N6-methyladenosine (m6A) modification is a widely present dynamic reversible modification in eukaryotic mRNA, which plays an important role in regulating lipid metabolism. m6A modification participates in biological processes such as adipocyte differentiation, lipid synthesis and breakdown, by regulating mRNA stability, splicing, transport, and translation. This article systematically reviews the mechanism of m6A modification in regulating adipocyte differentiation, lipid synthesis and breakdown, aiming to provide new strategies for regulating adipogenesis in animals, improving meat quality and promoting the development of animal husbandry.

The escalating global temperatures attributed to climate change have markedly amplified porcine susceptibility to heat stress, intensifying related agricultural challenges. The prenatal milieu constitutes a critical determinant for both sow health and offspring developmental trajectories. Contemporary investigations have prioritized delineating the biological consequences of in-utero thermal stress on porcine performance metrics, concurrently elucidating emerging genetic regulatory frameworks. Porcine thermoregulatory constraints manifest through restricted eccrine sweating capacity and elevated gestational metabolic requirements, culminating in exacerbated heat stress vulnerability. Such intrauterine thermal adversity in gravid sows jeopardizes fetal morphogenesis through uterine microenvironmental alterations, adversely affecting vital economic parameters including conceptus viability indices and neonatal anthropometrics, consequently precipitating substantial financial ramifications for international livestock production systems. This comprehensive analysis systematically evaluates the multigenerational impacts of prenatal heat stress on porcine postpartum characteristics, production efficiency, genetic expression patterns, and reproductive outcomes, while deciphering associated pathophysiological and epigenetic mechanisms. The synthesis ultimately seeks to establish an empirical foundation for advanced thermoregulatory intervention protocols and precision breeding initiatives targeting optimized porcine thermal tolerance.

Hatching is an important part of poultry production. Egg management and hatching technology directly affect the propagation efficiency and production efficiency of excellent seed sources. Under the current poultry production system, it is inevitable to extend the storage time of eggs in order to obtain enough eggs that can be hatched at the same time. Improper storage of eggs not only has a negative impact on their hatching performance, but also affects the embryonic development process and even leads to a decline in chick quality. Therefore, studying the changes of key indicators related to the storage period of eggs and developing corresponding technologies to improve hatchability have attracted widespread attention from the industry and researchers. This review will synthesize existing research findings to explore the latest advances in the relationship between changes during egg storage caused by environmental factors and the corresponding technologies for improving hatchability. It will also look forward to future research directions, providing a theoretical basis for improving the reproduction efficiency and economics profits of breeders production.

With the rapid advancement of high-throughput sequencing technologies and the deepening of genomic research, an increasing number of genomic data from livestock and poultry have been made publicly available, significantly enhancing the study of important economic traits at the whole-genome level. Whole-genome resequencing technology is extensively utilized in genomic selection, population structure analysis, genetic diversity assessment, and evolutionary mechanism studies, particularly demonstrating significant value in elucidating how genomic sequence variations influence economic traits. This article aims to review the application progress of whole-genome resequencing technology in the study of genetic structure, genetic diversity, origin and evolution, adaptive traits, and important economic traits in local yellow cattle populations, providing crucial references for the improvement and conservation of local yellow cattle genetic resources.

T-2 toxin is a mycotoxin widely found in grains and animal feed, posing a significant threat to human and animal health. The review aims to provide a comprehensive summary of the sources, physicochemical properties, and reproductive toxicity of T-2 toxin, with a particular focus on its mechanisms and effects on male reproductive systems. The review explores the toxicological pathways of T-2 toxin, including oxidative stress, mitochondrial damage, and cell apoptosis, and their impact on testicular function, sperm quality, and endocrine regulation of reproduction. Additionally, the review discusses the effects of T-2 toxin exposure on leydig cells, sertoli cells, and spermatogenesis, as well as examines the potential application of antioxidants and natural detoxifying agents in alleviating reproductive dysfunction induced by T-2 toxin. By analyzing the toxicity mechanisms of T-2 toxin, this review provides guidance for future research and aims to provide scientific evidence for developing effective prevention strategies and detoxification agents, thereby safeguarding livestock reproductive health and promoting sustainable development of the animal husbandry industry.

Superovulation serves as a pivotal technology in bovine embryo in vivo production, constituting a vital method for embryo in vivo procurement and forming the foundation for various other embryonic engineering techniques. During the process of superovulation, a plethora of reproductive hormones are employed to foster follicular development and ovulation, among which follicle-stimulating hormone (FSH) is one of the most crucial and central hormones. In recent years, there has been a decline in the production of pituitary-derived FSH while the market demand for FSH continues to escalate. This discrepancy has prompted an increasing number of researchers to focus on the development of recombinant FSH and its efficacy in bovine superovulation. This review is grounded in the theoretical foundations of FSH protein structure and protein engineering, presenting the evolution of recombinant FSH and summarizing the research advancements concerning the application of recombinant FSH in bovine superovulation over the past decade, both domestically and internationally. Furthermore, it delves into potential areas of interest within the field of bovine superovulation. The aim is to provide valuable insights for the establishment of an efficient superovulation protocol utilizing recombinant FSH.

Zearalenone(ZEN) and its derivatives pose a serious contamination issue for crops and their products globally, not only affects agricultural production and feed safety, but also threats the health of animals and humans, which leads to a decrease in immune function, stunted growth, and triggers estrogen-related diseases even cancer. Therefore, the toxicity of ZEN and its derivatives are receiving increasingly attention. However, there is limited research on ZEN derivatives so far. This article summarizes the existing toxicity and detoxification mechanisms of ZEN and its derivatives, including estrogen-like effects, induction of organ damage, triggering of cell apoptosis and autophagy, inflammation induction, activation of cancer-related genes, and DNA damage. And also categorizes the detoxification mechanisms using physical, chemical, and biological methods as a reference for the development of more efficient and safe detoxification drugs for ZEN and its derivatives.

Camellia seed meal, a by-product of Camellia oil extraction, is rich in protein, fat, sugars, and bioactive compounds such as saponins, flavonoids, protease, polysaccharides, and polyphenols. These components exhibit potent antioxidant, anti-inflammatory, antibacterial, immune-regulating, lipid metabolism-modulating, hypoglycemic, and anticancer properties. The inclusion of Camellia seed meal in livestock and poultry feed has shown significant potential in enhancing growth performance, immunity, and intestinal microbiota composition. This review systematically examines the bioactive components and biological functions of Camellia seed meal and explores its application potential in livestock and poultry production in depth, offering scientific insights for its broader utilization in the industry.

There are a large number and variety of microorganisms inhabiting the rumen tract of dairy cows, forming a 'mutually beneficial' symbiotic relationship with the body. Short-chain fatty acids (SCFAs), as metabolites of microorganisms, interact with microorganisms to provide nutrients to the body and maintain the stability of the internal environment. This paper describes the interactions between SCFAs and microorganisms in the rumen tract of dairy cows, focuses on their effects on the health and homeostasis of the rumen tract of dairy cows, and reviews the mechanisms of SCFAs in rumen of dairy cows regulating the internal environment, so as to provide theoretical basis for the nutritional regulation of the health of dairy cows by SCFAs.

With the rapid development of animal husbandry in China, the problems of large emissions of animal manure and the low treatment rate have become increasingly serious. Farming odors are mainly generated from the animal manure, sewage and bedding materials. These gases not only have a strong pungent smell, but also harm the animal health and production performance. Implementing effective measures to curtail the production of odors is of great significance for environmental protection, the healthy growth of animals, and the sustainable development of the breeding industry. Deodorization methods include physical, chemical and biological ones. Among them, microbial deodorization technology is a method of removing odors through the physiological metabolic activities of microorganisms. Compared with other deodorization technologies, it has the advantages of environmental friendliness, safety and high efficiency. Therefore, microbial deodorization technology has demonstrated substantial application potential and value. In this paper, the sources and hazards of odors in livestock and poultry farming, the characteristics and principles of different deodorization technologies were exponded, and the application of microbial deodorization technology in livestock and poultry breeding were reviewed and its perspects were explored, in order to provide a reference for further research for microbial deodorization technology.

Heart diseases have become a primary cause of mortality in dogs and cats. Imaging technologies, with notable advantages of non-invasiveness, convenience, and high resolution, have been extensively employed in diagnosing and treating cardiac diseases, However, when compared to humans, X-ray and ultrasound imaging still represent the predominant techniques for cardiac imaging in dogs and cats, with the development of cardiac imaging technologies for these species progressing relatively slowly. To contribute insights and guidance for diagnosing cardiac diseases in animals, and to drive the advancement of animal models for canine and feline heart diseases, thereby offering a preclinical research foundation for the diagnosis and treatment of human heart diseases, this article explores recent advances in using novel imaging techniques to diagnose common cardiac diseases in dogs and cats, offering an understanding of potential future developments.

