeres 1 mL×10¹⁰ CFU of B.S JZH 9, MBS), and high dose group (fed with basal diet and orally administeres 1 mL×10¹¹ CFU of B.S JZH 9, HBS). The model group (fed with basal diet and orally administered 1 mL×4.5×10¹² CFU of E.coli O78, Model), low dose prevention group (LBS + E.coli), medium dose prevention group (MBS + E.coli), and high dose prevention group (HBS + E.coli). The study analyzed the effects of the B.S JZH 9 on the growth performance and intestinal health of E.coli O78 challenged laying chicks. The experiment lasted for 21 days, with oral administration of B.S JZH 9 fod 1~18 days and oral administration of E.coli O78 for 19~21 days. The results showed that the isolated Gram-positive bacteria, identified as B.S by 16S rDNA, named B.S JZH 9. With the characteristics of acid and bile salt resistance, no hemolytic reaction, and tolerance to artificial gastrointestinal fluids. The results of animal testing showed that compared with the CON, the LBS, MBS, HBS had no significant effect on average daily gain (ADG), average daily feed intake (ADFI), and feed conversion rate (FCR) from 1 to 21 days. Compared with the Model, the LBS + E.coli showed a trend of reducing FCR from 1 to 21 days and the LBS + E.coli, MBS + E.coli and HBS + E.coli significantly increased the villus height and villus-crypt ratio, while the MBS + E.coli and HBS + E.coli significantly reduced the crypt depth of duodenal. Compared with the CON, the content of E.coli was significantly increased in the jejunum of the Model, while feed different concentrations of B.S JZH 9 reduced the relative content of E.coli to varying degrees. In summary, B.S JZH 9 isolated from the feces of healthy laying hens, promoted growth performance, increased the villus height and villus-crypt ratio of small intestinal, reduced crypt depth, increased the absorption area of the small intestine, and reduced the content of E.coli in the jejunum, thereby regulating intestinal health and slowing down the damage caused by E.coli O78 to the intestine.

This study aimed to isolate a lytic Klebsiella variicola bacteriophage, characterize its biological properties, and conduct whole-genome analysis. The phage was isolated and purified by the double-layer agar culture method from a dairy farm in Sichuan province. Its biological characteristics, including lysis spectrum, thermal stability, acid-base tolerance, optimal infection multiplicity, and one-step growth curve, were determined. Additionally, whole-genome analysis was conducted and phage morphology was observed by transmission electron microscopy (TEM). The results showed that, the isolated lytic Klebsiella variicola bacteriophage vB_Kvc_Y10. The morphology of plaque is round and transparent with clear boundaries, and the TEM shows that Y10 has a short, filamentous tail. The biological results showed that phage Y10 has high host specificity, and is not resistant to high temperatures and acidic environment. Meanwhile, the optimal multiplicity of infection (OMOI) is 0.01. The result of one-step growth curve assay indicates that the latency period is 40 minutes, and the lysis amount is 306. Whole-genome analysis indicates that bacteriophage Y10 belongs to the family of Autographiviridaes and is a dsDNA virus. The complete genome size is 43 281 bp, with a GC content of 56.19%. Among the 51 ORFs, 29 have putative functions and exhibit distinct functional partitions. No tRNA or toxin genes are found in the Y10 genome. Both the phylogenetic tree and ANI analysis indicate that bacteriophage Y10 is closely related to bacteriophage vB_Kpl_K59PH2 (accession number: OY757063). In conclusion, bacteriophage Y10 is a highly specific lytic bacteriophage without virulence genes, presenting potential for the prevention and treatment of bacterial infections.

