The extensive application of genomic selection has significantly accelerated genetic advancements in livestock and poultry. With the commercialization of livestock and poultry chips and the continuous reduction of sequencing costs, the available genomic information for livestock and poultry has become increasingly abundant. Many challenges have arisen in genomic selection, such as the number of genotypic markers far exceeds the number of samples with phenotype data, and the relationships between genomic information have become more complex. These problems greatly restrict the use of traditional evaluation models such as best linear unbiased prediction (BLUP) and Bayes. Machine learning algorithms, which do not rely on predetermined equation models, have demonstrated superior capability in handling nonlinear relationships. Machine learning methods can offer solutions to the aforementioned challenges, thus they are gradually being applied in genomic selection. This paper reviewed the developmental of genomic selection, elucidated the principles of several commonly used machine learning algorithms. Furthermore, the current status and implementation methods of machine learning in livestock and poultry genomic selection were summerized. Finally, the challenges faced by machine learning in livestock and poultry breeding, as well as its development prospects were discussed.

Feed efficiency is an important indicator for evaluating the efficiency of livestock production, and it is one of the important traits in livestock and poultry breeding research. In the past few decades, livestock and poultry production traits such as meat, egg and milk yields have made significant genetic progress. With the increase of feed cost year by year, feed efficiency trait has gradually received high attention from breeders in recent years. European and American countries have successively included feed efficiency as a breeding target in the comprehensive selection index, and China has also begun related research and recieved research progress. This paper introduces the concept of feed efficiency, commonly used indicators, statistical analysis models and their genetic parameter estimation, and summarizes the research progress of common livestock and poultry feed efficiency traits at home and abroad, in order to further analyze the genetic mechanism of livestock and poultry feed efficiency traits and optimize the breeding program to provide reference.

Selection and mating are the core contents of pig breeding work, and scientific and rational selection and mating are important means to accelerate genetic improvement. With the rapid development of biological breeding technology, genomic selection (GS) and genomic mating (GM) techniques have played significant roles in the genetic improvement of pigs. GS is a method that uses molecular markers covering the entire genome to evaluate and predict the genetic potential of breeding target individuals, which achieves early selection by constructing models to predict individual genomic breeding values. GM, on the other hand, is about optimizing mating combinations to produce superior offspring for the rapid transmission of excellent traits. This article reviews the development, application progress and future prospects of GS and GM techniques in pig breeding, aiming to provide a reference in pig breeding in China.

Chicken is one of the most widely distributed animals in the world.It has evolved a stable genetic mechanism that allows it to adapt well to various environments, leading chicken as to become an important model for studying the biological mechanism of environmental adaptability. The development of sequencing technology and global environmental databases has made it possible to analyze the association between genetics and the environment. Population genetics, landscape genomics, pan-genome, and other research methods constantly promote researches on the chicken environmental adaptability, identifying a series of candidate factors, such as EPAS1 and HIF1A related to hypoxia adaptability; av-UCP related to cold adaptability and HSP related to heat adaptability in chickens, which laid the foundation for further studies on the genetic mechanism of environmental adaptability. Nevertheless, the molecular mechanisms of chicken environmental adaptability are still elusive. In this review, we summarize the recent research progress that chicken adapted to high-altitude, cold, heat, drought, and comprehensive climate, and put forward the challenges faced and the future research spots in the field, aiming to provide theoretical support for the conservation and utilization of chicken genetic resources.

The ovary is an important component of the reproductive system of female livestock, and its function is regulated by many factors, among which oxidative stress, as an important cellular biological process, has been found to have a profound effect on the ovarian function. The balance between oxidation and antioxidants in the ovary plays a significant role in maintaining ovarian function. Reactive oxygen species (ROS) in the ovary participates in the regulation of oocyte growth, meiosis, ovulation, and other physiological processes. The accumulation of ROS can lead to ovarian dysfunction and affect the formation of dominant follicles. It has influence on oocyte maturation, fertilization, and subsequent embryo implantation and development. Aiming at the effects of oxidative stress on ovarian function of female animals, this paper further explored the effects of oxidative stress on oocytes, granulosa cells, follicular membrane cells and corpus luteum of female animals, and in-depth understanding of the mechanism will help reveal the internal relationship between ovarian physiology and pathology, and provide new ideas for the prevention and treatment of reproduction related diseases.

Acidity is one of the most important indicators for the freshness of raw milk in recent years. The level of acidity directly affects the freshness of raw milk, which in turn reflects the quality of raw milk. Acidity in raw milk is affected by natural acidity and fermentation acidity. Natural acidity originates from the acidity formed by the inherent ingredients in raw milk, such as casein, whey protein, phosphate, citrate, carbonate and other acidic substances. Fermentative acidity is the part of the acidity that the microorganism in raw milk makes use of and decomposes the nutrient substance in milk, and produces organic acid and makes the acidity increase. Species, variety, lactation stage, lactation parity, milking frequency, maternal health status, season, feeding level, and storage time and temperature of raw milk can all affect the acidity of raw milk, thereby affecting the quality of raw milk. This article mainly introduces the concept and expression of acidity, analysis the acidity components, and the factors of acidity of raw milk. It aims to provide theoretical basis and new ideas for the mechanism of acidity production and composition.

The intestinal microenvironment is a microscopic internal environment composed of intestinal cells, microorganisms and their metabolites, which is crucial to the structure and function shaping of intestinal mucosal immune system. This paper systematically reviews the mechanisms of intestinal microenvironment nutrition metabolism affecting the fate of intestinal mucosal immune cells, in order to better understanding the intestinal microenvironment and intestinal immunity, and providing references for the treatment of intestinal health and related diseases.

Animal diseases not only seriously jeopardize the health of animals and hinder the healthy development of the breeding industry, but also pose a potential threat to human health. Therefore, it is vital to develop sensitive, specific and rapid diagnostic methods for disease prevention and control. The current gold standard for pathogens detection, PCR, has limitations such as long operational cycles, high instrumentation and technician requirements, cannot be applied to point-of-care testing. Clustered regularly interspaced short palindromic repeats/clustered regularly interspaced short palindromic repeats-associated protein (CRISPR/Cas) have the advantages of high sensitivity, high specificity and easy operation, which can provide a new method and opportunity for the diagnosis of animal diseases. CRISPR-Cas9 specificity in targeting nucleic acids and "collateral cleavage" activity of CRISPR-Cas12/Cas13 show significant promise in nucleic acid detection. In this review, the classification of CRISPR/Cas systems, the strategies adopted by three commonly used CRISPR/Cas systems applied to nucleic acid detection were summarized, as well as the latest research progress of their application in animal disease diagnosis, finally their advantages, challenges, and future development were analyzed, in the hope of providing a reference for the application of CRISPR/Cas biosensing platforms in animal diseases diagnosis.

The study aimed to establish the system for the isolation, culture and adipogenic differentiation of Beijing Black pig preadipocytes, to provide experimental materials for studying the molecular mechanism of fat deposition in pigs, and technical guidance for the preparation of cultured meat. The neck subcutaneous fat of a healthy 7-day-old male Beijing Black pig was collected, and preadipocytes were isolated using mechanical method combined with type Ⅰ collagenase digestion. The obtained preadipocytes were subjected to morphological observation, growth curve drawing, immunofluorescence identification, adipogenic differentiation, Oil Red O staining, BODIPY staining, triacylglycerol content determination, and detection of the expression of adipogenic differentiation-related genes using real-time quantitative PCR and Western blot techniques. The results showed that the isolated Beijing Black pig preadipocytes were spindle-shaped after adherence and the cell growth curve showed S type. The immunofluorescence results showed that PREF1 was positive, indicating that the isolated cells were indeed preadipocytes. Under the same combination of induction reagents, 10% FBS exhibited a better adipogenic differentiation effect than 2% FBS. Under the same serum concentration, the combined use of the cocktail with sodium oleate and rosiglitazone displayed a better differentiation effect. A large number of lipid droplets appeared in cells differentiated for 8 days, and the lipid droplets became red after Oil Red O staining and green after BODIPY staining. Moreover, the triacylglycerol content in cells differentiated for 8 days was significantly increased (P < 0.001). The qPCR results showed that the expression of PPARγ, CEBPα, ACCα, LPL, SCD, and FABP4 were all significantly increased after 2 days of differentiation (P < 0.05), reached a peak at 4-6 days, and decreased at 8 days. The Western blot results showed that the expression of PPARγ, CEBPα, FABP4, and APN gradually increased during cell differentiation. Expression of PREF1 gradually decreased during cell differentiation. Under three-dimensional culture condition, the combination use of 10% FBS, the cocktail, sodium oleate and rosiglitazone could also induce preadipocytes into mature adipocytes. The lipid droplets in cells differentiated for 8 days became green and red after BODIPY and Oil Red O staining, respectively, and the expression levels of PPARγ, CEBPα, ACCα, LPL, SCD, and FABP4 in cells differentiated for 8 days increased significantly compared with the control group (P < 0.001). In summary, this study successfully established an isolation and culture system for Beijing Black pig preadipocytes and screened out adipogenic differentiation methods suitable for two-dimensional and three-dimensional culture, laying a foundation for further studying the molecular mechanism of fat deposition in pigs and improving meat quality, and also providing technical guidance for the preparation of cultured meat.