Coronavirus seriously endangers human and animal health, and its genome is prone to mutation and recombination, which brings great difficulties and challenges to the prevention and control of coronavirus disease. In addition to safe and effective vaccines, broad-spectrum antivirals are urgently needed. Antiviral drugs targeting host factors dependent on viral replication have the advantages of broad spectrum and being less susceptible to resistance. Lipid rafts are a kind of special lipid microdomain located on the surface of cell membrane, which is also the binding site for many viruses to invade host cells, and play a crucial role in the process of coronavirus infection. This review summarized the structure and functions of lipid rafts, coronavirus invasion and its correlation with lipid rafts, the research on lipid rafts ' involvement in coronavirus infection, and the recent application of lipid rafts in the prevention and treatment of coronavirus diseases, aiming to provide direction and reference for the research and development of therapeutic targets for coronaviruses.

This study aimed to investigate the molecular mechanisms by which photoperiod influences the laying performance of Taihe black-boned silky fowl. Eighty healthy Taihe black-boned silky fowl, approximately 132 days old with similar body weights, were selected for the experiment. They were randomly divided into a control group (CP group) and a long photoperiod group (LP group), with 4 replicates of 10 chickens each. The experiment lasted for 240 days. At the conclusion of the experiment, the chickens were slaughtered and dissected, and serum and ovarian tissues were collected. Serum was used for hormone determination, while ovarian tissues underwent HE staining, antioxidant level detection, and RNA-seq analysis. The results indicated that extending the photoperiod increased the number of primary follicles, secondary follicles, and total follicles (P < 0.01), as well as small white follicles (P < 0.05) and large white follicles (P < 0.05) in Taihe black-boned silky fowl. However, it did not significantly affect the number of small yellow follicles, large yellow follicles, hierarchical follicles, or post-ovulatory follicles (P>0.05). Simultaneously, extending the photoperiod significantly increased serum FSH and LH levels in the chickens (P < 0.05), but had no significant impact on E2, P4, PRL, and Mel levels (P>0.05). Furthermore, extending the photoperiod did not significantly influence ovarian free radical scavenging rate, MDA, CAT, and ROS levels (P>0.05). RNA-seq analysis of ovarian tissue revealed a total of 230 differentially expressed genes (DEGs) between the two groups, primarily enriched in signaling pathways such as neuroactive ligand-receptor interaction, tryptophan metabolism, circadian rhythm, and the GnRH signaling pathway. Notably, the expression of APOLD1, KCNK3, RORC, and SEMA6C was significantly upregulated. These findings suggest that extending the photoperiod activates primordial follicles, promotes follicle development, increases the number of follicles at all stages, and ultimately improves the laying performance of Taihe black-boned silky fowl.

The study aimed to explore the genetic characteristics of clutch traits and screen appropriate indicators to improve the reproductive performance of broiler breeders. In this study, a total of 4 273 hens from two generations of the Yellow-feathered broiler GF33A line were used as the research population, and the clutch traits were taken as the research object. By comprehensively applying methods such as the division of egg-laying time nodes, estimation of genetic parameters, and calculation of relative selection efficiency, the genetic characteristics of clutch traits and their genetic relationship with the egg number were systematically explored, and then the selection indicators of clutch traits for improving the egg number were screened out. Through an in-depth analysis of the changing trend of the egg-laying rate, 4 key egg-laying time nodes were divided, which were 35, 43, 51, and 63 weeks of age. Based on the above-mentioned egg-laying time nodes, 4 egg-laying periods were defined, and the genetic parameters of the clutch traits in these 4 periods were estimated. At the same time, the relative selection efficiency between these traits and the egg number at 63 weeks of age was calculated. The results showed that 3 indicators, namely the number of pauses in the first period (NP1), the number of clutches in the second period (NC2), and the number of clutches in the third period (NC3), had high heritability, a strong genetic relationship with the egg number at 63 weeks of age, and high relative selection efficiency. NP1, NC2, and NC3 can be used as candidate indicators for the selection of the egg number by hens. Through negative selection, the improvement of the egg number throughout the whole period can be indirectly achieved, providing a scientific basis for the improvement of the reproductive performance of broiler breeders.

This experiment aimed to investigate the changes of eggshell quality and its influencing factors in late-phase laying hens in order to improve eggshell quality and reduce the loss of eggs in the process of trading. In this experiment, 60 laying hens in peak laying period (D280) and 60 in late laying period (D580) were selected with similar weight and good health. Laying hens in each laying period were randomly divided into 6 replicates of 10 hens each for sampling, detection, and observation. Firstly, 60 eggs from each group were collected to detect the eggshell quality. And the differences of ultrastructure of eggshell were observed by scanning electron microscope. Subsequently, 6 chickens in each group were randomly selected to collect eggshell gland tissue, part of which was made into paraffin sections for observation of eggshell gland tissue structure, and part of which was extracted with RNA for detection of gene expression levels of OC-17, OC-116 and OCX-32 in eggshell gland tissue. The results showed that, compared with peak-phase laying hens, the following changes were observed in late-phase of laying hens: ①The thickness, strength, percentage and effective thickness of eggshell were significantly decreased by 8.33%, 9.98%, 5.15% and 4.17%. ②The ultrastructure of eggshell showed that the width of the mastoid gap, the width of the mammillary knobs and the effective thickness of the mastoid layer were significantly increased. ③The length of eggshell gland villi was significantly reduced, while the width between two adjacent villi showed a significant increase. ④The expression levels of OC-17, OC-116 and OCX-32 genes in eggshell glands were significantly down-regulated. The results indicated that the structural changes of eggshell gland in late-phase laying hens might lead to the decrease of eggshell gland secretion ability and affected the concentration of mineral ions in uterine fluid. The significantly down-regulated expression of OC-17, OC-116 and OCX-32 genes indicated the mineral deposit ability of eggshell gland was decreased. These two aspects together affected the mineralization process of eggshell in eggshell gland, changed the ultrastructure of eggshell, and led to the decline of eggshell quality.

This study aimed to evaluate the effectiveness of machine learning (ML) algorithms based on genomic markers in breed classification and to examine the performance of different ML algorithms in the classification of sheep breeds. In this study, 2 methods were used to select single nucleotide polymorphisms (SNPs) sites for 10 sheep breeds. The first method used the fixation index (F_ST) for selection, and the second method used the Boruta feature selection algorithm based on F_ST to further screen the SNPs sites. The 8 different types of ML algorithms, including K-nearest neighbor, support vector machines (SVM) and adaptive boosting (AdaBoost), were used to classify sheep breeds. The accuracy was used to evaluate the differences between different SNPs selection methods and different ML algorithms in breed identification, and the best combination method for sheep breed classification was identified. The data used in this study included both genetically distantly related varieties and genetically similar varieties, ensuring the reliability of subsequent analysis. Based on the top 1% selection criteria, the F_ST analysis identified 5 361 SNPs loci in each run, while the Boruta algorithm ultimately retained (328±11.7) SNPs loci for ML-based breed classification. After multiple iterations, SNPs loci marked as "confirmed" by the Boruta algorithm consistently scored higher than shadow features and the other two categories of SNPs loci. The number of SNPs loci retained by the Boruta algorithm was significantly lower than that of the F_ST analysis. When ML models were used for breed classification, most models achieved an accuracy above 0.9. The SVM model, using SNPs loci selected by the Boruta algorithm, achieved the highest classification accuracy (0.953), followed closely by AdaBoost (0.947). In contrast, the NB model, using SNPs loci selected solely by the F_ST analysis, showed the lowest classification accuracy (0.601). Except for NB, the area under the receiver operating characteristic curve (AUC) for the other models was close to 1. Regardless of the SNPs selection method, both approaches demonstrated strong discriminatory power, with slightly better performance observed after applying the Boruta algorithm. The results indicate that the implementation of ML methods effectively improves the accuracy of breed classification and demonstrates strong potential for application in sheep breed identification.