This study focuses on female adult Hyalomma asiaticum, aiming to thoroughly investigate the microbial community composition in certain internal organs during different blood-feeding stages and the fully engorged state. By analyzing the impact of the blood-feeding period on the differences in microbial abundance, the key data on the changes in the microbial community within female H. asiaticum were obtained, which was used to provide a data reference for relevant research. In this study, the Illumina Novaseq sequencing platform was employed to conduct high-throughput sequencing of the 16S rRNA V3 - V4 region in female H. asiaticum at different blood-feeding stages and in multiple organs (including the midgut, Malpighian tubules, ovaries, and salivary glands) of fully engorged female H. asiaticum. Through comparative analysis, the dynamic changes in the microbial communities in female ticks at different stages and in the organs of fully engorged ticks were revealed. As the blood-feeding process of H. asiaticum progressed, the Shannon index of microbial community diversity showed a significant decreasing trend, dropping from 4.2±0.3 in the starved state to 1.8±0.2 in the fully engorged state (P<0.01). During this process, Proteobacteria gradually became the absolutely dominant phylum, with a relative abundance exceeding 99%. Analysis of the fully engorged organs indicated that Francisella was highly enriched in the engorged ovaries, with a relative abundance as high as 99.75%, and there was an extremely significant difference compared to Francisella in the midgut (P<0.01). In addition, the abundance of this genus exhibited a trend of first increasing and then decreasing with blood-feeding time. Specifically, the abundance was 12.3% at 24 h of blood-feeding, reached a peak of 89.7% at 96 h, and then decreased to 75.3% in the fully engorged state. The results of this study fully reveal the complexity and diversity of the microbial communities in the internal organs of female H. asiaticum at different blood-feeding stages and in the fully engorged state. This discovery not only enriches our understanding of the microbial communities associated with H. asiaticum but provides an important data foundation for further in-depth comprehension of the interactions between the microbial communities and the host within H. asiaticum adults.

This study aimed to investigate the microflora structural characteristics of the midgut of different engorged Haemaphysalis longicornis(H. longicornis) females collected from Panthera tigris tigris. Female H. longicornis in fully engorged (FF) and half engorged (HF) were sampled, and midgut contents were collected from each group of ticks under aseptic conditions. Total bacterial DNA was extracted from each group of samples. The V3-V4 areas of 16S rRNA was amplified by PCR, and the libraries were constructed for each group. Paired-end sequencing was performed using the Illumina NovaSeq platform. Sequencing data were processed by merging, filtering, and denoising, followed by Amplicon Sequence Variant (ASV) clustering and diversity analysis. The results showed that 100 021 and 82 657 effective tags were obtained from FF and HF, respectively. The number of ASVs obtained from FF and HF after clustering was 259 and 541, respectively, of which 116 were common between the two groups. At the phylum level, the dominant common phyla were Proteobacteria, Firmicutes, Bacteroidota, and Actinobacteriota in both groups, with Proteobacteria being more abundant in FF, while the other three dominant phyla were relatively more abundant in HF. At the genus level, the dominant common genera were Coxiella, Muribaculaceae, Corynebacterium, Delftia, and Staphylococcus in both groups. Coxiella had the highest relative abundance, and the relative abundance of Coxiella was higher in FF than in HF. In FF, genera such as Schlegelella were unique and relatively abundant, while in HF, genera such as Prevotella were unique and relatively abundant. At the species level, Coxiella endosymbionts were the most abundant taxa in both groups, with distribution patterns consistent with Coxiella. The results suggested that the midgut of female H. longicornis has a complex microflora. There were certain differences in the midgut microflora of different engorged ticks, and the relative abundance of the same genus varied in different samples.