This study aimed to analyze the genetic parameters of feed intake traits in lactating sows and conduct a genome-wide association study (GWAS) to evaluate the impact of intelligent feeding technology on these traits during lactation. Feed intake data from 922 first to fourth parity Large White pigs, born between July 2022 and May 2023, were collected to analyze feeding conditions during lactation. The Hiblup software estimated the genetic parameters of feeding traits during lactation, while Plink, GEMMA, and R software conducted a genome-wide association study to identify significant SNP loci and candidate genes associated with feed intake traits. Functional enrichment analysis was then performed on annotated genes within the 420 kb range surrounding these SNP loci. The results indicated that the heritability of feeding traits in lactating Large White pigs ranged from 0.20 to 0.27, and 28 SNP sites were significantly correlated with these traits. Annotation of the associated SNP sites identified 15 candidate genes related to lactation feeding traits, which were significantly enriched in anabolic processes including glucose, fatty acids, hormones, and coenzymes (P < 0.05). HAMP, NDUFB8, UPK1A, CHSY1, U6, and COX7A1 were identified as key candidate genes for feeding traits in Large White pigs during lactation. The above research results provide crucial genetic variation sites and associated genes linked to feeding traits in Large White pigs during lactation, establishing a foundation for further enhancement of feed intake traits by utilizing these genetic markers in future breeding programs, and also offers theoretical support for the breeding of lactating Large White sows with enhanced fertility and lactation performance.

The aim of this study was to employ ultrasound technology to determine the breast muscle thickness of Tianfu broiler chickens (TF) so as to establish efficient selection method for broiler chickens based on their muscle development. In this present study, the breast muscle thickness, body measurements, and carcass traits of 31 healthy ((2.1±0.5) kg) Tianfu broiler cock chickens (65 days old) were measured, as well as the correlation and path coefficients between the breast muscle thickness, body measurements and carcass traits were analyzed using SPSS 26.0. The optimal regression models for the breast muscle thickness and carcass traits, as well as the lean meat weight and body measurements, were established. Furthermore, the principal component analysis was performed on the body measurements using the JMP. The ultrasound results showed that the breast muscle thickness of the Tianfu broiler male chickens followed a normal distribution. The coefficient of variation in the body slope length, chest depth, chest angle, shank girth, and crown thickness were greater than 10%, and the breast muscle thickness was significantly positively correlated with the chest circumference, chest depth, shank girth, crowm thickness, and breast muscle weight (P < 0.01), and was significantly positively correlated with the keel length, chest girth, live weight, half-carcass weight, and lean meat weight (P < 0.05). Taken together, it was revealed in this study that the lean meat weight of the Tianfu broiler male chickens were closely related to the chest girth and body slope length. Furthermore, the meat production performance were indirectly selected through optimal linear regression models of body size traits for lean meat weight and carcass traits for breast muscle thickness, whereas, live weight, chest circumference, chest girth, shank girth, and crown thickness are also traits of importance to be considered in selection of Tianfu broiler chickens. The results of this study will provide a theoretical basis for further selection of high-quality meat chickens.

The aim of this study was to understand the genetic structure and the genetic source of the dominant performance of 'Tezanghan' sheep, to provide the basis for the breeding plan and cross utilization, and to help the formation of the specific performance in different families. In this study, sheep SNP 50K v3 chip was used to detect the single nucleotide polymorphism of 50 'Tezanghan' sheep (18 rams, 32 ewes), 24 Texel sheep (12 rams, 12 ewes) and 48 Euler Tibetan sheep (20 rams, 28 ewes).Plink, GCTA and MegaX softwares were used to analyze the genetic structure of 'Tezanghan' sheep; Tajima'D method was used to analyze the intragroup genome selection signals of 'Tezanghan' sheep, Texel sheep and Euler Tibetan sheep; The selection signals of 'Tezanghan' sheep were intersected with those of Texel sheep and Ola sheep (Tibetan sheep) respectively for GO and KEGG functional enrichment analysis which was aimed to find out the parental origin and related genes that may affect the dominant performance of 'Tezanghan' sheep. The results showed that the average polymorphism information content (PIC), the average expected heterozygosity (He), the average observed heterozygosity (Ho), the average effective allele number (Ne) and the average minimum allele frequency (MAF) of 'Tezanghan' sheep were 0.26±0.12, 0.36±0.14, 0.37±0.16, 1.59±0.07 and 0.25±0.15, respectively. The length of ROHs mainly distributed in 1-5 Mb, accounting for 50.4%, the length of 20 Mb⁺ ROH accounted for 9.52%, and the median F_ROH was 0.085. The D values of genetic distance ranged from 0.152 7 to 0.349 1, with an average genetic distance of 0.279 6. Five families were divided by NJ cluster analysis, and the distribution of individuals within the families was uneven. The localized genes according to intersected signals between 'Tezanghan' sheep and Euler Tibetan sheep were related to immunity, including primary immune deficiency, hematopoietic cell line and T cell receptor signaling pathway. The localized genes according to intersected signals between 'Tezanghan' sheep and Texel sheep were related to biological metabolism. In summary, 'Tezanghan' sheep had a certain degree of inbreeding, which indicated the needs of reinforced management; Euler Tibetan sheep had more influence on the immune function of 'Tezanghan' sheep, and Texel sheep more affected the growth and meat quality traits of 'Tezanghan' sheep.

The study aimed to verify the functional role of bovine INTS11 gene in myogenic cells, explore the sequence of the CDS region of INTS11 and the characteristics of its encoded protein by bioinformatics analysis, and verify its role in the proliferation process of bovine myogenic cells using different molecular test techniques. The bovine INTS11 gene sequence was compared with other species. This involved the construction of a phylogenetic tree and the examination of the physicochemical properties, functional structure, and subcellular localization of the encoded protein. The test material comprised 3-month-old healthy fetal bovine myoblasts isolated and cultured in vitro. The sequences of the coding sequence (CDS) region of INTS11 in myoblasts were cloned. An overexpression vector of INTS11 was constructed, and interfering sequences of the gene were designed. This was done in order to investigate its effect on the proliferation of myoblasts by CCK8, Edu, RT-qPCR and other techniques. Bioinformatic analysis showed that INTS11 had a CDS region of 1 800 bp, two functional domains (MBL-foldmetallo-hydro and β-CASP motif), which belonged to the non-transmembrane proteins and the cellular localization was mainly distributed in the cytoplasm. Transfection of pcDNA3.1-INTS11 plasmid showed that the proliferation rate of myoblasts in the test group was significantly increased compared with that in the control group at 72-96 h (P<0.05); Edu assay revealed a significant increase in the number of positive cells (P<0.05); and the RT-qPCR results showed that the mRNA expression levels of proliferation marker factors CDK2 and CYCLIND1 were significantly increased (P<0.05) compared with that in the control group. After transferring the interference sequence, the cell proliferation rate was significantly decreased (P<0.05), while the mRNA expression levels of proliferation marker factors CDK2 and CYCLIND1 were significantly decreased (P<0.05) compared with the control group. In this study, the conservation of INTS11 was predicted in the domesticated animals, and that INTS11 protein has two functional structural domains, and it promotes the proliferation of bovine myoblasts by mediating the transcription of CDK2 and CYCLIND1. This research has perfected the gene network regulating skeletal muscle growth, provided theoretical support for further exploration of the regulating mechanisms of skeletal muscle growth in the post-sequence.

The present study aimed to establish a method for early genomic detection and genetic evaluation of embryos in Holstein cattle in order to select excellent breeding embryos, and further breed outstanding bulls through embryo transfer technology thereby shortening the generation interval and accelerating genetic progress. Firstly, the cloned somatic cells from 4 Holstein bulls was utilized to compare the efficiency of genomic DNA extraction, and call rate and mismatch rate for genotypic data with genome-wide SNP chip from 2-cell and 5-cell samples. Subsequently, the genomic DNA was extracted from 24 early-stage embryo samples of Holstein cattle with 5 cells in each sample, and then genotyped using 150K chip. Further, the genomic estimated breeding values for 9 traits of each embryo were calculated with the GBLUP method based on a large-scale genomic selection reference population. The results indicated that the concentration, purity and relative integrity of genomic DNA of both 2-cell and 5-cell samples were good. Of them, the call rates of 150K chip genotypic data from 5-cell samples were higher (ranging from 89.54% to 95.07%) than those of 2-cell, the mismatch rates were lower (1.02%-4.67%). The quality, quantity, and relative integrity of the genomic DNA from 5-cell samples of 24 embryos were also usable, which call rate of 150K chip data ranged from 92.28% to 98.51%. The genomic evaluation accuracy for the 9 traits (including milk yield, type and somatic cell score) reached 55%-79%, with an average accuracy of not less than 65%. In conclusion, the present study optimized and established an available method for genomic DNA extraction and genomic selection from a small number of cells in Holstein cattle embryos. The genomic call rate of the embryo samples was not less than 90%, and the average accuracy of genomic evaluation was not less than 0.65.