The purpose of this study was to detect the selection signal of SNPs microarray genotyping results of Hu sheep, explore the population structure of Hu sheep, and mine candidate genes related to important economic traits of Hu sheep, so as to provide a theoretical basis for further innovation and utilization of Hu sheep breeds. The total of 396 individuals (320 ewes and 76 rams) from the high breeding Hu sheep population were genotyped using sheep 50K SNP chip. After quality control, the selected Hu sheep populations were analyzed by principal component analysis and genetic relationship evaluation. The candidate regions were comprehensively screened and annotated by integrating haplotype score (IHS), neutral test statistic (Tajima 's D), composite likelihood ratio test (CLR) and continuous homozygous fragment (ROH). The functions of candidate genes were determined by GO-KEGG enrichment analysis, NCBI database, literature retrieval and GWAS ATLAS database. The results of principal component analysis showed that most Hu sheep individuals were concentrated in distribution, and the results of genetic relationship showed that the relationship between Hu sheep individuals was far, and the average inbreeding index of Hu sheep population was F_ROH=0.010 3, indicating that the inbreeding level of Hu sheep population was low. At the same time, the results of ROH analysis showed that the vast majority of ROH fragments were short ROH fragments < 10 Mb, indicating that Hu sheep had been inbred in the distant generation. Set the top 5% of IHS, Tajima 's D, CLR and ROH results in the population as the selected region, and a total of 45.86 MB candidate regions were found, accounting for about 1.75% of the sheep reference genome. After gene annotation, 1 424 genes were detected by iHS, 760 genes were detected by Tajima 's D, 964 genes were detected by CLR, 647 genes were detected by ROH, and 337 candidate genes were mined from the gene intersection obtained by two or more methods. GO-KEGG enrichment results showed that GO analysis enriched (P < 0.05) to items such as keratinization, intermediate filament tissue and skeletal system development, and KEGG analysis enriched (P < 0.05) to pathways such as ECM receptor interaction, arrhythmogenic right ventricular cardiomyopathy and GnRH secretion. Functional annotation found 65 candidate genes related to important economic traits, most of which were related to growth and reproduction traits. This experiment analyzed the high breeding single population of Hu sheep, and screened out the genes related to the growth traits of Hu sheep, including birth weight (CAPN3、ERLEC1、LAP3), bone development (ACAN、COL5A2、HAPLN1、HAPLN3), muscle development (FLVCR1、MUSTN1、PDLIM5), feed conversion rate and feed intake (PLIN1、CCSER1、LAP3) and body size (CSMD3、GPC6、LCORL). The genes related to the reproductive traits of Hu sheep included litter size (BMPR1B、GPRIN3、HCRTR1、MEPE、MAST4).And the genes related to other economic traits provide a certain theoretical basis and technical support for improving the breeding efficiency of Hu sheep breeds and optimizing the breeding methods of sheep breeds.

The study aimed to explore the molecular mechanism of foie gras formation, liver samples of Landes geese at 3 different feeding stages were analyzed by 4D-DIA quantitative proteomics. The 70-day-old Landes geese of the same batch with similar body condition were randomly selected and slaughtered at the early stage (7 d), middle stage (16 d) and final stage (25 d), 3 in each stage, respectively. The liver lobe and liver apex tissue samples were collected for 4D-DIA quantitative proteomic analysis. There were 3 groups with 3 biological replicates each. Result: 1) A total of 5 208 proteins were identified by comparison of the Landes geese liver at three different feeding stages. 2) Principal component analysis revealed significant differences in liver protein profiles across the 3 over-feeding stages, with 449 differential proteins identified in the comparison of early stage vs. middle stage, and 303 differential proteins identified in the comparison of middle stage vs. final stage. 3) Through pathway analysis of the differential metabolites, the enriched pathways were observed to be mainly involved in steroid hormone biosynthesis, ADIPOQ signaling pathway, amino sugar and nucleotide sugar metabolism, cysteine and methionine metabolism, arachidonic acid metabolism, and amino acid biosynthesis. The key differential genes ADIPOQ and PASK were identified based on RT-qPCR. The proteomic profile of liver of geese was significantly changed after over-feeding. The proteins related to steroid hormone biosynthesis and adipocytokine signaling pathway may be involved in the molecular regulation of foie gras formation.

This study aimed to explore the genetic diversity of the mitochondrial genome and maternal origin of cattle in Tibet Autonomous Region. In this study, the genetic diversity and their maternal origin of 167 individuals from 10 populations in Tibet Autonomous Region were analyzed by comparing mitochondrial genome. Through phylogenetic and genetic diversity analysis, 120 haplotypes from 167 individuals were detected including the T2, T3 and T4 haplotypes of taurine cattle; the Q haplotype of Bos primigenius; the I1 and I2 haplotypes of indicine cattle; and the haplotype of yak. The T3 haplotype also included T3₁₁₉ and T3₀₅₅, two subhaplotypes specific to East Asian taurine cattle. The results of genetic diversity analysis revealed that the haplotype diversity of Basang cattle was the highest (0.990±0.028), while the lowest was Bailang cattle (0.867±0.107), the highest nucleotide diversity was Jilong cattle (0.030±0.000 5), and the lowest was Shigatse cattle (0.001±0.000 2). The phylogenetic tree and haplotype network diagram revealed that the 10 cattle groups were mainly of taurine origin, and a small number of yak and indicine origin. Overall, the 10 cattle populations all presented high genetic diversity, with the maternal origins of taurine cattle, indicine cattle and yak, indicating that the maternal genetic diversity of cattle was high and that there was genetic exchange with yak. These results are important for understanding the genetic evolution, protection and utilization of valuable genetic resource of cattle in Tibet Autonomous Region

This study aimed to analyze the effect of health events on fertility, milk production and culling rates in dairy cows by using large-scale population data. Data were collected from 58 549 lactating cows calved between 2015 and 2021 in 13 farms in Ningxia, included 25 health events (6 composite traits and 19 independent traits), 4 fertility traits, 5 milk production traits and culling rates data. The GLM process of SAS 9.2 was used to analyze the impact of health events on economically important traits; Logistic regression models were used to analyze the effect of health events on culling rate of cows. The occurrence of health events lengthened the interval from calving to first insemination (ICF) by 1.51 d, increased the interval from first to last insemination (IFL) by 13.24 d, increased the days open (DO) by 15.42 d and the calving interval (CI) by 14.92 d. In independent traits, endometritis, enteritis and hoof disease had the greater impact on reproductive performance. The increased frequency of health events had a significant impact on the reproductive performance of cows. At the same level of health, reproductive performance of primiparous cows was better than that of multiparous cows. The sensitivity of ICF and DO to health events was higher in primiparous cows than in multiparous cows. Clinical mastitis, abortion, diarrhoea and displacement of abomasum significantly affected milk production traits in lactating cows, reducing 305-d milk yield (M305) by 1 225.99-2 416.48 kg, peak milk yield (PY) by 6.29-8.52 kg, protein yield (Pday) by 0.14-0.22 kg·d^-1 and fat yield (Fday) by 0.16-0.22 kg·d^-1. The increased frequency and class of health events had a significant impact on milk production performance of dairy cows. The M305, PY, Pday and Fday of multiparous cows were more sensitive to health events than that of primiparous cows. All types of health events significantly increased the culling risk of cows (1.45-17.81-fold), and cow with hoof sole cuticle splitting (HSCS) increased the culling risk a 17.81-fold. Based on large-scale herd health records, this study found that all types of health evens seriously affect the production performance of dairy cows, and the more frequency and type of events, the greater the impact. The sensitivity of primiparous and parturient cows to health events is different. The results of this study investigated the importance health events, clarified the specific degree of influence on important economic traits, provided theoretical references for farm management, and provided theoretical support for research and disease-resistant selection of dairy cow.

This study aimed to explore the mechanism of glutamine action during the cryopreservation process of sheep sperm. Semen samples were collected from 6 healthy adult Hu sheep aged 2-4 years. Frozen-thawed sperm motility, plasma membrane and acrosome integrity, mitochondrial membrane potential, ATP content, α-ketoglutarate dehydrogenase (α-KGDH) activity, isocitrate dehydrogenase (IDH) activity, and the expression levels of HSP70, HSF1, and phosphorylated HSF1 (pHSF1) proteins were evaluated. The results demonstrated that the addition of glutamine to the freezing extender significantly improved the total motility of sheep sperm (P < 0.05). The 5 mmol·L^-1 glutamine group exhibited significantly higher frozen-thawed sperm motility, plasma membrane and acrosome integrity, mitochondrial membrane potential, ATP content, α-KGDH activity, and IDH activity compared to the control group (P < 0.05). Glutamine significantly upregulated the expression of HSP70 and pHSF1 in frozen-thawed sperm (P < 0.05). The HSF1 inhibitor DTHIB suppressed this process and significantly reduced the total motility of frozen-thawed sperm (P < 0.05). Exogenous supplementation of HSP70 protein counteracted the inhibitory effect of DTHIB and significantly alleviated the reduction in sperm total motility induced by DTHIB treatment (P < 0.05). In this study, various quality parameters of frozen-thawed semen from sheep were examined and found that glutamine could improve the cryopreservation of sheep sperm by upregulating the phosphorylation level of HSF1 protein, increasing the expression of HSP70 protein, and protecting the mitochondrial function in frozen-thawed sperm, thereby enhancing the cryopreservation effect of sheep sperm.