The aim of this study is exploring the mechanism of Yupingfeng oral liquid combined Maxingshigan oral liquid and Shuanghuanglian oral liquid to treat avian respiratory tract infection via network pharmacology, and screen potential anti-IBV drugs by molecular docking technique, as well verifying the anti-IBV activity of Luteolin. The blood entry components and potential targets in three traditional Chinese medicine compounds were retrieved in TCMSP and Uniprot database. The related targets of diseases (respiratory tract infection) were discovered in DrugBank, GeneCards, OMIM and PharmGkb databases, the DAVID database was used to analyze the GO enrichment and KEGG pathway annotation. Then, the key compounds were docked with IBV-nsp9, S1 and main proteases (M^pro) by PDBQT and AutoDock vina software. The anti-IBV activity of Luteolin was validated in Vero cell. The result exhibited that Baicalein, Phillygenin, Luteolin, Formononetin, Kaempferol and Quercetin in three traditional Chinese medicine prescriptions might play a role in the treatment of avian respiratory tract infection through key targets such as FN1, IL6 and PTGS2. Molecular docking result indicated that Phillygenin, Kaempferol, Glycyrrhizin, Luteolin, Quercetin and Isovitexin showed the strongest binding ability with IBV-nsp9, S1 and M^pro. The effect of Luteolin inhibiting IBV replication and the number of viral plaques were verified in Vero cell. The study preliminarily revealed the basic pharmacological effects mechanism of three kinds of traditional Chinese medicine combinations in the treatment of avian respiratory tract infection, and confirmed the anti-IBV effect of Luteolin in vitro.

This study aims to evaluate the protective effects of curcumin (Cur) on oxidative stress and apoptosis induced by heat stress (HS) in bovine mammary epithelial cells and to explore its potential mechanism via the regulation of related gene expression through the Nrf2 signaling pathway. Different concentrations of curcumin (0, 5, 10, 20, 50 nmol·L^-1) were used to screen the concentration which doesn't have cytotoxic effect on the cells. Then the concentration that showed the most significant recovery in cell viability under heat stress was selected as the treatment concentration for subsequent experiment. Bovine mammary epithelial cells were divided into four groups: control group (NC), curcumin-treated group (Cur), heat stress group (HS), and curcumin-treated heat stress group (HS+Cur). And oxidative stress response, apoptosis, and lactation-related gene expression were assessed using immunofluorescence, Western blot, and RT-qPCR. The results showed that, curcumin at concentrations of 5 and 10 nmol·L^-1 did not significantly affect the viability of bovine mammary epithelial cells (P>0.05). However, 10 nmol·L^-1 curcumin significantly improved cell viability under heat stress conditions (P<0.001). Curcumin increased mitochondrial membrane potential, reduced ROS and H₂O₂ levels, enhanced the activity of antioxidant enzymes (SOD and GSH), decreased malondialdehyde (MDA) levels, and significantly inhibited the Bax/Bcl-2 ratio (P<0.05), thereby reducing apoptosis. Western blot results showed that curcumin reduced Keap1 expression while upregulating Nrf2, HO-1 (P<0.01) and NQO1 (P<0.05) downstream antioxidant gene expression. RT-qPCR results demonstrated that curcumin significantly restored the expression of lactation-related genes (CSN1S1, CSN1S2, CSN2) under heat stress conditions (P<0.05).In conclusion, curcumin alleviates oxidative stress and apoptosis induced by heat stress in bovine mammary epithelial cells by modulating the Nrf2 signaling pathway. It restores cell viability and upregulates lactation-related gene expression, providing a theoretical basis for the application of curcumin in mitigating heat stress in dairy cows.

