Model animals are widely used in human diseases research. With the development of research on this field, classical model animals, such as rodents, dogs, monkeys, can not meet the requirements for human medical research, then miniature pigs have gradually been recognized as an effective supplement to medical model animals. This review introduces the basic biological characteristics of miniature pigs, including anatomical and physiological characteristics, and discusses the advantages of them as medical model animals. Moreover, the main characteristics, origins, breeding methods, and application progress of 18 different breeds of miniature pigs, both in domestic and abroad, are introduced. In addition, this paper also reviews the current research and application situation of miniature pigs as model animals in the fields such as cardiovascular diseases, metabolism, xenotransplantation, and so on. This review is expected to provide reference for the conservation, development and utilization of the miniature pigs' germplasm resources in China, to benefit for the further research on human diseases.

Avian leukemia virus E subgroup (ALV-E) is an endogenous retroviral element integrated in chicken genome. In recent years, many new ALVEs have been described with the develop-ment of genome sequencing projects. However, the ALV-E effects on host have not been fully understood. In this review, the structure and function of ALV-E were summarized, and the regu-lation of transcription initiation activity of ALV-E was elaborated, and the structure characteristics of ALV-E location in host genome was expounded. Finally, the effects of ALV-E on host function was discussed, which would provide reference for further understanding the function and expression regulation mechanism of avian leukemia virus in the genome of host.

Heat stress of dairy cows is a non-specific physiological reaction caused by high tempera-ture environment, which can seriously affect the physiological state and embryonic development of dairy cows, and then cause huge economic losses. Therefore, how to improve the development ability of dairy cow embryos under heat stress is of great significance to promote the healthy development of dairy cattle breeding industry. In this paper, the negative effects of heat stress on endocrine, oocytes and embryos of dairy cows, as well as the solution measures such as physical cooling, hormone therapy, addition of IGF1, modification of HSPA1L and PRLR genes by gene editing technology were reviewed, so as to provide some reference for improving the develop-ment ability of dairy cow embryos under heat stress.

A variety of microorganisms exist in the intestinal tract of ruminants, which are interdependent with the organism and constitute a dynamic equilibrium that has an impact on the host's nutrient absorption, metabolism and immune system development. The intestinal microorganisms of ruminants mainly originate from colonization at a young age and gradually develop into a complex microbial community. Therefore, exploring the intestinal microbial colonization of young ruminants has a positive impact on improving the survival rate and growth quality of ruminants. In this review, patterns of gut microbial colonization in young ruminants were summerized and some regulatory tools were introduced. However, the optimal time of implementation of these regulatory tools and the specific key microorganisms need to be further researched.

Sperm motility is a necessary condition for natural fertilization and achieving sperm-oocyte fusion under natural conditions. Sperm exhibit different motility patterns during the passage from the male epididymis to the female reproductive tract, which could be altered within a short period of time depending on changes in the environment, demonstrating that the sperm membrane proteins (ion channels) are involved in the regulation of sperm motility, and the normal expression of ion channels is closely related to the process of fertilization. Current studies have found that Ca²⁺, K⁺ and Na⁺ plasma channels are involved in physiological processes such as sperm membrane polarization, movement, capacitation and acrosomal reaction. The molecular mechanism and topological structure of sperm ion channels are preliminary understood through patch clamp and electrophysiology techniques. The physiological functions and topological structures of Ca²⁺, pH, voltage-gated Na⁺, K⁺, Cl^- channels involved in the regulation of sperm motility were summarized in this paper, in the hope of providing the comprehensive information on regulating mechanism and function of ion channel research, and promoting the development of animal reproductive biology.

The unconventional feed resources such as pomace, oilseed meal and bran are abundant in China, but the high non-starch polysaccharides content affects their application in animal production. The pectin component, which is as high as 20%, is not only poorly utilized but also affects the digestion and absorption of other nutrients, and is regarded as an anti-nutritional factor in feed of monogastric animals. In recent years, there have been more studies on the cellulose and hemicellulose components of non-starch polysaccharides, but little research has been reported on the biodegradation of pectin in feeds. This paper reviewed the structure, distribution and nutritional properties of pectin in feed and the progress of its microbial degradation in order to provide a new idea and method for reduction and substitution of corn and soybean meal.

Nucleic acid vaccines based on messenger RNA (mRNA) are a new technology that has emerged in recent years. Compared with traditional vaccines, mRNA vaccines are characterized by strong immunogenicity, good safety, fast development cycle and easy large-scale production. In the past few years, kinds of mRNA vaccines for rabies and influenza and other applications, have entered clinical trials and shown good application prospects. The recent successful application of mRNA vaccines against COVID-19 further validates this platform. It also opens the floodgates to the application of mRNA vaccines for infectious disease prevention, particularly in veterinary medicine. In this paper, mRNA vaccine research, functional structures, delivery systems, the mechanism of action, and clinical applications, are reviewed in detail to provide references for research and development of mRNA vaccines in the field of animal disease prevention and control.