The experiment aimed to investigate the effects of interfering and overexpressing GnIH on the proliferation, apoptosis and estrogen secretion in mouse ovarian granulosa cells. In this study, healthy female ICR mouse of 21-23 days old were selected. GCs were isolated from the mouse ovaries and identified using follicle-stimulating hormone receptor (FSHR). qRT-PCR was used to detect the expression of GnIH and GnIH-R genes in mouse granulosa cells. pshRNA-1, 2, 3, 4 interfering plasmid vectors and pLV-GnIH overexpressed plasmid vectors were constructed to overexpress and interfere with GnIH gene in mouse granulosa cells, respectively (GnIH interference groups were divided into pshNC group and pshRNA1-4 groups, and GnIH overexpression groups were divided into PLV-NC group and PLV-GnIH group). The interference and overexpression efficiency were detected by lentivirus titer detection and RT-qPCR, and the effects of psh-RNA and pLV-RNA on the proliferation and apoptosis of granulosa cells were detected by MTT and Annexin V-FITC/PI flow cytometry. The concentrations of progesterone and estradiol were detected by ELISA, all the above results were repeated 3 times. The results showed that pshRNA-1, 2, 3, 4 significantly interfered with GnIH expression at 48 h and 96 h (P < 0.01), but only pshRNA-4 significantly decreased GnIH expression at 72 h (P < 0.01). At 72 h, pLV-GnIH significantly up-regulated the expression of GnIH (P < 0.001). Overexpression of GnIH significantly down-regulated the proliferation activity of granulosa cells and promoted cell apoptosis (P < 0.001), and significantly inhibited granulosa cell estrogen secretion (P < 0.001). After transfection with pshRNA-2, 4 for 72 and 96 h, the proliferative activity of granulosa cells was extremely significantly increased (P < 0.01), the secretion of estrogen was significantly enhanced (P < 0.05), and apoptosis was significantly decreased(P < 0.05 and P < 0.01). In conclusion, inhibition of GnIH expression can promote the proliferation of granulosa cells and the secretion of estrogen, and down-regulate cell apoptosis, while promotion of GnIH expression can inhibit the proliferation of granulosa cells and the secretion of estrogen, and promote cell apoptosis.

Co-administration of porcine epidermal growth factor (pEGF) and trefoil factor 3 has been shown to be effective in promoting intestinal proliferation and repair in pigs. Herein, this study was to construct a food-grade recombinant Lactococcus lactis strong constitutive expression system for the fusion expression of pEGF as well as a porcine gut beneficial protein p40, derived from Lactobacillus rhamnosus. As such, the researchers sought to overcome the current status quo of the low levels of L. lactis recombinant constitutive expression as well as to avoid the use of antibiotics, and to preliminarily develop a more potent porcine gut beneficial agent. Firstly, using the commonly used L. lactis constitutive promoter P32 as a control and the Staphylococcus aureus nuclease reporter system, this study visually screened out the promoter of the elongation factor Tu of L. lactis NZ9000 (P1) outperformed the other promoters of L. lactis NZ9000 constitutively highly expressed proteins and the previously found strong constitutive promoters of L. lactis. Furthermore, by replacing the chloramphenicol resistance gene of pNZ8148 with the nisin resistance gene, a food-grade recombinant L. lactis strong constitutive expression system named NZ9000/pP1NR was successfully constructed, which possessed good nisin resistance and completely lost the chloramphenicol resistance. Using the system, pEGF and the fusion protein pEGF-p40 were successfully expressed. Cell proliferation assays on IPEC-J2 cells showed that the culture supernatant containing pEGF-p40 was more effectively in promoting the proliferation. SGLT-1, GLUT-2, SUC, EGFR and GLP2R were detected in IPEC-J2 cells after stimulation by qRT-PCR, indicating that the culture supernatant containing pEGF-p40 regulated the expression of the relevant genes differently from that of the culture supernatant containing pEGF alone. In this study, a food-grade recombinant L. lactis strong constitutive expression system was constructed. By which, pEGF-p40 with a beneficial effect on porcine intestines, was successfully expressed. This study provides a reference for the safe production of other probiotic substances, and preliminarily explored a potent beneficial preparation for the porcine intestine.

This study aims to isolate and identify a strain of PoRV, and analyze the genetic variation of its genome sequence and the pathogenicity of the virus. Small intestinal samples of diarrheal piglets were selected and processed, then inoculated into MA104 cells for isolation and culture. The cell cultures were identified through viral nucleic acid test, serological detection and morphological observation. RT-PCR and sequencing were used to obtain the whole genome sequence of the isolate, genetic evolutionary relationship and pathogenicity were evaluated. The results showed that a PoRV strain were successfully isolated and can be cultured in vitro, named HLJ/2021, and its genotype is G5-P[7]-I5-R1-C1-M1-A1-N1-T1-E1-H1 type. Pathogenicity experimental results showed that the isolate can cause watery diarrhea, emaciation and other symptoms in piglets. Virus shedding can be detected in piglet feces 60~174 h after infection. Necropsy revealed that the intestinal wall of the infected piglets became thinner and the contents were watery. Histopathological examination showed that the intestinal villi of the ileum of the infected piglets were highly atrophied. Viral load detection results in intestinal tissues showed that virus can be detected in the ileum, cecum, colon and rectum, with the highest viral load in the ileum. In summary, this study successfully isolated a G5P[7] type PoRV strain, which is pathogenic to piglets. The results provide important information for understanding the prevalence of PoRV.

The aim of this study was to investigate the inhibitory effect of baicalin on porcine reproductive and respiratory syndrome virus (PRRSV) in vitro.Using real-time quantitative polymerase chain reaction (RT-qPCR), indirect immunofluorescence (IFA), half tissue culture infection dose (TCID₅₀) test to observe the three modes of action of baicalin on PRRSV, including infection blocking, direct killing and proliferation inhibition; and four steps including adsorption, internalization, replication and release; as well as the direct interaction with PRRSV virus particles. The results found that baicalin could inhibit PRRSV replication under different modes of action, and the inhibition effect was significantly at 4 h of infection blockade (P < 0.05); baicalin could inhibit the PRRSV replication process in the adsorption and internalization stages (P < 0.05) and was dose-dependently; baicalin could directly interact with PRRSV virions and inhibit PRRSV replication (P < 0.05). In conclusion, baicalin can inhibit PRRSV replication in vitro.

The aim of this study was to screen for the binding of the 3′UTR to the negative-strand RNA during replication of the foot and mouth disease virus (FMDV) negative-strand RNA. Firstly, we identified the host proteins that bind to the 3′UTR negative-strand by RNA pull down combined with mass spectrometry, and then we performed GO functional annotation analysis, KEGG pathway enrichment analysis, and screened of the interaction network on the identified proteins. We constructed eukaryotic plasmids as well as prokaryotic plasmids to express the proteins to validate the specificity of the interactions between the RNAs and the proteins. Secondly, we truncated the 3′UTR to explore the interaction region between RNA and protein. Finally, we used Co-IP to explore the interaction relationship between the interaction protein and viral protein. It was found that most of the proteins bound to the 3′UTR interactions were nucleolar proteins, among which the Ddx18 protein of the DEAD-box RNA helicase protein family was the focus of the study. By constructing the eukaryotic expression vector of this protein, we confirmed that Ddx18 did interact with the 3′UTR negative-strand using RNA pull down, RIP, and EMSA. A truncated 3′UTR negative-strand RNA was transcribed, and the RNA pull down assay was used to conclude that Ddx18 interact with the 3′UTR negative-strand. It does not depend on poly(U) and not interact with the truncated SL1 and SL2. Using Co-IP assay, it was demonstrated that 2C viral proteins can interact with Ddx18 proteins. In summary, the interactions between 3′UTR and Ddx18 protein may be based on spatial structure, whereas the 2C viral proteins bind to the Ddx18 protein and may indirectly bind to the 3′UTR and together form a replication complex that is involved in the replication of FMDV. By screening the host proteins that interact with the negative-strand of the 3′UTR, we can lay the experimental foundation for further investigating the mechanism related to FMDV replication and thus the related drug targets.

This study aims to evaluate aluminum-manganese bimetallic organic framework nanoparticles modified with Eudragit L100 as an oral delivery vehicle for inactivated porcine epidemic diarrhea virus (PEDV). By utilizing the coordination interaction between metal ions (Al³⁺, Mn²⁺) and the organic ligand 2-aminoterephthalic acid, inactivated PEDV is encapsulated in situ within Al/Mn-MOF to form Al/Mn-MOF@PEDV nanoparticles. An anionic polymer, Eudragit L100, is then electrostatically attached to the surface of Al/Mn-MOF@PEDV nanoparticles, producing L100@Al/Mn-MOF@PEDV nanoparticles. The morphology is examined using scanning electron microscopy, the surface charge is measured by zeta potential meter, the composition is analyzed by fourier transform infrared spectroscopy, and the crystal structure is determined by X-ray Diffraction. The loading efficiency of inactivated PEDV is characterized using ELISA and antigen detection cards. The stability of L100@Al/Mn-MOF@PEDV nanoparticles in simulated gastric fluid and the release rate of inactivated PEDV are examined using ELISA. Macrophage uptake of the nanoparticles is observed using confocal laser scanning microscopy. The cytotoxicity of nanoparticles with different concentrations is determined by CCK8 method, and the effect on the secretion of immunomodulatory factors by macrophages is evaluated using ELISA kits. Results indicate successful preparation of L100@Al/Mn-MOF@PEDV nanoparticles, with sizes under 1 μm. At a PEDV/metal ion/organic ligand ratio of 0.42 ∶1 ∶1, the loading rate of inactivated PEDV reaches 99.6%. After 2 hours in simulated gastric fluid, L100@Al/Mn-MOF@PEDV nanoparticles maintained high stability with only about 12.73% release rate of inactivated PEDV. Uptake experiments in mouse RAW264.7 cells show enhanced uptake of Al/Mn-MOF@PEDV compared to unencapsulated inactivated PEDV. Additionally, Al/Mn-MOF@PEDV significantly boosts the expression of Th1-type factor IFN-γ and Th2-type factor IL-4 in RAW264.7 cells. This research offers new insights into the design of oral carriers for inactivated PED vaccines and lays the foundation for future applications in animal immunization.