The aim of this study was to reveal and validate the improvement effect of bilobalide (BB) on cartilage degeneration in a rat osteoarthritis (OA) model induced by anterior cruciate ligament transection (ACLT) via the Hippo-YAP/TAZ pathway. Forty-five male Sprague-Dawley (SD) rats were randomly divided into the control group, model group (OA group) and drug intervention group (BB group). In addition to the control group, the rat OA model was constructed by ACLT method. The rats in the BB group were given 10 mg·kg^-1 BB daily for 6 weeks, after that, knee joint and cartilage tissue samples were taken for consequtive examination. After the operation, the rats in each group were examined once a week for joint swelling. The degree of cartilage degeneration of rats was evaluated by gross observation, Pelletier score, histological observation and the OARSI score. The degree of subchondral bone injury was observed by Micro-CT, and the levels of Hippo-YAP/TAZ pathway related proteins MST1, LATS1, YAP and TAZ, as well as the cartilage extracellular matrix (ECM) degradation protein MMP-3 were detected by Western blot.Computer molecular docking was used to predict the binding affinity of BB to potential targets. The results showed that in an ACLT-induced OA model, BB could inhibit joint swelling, alleviate cartilage pathological features and highly significantly reduce OARSI score (P<0.01). It also mitigated damage to the subchondral trabecular structure and ameliorated OA-related cartilage degeneration. In terms of mechanism, BB highly significantly increased the levels of MST1 and LATS1 in Hippo-YAP/TAZ pathway in rat cartilage (P<0.01), inhibited the expression of YAP and TAZ in the nucleus (P<0.01), and decreased the expression of MMP-3 (P<0.01), thus playing an anti-ECM degradation role. In addition, molecular docking showed that BB had good affinity with YAP and TAZ (binding energy<-20.92 kJ·mol^-1). The results suggested that BB inhibited the degradation of cartilage ECM, alleviated joint swelling and inhibited cartilage degeneration through Hippo-YAP/TAZ pathway, and played a role in improving OA.

The aim of this study was to investigate the effects of unfermented Astragali Radix (AR) and fermented AR on the growth performance, serum biochemical parameters, antioxidant capacity, and immune function of weaned piglets. One hundred healthy Landrace×Large crossbred weaned piglets with similar body weight (8.77±0.68 kg) at 28 days of age were assigned randomly into four treatment groups. Each treatment group had five replicate pens per group and five piglets per pen. The dietary treatments included basal diet (Control group), basal diet + amoxicillin 800 mg·kg^-1 (Amoxicillin group), basal diet + unfermented AR 1 g·kg^-1 (AR group), and basal diet + fermented AR 1 g·kg^-1 (Fermented AR Group). The experiment lasted 42 days. Results showed that: 1) Compared with the control group, dietary fermented AR supplementation significantly increased the average daily gain and final weight of weaned piglets (P<0.05, P<0.01). Compared with the AR group and the amoxicillin group, the final weight of weaned piglets in the fermented AR group was significantly increased (P<0.05, P<0.01). 2) Compared with the control group, dietary fermented AR supplementation significantly increased the apparent digestibility of dry matter, organic matter, crude protein and phosphorus in weaned piglets (P<0.01 or P<0.05), and dietary AR supplementation significantly increased the apparent digestibility of crude protein and phosphorus (P<0.05, P<0.01). 3) The total protein (TP) and albumin content in the serum of weaned piglets in the fermented AR group was significantly higher than that of the control group (P<0.05), and the TP content was significantly higher than that of the amoxicillin group and the AR group (P<0.05). 4) Compared with the control group, dietary fermented AR supplementation significantly increased the total antioxidant capacity and catalase activity in the serum of weaned piglets (P<0.05). 5) The content of immunoglobulin G in the serum of weaned piglets in the fermented AR group was significantly higher than that of the control group and the amoxicillin group (P<0.05), and the contents of interleukin 1β and interleukin 6 were significantly lower than those of the control group (P<0.05). In conclusion, under the condition of the current experiment, the effect of adding fermented AR to the basal diet is better than that of AR. The addition of fermented AR can improve the apparent digestibility of nutrients, improve the antioxidant, immune and biochemical indicators in the serum of weaned piglets, thereby promoting the growth performance and health status.

This study aimed to establish a DF-1 cell line stably overexpressing chicken mmp7 gene, providing a foundation for investigating the role of mmp7 in related diseases at the cellular level. The chicken mmp7 gene sequence was synthesized and cloned into the lentiviral vector pLVX-MMP7-IRES-puro. Using a three-plasmid lentiviral packaging system, the recombinant plasmid pLVX-MMP7-puro was co-transfected with the helper plasmids pCMV-VSV-G and psPAX2 into 293T cells at the logarithmic growth phase to produce mmp7-carrying lentiviral particles. DF-1 cells were infected with the recombinant lentivirus for 72 h, followed by puromycin selection (1.5 μg·mL^-1). Integration of mmp7 into the DF-1 genome was confirmed by PCR, while its transcriptional and protein expression levels were assessed via RT-PCR and Western blot, respectively. The CCK-8 assay was used to evaluate the effect of MMP7 overexpression on DF-1 cell proliferation. A DF-1 cell line stably overexpressing chicken MMP7 was successfully generated. Overexpression of MMP7 showed no significant impact on cell viability at 24-48 h but led to a notable reduction at 120-144 h (P<0.05). This cell line serves as a valuable tool for further exploration of MMP7’s biological functions.