The aim of this study was to screen and analyze the differentially expressed genes by studying the blood transcriptome of different meat performance of Yili horses, so as to provide molecular basis for subsequent research on meat performance of Yili horses. The 12 healthy Yili mares aged 4-5 were selected from the core group of 112 meat type Yili horses. The horses with the backfat thickness higher than 4.4 mm and eye muscle area higher than 31.5 cm² were from the (HW group) with better meat performance, with an average weight of 528.67 kg; The horses with the backfat thickness lower than 2.2 mm, eye muscle area lower than 19.5 cm² were from the common group (LW group) with an average weight of 327.00 kg, with 6 horses in each group. Blood samples were collected for transcriptome sequencing. The results of bioinformatics analysis showed that a total of 370 differentially expressed genes were screened (P<0.05), including 158 upregulated genes and 212 downregulated genes. GO functional enrichment analysis showed that the meat performance of Yili horses might be related to the G-protein coupled receptor signaling pathway process, extracellular regions, and transmembrane signal receptor activity. KEGG pathway analysis showed that the meat performance of Yili horses might be related to pathways such as protein digestion and absorption, thyroid hormone synthesis, and cortisol synthesis and secretion. CTSH, SSTR1, APOA1, and ITM2A could be candidate genes for meat performance of Yili horse breeds, and the research results are intended to provide reference for a better understanding of the molecular regulatory mechanism of growth and development of meat-producing Yili horses.

The study aimed to explore the internal relationship between the racing performance and the cardiac dimension of 2 years old speed racing Yili horse in 2 000 m and improve the breeding efficiency of Yili sports horses. The 30 speed racing Yili horses aged 2 years old in 2 000 m were selected as the research objects, including 21 male horses and 9 female horses. The top 10 horses were ranked according to their best race time, and the top 10 were classified as excellent group, the ranked 11-20 were classified as ordinary group, and the ranked 21-30 were classified as poor group. There was no significant difference in the body weight of the horses (P>0.05), fitness was good and jockeys were of similar calibre. Cardiac dimensions were measured in horses using echocardiography, and the correlation between cardiac dimensions and performances such as race time, rank, GPA, and bonuses was analyzed, principal component analysis was performed on the 19 cardiac dimension indicators, and the prediction equation was fitted. The results showed that in the case of no significant differences in body size, in the 2 years old speed racing Yili horse in 2 000 m, the GPA and bonuses of the excellent group were significantly higher than those of the ordinary group and the poor group (P<0.01), the race time and ranking of the competition were significantly lower than those of the poor group (P<0.01); end-systolic left ventricular free wall thickness (LVFWs) was significantly greater in the excellent group than in the poor group (P<0.01). The left ventricular long axis diameter (LVLD) was highly significantly larger in the excellent group and the ordinary group than in the poor group (P<0.01). The race time of the 2 years old speed racing Yili horse in 2 000 m was highly significantly negatively correlated with the end-diastolic left ventricular diameter (LVIDd) and LVLD (P<0.01);the GPA was highly significantly positively correlated with the end-systolic pulmonary artery diameter (PADs) (P<0.01); the ranking was highly significantly negatively correlated with LVIDd and LVLD (P<0.01); there is a highly significant positive correlation (P<0.01) between bonus and LVIDd. A total of 5 principal components were obtained, which comprehensively reflect the cardiac dimension information of the 2 years old speed racing Yili horse in 2 000 m. By fitting 4 prediction equations, the coefficient of determination for both race time and rankings is higher than 0.6. Cardiac dimension indicators such as LVIDd and LVLD have a significant impact on racing performance in Yili horses. Conducting principal component analysis on the cardiac dimension and constructing a multiple stepwise regression equation between racing performance and cardiac dimension is of great significance for evaluating the racing performance of 2 years old speed racing Yili horse in 2 000 m.

The aim of this study was to investigate the effects of heat stress on the structure and function of blood-testis barrier (BTB) in goats, and to provide a theoretical basis for the prevention and treatment of heat stress in goats. Six healthy male Guanzhong goats aged 1 to 1.5 years old were selected and randomly divided into two groups with three goats in each group. Goats in the control group were fed normally, and the heat stress model was established in the experimental group by the modified scrotal heat stimulation method. The semen quality was detected by CASA, the testicular structure was observed by light microscopy and transmission electron microscopy, the permeability of BTB was detected by biotin tracer, and the expression and localization of key proteins of BTB were detected by Western blot and immunohistochemistry. In heat stress group, the testis index, spermatozoa count, spermatozoa viability, spermatozoa total motility and progressive motility were significantly decreased (P < 0.05). Heat stress caused obvious morphological damage of the testis, and the number of spermatogenic cells decreased and the arrangement was disordered. Endoplasmic reticulum of sertoli cells was expanded and a large number of vacuoles appeared. The tight junction between sertoli cells was loose and discontinuous. The permeability of BTB increased, and the expression level of key junction proteins decreased significantly (P < 0.05), but the distribution and location of BTB were not affected in heat stress group. Heat stress damages the morphology of goat testis, destroys the structure of sertoli cells and reduces the expression of key BTB proteins, thereby impairing the structural and functional integrity of BTB, resulting in the decline of spermatozoa quality and affect the reproductive capacity of male goats.

The study aimed to investigate the effect of HIG1 hypoxia inducible domain family member 1C (HIGD1C) on apoptosis of sertoli cells in yak testis. In this study, 3 healthy male yaks aged 3-6 years and male yaks with cryptorchidism were selected, respectively as research materials, and their testis was collected. The regulation mechanism of HIGD1C on apoptosis of yak testicular sertoli cells was explored by HE staining, isolation and culture of yak testicular sertoli cells, indirect immunofluorescence, RT-qPCR, Western blot, HIGD1C overexpression and interference vector construction and in vitro cell transfection. The results showed that the basement membrane and interstitial tissue of normal testis were intact, the seminiferous tubules, peritubular muscle cells and sertoli cells were closely arranged, the seminiferous tubules in cryptorchidism were broken and atrophied, and the transcription and protein expression level of HIGD1C in yak cryptorchidism was significantly higher than that in normal testis (P < 0.01); HIGD1C protein was mainly distributed in leydig cells and sertoli cells; yak sertoli cells were successfully isolated and cultured; and HIGD1C protein was located in the nucleus of primary sertoli cells; after overexpression of HIGD1C, the mRNA expression level of Bcl-2 was up-regulated (P < 0.05), the protein expression level was significantly up-regulated (P < 0.01), and the mRNA expression levels and protein expression levels of Bax and Caspase-3 were significantly up-regulated (P < 0.01), the results of flow cytometry showed that the apoptosis rate of sertoli cells in HIGD1C overexpression group was significantly increased (P < 0.01). On the contrary, after HIGD1C knockout, the mRNA and protein expression levels of Bax, Bcl-2 and Caspase-3 were significantly down-regulated (P < 0.01), the results of flow cytometry showed that the apoptosis rate of sertoli cells in HIGD1C knockout group was significantly down-regulated (P < 0.01). The results of this study show that the normal testicular tissue structure is complete, the cryptorchidism stroma is loose, and HIGD1C is highly expressed in the cryptorchidism of yak. In vitro cell experiments showe that up-regulation of HIGD1C could activate pro-apoptotic proteins and promote apoptosis of yak sertoli cells. On the contrary, down-regulation of HIGD1C expression level will inhibit the apoptosis of sertoli cells. It is suggested that HIGD1C is involved in the regulation of apoptosis of yak testicular sertoli cells, which provides a reference for revealing the mechanism of cryptorchidism in yak.

The study aimed to investigate the role and mechanism of ADP-ribosyl transferase 3 (ART3) in spermatogenesis and deformation. In this study, 21 healthy ICR male mices at 6-8 weeks were randomly selected and divided into control group, ART3 inhibitor 3-MBA treatment groups, with 3 replicates in each group. Testicular and epididymis tissues were collected for 12 h, 24 h, 48 h, 5 d, 7 d, and 14 d after testicular injection of 3-MBA in the treatment groups. Analysis of spermatogenesis in them showed that inhibition of ART3 resulted in testicular atrophy, decreased sperm density in the cauda epididymidis, and increased sperm deformity rate. Histological observation showed that the spermatogenic cells in the seminiferous tubules were gradually reduced and loosely arranged, and some of them were detached to the lumen; non-sperms components appeared in the caput and cauda epididymidis.The components were identified as round spermatids by immunofluorescence of acrosomal proteins. Further immunofluorescence showed that the inhibitor treatment reduced the level of Nectin-3, a cell adhesion protein between spermatids and sertoli cells, led to the apoptosis of spermatids and the expression of AKT signaling pathway-related proteins decreased. It is hypothesised that ART3 promotes round spermatids deformation by modulating spermatids-sertoli cells junctions and affecting the anti-apoptotic effect of AKT pathway.