The aim of this study was to develop a nanoparticle vaccine targeting the E2 protein of classical swine fever virus (CSFV) and analyze its immunogenicity. Using the E2 gene sequence of Shimen strain of CSFV as template, the human codon was optimized in its extracellular domain. E2, E2-pFc (fusion expression of E2 with Fc fragment of porcine antibody), SpyTag-E2 (fusion expression of E2 with SpyTag) and the nanoskeleton protein LS (Lumazine Synthase-protein A) were prepared by 293F mammal cell eukaryotic expression system. Nanoskeleton protein mi3 (mi3-SpyCatcher) was purified by prokaryotic expression system and then LS and mi3 assembled into nanoparticles LS-E2-pFc NP and mi3-E2 NP, which were identified by transmission electron microscopy and dynamic light scattering test, and each protein was emulsified with ISA201 adjuvant to prepare subunit vaccines. Thirty-five Japanese white rabbits (1.5-2.0 kg) were randomly divided into 7 groups. Blank control group (n=3), PBS negative control group (n=3), E2 group (n=5), E2-pFc group (n=5), LS-E2-pFc NP group (n=5), mi3-E2 NP group (n=5), and CSFV E2 commercial subunit vaccine group (n=3). On 0 d and 21 d, intramuscular immunization was administered, and on 42 d, CSFV C strain was challenged with 100 RID₅₀ via ear vein. The specific antibody level, neutralizing antibody level, body temperature response and spleen viral load of rabbits were evaluated by blocking ELISA, neutralizing test, temperature monitoring and RT-qPCR. The results showed the successful expression of all component proteins, with single-band profiles and high purity. The average particle size of mi3-E2 nanoparticles (68 nm) was larger than mi3 (42.73 nm), and LS-E2-pFc nanoparticles (42.4 nm) was larger than LS (13 nm). Immunization results showed that E2, E2-pFc, LS-E2-pFc NP, mi3-E2 NP vaccine and commercial subunit vaccine all had good immune effect, and 7 days before challenge, the specific antibody blocking rate in serum was above 89%, with no significant difference between immunized groups (P>0.05). After challenge, the PBS negative control group showed typical stereotyped heat reaction, while the other immune groups showed no obvious changes in body temperature. Seven days post-challenge, the titers of neutralizing antibodies of E2, E2-pFc, LS-E2-pFc NP, mi3-E2 NP vaccine and commercial subunit vaccine groups were 1:10 392.4, 1:37 168.4, 1:75 473.6, 1:20 201.8 and 1:2 407, respectively, with the LS-E2-pFc NP group showing the highest neutralizing antibody levels. RT-qPCR results showed that one rabbit in the E2 group had a low viral load of 5.83 copies ·μL^-1, while no viral load was detected in other immunized groups. The above results showed that LS-E2-pFc NP immunized group had the best effect and the highest titer of neutralizing antibody. This "marker" vaccine allows for differential diagnosis of naturally infected and vaccinated animals, helping to eradicate CSF in swine farms.

Mannheimia haemolytica (Mh) is one of the most important pathogens causing bovine respiratory disease (BRD), which seriously affects the healthy breeding and food safety of cattle. In this study, nasal swabs of calves with respiratory symptoms such as fever and runny nose were collected for bacterial isolation and culture, and serotype identification, 16S rRNA sequencing analysis and PCR typing of the isolates were performed. The drug-resistant phenotype of the isolates to 15 antibiotics was tested, the possibility of mice as Mh replacement animal infection models was evaluated by mortality and pathological changes of the mice. A mouse bacterial infection model was constructed, and the pathogenicity and immunogenicity of the isolates were evaluated. The results showed that the purified strain was identified as Mh and named KQ-Mh-1.The strain was resistant to amicacin, melocillin, gentamicin, streptomycin, amoxicillin and sulfamethoxazole, and showed high sensitivity to cefotaxime, norfloxacin, cefoperazone, ciprofloxacin, polymyxin, neomycin and doxycycline. The median lethal dose (LD₅₀) of KQ-Mh-1 on BALB/c mice was 7.29×10⁹ CFU ·mL^-1, and the infected mice all showed severe hemorrhages in the lung and spleen. KQ-Mh-1 was prepared into inactivated vaccines with different antigen content to evaluate its immunogenicity, in which the inactivated vaccine with high antigen content (2.5×10¹⁰ CFU ·mL^-1) had a 70% protection rate against KQ-Mh-1 after immunizing mice. In this study, a Mh strain that can cause bovine BRD was successfully isolated and its biological characteristics, pathogenicity and immunogenicity were investigated, which provided a good vaccine candidate strain for the research and development of bovine BRD vaccine.

This study conducted drug susceptibility tests, as well as resistance and virulence gene detection, on 107 Enterococcus faecalis strains isolated from mastitis cases to understand the drug-resistance characteristics and virulence of E. faecalis from bovine mastitis. E. faecalis strain F2 was selected as a representative strain for whole-genome sequencing and biological characterization. Further, bovine mammary epithelial cells (BMECs) were infected with Enterococcus JH2-2 and E. faecalis F2 at a multiplicity of infection (MOI) of 1 000 to assess the adhesive and invasive abilities of strain F2, as well as its impact on BMECs damage and proliferation. A larval infection model using Galleria mellonella was established to verify the pathological features of tissue damage caused by this strain through larval survival rates and histopathological analyses. The results indicated that E. faecalis strains from bovine mastitis exhibited the highest resistance rates to lincomycin, tetracycline, and erythromycin, ranging between 90% and 100%. Among the isolates, 95 strains carried resistance genes, and 84 strains carried virulence genes. The most frequently detected resistance gene was ermB (91.6%), and the most common virulence gene was esp (78.6%). Whole-genome sequencing of strain F2 revealed resistance to 23 classes of antibiotics, including lincosamides, macrolides, and quinolones. Additionally, three integrative and conjugative elements (ICEs) carrying resistance genes and 91 associated virulence factors, such as efaA, gelE, ace, and AS, were identified. Strain F2 demonstrated strong environmental tolerance, surviving under high temperatures, acidic, and alkaline conditions, and exhibited motility and robust biofilm formation capabilities (OD_{570 nm}>2 ODc). Six hours post-infection of BMECs, strain F2 showed significantly greater adhesion and invasion abilities (P < 0.001) compared to the control group. Over time, strain F2 exhibited increased toxicity to BMECs, leading to severe cell damage and death. Following infection of G. mellonella larvae with strain F2 for one day, larval mortality reached 100%, accompanied by extensive inflammatory cell infiltration and severe disruption of the larval body cavity. This study reveals that E. faecalis isolates from bovine mastitis exhibit severe multidrug resistance, harbor multiple ICEs carrying resistance genes, and possess numerous virulence factors, contributing to BMECs damage. These findings provide an experimental foundation for further research on the transmission of antimicrobial resistance and pathogenicity of E. faecalis from bovine sources.

This study aimed to determine the molecular basis of the pathogenicity of Pasteurella multocida(P. multocida). The pathogenicity and whole genomic sequences of 2 rabbit sourced serogroup A P. multocida isolates (Pm3 and Pm6) were determined, and then comparative genomics analysis between the 2 isolates and other P. multocida strains was performed. The intranasal inoculation of Pm3 and Pm6 caused severe pathologic lesions and high mortality in inoculated rabbits. By comparison with the low-virulent rabbit-sourced P. multocida strains s4 and AH09, about 120 specific functional genes were identified in the genomes of Pm3 and Pm6, and about 100 of these specific functional genes could also be detected in the genomes of high-virulent rabbit-sourced P. multocida strains. Moreover, different nucleotide polymorphism profiles were identified in the lipopolysaccharide (LPS) outer core biosynthetic genes natC and gatF among the P. multocida strains of L3 genotype isolated from different hosts. Nevertheless, the sequences of the natC and gatF were identical among the rabbit-sourced P. multocida strains of LPS L3 genotype (except for SD11, CIRMBP-0873 and CIRMBP-0884), which suggested that the potential species specificity of the natC and gatF. The Pm3 and Pm6 were highly pathogenic for rabbits, which indicated that the 2 isolates were high-virulent strains. The specific functional genes that identified in the genomes of Pm3, Pm6 and the previously reported high-virulent rabbit-sourced P. multocida strains but not in the genomes of low-virulent rabbit-sourced P. multocida strains s4 and AH09 might contribute to the pathogenicity of the Pm3 and Pm6.