To thoroughly evaluate the pathogenic characteristics of Bibersteinia trehalosi (B. trehalosi) in experimental animals (mice) and target animals (sheep), this study will inoculate mice and sheep with varying doses of the B. trehalosi GD01 strain, which was isolated from a sheep that died of pneumonia. Key indicators, including clinical symptoms, pathological changes, and pathogen load post-inoculation, will be assessed to provide a detailed understanding of the virulence of the strain. In the mouse experiment, intraperitoneal inoculation of the bacteria resulted in a dose-dependent lethal effect. As the dose increased, the mortality rate gradually rose, accompanied by severe pulmonary lesions and a significant increase in bacterial load. In the sheep experiment, intratracheal inoculation of the bacterium at different doses induced acute fever, coughing, nasal discharge, and anorexia, with more severe symptoms at higher doses. Necropsy results showed varying degrees of congestion, consolidation, fibrinous exudation, pleuropulmonary adhesions, and purulence in the lungs, with pathological changes typical of fibrinous pneumonia and pleuropneumonia. The severity of these changes was positively correlated with the infection dose. Histopathological observations revealed fibroblast proliferation and inflammatory cell infiltration, and immunohistochemistry showed significant pathogen aggregation within the alveoli. qPCR analysis showed that sheep continued to shed bacteria through the nasal cavity after infection, with high viral loads detected in the lungs, indicating that the bacterium successfully colonized the sheep’s lungs and caused disease. In conclusion, this study characterized the clinical symptoms, pathological changes, and pathogen load after B. trehalosi infection in mice and sheep, confirming that B. trehalosi induces a dose-dependent pathogenic response in both species. These findings provide important experimental data for further understanding the pathogenic mechanisms of this bacterium, establishing animal infection models, and developing prevention and control strategies.

This study was conducted to investigate the effects of baicalein (Bai) on inflammation, intestinal barrier and intestinal flora in mice with ulcerative colitis (UC). After 7 days of adaptive feeding, 60 BALB/c male mice aged 5 weeks were randomly divided into 6 groups: blank group (NC group), dextran sodium sulfate group (DSS group), mixed antibiotics + dextran sodium sulfate group (ABX+DSS group), dextran sodium sulfate + baicalein group (DSS+Bai group). Mixed antibiotics + dextran sodium sulfate + fecal microbiota transplantation group (ABX+DSS+FMT group, ADF group) and baicalein administration group (Bai group). Animal models of acute ulcerative colitis were replicated by dextran sulfate sodium (DSS), and the therapeutic effect of baicalein in inflammatory bowel disease was observed, and the effect of baicalein on intestinal flora in inflammatory state was analyzed, so as to explore the mechanism of baicalein in the treatment of inflammatory bowel disease. The results showed that baicalein could alleviate intestinal inflammation in colitis mice through PPAR-γ/NF-κB pathway, significantly down-regulated the expression of IL-1β and TNF-α in DSS group (P<0.001), and up-regulated the expression of IL-10 and colonic protein (P<0.001). Histopathological sections showed that colon epithelial cells in the DSS group had a large number of exfoliation, necrosis and disintegration, intestinal villi structure was blurred, crypts were missing, mucosa and submucosa were infiltrated by a large number of inflammatory cells, fibrotic structures appeared in the submucosa, glandular structures in the mucosa were destroyed, and goblet cells were greatly reduced. The colon structure of Bai treatment group was intact, there were inflammatory cells in the mucosa and submucosa, and the degree of intestinal epithelial exfoliation was significantly reduced than that of DSS group. Further evaluation of intestinal flora showed that baicalein could regulate intestinal flora compared with DSS group, mainly Firmicutes and Proteobacteria, and baicalein administration group could significantly increase the abundance of Firmicutes, reduce the abundance of Proteobacteria, promote the growth of intestinal probiotics and inhibit pathogenic bacteria, thus achieving the therapeutic effect of colitis. In conclusion, baicalein can relieve colitis induced by sodium glucan sulfate, and play a therapeutic effect by regulating intestinal flora through PPAR-γ/NF-κB pathway.