This experiment was conducted to investigate the effects of dietary energy level on rumen fermentation characteristics and microbial composition of weaned male Shaanbei white cashmere goats. A total of 40 healthy Shaanbei white cashmere goats with similar body weight (approximate (13.80±1.60) kg) were selected and randomly divided into 4 groups (Ⅰ, Ⅱ, Ⅲ and Ⅳ). The crude protein (CP) level was 11.8%, and group Ⅰ, Ⅱ, Ⅲ and Ⅳ were fed four diets with different digestible energy (DE) of 9.16, 9.66, 10.16 and 10.66 MJ ·kg^－1, respectively. The feeding experiment lasted for 60 days, including 7 days of pre-experiment and 53 days of formal experiment. At the end of experiment, rumen fluid was collected to investigate rumen fermentation parameters, and total DNA was extracted from rumen fluid. After PCR amplification, high-throughput sequencing and analysis were performed on the V3-V4 regions of 16S rRNA to study the rumen microbial composition. The results were showed as follows: 1) As the energy level increasing, pH, ammonia nitrogen (NH₃-N) content, and acetate to propionate ratio of rumen fluid decreased gradually. Total volatile fatty acid, acetate, and propionate content showed a trend of first increasing and then decreasing, while microbial crude protein (MCP) concentrations, butyrate and valerate content gradually increased; The pH of group Ⅳ was significantly lower than the other three groups (P < 0.05), the propionate in group Ⅰ was significantly lower than that in groups Ⅲ and Ⅳ (P < 0.05), and the acetate to propionate ratio in group Ⅰ was extremely significantly higher than that in groups Ⅲ and Ⅳ (P < 0.01). 2) Rumen microbial alpha diversity showed that with the increase of dietary energy levels, the Shannon and Simpson indices gradually decreased, while Chao1、faith_pd and observed_OTUs index showed a trend of first increasing and then decreasing, with Chao1 and observed_OTUs index in groups Ⅰ and Ⅱ were significantly higher than that of groups Ⅲ and Ⅳ (P < 0.05). 3) The analysis of rumen microbial community composition showed that the dominant bacterial phyla included Bacteroidetes and Firmicutes, followed by Proteobacteria; With the energy level increasing, the relative abundance of Bacteroidea gradually decreased, while the relative abundance of Firmicutes and Proteobacteria gradually increased; The dominant bacterial genera were Prevotella and Ruminococcus, with the energy level increasing, the relative abundance of Prevotella gradually decreased, while the abundance of Ruminococcus significantly increased. 4) The analysis of differential abundance comparison of rumen microbiota showed that: At the phylum level, the relative abundance of Proteobacteria in group Ⅳ was the highest and significantly higher than that of the other three groups (P < 0.05). The relative abundance of TM7 in group Ⅱ and Ⅲ was significantly higher than that in groups Ⅰ and Ⅳ (P < 0.05), while the relative abundance of Synergistetes in group Ⅰ was significantly higher than that in the other three groups (P < 0.05). At the genus level, the relative abundance of Ruminococcus in group Ⅰ was extremely significantly lower than that of the other three groups (P < 0.01), while the relative abundance of Oscillospira in groups Ⅰ and Ⅱ were extremely significantly lower than that of groups Ⅲ and Ⅳ (P < 0.01). The relative abundance of Unspecified Bacteroides in groups Ⅰ and Ⅲ was significantly higher than that of groups Ⅱ and Ⅳ (P < 0.05), the relative abundance of Succinomycetes, Unspecified Veillonellaceae, Unspecified Lachnospiraceae, and Unspecified Clostridials in groups Ⅰ and Ⅱ were all significantly higher than that of groups Ⅲ and Ⅳ (P < 0.05), the relative abundance of Unspecified_RF32 in groups Ⅱ and Ⅳ was significantly higher than that of groups Ⅰ and Ⅲ (P < 0.05), and the relative abundance of Unspecified_F16 in group Ⅱ and Ⅲ was significantly higher than that of groups Ⅰ and Ⅳ (P < 0.05). In conclusion, the increased dietary energy level affected the rumen fermentation characteristics of weaned male Shaanbei white cashmere goats, with a trend towards reducing the richness of rumen bacteria, as well as significantly changed the relative abundance of Proteobacteria, Synergites, Ruminococcus and Succinococcus. And the diet with digestible energy level of 9.66 MJ ·kg^－1 can effectively improve the rumen fermentation characteristics and bacterial composition of weaned male Shaanbei white cashmere goats.

This study was conducted to investigate the effects of different levels of methionine supplementation on wool production performance and hair follicle development of Angora rabbits. A total of 200 9-month-old Angora rabbits, half male and half female, with similar body weight and good growth condition, were randomly divided into five groups with 40 replicates per group and one rabbit per replicate. Rabbits in the control group were fed a basal diet, and those in the experimental groups were fed the basal diet supplemented with 0.15%, 0.30%, 0.45% and 0.60% methionine, respectively (the overall dietary methionine levels were 0.41%, 0.56%, 0.71% and 0.86%, respectively). This trial lasted for 70 days. After the experiment, indexes related to the production performance were calculated, hair and fur samples were collected to detect the hair quality, hair follicle density and the expression of genes related to hair follicle development. The results showed that, compared with the control group, 0.30% and 0.45% methionine supplementation significantly improved the average daily feed intake (P < 0.05). Dietary supplemented with 0.15% methionine significantly increased elongation length and elongation rate of hair (P < 0.05) of rabbits and significantly reduced primary hair follicle density (P < 0.05) of rabbits. Moreover, supplementation of 0.30% methionine significantly increased the hair production rate and secondary primary ratio (P < 0.05) of rabbits and significantly reduced the more than 25 μm fiber content, coarse hair rate and primary hair follicle density (P < 0.05) of rabbits. Meanwhile, dietary supplemented with 0.45% methionine significantly increased (P < 0.05) the ratio of feed to hair, elongation length, elongation and secondary primary ratio and significantly decreased the fiber content above 25 μm, coarse hair percentage and primary hair follicle density (P < 0.05) of rabbits. In addition, dietary supplemented with 0.30% methionine significantly upregulated the mRNA expression of epidermal growth factor (EGF) (P < 0.05) in the skin of rabbits. In conclusion, the appropriate dietary methionine supplemental level can improve the hair production performance and hair quality of Angora rabbits. EGF signal may be involved in promoting rabbit hair follicle development. Under the condition of the current experiment, it is suggested that the optimal supplemental level of methionine is 0.30% in dietary of Angora rabbits (the overall dietary methionine level is 0.56%).

This experiment was conducted to study the effects of different levels of dietary copper on production performance and hair follicle development of Angora rabbits. A total of 180 Angora rabbits (5 months old) were randomly divided into 5 groups (36 replicates per group, one replicate per group) and fed with a basal diet containing 0, 5, 10, 20 and 40 mg·kg^－1 copper, respectively. The prefeeding period was 3 days, and the experimental period was 76 days. The results showed that: 1) The hair yield reached the maximum and the feed/wool ratio was relatively low when Cu was added at the level of 5 mg·kg^-1 (P < 0.01), but it had no significant effect on the average daily feed intake of Angora rabbits (P>0.05). 2) Compared with the control group, the addition of 40 mg·kg^-1 copper significantly increased the breaking strength F, breaking strength P and breaking work W (P < 0.01), significantly affected the elongation and elongation rate (P < 0.05), and improved the quality of rabbit hair. 3) Dietary copper concentration significantly increased the secondary hair follicle density (P < 0.01) and total hair follicle density (P < 0.05), and the total hair follicle density and secondary hair follicle density reached the maximum when the amount of copper was 5 mg·kg^-1. Dietary copper (5 mg·kg^-1) significantly increased the expression of Wnt10b and mammalian target of rapamycin (mTOR) (P < 0.01), and significantly increased the expression of keratin-related protein 6-1 gene (Kap6.1) (P < 0.05). The expression of mTOR reached the maximum at the copper addition of 40 mg·kg^-1. Copper addition had no significant effect on the gene expression of epidermal growth factor (EGF), transforming growth factor-β1 (TGF-β1) and transforming growth factor-β2 (TGF-β2) (P>0.05). In conclusion, dietary Cu supplementation can improve hair production performance, improve hair quality, and promote hair follicle development in Angora rabbits, which may be regulated by Wnt10b, mTOR and Kap6.1 genes. Under the conditions of this experiment, 13.72 mg·kg^-1 copper level in the diet of Angora rabbits is the most suitable.

This study aimed to obtain feline herpesivirus-1 (FHV-1) isolates from clinical samples through subculture and systematic identification, which laid the foundation for the study of FHV-1 vaccine. Ocular and nasal swabs of suspected FHV-1 infection cats were collected from pet hospitals. Crandell reese feline kidney (CRFK) were used to isolate virus, and evolutionary genetics analysis, ultrastructural observation via electron microscopy and animal regression test were performed on the isolates. The samples were isolated and sub cultured with CRFK cells, followed by identification as FHV-1 by PCR and immunofluorescence, and finally named as "FHV-ZH2202" strain. After the complete gene sequence of the virus was obtained by High-throughput sequencing and gene reference assembly, SNP analysis was conducted between the virus and domestic endemic strains. It was found that the non-synonymous mutant SNPS were mainly distributed in UL22 and US7 genes. Through phylogenetic analysis, FHV-ZH2202 and domestic endemic strains shared high homology in UL22 gene. However, US7 gene harbered by FHV-ZH2202 was with a relatively far relationship with the other domestic endemic strains. The results of animal regression test showed that all cats in infection groups were with typically clinical symptoms such as sneezing, ocular and nasal discharge. However, there was no death case in infection groups. Cats in the FHV-ZH2202 group shed viral DNA in ocular and nasal secretions to from the second day post infection (dpi) to 6-8 days post infection. One FHV-1 virus strain was successfully isolated and part of biological characteristics were analyzed. It was proved that FHV-ZH2202 strain had certain pathogenicity, which laid a foundation for the study of FHV-1 vaccine.