In recent years, the risk factors for Toxoplasma gondii infection in humans and animals have increased, so the investigation of the prevalence of T. gondii has become more important. In view of the fluorescence characteristics of lanthanide microspheres, such as long life span, large stockes displacement and excellent photostability, this study applied the fluorescent microspheres as probes for the detection of T. gondii antibodies in serum. In order to improve the accuracy of the test, a mixture of three antigens of soluble expressed SAG1, SAG2, and GRA1 was used as the coating antigens, and fluorescent microsphere-labelled SPA was used as the probe to develop test strips for the detection of T. gondii antibodies by indirect immuno-lateral chromatography, and the conditions of the sampling method, the co-incubation time of the microsphere and sera, the reaction time were optimized, meanwhile, the specificity, sensitivity, reproducibility and reliability of the test strips were investigated. The results showed that the prepared lanthanide microsphere immunochromatographic test strips have the following characteristics: good specificity, no cross-reactivity with other pathogenic sera; high sensitivity, the positive sera can still be detected at a dilution of 1 600 times; fast detection speed, the results can be obtained in as fast as 10 minutes; stable and reliable, the Kappa analyses of serum samples show that the lanthanide microsphere test strips have good consistency with the two ELISA assays. The present study provides a rapid, sensitive and quantitative immunochromatographic method for the detection of T. gondii antibodies.

The aim of this study was to prepare a monoclonal antibody to avian adenovirus type 4 (FAdV-4), to characterize its specific binding site, and to evaluate its value for use in immunohistochemistry (IHC) and immunoprecipitation reactions. Mice were immunized with purified FAdV-4-AH-F41 whole virus, a positive hybridoma cell line was screened and subcloned by indirect enzyme linked immunosorbent assay (ELISA), and the prepared monoclonal antibody was tested for potency. Indirect immunofluorescence assay (IFA) and protein immunoblotting (Western blot) were used to characterize the biological properties of the monoclonal antibodies. pCold TF-Hexon, pET-32a(+)-Fiber1, pCold TF-Penton, pCold TF-Fiber2 prokaryotic expression vectors, and the specific binding sites of the resulting monoclonal antibodies were preliminarily characterized by Western blot, and after the subtypes of the antibodies were detected, they were initially applied to IHC and immunoprecipitation experiments. The results showed that a monoclonal antibody strain with stable secretion of anti-Fiber1 protein was obtained in this experiment, and it was named 5C7. The antibody had a potency of 1∶102 400, and its subtype was IgG-2b, which showed good biological properties. In IHC, obvious lesion characteristics could be observed; in immunoprecipitation experiments, 5C7 could act as a capture antibody and bind specifically to the Fiber1 protein in virus-infected cells. The results suggest that the monoclonal antibody prepared in this experiment by immunizing mice with whole virus particles can specifically recognize the Fiber1 protein and shows good application prospects in IHC and immunoprecipitation reactions, which lays the foundation for the establishment of laboratory pathological diagnosis methods of FAdV-4 and the functional study of Fiber1 protein.

There is widespread misuse of antibiotics in pigeon breeding, leading to an increase in antibiotic-resistant strains and exacerbating multidrug resistance. The present study aimed to understand the prevalence of antibiotic resistance and resistance genes in Escherichia coli sourced from racing pigeons in Tongliao City, and to provide reference information for clinical treatment of E. coli infections in racing pigeons. Using E. coli selective culture media and methods such as 16S rRNA sequencing, pathogenic bacteria from diarrheic racing pigeons in Tongliao were isolated and purified. Drug susceptibility testing and PCR detection of resistance genes were conducted to assess the resistance profiles and distribution of resistance genes in the isolated strains. Whole genome sequencing analysis was performed on two highly resistant E. coli strains. Eight strains of E.coli sourced from racing pigeons were isolated, and all of them exhibiting multidrug resistance. The resistance rates to kanamycin and streptomycin were both 100%, while all strains were sensitive to ceftriaxone-sulbactam. Strains E-ge1 and E-ge2 showed severe resistance profiles. Whole genome sequencing analysis of these two strains revealed complete genome sequences, with lengths of 4 680 817 bp and 4 691 799 bp respectively. They encoded 4 888 and 5 070 genes respectively, and both strains carried three plasmids.E-ge1 carries a total of 81 antibiotic resistance genes, whereas E-ge2 harbors 74 such genes. The situation of antibiotic resistance in E.coli sourced from racing pigeons in Tongliao is severe, with strains carrying a large number of resistance genes. It is imperative to optimize treatment strategies to control the exacerbation of resistance and the spread of resistance genes.

African swine fever (ASF) is an acute, virulent and highly contagious disease of pigs caused by African swine fever virus (ASFV). In August 2018, ASFV was introduced into China for the first time, bringing a disastrous blow to the pig industry. In order to clarify the pathological characteristics of ASFV CN2018 strain, a case of pig artificially infected with ASFV CN2018 strain was studied. The experimental pig in this case died on day 11 after being artificially infected with ASFV CN2018 strain. Systematic necropsy and histopathological examination were performed. Necropsy showed systemic hemorrhagic septicemia. There were petechiae and ecchymosis in the skin, lymph nodes, kidneys, bladder, cecum, gallbladder, heart and brain, with hemathorax, hematocelia and pulmonary congestion and edema. Microscopic lesions were characterized by diffuse hemorrhage and congestion with microthrombus, and severe lymphocyte depletion in lymph nodes and spleen. Immunohistochemical staining showed that the ASFV antigen was positive in the cytoplasm of macrophages, renal tubular epithelial cells and hepatocytes. In summary, in this case, the pig artificially infected with ASFV CN2018 strain developed extensive hemorrhagic lesions, and the main target organs of the lesions and antigen distribution were similar to those reported previously. It is speculated that the extensive hemorrhage caused by ASFV infection is mainly related to the inflammatory cytokine storm caused by the activation of mononuclear macrophage system and the injury of vascular endothelial cells. Our results provide theoretical data for further study of the pathogenic mechanism of the prevalent ASFV strain in China.

The aim of this study was to carry out pathological diagnosis of different types of canine ovarian tumors, summarize and analyze their histopathological features, and provide reference for precise diagnosis of canine ovarian tumors. Cases of canine ovarian tumors diagnosed at the National Laboratory for Detection of Bovine Transmissible Spongiform Encephalopathy of the School of Zoology, College of Agricultural Sciences, China Agricultural University, and their detailed case information were retrospectively selected for histopathological diagnosis, and the characteristics of the cases and their morbidity were summarized. Results were as follows: Seven cases of ovarian epithelial tumors were diagnosed, including four cases of ovarian papillary adenoma, one case of ovarian papillary adenocarcinoma, one case of ovarian reticulum adenoma, and one case of ovarian reticulum cystadenoma; and eight cases of ovarian gonadal mesenchymal tumors were diagnosed, including three cases of ovarian granulosa cell tumor, one case of follicular membranous cell tumor, and four cases of luteoma. Dogs over 11 years of age accounted for 33.3% of all cases. These results showed that the incidence of ovarian epithelial tumors and gonadal mesenchymal stromal tumors were comparable, and the epithelial tumor with the highest incidence rate was ovarian papillary adenoma, while luteoma and granulosa cell tumors were frequent among gonadal mesenchymal stromal tumors; the prevalence rate of the old dogs was high, and the rate of prevalence increased with the increase of age; the occurrence of ovarian tumors had no significant correlation with the breeds of the dogs.

This study explored the reparative effect and mechanism of glutathione (GSH) on cadmium (Cd)-induced PK-15 cell injury. In the present study, we used MTT assay, cellular microscopy observation, JC-1 staining, comet assay, and flow cytometry to investigate the effects of GSH on Cd induced oxidative damage and apoptosis in PK-15 cells. Results showed that GSH significantly alleviated the decrease in cell viability and increase in ROS, attenuated DNA damage, mitochondrial damage, and apoptosis in Cd-treated PK-15 cell, and western blot analysis showed that GSH may inhibit Cd induced PK-15 cell apoptosis by upregulating the expression of anti-apoptotic protein Bcl-2 and downregulating the expression of pro-apoptotic protein Bax. Taken together, these data suggest that GSH can alleviate Cd induced PK-15 cell damage by inhibiting oxidative stress and cell apoptosis.

Porcine α interferon (PoIFN-α) can be used to treat and prevent viral infection. However, the natural pig α interferon has low antiviral activity, so it is of great significance to find the mutant sequence of high biological activity for the prevention and control measures of swine infectious diseases. After using bioinformatics methods to analyze and compare the amino acid sequences of various subtypes of PoIFN-α, PoIFN-α 8s mutants were constructed by site-directed mutagenesis of eight sites in the native PoIFN-α sequence and purified by prokaruclear expression. After obtaining the recombinant protein, activity assays were conducted against the vesicular stomatitis virus (VSV) in PK-15 cells and validated the inhibitory effect of the PoIFN-α 8s mutant on the pseudorabies virus (PRV) in mice. The results showed that, antiviral activity of recombinant PoIFN-α 8s was 1.13×10⁷ IU·mg^-1, which reduced the viral load and gene copy number of VSV on PK-15 cells by more than 10 times compared to the PoIFN-α product. Animal experiments indicated that PoIFN-α 8s can reduce the viral load in various organs of mice infected with PRV. Thus, this study successfully constructed an efficient PoIFN-α 8s mutant and verified its good antiviral activity both in vivo and in vitro.