Zearalenone (ZEA) is a common mycotoxin that can cause severe reproductive toxicity. In our previous study, we found that chlorogenic acid could alleviate ZEA-induced ovarian granulosa cell toxicity, however, the mechanism of action is not clear. The aim of this study was to investigate the molecular mechanism by which chlorogenic acid antagonizes ZEA-induced follicular granulosa cell death in mice using proteomics techniques. Primary isolated and cultured mouse ovarian granulosa cells were divided into two groups: the ZEA group and the chlorogenic acid intervention group. After 24 h of treatment, the regulatory network of chlorogenic acid was systematically analyzed using quantitative proteomics technology combined with bioinformatics analysis and RT-qPCR validation. The results showed that a total of 32 differentially expressed proteins (DEPs) were identified in the chlorogenic acid-treated group compared with the ZEA-treated group, of which 15 were significantly up-regulated and 17 were significantly down-regulated. Through GO functional annotation and KEGG pathway enrichment analysis, the DEPs were mainly involved in biological processes such as mitotic regulation and cytoskeletal remodeling, and were significantly enriched in key regulatory pathways such as apoptosis, focal adhesion, Toll-like receptor signaling pathway and NF-κB signaling pathway. The changes in the expression of the key regulator RIPK1 were further verified by RT-qPCR and it was found that ZEA treatment significantly inhibited the expression of RIPK1, whereas chlorogenic acid intervention partially restored its expression level. Chlorogenic acid acts synergistically through multi-targets and multi-pathways such as regulation of cell cycle progression and apoptosis and maintenance of cellular structural homeostasis to antagonize ZEA-induced reproductive toxicity. In addition, RIPK1 may play a critical role as a key regulatory switch for cell survival and death. The present study reveals the mechanism of action of chlorogenic acid in antagonizing reproductive toxicity of ZEA at the protein level, providing new molecular targets and theoretical basis for the development of chlorogenic acid-based ZEA detoxifying agents or functional feed additives.