To understand the pathogenic characteristics and pathogenicity of the latest epidemic strain of porcine epidemic diarrhea virus (PEDV), isolation and identification of PEDV were conducted this study. PEDV positive samples were inoculated into Vero cells for continuous transmission for isolating PEDV epidemic strains. Then the culture characteristics, whole genome sequence of the isolated strains were analyzed, and their pathogenicity in piglets were evaluated. Results showed that a PEDV strain (named CHN-SC2021) was isolated from the 3rd passage (P3) Vero cells of continuous inoculation of PEDV positive sample. Under the electron microscope, the diameter of the virus was about 60-110 nm, forming classic "syncytium" on the cell; The virus titer from P10 to P35 generations increased from 10^4.7 TCID₅₀ ·mL^-1 to 10^6.43 TCID₅₀ ·mL^-1; The whole genome sequence of CHN-SC2021 (P5) (GenBank accession number: OM505025.1) is 28 014 bp, and its nucleotide sequence homology about 99.16%-96.32%, 99.01%-96.57% and 98.94%-96.33% compared with the reference strains from China, Europe and America, and other Asian countries, respectively; The phylogenetic analysis showed that the CHN-SC2021 strain is closely related to Jiangxi CH/JXJA/2017 (MF375374.1), Henan CH/HNAY/2015 (KT199103.1), and CH/HNKF-16 (KY649107.1) strains, belonging to the GⅡ a genotype type. The similarity between the S gene sequence and other strains ranges from 98.7% to 93.4%. Oral infection of 6-day-old suckling piglets with 3 mL of 10⁵ TCID₅₀ ·mL^-1 per pig of CHN-SC2021 virus solution resulted in loss of appetite at 12 hours, vomiting at 20 hours, and severe vomiting and yellow watery diarrhea at 24 hours; The main pathological changes during dissection are gastrointestinal distension, transparent jejunum and ileum; Histopathological changes showed that each segment of the intestine had varying degrees of intestinal villus lesions, shedding, loose connective tissue, and widened space between the muscularis mucosae and the muscle layer; Immunohistochemistry showed that viruses were present in all segments of the small intestine, with the most viruses present in the ileum. In conclusion, a genotype GⅡ a PEDV virulent strain was isolated, which will contribute to enriching the information of PEDV circulating strains in China and be helpful for clinical prevention and control of PED.

Mycoplasma hyopneumoniae (Mhp) is the primary pathogen of endemic pneumonia (EP) in pigs, which causes coughing, shortness of breath, and wasting, resulting in higher feed conversion ratio, lower daily weight gain, longer farrowing time, and much higher farming costs, which cause significant economic losses to the pig farming industry. The aim of this study was to investigate and analyze the infection status of Mycoplasma hyopneumoniae in selected areas of southern China. A total of 21 large-scale pig farms in southern China were fixed-point sampled, and the sampling efficiency of nasal swabs and laryngeal swabs was compared through pathogen detection. Subsequently, the sample size for each farm was determined using simple random sampling combined with stratified sampling. A total of 1 478 laryngeal swab samples from sows were collected, and the surveyed pig populations were divided into "0-1 parity sows" and "≥2 parity sows". Pathogen detection was performed using real-time fluorescence quantitative PCR, and molecular epidemiological analysis was conducted using multi-locus sequence typing (MLST) combined with conserved p36 gene and non-conserved p97 and p146 genes. The results showed that the detection rate of nasal swab samples was 16.67%, and the detection rate of laryngeal swab samples was 56.67%, with lower Ct values. The detection rate of Mycoplasma hyopneumoniae infection in the field was 66.67%, but the infection pressure in each field was relatively low, with detection rates ranging from 1.39% to 18.57%. Among them, the detection rate of "0-1 parity sows" was 10.05%; the detection rate of "≥2 parity sows" was 2.17%. Molecular epidemiological results showed that the prevalent strains in 7 farms were of the same sequence type (ST128), and the genetic variation of the prevalent strains among the different farms was small and highly homogeneous. For the pathogen detection of Mycoplasma hyopneumoniae, laryngeal swabs are a more sensitive and effective sample type in vivo than nasal swabs. Parity of sows and the transportation of pigs are important factors for the infection of Mycoplasma hyopneumoniae in large-scale breeding farms. This study provides a theoretical basis for the prevention and control of Mycoplasma hyopneumoniae.

The aim of this study was to isolate novel duck-derived picornavirus for whole genome sequencing and genetic in order to perform whole genome sequencing and genetic evolutionary study. In 2021, we identified the presence of an unknown class of novel picornavirus infection in the diseased materials of breeding ducks and meat ducks submitted for inspection. By inoculating the diseased material into specific pathogen-free (SPF) chicken embryos, we isolated the virus. And the isolated viruses were further detected by PCR with primers designed from sequences available and sequenced the whole genome by overlapping PCR method. The amino acid sequences of each protein of the isolated virus were compared. At the same time, the sequences of the representative strains of small RNA viruses in the GenBank database were selected to draw the phylogenetic tree, and the sequences of the main proteins P1, 2C and 3D were analysed in comparison with each other. Three small RNA virus strains were isolated and named 21101, 21016 and 21075 (GenBank ID: OQ927377-OQ927379), with genome lengths of 7 445, 7 445, and 7 447 bp. These sequence encompasses an open reading frame (ORF) that encodes a total of 2 141 amino acids. The ORF can be divided into three distinct parts: P1, P2 and P3. These protein divisions align with the characteristic features observed in picornaviruses. The genetic evolutionary analysis, conducted through the utilization of whole genome sequences, demonstrated that the three isolates were positioned within the same branch as the Duck/FC22/China/2017 strain (GenBank ID: MN102111) previously separate inour laboratory and Duck/AH15/CHN/2015 strain (GenBank ID: MT681985) at the Shanghai Veterinary Research Institute. Furthermore, these isolates exhibited the highest degree of genetic similarity to the Duck hepatitis A virus (DHAV). Three strains of duck-derived picornavirus were isolated, and whole genome sequencing and genetic evolution analysis revealed that the three isolated viruses belong to the same class of the two currently known novel duck-derived picornavirus.

To identify specific expressed miRNAs of Neospora caninum and to investigate their expression in host cells, the samples of caprine endometrial epithelial cells (EECs) infected with tachyzoites of N. caninum at 24 and 48 h were collected. Specific expressed miRNAs of N. caninum and their in vitro expression were analyzed by using Illumina highthroughput sequencing technology and bioinformatics analysis. The effect of an up-regulated miRNA, novel3_mature, on the propagation of N. caninum in caprine EECs was also studied. The results showed that a total of 3 404 specific expressed miRNAs (155 novel predicted miRNAs) of N. caninum were identified. Of them, 77 (47 up- and 30 down-regulated) miRNAs were significantly differentially expressed. The consistent results were found with highthroughput sequencing by using quantitative real-time PCR analysis of seven significantly differentially expressed miRNAs randomly selected. Predictive analysis found 3 978 target genes of 61 miRNAs. Functional enrichment analysis showed that these target genes were possible involved in MAPK signaling pathway, axon guidance, focal adhesion, cAMP signaling pathway, calcium signaling pathway. Transfection of novel3_mature mimics significantly promoted the propagation of N. caninum in caprine EECs. Many specific expressed miRNAs were encoded by N. caninum and they played important roles in the propagation of N. caninum, providing a novel way to understand the intracellular survival of N. caninum.

Bovine coronavirus (BCoV) is one of the important pathogens causing respiratory and digestive tract diseases in cattle. The objective of this study was to isolate and investigate its partial biological characteristics of the bovine respiratory coronavirus (BRCV). Nasal swabs were collected from beef cattle with outbreak of respiratory diseases, and the samples tested positive for BRCV by real-time qPCR were inoculated into human cecal adenocarcinoma cells (HCT-8) for virus isolation and purification. Then sequencing and phylogenetic analysis were performed on the S and HE gene of the isolates which could cause cytopathic effect (CPE). Furthermore, the pathogenicity of the BRCV isolate was evaluated in BALB/c mice. The results showed that one strain exhibiting noticeable CPE was successfully isolated, and was confirmed as BRCV through real-time qPCR, indirect immunofluorescence (IF) assay and transmission electron microscopy (EM). After plaque purification, the TCID₅₀ of the BRCV isolate was 10^-8.46·0.1mL^-1. The complete S and HE gene sequences of the isolate were successfully obtained, and recombination events were observed in both genes. Comparative analysis with all available S and HE genes in GenBank revealed two unique amino acid (aa) mutations (A33G and A975V) in the S protein, which were located in the S1 subunit receptor binding domain and S2 subunit, and HE gene had two unique amino acid mutations (L235R and L365F), which were located in the lectin domain and near membrane terminal domain, respectively. The BRCV isolate demonstrated strong pathogenicity in 4-week-old BALB/c mice, causing clinical symptoms such as accelerated respiration and diarrhea. Postmortem examination and histopathological analysis revealed hemorrhage in lung and intestinal tissues. Tissue distribution analysis revealed that the virus predominantly localized in the lung and intestinal tissues, with the highest viral load observed in the lung tissue. This study reports the first isolation of a BRCV from beef cattle with respiratory disease outbreak in China. The S and HE proteins of the isolate exhibit unique amino acid mutations. Moreover, a mouse infection model was successfully established, providing a valuable reference for further investigation into the biological characteristics and genetic evolution of BCoV.