The study aims to improve the diagnostic criteria of cat acute stress (CAS). The criteria were then applied to collect cats in the efficacy and safety trial of gabapentin tablets for CAS prevention. CAS was diagnosed by cat stress score (CSS). Fifteen healthy cats and fifteen acute stress cats were collected and divided into the healthy group and the acute stress group, respectively. Through the comparison of clinical data, CSS assessment, and the results of corticotropin releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and epinephrine (EPI) detection, so as to evaluate the diagnostic value for CAS of three kinds of hormone and improve the CAS diagnostic criteria. Then, using the improved diagnostic criteria for CAS in this study, 106 acute stress cats were collected and randomly divided into the placebo group and the gabapentin group. A total of three visits were scheduled, and 1.5 h before the second visit, the cats in the two groups received the recommended dose of placebo tablets and gabapentin tablets orally. The efficacy and safety of gabapentin tablets were evaluated by comparison of clinical data, basic physiological indexes examination, behavioral evaluation, blood routine examination, blood biochemical examination, hormone examination, and adverse reaction monitoring. The results showed that the clinical data of the healthy group and the acute stress group were homogeneous. When CRH, ACTH, and EPI were combined in the diagnosis of CAS, the best critical value was 0.212, the area under the curve was 0.973, the sensitivity was 100.00%, and the specificity was 86.67%. The results of the clinical trial of gabapentin tablets showed that the clinical data of the placebo group and the gabapentin group were homogeneous. Compared with the placebo group, the gabapentin group had significantly higher ΔCSS, Δtemperature, Δrespiratory rate, Δheart rate, ΔACTH, ΔEPI, and incidence of adverse effects (weakness, ataxia, and lethargy), while the Δglobal sedation score was significantly lower. The above adverse reactions occurred about 2-3 h after the drug administration and lasted about 1.5-3.0 h. Gabapentin tablets did not cause changes in blood routine and blood biochemical indicators. In conclusion, this study suggests that the diagnostic criteria of CAS can be improved as follows: when the cat is stimulated by a stressor within a few hours before the test, the predictive probability value of the combination of serum CRH (pg·mL^-1), ACTH (pg·mL^-1) and EPI (ng·mL^-1) concentrations during the test is greater than 0.212 and the CSS score is greater than 3. A single oral administration of gabapentin tablets at the recommended dose 1.5 h before a stressful event has a certain preventive effect on CAS, but adverse reaction such as atony, ataxia and somnolence may occur.

The study aims to explore the impact and mechanism of Radix pseudostellariae polysaccharide (RPP) in regulating miRNA Let-7d-3p on the inflammation caused by pseudorabies virus (PRV) infection. PK-15 cells were cultured in vitro and treated with various concentrations of RPP (0.5-32.0 mg·mL^-1). The cell viability was then assessed using the CCK-8 assay. And PK-15 cells were divided into the control group, PRV group, 0.5 mg·mL^-1 RPP group, 1.0 mg·mL^-1 RPP group, and 2.0 mg·mL^-1 RPP group. The expression levels of inflammatory cytokines IL-6, TNF-α, and IL-17 in cells from each group were detected using real-time fluorescent qPCR. Let-7d-3p inhibitors were transfected into PK-15 cells and then subjected to intervention with 0 or 0.5 mg·mL^-1 RPP. The expression levels of Let-7d-3p and inflammatory factors were examined using qPCR. In the vivo experiment, 15 healthy mice were randomly assigned to the control group, PRV group, and RPP (10.0 mg·kg^-1) group, with 5 mice in each group. The expression levels of Let-7d-3p and inflammatory factors in mouse kidney tissues were detected using qPCR. The results showed that within the concentration range of 0 to 4.0 mg·mL^-1, RPP has no cytotoxicity. Compared to the control group, the transcription levels of the inflammatory factors IL-6, TNF-α, and IL-17 mRNA in PK-15 cells infected by PRV are significantly up regulated (P < 0.01), while the transcription level of Let-7d-3p is significantly down regulated (P < 0.01). In comparison with the PRV group, the expression of Let-7d-3p was significantly up regulated (P < 0.01) after the cells were treated with three concentrations of RPP, while the transcription levels of the three inflammatory factors were concurrently and significantly down regulated (P < 0.01). After the inhibition of Let-7d-3p, compared with the PRV group, the transcription levels of IL-6, TNF-α, and IL-17 in the Let-7d-3p inhibition group were significantly up regulated (P < 0.01). Compared with the PRV+RPP group, after the suppression of Let-7d-3p and subsequent intervention with RPP, the transcription levels of cellular inflammatory factors were significantly up regulated (P < 0.01). In vivo results indicate that, in comparison with the control group, the transcriptional level of Let-7d-3p in the kidneys of mice in the PRV group was significantly decreased (P < 0.01), and the transcriptional levels of inflammatory factors IL-6, TNF-α, and IL-17 were significantly up regulated (P < 0.01). Compared with the PRV group, the expression of Let-7d-3p in the kidneys of mice treated with RPP was significantly increased (P < 0.01), and the transcription levels of the inflammatory factors IL-6, TNF-α, and IL-17 were significantly decreased (P < 0.01). Results from both in vivo and in vitro experiments indicate that RPP can downregulate the transcription levels of inflammatory genes induced by PRV infection through Let-7d-3p, providing a rationale for subsequent studies on the alleviation of PRV-induced inflammation by RPP.

The study aimed to investigate the bacteriostatic activity and the underlying bacteriostatic mechanism of tea saponins against a strain of porcine-originated multidrug-resistant enterotoxigenic Escherichia coli (ETEC), ultimately providing novel perspectives for the prevention and control of piglet diarrhea caused by ETEC infection. The Kirby-Bauer disk diffusion method was utilized to assess a series of antibiotics sensitivity of the porcine-originated ETEC strain. Furthermore, the minimum inhibitory concentration (MIC) of antibiotics and tea saponins against the ETEC strain was determined through the 96-well plate broth microdilution method. To elucidate the bacteriostatic effect of tea saponins against the ETEC strain, a comparative analysis was conducted on the growth curve, ultrastructure, nucleic acid and protein leakage, alkaline phosphatase (AKP) activity between the tea saponins treated and untreated samples. Meanwhile, transcriptomic technology was utilized to analyze the differences in bacterial gene expression profiles under the conditions of tea saponins treatment and non-tea saponins treatment, revealing the bacteriostatic mechanism of tea saponins on this porcine ETEC. Additionally, the safety and efficacy of tea saponins on IPEC-J2 cells were assessed utilizing the Cell Counting kit-8 assay. The drug sensitivity results indicated that the porcine-originated ETEC strain exhibited resistance to 9 out of the 14 commonly used antibiotics in clinical practice, classifying it as a multidrug-resistant strains. The MIC of tea saponins was 50 mg·mL^-1. At the tea saponins concentrations of 1×MIC and 2×MIC, it exhibited strong bacteriostatic activity against the porcine-originated multidrug-resistant ETEC strain, effectively inhibiting the growth and reproduction of the ETEC strain within 12 h. After treatment with tea saponins at 1×MIC and 2×MIC concentrations, the bacteria morphological structure was significant damaged, resulting in a significant increase in the concentration of nucleic acid, soluble protein, and AKP in the extracellular bacterial cells within the initial 2 h. Moreover, during the entire 12 h observation period, the concentrations of these substances were significantly higher than those in the control group, ultimately triggering to the death of bacteria. SDS-PAGE electrophoresis result revealed that the protein bands within the molecular weight range of 15-25 ku in the ETEC strain treated with tea saponins were significantly reduced. Transcriptome analysis further indicated that the bacteriostatic mechanisms of tea saponins might involve multiple levels, including the suppression of lipopolysaccharide biosynthesis, ribosomal function, protein synthesis processes and other related pathways. In addition, the study discovered that within the concentration range of 0.781-12.5 μg·mL^-1, tea saponins exhibited no discernible toxic side effects on IPEC-J2 cells. Particularly, at a concentration of 12.5 μg·mL^-1, tea saponins significantly enhanced the survival rate of IPEC-J2 cells after ETEC infection. In conclusion, tea saponins exhibited significant bacteriostatic effects against porcine-originated multidrug-resistant ETEC strain, with its bacteriostatic mechanisms being intricated and multifaceted. The results provide theoretical support for the application of tea saponins as a natural bacteriostatic agent in animal husbandry.