The objective of this study was to explore how Sijunzi San influences the rumen microbial community and fermentation processes in postpartum dairy cows, to investigate the potential mechanisms of its spleen-invigorating effects. Sixteen multiparous Holstein cows with similar body weight (600 kg±30 kg) and body condition score (3.0-3.5) were randomly assigned to either an experimental or a control group. The experimental group received an oral administration of 300 grams of Sijunzi San following calving, whereas the control group was given an equal volume of clean drinking water, both for a duration of 7 consecutive days. On the eighth day, rumen fluid was collected from both groups prior to morning feeding. Rumen fermentation parameters were measured using ELISA, the content of volatile fatty acids was determined by gas chromatography, and changes in the quantity and structure of the rumen microbiota were analyzed using 16S rRNA amplicon high-throughput sequencing. The results indicated that following the administration of Sijunzi San, the rumen pH remained relatively stable and did not exhibit a significant change (P>0.05). However, the levels of ammonia nitrogen (NH₃-N), microbial protein (MCP), cellulase, and α-amylase experienced a notable increase (P<0.01), and urease activity also rose (P<0.05). Conversely, the concentrations of propionic acid and isobutyric acid in the rumen saw a decrease (P<0.05), whereas the content of n-butyric acid surged (P<0.01). At the phylum level, Bacteroidota and Firmicutes maintained their predominance as the two dominant bacterial phyla, showing no statistically significant alterations in their relative abundances (P>0.05). However, genus-level analysis revealed distinct shifts in rumen microbiota composition: the relative abundances of Prevotella and Rikenellaceae_RC9_gut_group exhibited a significant increase, while Muribaculaceae showed a marked reduction in abundance. Additionally, the abundance of 10 non-dominant microbiota changed significantly, with 6 beneficial genera significantly upregulated (P<0.05), including the Eubacterium ruminantium group, Rothia, Bacteroidales RF16 group, Limnohabitans, hgcI clade, and Escherichia Shigella. Four harmful genera were significantly downregulated (P<0.05), including UCG-001, Clostridium_sensu_stricto_1, Dubosiella, and Parvimonas. Rothia exhibited a positive correlation with α-amylase, propionic acid, and isobutyric acid (P<0.05); Limnohabitans was positively associated with urease, cellulase, α-amylase, NH₃-N, MCP, propionic acid, and isobutyric acid (P<0.05), and negatively correlated with pH (P<0.001); HgcI_clade had a positive correlation with NH₃-N (P<0.05); Clostridium_sensu_stricto_1 was negatively correlated with pH (P<0.001); Dubosiella was negatively associated with MCP, NH₃-N, and propionic acid (P<0.05); and Parvimonas was negatively correlated with MCP and NH₃-N (P<0.05). The results suggest that the spleen-invigorating effect of Sijunzi San may be closely related to the regulation of rumen microbiota abundance and the enhancement of digestive enzyme activity in the rumen.

The aim of this study was to investigate the protective effect of chlorogenic acid (CGA) against chronic stress-induced intestinal injury in rats and its regulatory role in the nuclear factor-kappa B/NOD-like receptor protein 3 (NF-κB/NLRP3) pathway-mediated pyroptosis, in order to provide new strategies for preventing stress-related intestinal diseases in livestock. Thirty-two Sprague-Dawley (SD) rats with an initial body weight of 200±20 g were randomly divided into four groups: control group (CON group), chlorogenic acid group (CGA group), chronic restraint stress group (CRS group), and chlorogenic acid intervention group (CRS+CGA group). The CON group received no intervention. The with CGA group was administered CGA via gavage at a dose of 100 mg·kg^-1. The CRS group was subjected to 6 h of daily restraint for 21 d consecutively. The CRS+CGA group was gavaged with an equivalent dose of CGA and followed by the same restraint procedures as the CRS group. Open-field test was conducted on day 22 to evaluate the model establishment. Rats were euthanized on day 23 for blood and ileum collection. Serum corticosterone (CORT) levels were measured, ileum histopathology was observed by hematoxylin-eosin (HE) staining, inflammatory cytokine levels in the ileum were detected by enzyme-linked immunosorbent assay (ELISA), tight junction (TJs) proteins and gene expression were analyzed by immunohistochemistry (IHC) and real-time quantitative PCR (RT-qPCR), and pyroptosis-related proteins were assayed by Western blot. The results showed that the chronic stress model was successfully established. CGA intervention significantly promoted average daily body weight gain of rats under chronic stress (P<0.01), alleviated ileum structural damage, reduced inflammatory cytokine levels (P<0.01), upregulated TJs-related proteins and gene Occludin, Claudin3, Zonula occludens-1 (ZO-1) expression (P<0.01), and inhibited pyroptosis-related proteins nuclear factor-kappa B p65 (NF-κB p65), NLRP3, cysteinyl aspartate specific proteinase-1 p20 (Caspase-1 p20), and N-gasdermin D (N-GSDMD) expression (P<0.01). Collectively, CGA alleviates chronic stress-induced ileum injury in rats, possibly by inhibiting the NF-κB/NLRP3 pathway, attenuating pyroptosis, reducing inflammatory cytokine levels, and thereby enhancing intestinal barrier function.