The aim of this study was to isolate and identify Pasteurella multocida which caused respiratory tract infection in Tibetan yaks, and to identify the biological characteristics, such as serotype, drug resistance, pathogenicity, genome characteristics and genetic evolution of the pathogenic bacteria. In this study, 498 nasopharyngeal swabs and tissue samples of yaks were collected from Nyingchi, Lhasa and Nakchu regions of Tibet. Epidemiological investigation, bacteriological isolation and purification, biochemical tests, PCR identification, K-B susceptibility test, animal pathogenicity test and whole genome sequencing of the isolates were conducted. Gene collinearity and phylogenetic analysis were carried out by Mauve software and MEGA7 software. The results showed that the positive rate of Pasteurella multocida was 2.61% in Nyingchi, Lhasa and Nakchu regions of Tibet, and a strain of Pasteurella multocida type B was isolated from the lung tissue of dead yaks, named "Pm XZ01" strain. Drug susceptibility tests showed that Pm XZ01 strain were sensitive to cephalosporins, quinolones, carbenicillin, bucaricana, cotrimoxazole and chloramphenicol antibiotics; Animal experiments showed that the mortality of rabbits (1.2×10⁸ CFU·mL^－1) and mice (1×10⁷ CFU·mL^－1) infected with Pm XZ01 strain was as high as 100%, indicating that Pm XZ01 strain had strong pathogenicity and lethality at this concentration. The total genome size of the Pm XZ01 strain obtained by whole genome sequencing was 2 331 787 bp, the GC content was 40.47%, and the total length of the repeats was 3 266 bp. It contained 121 non-coding RNA genes, including 59 tRNA genes, 19 rRNA genes and 43 ncRNA genes. There were 2 pseudogenes, 5 gene islands, 3 prophage, 68 carbohydrate active enzyme genes; CARD annotation analysis found out one PBP3 resistance gene and two EF-Tu resistance genes. VFDB and PHI-base annotation analysis revealed 101 virulence factor related genes and 663 phenotypic mutant genes. The collinearity analysis showed that Pm XZ01 strain had the best collinearity with pig HB03 (CP003328.1), HN07 (CP007040.1) and avian P1702 (CP097616.1) strains, but poor collinearity with yak Tibet-Pm1 (CP072655.1) strain. Phylogenetic tree analysis showed that Pm XZ01 strain belonged to the same branch as GDZQ201401 strain (KP083466.1) from Guangdong cattle in China, and had the closest evolutionary relationship. This study completed the epidemiological investigation of pasteurellosis in Tibetan yaks, as well as the isolation, identification, biological characteristics and whole genome sequencing analysis of Pasteurellosis multocida type B of yak origin, providing a reference for exploring the epidemic rule, pathogenic mechanism and prevention and control of type B pasteurellosis of Tibetan yaks.

This experiment was conducted to study the in vitro anti-Toxoplasma gondii (T. gondii) effect of rutin and its mechanism in BV2 cells. Firstly, the anti-T. gondii effect of rutin in BV2 cells was explored by detecting rutin cytotoxicity, T. gondii inhibition rate and proliferation by CCK8 and indirect fluorescence staining, etc. Subsequently, the cells were divided into four groups: normal group (Normal), T. gondii infected control group (T. gondii), and T. gondii infected groups with low-dose rutin (Rut 50) and T. gondii infected groups with high-dose Rutin (rut 100). Then T. gondii infection and rutin treatment experiments were performed, and the levels of antioxidant enzymes SOD, GSH and lipid oxidation product MDA, the levels of inflammatory factors TNF-α, IL-6 and IL-1β in the supernatant were detected by ELISA methods. The expression of key proteins in PI3K/AKT/Nrf2/HO-1, TLR4/NF-κB and P2X7R/NLRP3 signaling pathways in BV2 cells were detected by Western blot. The results showed that 50 and 100 μg·mL^－1 of rutin had a significant anti-T. gondii effect in BV2 cells, as the T. gondii infection rate, invasion ability and proliferation were significantly inhibited (P < 0.05). Rutin also significantly increased the SOD and GSH activities and decreased the MDA content (P < 0.05), and decreased TNF-α, IL-6 and IL-1β levels in the supernatant (P < 0.05). And it also significantly up-regulated the expression of key proteins in the antioxidant pathway PI3K/AKT/Nrf2/HO-1, and down-regulated the inflammatory pathway TLR4/NF-κB and P2X7R/NLRP3 (P < 0.05). These results indicated that rutin has a significant anti-T. gondii effect in BV2 cells in vitro, which may be related to its ability to enhance cellular antioxidant capacity through activation of the PI3K/AKT/Nrf2/HO-1 pathway, as well as inhibit the TLR4/NF-κB and P2X7R/NLRP3 pathways to attenuate inflammatory responses. These findings may provide some theoretical references for further research on the anti-T. gondii effect of rutin in vivo, as well as its protective effect in T. gondii-induced brain damage. It may also provide some theoretical basis for the development of rutin as a potential therapeutic agent for the treatment of toxoplasmosis in humans and animals.

MHC B2 haplotype chickens are highly resistant to avian leukosis virus infection, and this resistance may be related to the activated T cell immune response stimulated by MHC molecule presented epitopes. This study aims to elucidate the structure and mechanism of ALV-J epitope presentation by the MHC class Ⅰ molecule BF2*0201. Peptides that can bind stably to MHC Ⅰ were screened by gel filtration chromatography. The complex crystals of the peptide and MHC Ⅰ were grown in vitro using a crystal primary screening kit, and the crystal structure was resolved and analyzed using COOT, CCP4 and PYMOL. In this study, we identified two ALV-J CTL epitopes FVDFANRLI and SALQAFREV that bind stably to BF2*0201. We obtained a high quality crystal BF2*0201-SALQAFREV (SV9) grown under the conditions of 1.0 mol ·L^-1 succinic acid (pH 7.0), 0.1 mol ·L^-1 HEPES (pH 7.0), and 1% polyethylene glycol monomethyl ether 2000. We resolved the structure of the high-resolution (1.72 Å) BF2*0201-SV9 complex and found that the BF2*0201 antigen-binding groove exhibits weak electronegativity overall. Conserved residues were located at both ends of the antigen-binding groove, and specific amino acids gave BF2*0201 a medium-sized peptide-binding groove. The B, C, and F pockets played an important anchoring role. The residues in pockets and water molecules formed hydrogen bonds with the antigenic polypeptide to anchor it to the peptide-binding groove, and the antigenic polypeptide was in an "M"-shaped conformation. The conformational features of the prominent Gln-4 and Arg-7 residues in the SV9 epitope conformation suggested potential recognition of this site by specific TCRs. The crystal structure of BF2*0201-SV9 complex can help to elucidate the mechanism of ALV-J epitope presented by BF2*0201, which can help to explain the reason for the resistance of B2 haplotype chickens to ALV, and lay a theoretical foundation for the breeding of resistance to the disease.

To prevent allergic reactions induced by cats and dogs, we purified rFel d 1-Can f 1 fusion allergen by fusion prokaryotic expression of cat primary allergen Fel d 1 and dog primary allergen Can f 1. We mixed Layered double hydroxides (LDH) with rFel d 1-Can f 1 to prepare LDH-rFel d 1-Can f 1 and immunized BALB/c mice by subcutaneous injection. Subsequently, we used rFel d 1-can f 1 fusion allergen to induce allergic mice models. We evaluated the preventive effect of LDH-rFel d 1-Can f 1 on allergic reactions caused by cats and dogs through anaphylaxis, ear prick test, H&E staining of lung tissue sections, and serum IgE level. The results showed that LDH could completely adsorb the fusion protein of rFel d 1-Can f 1 when the mass ratio of LDH to rFel d 1-Can f 1 was 7 ∶1. The LDH-rFel d 1-Can f 1 can effectively relieve the hypothermia caused by stress allergic reaction in mice, and significantly reduce the dye penetration area caused by allergy. H&E staining of lung tissue sections showed that LDH-rFel d 1-Can f 1 could relieve inflammatory cell invasion caused by the allergy. In addition, the LDH-rFel d 1-Can f 1 can induce mice to produce higher levels of IgG, which lays the foundation for the development of vaccines that inhibit allergic reactions induced by pet cats and dogs.

The aim of this experiment was to prepare a hydrogel that can be used for local delivery of esculin. Carboxymethyl cellulose sodium (CMC-Na) and carboxymethyl chitosan (CMCS) were selected as the raw materials, to which esculin (ESC) was added to prepare Esculin-loaded CMC-Na/CMCS hydrogel (CCE) for topical drug delivery, and its properties and functions were investigated. In this experiment, the CCE hydrogel was successfully prepared, and orthogonal tests were designed with different ratios of CMC-Na/CMCS, based on the screening of drug loading and encapsulation rate, and the formulation of the hydrogel was finally determined as 2% CMC-Na, 2% CMCS, 200 μL of Genipin, and the rotational speed was 800 r·min^-1, and the appearance of the hydrogel was a blue solution, with no precipitation and impurity-free transparency and uniformity; Scanning electron microscopy showed that it had a three-dimensional structure and many smooth pores; the particle size was 0.841 5 nm, and the zeta potential was -32.1 mV; Fourier infrared spectroscopy and X-ray diffraction showed that esculin was successfully loaded into the CMC-Na/CMCS hydrogel without affecting its original functional groups; the solubilization rate was 234%; the in vitro release time was significantly prolonged compared with that of raw materials; the antibacterial effect was prolonged compared with raw materials at the same concentration of loaded gel. The antimicrobial effect was prolonged and better than that of raw materials. In this experiment, we successfully loaded esculin into CMC-Na/CMCS hydrogel and proved through a series of experiments that esculin has a certain slow-release effect compared with the API, which can prolong the action time of the drug and reduce the number of times of administration, providing a new way of administration for the treatment of diseases with Esculin in clinical practice.