Chelidonium majus can be used to treat various digestive and respiratory tract inflammations, but its mechanism of action is still unclear. This study predicts and analyzes the pharmacodynamic components and mechanisms of C. majus using fingerprint and network pharmacology. A total of 15 batches of C. majus fingerprint were established using HPLC and evaluated for similarity and principal component analysis. A network of active components and targets of C. majus was established using network pharmacology methods. The core anti-inflammatory components were analyzed, along with the core targets through protein interaction network analysis, followed by GO enrichment and KEGG enrichment analysis. The common peaks identified by fingerprint and the core components filtered by network pharmacology were intersected. The effects of C. majus on IPEC-J2 cell inflammation damage were studied using MTT and ELISA methods. The results showed that 12 out of 15 batches of C. majus had a similarity >0.9, with 21 common peaks identified, among which 6 were identified as protopine, chelidonine, coptisine, sanguinarine, berberine, and chelerythrine. Network pharmacology filtered out 8 core anti-inflammatory components, namely thalictriline, (s)-thalictroidine, (s)-tetrahydropalmatine, berberine, chelidonine, chelerythrine, oxidized sanguinarine, and (+/-)-thalicarpine. The intersection of these two sets of components was chelidonine, berberine, and chelerythrine. Referring to the regulations of the Chinese Pharmacopoeia, chelerythrine was selected for a study on its effect on IPEC-J2 cell inflammation. The study found that chelerythrine at concentrations of 2.5 to 10 μg·mL^-1 could significantly improve cell survival rate, increase the content of IL-4 and IL-10, and decrease the content of NO, IL-1β, IL-6, IL-8, TNF-α, and TGF-β1. This study has established the HPLC fingerprint of C. majus and preliminarily revealed its pharmacodynamic components and mechanisms of action using network pharmacology methods. This provides a reference for its quality control and further development.

The purpose of this study was to prepare a milk-derived exosome (miEVs) delivery system loaded with forsythiaside A (FTA) and kaempferol (KPF) and evaluate its effect on lipopolysaccharide (LPS)-induced bovine mammary epithelial cells (MAC-T) in vitro Effects of cell migration and related inflammatory factors in inflammation models. FTA-loaded milk-derived exosomes (FTA-miEVs) and KPF-loaded milk-derived exosomes (KPF-miEVs) were prepared by using the acetic acid method established and optimized in the early stage, and the low-temperature ultrasonic co-incubation method was usedAs well as the establishment of inflammation models, and the evaluation of the preparation effect by transmission electron microscopy (TEM) and immunoblotting (Western blot) methods, the liquid chromatography-mass spectrometry method for drug loading determination, immunofluorescence and scratch experiments to observe the effect of inflammation models on the uptake of milk-derived exosomes loaded with anti-inflammatory drugs, and the effect of cell migration in the inflammation model, enzyme-linked immunosorbent assay (ELISA) and real-time quantitative PCR (RT-qPCR) were used to verify the effects of milk-derived exosomes loaded with anti-inflammatory drugs on the secretion of related inflammatory factors and gene expression levels in inflammation models. The results showed that FTA-miEVs and KPF-miEVs both had typical exosome structures, with circular shapes and partial depressions, similar to the "saucer" structure. Both surface and internal specific marker proteins leukocyte differentiation antigen 81 (CD-81) and heat shock protein 70 (HSP70) can be expressed. The drug loading was 0.78% and 9.79%, respectively. Within 8 hours, the LPS-induced MAC-T inflammation model had a good uptake effect; At 24 h, the inhibition rates of FTA-miEVs and KPF-miEVs on cell migration in inflammation models were 608.48% and 522.88%, respectively, which were significantly higher than those of miEVs, FTA and KPF alone. It has a positive effect on interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor α (TNF-α) in models of inflammation. The secretion and gene expression levels were better than those of miEVs, FTA and KPF alone. In summary, the FTA-miEVs and KPF-miEVs drug delivery systems were successfully prepared in this experiment, and the comprehensive results showed that the anti-inflammatory effects of FTA and KPF could be improved at the same dosage, which provided a new idea for the preparation and clinical application of exosomes in drug delivery systems.

This study aims to investigate the effects and mechanisms of salidroside on bone metabolism in a methylprednisolone-induced animal model of femoral head necrosis in broilers. Thirty-two 1-day-old Arbor Acres broiler chickens were randomly divided into a control group, a methylprednisolone group, a salidroside intervention group, and a salidroside control group, with 8 chickens in each group. At 42 days of age, the chickens were subjected to gait scoring and weighing, followed by the collection of blood and liver samples for triglyceride and total cholesterol analysis. Femoral head tissue samples were collected for bone metabolism marker analysis, and femur and tibia samples were taken for bone density and strength testing. Results were as follows: Compared to the methylprednisolone group, the salidroside intervention group showed a significant increase in body weight, femur weight, length, Young's modulus, and fracture load, as well as a significant increase in tibial stiffness; The transcription of Collagen-1 and Runx2 genes in the femoral head was significantly upregulated; There was a reduction in cartilage lacuna vacuolation, with significant upregulation of Collagen-2, Aggrecan, and BMP2 gene transcription in cartilage; a significant downregulation of MMP13 gene transcription; the levels of triglycerides and total cholesterol in the liver and serum significantly decreased; the expression of FASN and SCD1 genes in the femoral head was downregulated. The results indicate that salidroside can regulate lipid metabolism and cartilage matrix synthesis, and alleviate methylprednisolone induced femoral head necrosis in broiler chickens.

The aim of this study was to develop a new type of traditional Chinese medicine polysaccharide mucosal immune adjuvant by combining the advantages of polysaccharide from Atractylodes macrocephala Koids and ultra-large mesoporous silica. First, ultra-large mesoporous silica nanoparticles loading polysaccharide from Atractylodes macrocephala Koidz (AMP-UCMS) was prepared and a series of characterizations were carried out on it, including transmission electron microscopy, particle size, potential, polymer dispersity index (PDI) and infrared spectroscopy. Then, the mucosal immune adjuvant activity of oral AMP-UCMS as an inactivated H9N2 avian influenza virus was further studied. In the experiment, 150 chicks were randomly divided into 6 groups and received different immunization treatments, namely negative control (Control), whole inactivated virus (WIV), WIV+AMP, WIV+UCMS, WIV+AMP-UCMS, and commercially available vaccine (Vaccine). Among them, 6 chicks from each group were intranasally challenged with H9N2 avian influenza virus. The hemagglutination inhibition (HI) test was used to determine the HI antibody titer of chicken serum in each group; the ELISA method was used to detect the specific sIgA antibody level of H9N2 avian influenza virus in the jejunum; the HE staining was used to observed the histomorphology changes of the jejunum and lung; real-time fluorescent quantitative PCR (RT-qPCR) was used to determine the mRNA expression levels of IgA J-chain, pIgR, TNF-α and IL-1β in the jejunum and various indicators such as the viral load in the lungs of chickens after challenge. The results showed that compared with the Control group, WIV+AMP-UCMS group could highly significantly increase the HI antibody titer of chicken serum (P < 0.000 1) and the specific sIgA antibody level in the jejunum (P < 0.01), and highly significantly reduce the viral load in the lungs (P < 0.000 1), and the mRNA expression levels of chicken jejunum IgA J-chain(P < 0.001), pIgR(P < 0.000 1), TNF-α and IL-1β (P < 0.000 1) were all highly significantly increased. Additionally, it could increase the number of intraepithelial lymphocytes (iIELs) in the chicken jejunum and reduce lung tissue damage following viral challenge. In conclusion, polysaccharide from Atractylodes macrocephala Koidz was successfully loaded into ultra-large mesoporous silica in the current study, and AMP-UCMS could effectively induce systemic immunity and mucosal immune responses in chickens. These research results provide a reference for the development and application of new traditional Chinese medicine polysaccharide mucosal immune adjuvants in the field of oral vaccines for livestock and poultry.

The aim of this study was to investigate the epidemiological status and genetic variation of novel duck reovirus (NDRV) in the Anhui region. A strain of NDRV was isolated from a flock of muscovy ducks exhibiting splenic infarction at a farm in Anhui Province. The strain was designated as AH812. Identification was conducted through RT-PCR, duck embryo isolation, pathogenicity assays, and σC gene sequence analysis. The results indicated that the isolated virus was confirmed as NDRV. Purity tests showed negative results for other common pathogens, including duck hepatitis virus, duck Tembusu virus, Muscovy duck parvovirus, adenovirus, and duck circovirus.Pathogenicity tests were performed by intramuscular injection of 0.5 mL of the AH812 isolate (TCID50=10^-4.25/0.1 mL) into ducklings leg muscle. The inoculated ducklings exhibited typical symptoms, such as splenomegaly and hemorrhage. Sequence analysis revealed that the AH812 strain shares the closest phylogenetic relationship with the 2020 NDRV isolate N7 from Shandong Province (GenBank accession number: MW030529.1).In conclusion, this study successfully isolated a novel NDRV strain, AH812, which demonstrates significant pathogenicity in muscovy ducks.