To systematically analyze the antimicrobial resistance of Streptococcus suis (S. suis) to commonly used antimicrobials in China over the past decade, and analyze their temporal changes and geographic characteristics, so as to provide scientific guidance for monitoring the resistance trends of S. suis, rational use of antimicrobials, and development of new drugs. In this project, the existing literature was collected and organized, and Meta-analysis was performed. Inclusion and exclusion criteria were set up, relevant literature was screened, and a database on the resistance status of S. suis to commonly used antimicrobial drugs in China was established, and the literature in the database was subjected to the steps of quality evaluation, heterogeneity test, publication bias test, and subgroup analysis as well as statistical calculations. The results showed that different resistance status of S. suis to 20 commonly used antimicrobial drugs, with the highest resistant rates to tetracyclines and lincosamides (> 80%, 91% for tetracycline, 84% for lincomycin, 81% for doxycycline and 81% for clindamycin), followed by aminoglycosides (50% to 80%, 76% for erythromycin, 76% for kanamycin 75%, azithromycin 74%, streptomycin 70%, cotrimoxazole 65%, gentamicin 64%, amikacin 55%); penicillins, cephalosporins, phenicols, fluoroquinolones (10% to 40%; enrofloxacin 37%, florfenicol 31%, ciprofloxacin 30%, penicillin 24%, chloramphenicol 24%, ampicillin 13%, ceftiofur 12%), and vancomycin, with the lowest resistant rate of 4%. Subgroup analysis showed that the resistant rate varied depending on their isolation year and area. Increasing resistance of S. suis to commonly used antimicrobials were observed during the past decade. Strategies are urgently needed to tackle antimicrobial resistance of S. suis in China in premise of strengthen surveillance, reduce the irrational use and explore the resistance mechanism.

As the level of pet medical care continues to improve, the average life span of domestic animals has been extended. This also led to a significant increase in the detection and diagnosis rates of tumors. For better diagnosis and treatment of canine and feline tumors, it is necessary to understand the characteristics of different tumors and expand their epidemiological database through investigation. Canine testicular tumor is one of the common clinical tumors in male dogs. It generally arises from either the sex cord stromal elements of the gonad or from germ cells. Our study collected 119 canine testicular tumor cases among 2 803 canine tumors sent to our laboratory from multiple hospitals from 2013 to 2021 for retrospective pathological analysis. We listed the gross pathological and histopathological features of four types of canine testicular tumors diagnosed and explored the risk factors for canine testicular tumors. For mixed germ cell-sex cord stromal tumors, which are difficult to diagnose by HE staining, three immunohistochemical markers are combined for accurate diagnosis. The results showed that the detection rate of testicular tumors in male dogs was 10.8%, accounting for 4.25% of all cases submitted for examination. Among them, 28 cases (23.5%) were Sertoli cell tumors, 37 cases (31.1%) Leydig cell tumors, 42 cases (35.3%) seminoma, and 12 cases (10.1%) mixed germ cell-sex cord stromal tumors. Our study provides the latest information on the epidemiology of primary testicular tumors in dogs in China. It confirmed that cryptorchidism is an important risk factor for Sertoli cell tumor and seminoma through statistical analysis of the background information of the affected dogs. In addition, we preliminarily explored possible correlations between breed, cryptorchidism location, and detection rates of different types of testicular tumors.

Amylin acts on the lateral dorsal tegmental nucleus (LDT), induces weight loss and diet-suppressive effects, and activates dorsal scapular brown adipose tissue (IBAT) to produce heat. Multiple appetite hormone receptors colocalized with choline acetyltransferase (ChAT) neurons in LDT to modulate saturation and energy expenditure. However, it remains to be determined whether ChAT neurons in LDT are involved in amylin-mediated weight regulation. Immunofluorescence was conducted to detect the expression of c-Fos and ChAT neurons in mouse LDT, retrograde tracer analysis was selected to confirme the downstream nucleus mass of LDT, and neuronal damage assays was conducted to analyze the neuromechanism of amylin action. Results showed that intraperitoneal administration of amylin significantly reduced the body weight of mice and significantly increased the expression of c-Fos and ChAT neurons in both the LDT and parabrachial nucleus (PBN). ChAT and neuronal nitric oxide synthase (nNOS) neurons were co-expressed in the dorsal motor nucleus of the vagus (DMV). Retrograde tracer experiments demonstrated that LDTs send nerve projections to PBN neurons. Furthermore, injury of LDT and PBN neurons reduced the weight loss of amylin. LDT is the center of energy balance regulation connected to PBN, and the ChAT neurons of this circuit are involved in the process of weight regulation of amylin.

This study aims to use adipose derived mesenchymal stem cells (ADSCs) to intervene in an experimental animal model of autologous skin transplantation in small pigs, and observe their effects on the healing process of autologous skin transplantation. The animal model of autologous skin transplantation of 9 miniature pigs was established, and the miniature pigs were randomly divided into 3 groups: model group (Model), phosphate buffer group (PBS), and adipose mesenchymal stem cell group (ADSCs). Blood and skin tissue samples were collected before surgery and 7, 14, 21, and 28 days after surgery, respectively, and clinical changes of grafts and surrounding tissues were observed, photographed, and recorded. The effect of ADSCs on wound healing of mini pig autologous skin transplantation was comprehensively evaluated through skin graft apparent examination, histopathological examination, blood routine examination and serum biochemical examination, skin graft tissue oxidative stress, inflammation, regeneration and blood flow recovery. The researchers observed that the skin recovery speed of ADSCs group was better than that of the other two groups, and the ADSCs group basically returned to normal skin status 28 days after surgery. In terms of oxidative stress, SOD and GSH contents in ADSCs group were significantly increased (0.01 < P < 0.05), while MDA contents were significantly decreased (0.01 < P < 0.05). In terms of inflammation, the contents of TNF-α and IL-1β in ADSCs group were significantly decreased, and IL-10 was significantly increased on 7 and 14 days after surgery (P < 0.01). In terms of skin tissue regeneration, the expression levels of VEGF, bFGF, TGFβ3 and CD31 in ADSCs group were significantly increased (0.01 < P < 0.05). In terms of blood flow recovery, the percentage of blood perfusion of ADSCs during healing was significantly higher than that of the other two groups (0.01 < P < 0.05). To sum up, ADSCs can accelerate the recovery of skin tissue pathological damage, reduce oxidative stress reaction promote skin tissue regeneration and Angiogenesis in the miniature pig autologous skin transplantation animal model. The results indicated that ADSCs had the functions of anti-oxidation, anti-inflammation and promoting tissue regeneration, and had good therapeutic effect on autologous skin transplantation of miniature pigs. This experiment provided experimental support and scientific basis for the clinical application of stem cells in autologous skin transplantation and allogeneic skin transplantation.

The purpose of this study is to use transgenic technology to induce Adipose-derived mesenchymal stem cells (AMSCs) to differentiate into insulin secured cells (IPCs), and to achieve the purpose of clinical treatment of diabetes through stem cell therapy. This study first verified the role of Hnf1b gene in inducing the differentiation of canine AMSCs into IPCs in vitro, and on this basis combined with the cascade regulatory factors Pdx1, Ngn3, Pax4 and Nkx6.1, which play a key role in the development of islet cells, three different polygenic co-expressing adenovirus infected canine AMSCs. It was reprogrammed to differentiate into IPCs, and the optimal induction combination was selected by detection. The results showed that Hnf1b gene could promote the differentiation of canine AMSCs into IPCs in vitro. Three combinations of multi-gene co-expression vectors were constructed, pAdEasy-Hnf1b-Pdx1-Ngn3 (A), pAdEasy-Hnf1b-Pdx1-Ngn3-Pax4 (B), pAdEasy-Hnf1b-Pdx1-Ngn3-Nkx6.1 (C). After 25 days infected with canine AMSCs, pancreatic islet-like cell clusters were formed in group A, B, and C. Group B had the largest number, and dithizone staining was the deepest. The amount of insulin secretion in group B under high glucose stimulation was significantly higher than group A and group C. The expressions of genes related to islet cell development in groups A, B and C were significantly increased. The expression of Pdx1 gene in group B was significantly higher than group A and group C; The Pcsk1 gene expression in group B was significantly higher than that in group C; The expression of Ins gene in group B was higher than that in group A. In summary, Hnf1b gene can promote the differentiation of canine AMSCs into IPCs in vitro. It is shown that the combination of Hnf1b, Pdx1, Ngn3 and Pax4 genes has the best induction efficiency, which provides a new reference for exploring more efficient methods of stem cell differentiation into IPCs, and provides new ideas for clinical treatment of diabetes.

This experiment aims to investigate the role of PINK1/Parkin-mediated mitochondrial autophagy in the alleviation of fluoride-induced depressive-like behavior by selenomethionine (Se-Met). Forty BALB/c mice were selected and randomly divided into five groups: the blank control group (Group C), the fluorine group (NaF 150 mg·L^-1, Group F), the fluorine+low selenium group (NaF 150 mg·L^-1+1.5 mg·L^-1, Group F+LSe), the fluorine+medium selenium group (NaF 150 mg·L^-1+3.0 mg·L^-1, Group F+MSe), the fluorine+high selenium group (NaF 150 mg·L^-1+6.0 mg·L^-1, Group F+HSe). After 90 d of sodium fluoride exposure, the behavioral performance of mice was assessed by means of an elevated O-maze and hanging tail experiments; HE staining was used to evaluate the damage of the mouse cortex; biochemical kits were used to measure the content of oxidative stress-related indices; and real-time fluorescence quantitative PCR was utilized to determine the expression levels of mitochondrial autophagy-related genes. Animal behavioral results showed: the results indicated that mice in group F had a significantly longer "resting" time in the tail suspension test compared to group C (P<0.05) and a tendency to take more time to enter the closed arm of the elevated O-maze (P<0.05). After Se-Met supplementation, the "resting" time of mice was reduced significantly (P<0.05), and the F+LSe and F+MSe mice took significantly less time to enter the closed arm (P<0.05), compared to the F group. The results of the histopathological sections showed: the mice in group F had disorganized cortical cells, reduced cell volume, and fewer vertebral cells. Following Se-Met enhancement, the F+LSe group showed distinct nuclei and more uniform cytoplasm, and the improvement was notably more pronounced. The results of oxidative stress related indicators showed: group F had a significantly higher level of reactive oxygen species (ROS) (P<0.05), and a significantly reduced content of glutathione peroxidase (GSH-PX) (P<0.05), when compared to group C. Groups F+LSe, F+HSe, and F+MSe showed significantly lower levels of ROS (P<0.05). Compared to group F, the ROS content was considerably lower (P<0.05), in groups F+LSe, F+HSe, and F+MSe, with the content decreasing in the order of F+LSe<F+HSe<F+MSe; the GSH-PX content, on the other hand, was significantly higher (P<0.05), in groups F+LSe and F+HSe, compared to group F. Results of mRNA expression levels of autophagy-related genes were shown: group F showed significantly higher mRNA expression levels of autophagy-related genes Parkin, PINK1, LC3, OPTN, NBR1, Drp1, and Fis1 (P<0.05), compared to group C, and significantly lower mRNA expression levels of OPA1, Mfn1, and Mfn2 (P<0.05). Compared to group F, the mRNA expression levels of Parkin, PINK1, P62, NBR1, OPTN, and Fis1 were significantly lower in the F+LSe and F+HSe groups (P<0.05), LC3, Parkin, P62 and Drp1 were significantly decreased in the F+LSe and F+MSe groups (P<0.05), OPA1 was significantly increased (P<0.05), and the F+LSe group showed a significant increase in the mRNA expression level of Mfn2 (P<0.05). In summary, 1.5 mg·L^-1 of Se-Met can alleviate cortical oxidative stress by affecting the expression of mitochondrial autophagy-related genes, restoring the dynamic equilibrium state of mitochondrial fusion and fission, and thus ameliorating cerebral cortex damage and the emergence of depressive-like behaviors.

To explore the mechanism of Coptidis Rhizoma treating duck viral enteritis (DVE), the active ingredients and targets of Coptidis Rhizoma were firstly screened by the Traditional Chinese Medicine System Pharmacology Analysis Platform (TCMSP), and the DVE related targets and their intersection targets with Coptidis Rhizoma were analyzed and drew using the GeneCards database and Venny and Cytoscape 3.7.2 software, which constructed a "drug component disease target" network. And the protein interaction network (PPI) were established by STRING databases and their GO function and KEGG pathway were analyzed with the DAVID database. Then, the results of network pharmacology were verified by animal experiments which narmol group, virus infection group and drug intervention group, and their tissue samples were collected for histopathological observation and related cytokine determination. The results showed that 14 active components and 212 action targets of Coptidis Rhizoma were selected, including 25 targets that intersect with DVE, involving biological processes such as RNA polymerase Ⅱ promoter transcription, sequence specific DNA binding transcription factor activity, immune response, inflammatory response, and cell apoptosis, enriched in signaling pathways such as Toll like receptor signaling, inflammatory bowel disease, cytokine-cytokine receptor interaction, and herpes simplex virus infection, the core targets are IL-6, IL-10, and STAT1. By animal experiments, there were the varting degrees of pathological damage in liver, spleen and duodenum in virus infection group ducks, and IL-6 content was significantly increased compared to the normal control group ducks (P≤0.05), while the IL-10 and STAT1 contents were significantly reduced (P≤0.05). However, the pathological damage was significantly reduced in drug intervention group ducks, the levels of IL-6, IL-10, and SYAT1 tended to normal levels. These results indicated that Coptidis can effectively alleviate the pathological damage of duck tissues caused by duck viral enteritis, and its mechanism may involves various inflammatory response signaling pathways.

The aim of this study was to explore the in vitro antioxidant property and intestinal microbiota modulation effect of Schisandra Chinensis polysaccharide-selenium nanoparticles (SCP-SeNPs). The in vitro antioxidant property of SCP-SeNPs was evaluated using ABTS⁺, DPPH radicals, OH^- and O₂^- scavenging assays, and reducing power assays. Furthermore, the impact of SCP-SeNPs on the intestinal microbiota of mice was investigated using 16S rRNA high-throughput sequencing technology. The results showed that, SCP-SeNPs exhibited significant in vitro antioxidant property, with better scavenging effects on OH^- and O₂^- radicals compared to SCP. The 16S rRNA analysis indicated that the α-diversity index of cecal microbiota in the SCP-SeNPs group was higher than that of the Blank control (BC) group (P>0.05). The β-diversity index suggested notable differences in microbial composition between the SCP-SeNPs and BC groups. Intestinal microbiota structure analysis revealed that Bacteroidota and Firmicutes were the major constituents of the mouse intestinal microbiota. Differential analysis of microbial composition indicated that at the genus level, beneficial bacteria such as Prevotellaceae, Muribaculum, Dubosiella, and Faecalibaculum and Clostridia_UCG_014 were significantly enriched in the SCP-SeNPs group (P < 0.05). PICRUSt 2 functional prediction analysis revealed that the pathways of Proteasome, Peroxisome, Phenylalanine metabolism, and biosynthesis of Tropane, piperidine, and pyridine alkaloids were significantly enriched in the SCP-SeNPs group (P < 0.05). In conclusion, SCP-SeNPs has certain antioxidant capacity and may have the potential to regulate intestinal microbiota composition and impact microbial metabolism through the enhancement of beneficial bacteria. These results suggest that SCP-SeNPs can serve as a novel functional trace element supplement.

This study aimed to investigate the effects of glucosamine (GlcN) on the growth performance, antioxidant capacity and immune function of weaned piglets. Two hundred and fifty 21-day-old healthy Duroc×Landrance×Yorkshire weaned piglets of similar weight were selected and randomly divided into 5 treatment groups with 5 replicates in each group. The basal diet (NC group), the basal diet+chlortetracycline 75 g·kg^-1 (PC group), the basal diet+GlcN 1 g·kg^-1 (G1 group), the basal diet+GlcN 2 g·kg^-1 (G2 group) and the basal diet+GlcN 3 g·kg^-1 (G3 group) were fed respectively for 35 days. The results showed that: 1) Compared with the NC group, the ADFI and ADG of day 15 to 35 and day 1 to 35 of the other four groups were increased significantly, and the diarrhea rate of day 1 to 35 in G1, G2 and G3 groups were decreased significantly (P < 0.05). 2) Compared with the NC group, the serum MDA levels of the other four groups were significantly reduced, the serum CAT activity in the G1, G2 and G3 groups were significantly increased (P < 0.05); the serum IL-1β levels of the G2 group and the serum TNF-α levels of the G1 group were significantly reduced (P < 0.05) at day 14. The serum MDA levels were significantly decreased and the serum CAT activity were significantly increased in the other four groups, and the serum T-AOC in the G2 and G3 groups were significantly increased (P < 0.05); the serum IL-10 levels in the G1 group was significantly increased, and the serum IL-1β levels in the PC group and the G1 group were significantly reduced (P < 0.05) at day 35. 3) Compared with the NC group, the serum A/G in the PC, G1 and G3 groups were significantly increased, and the serum AST levels in the other four groups were decreased significantly (P < 0.05) at day 35. In summary, GlcN can improve the growth performance, reduce diarrhea rate, enhance antioxidant capacity, and immune function in weaned piglets. The appropriate dosage of GlcN is 1 g·kg^-1, and its effect is better than 75 mg·kg^-1 chlortetracycline.

To establish a rapid and accurate method for detection of Mycobacterium avium subsp. paratuberculosis (MAP), primers and probes were designed according to the specific multicopy insert element IS900 of MAP. A recombinase-aid amplification combined with lateral flow dipstick (RAA-LFD) method was established for visual detection of MAP in the field within 30 min at 35 ℃ after being optimized. The assay was specific with the target sequence of MAP with a detection limit up to 10 fg·μL^-1. One hundred and forty-nine stool samples were tested by this method, showing 100% sensitivity with the PCR. In short, the RAA-LFD assay developed in this study was a rapid, reliable method which can be broadly used.

The aim of this study was to investigate the prevalence and virulence factors of Escherichia coli (E. coli) in calves in Ningxia region. A total of 332 samples of diarrhea calf rectal feces were collected in Ningxia region for pathogen isolation, and 156 strains of E. coli were detected for 13 related virulence genes using PCR method. The results show that a total of 156 strains (46.99%) of E. coli were isolated, and a total of 136 strains containing virulence factors were detected. This indicates that the current positive rate of E. coli in diarrhea calves in Ningxia is relatively high, and the combination of virulence factors is complex, with irp2 being the dominant virulence factor.