Organoids are defined as three-dimensional (3D) multicellular self-assembled tissue structures in vitro, which could simulate corresponding organs in vivo in terms of cell types, structural architecture and function. They have obvious tissue specificity and can be used as a novel experimental model in vitro. With the development of technology, organoids have been applied in many fields including organ morphogenesis, organ function, organ repair and transplantation, disease diagnosis, drugs screening, etc. Currently, other than murine intestinal organoid culture, only few studies on animal intestinal organoid culture were reported. However, other animals, especially domestic animals, still suffer from infectious and non-infectious intestinal diseases that urgently need a new study model for pathogenesis or new therapy. Therefore, this research progress focused on animal intestinal organoids culture in the last ten years with the advantages and the disadvantages of it were being summarized. This review may provide new ideas for the related research of animal intestinal organoids.

The changes of gastrointestinal flora play an important role in animal health and disease, and more and more research evidence has linked the immune system with gastrointestinal flora. The main mechanism may be that the disturbance of flora leads to the imbalance of flora-immune interaction, the imbalance of nutritional metabolism and energy regulation, the damage of immune system, and finally induces disease. Perinatal dairy cows face the severe challenge of maintaining normal physiological metabolism. Dairy cows are easy to be infected with a variety of diseases during the perinatal period, which brings serious economic losses to the pasture. Recent studies have shown that the disturbance of rumen flora in perinatal dairy cows is an important cause of productive diseases, and the dynamic interaction between gastrointestinal flora and host mucosal immune system plays a key role in maintaining gastrointestinal dynamic balance and inhibiting inflammation. In this paper, the changes of gastrointestinal microflora and the composition of gastrointestinal mucosal immune system in perinatal dairy cows were reviewed, and the important role of the interaction mechanism of microflora and mucosal immunity in maintaining the health of dairy cows was discussed. Finally, the productive diseases of dairy cows mediated by flora disorder and immune imbalance were introduced in order to provide new ideas for perinatal cow feeding and management and disease prevention and control.

The aim of this study was to identify and analyze the single nucleotide polymorphism (SNP) and screen the specific SNPs in Jinfen White pigs. In this study, the whole genome of 30 ear DNA samples from 9 male and 21 female Jinfen White pigs were resequenced. The GATK software was used to detect the mutation sites, and VEP software was used for SNPs classification and annotation. The genome resequencing data of its parents, including Mashen pig, Taihu (Erhualian) pig, Landrace and Large white pig, were integrated to detect the specific SNPs loci of Jinfen White pig based on allele frequency. The biological functions of the genes including SNP loci were annotated by enrichment analysis. The results of whole genome resequencing of 30 Jinfen White pigs showed that about 1 761.73 Gb Clean reads were obtained by sequencing and data filtering, among which the average values of Q30 reached 91.77%. The average sample alignment rate was 75.61%, the average coverage depth was 14.7 X. A total of 14 680 807 SNPs were identified after filtering, most SNPs were distributed on Chr1 and in the non-coding region of the gene. Subsequently, 87 366 specific SNPs of Jinfen White pigs were screened based on allele frequencies, most of which were located on Chr2 and distributed in the intron. In addition, the missense mutant SNPs located in the exon region belonged to 343 genes, which were significantly enriched in organelles, organelle components, intestinal immune, fatty acid degradation pathway, et al. The SNPs were analyzed and 87 366 specific SNPs loci were also screened through whole genome resequencing and bioinformatics techniques. This study will provide a research basis for the development of molecular markers, pure breed identification and molecular breeding of Jinfen White pigs.

This study aimed to compare the effects of SNP genotyping techniques (array and sequencing) and marker densities on the analysis results of genetic diversity, phylogenetic tree and genomic inbreeding coefficient, and explore low-cost and efficient genotyping method and appropriate SNP density in genetic structure study. The data of the CAUPorcine SNP50 array and the resequencing data of 35 Zaozhuang Heigai pigs were used in this study. In this study, using resequencing data as "raw materials", 4 SNP panels of random 34K, even 34K, even 340K and even 3 400K were constructed. The genetic diversity, phylogenetic tree and genomic inbreeding coefficient of Zaozhuang Heigai pigs were analyzed using SNP data of CAUPorcine SNP50 array and several sequencing panels. The results showed that:1) The index values of genetic diversity, including observed heterozygosity (H_O) (0.385 9 vs. 0.320 0-0.324 1), expected heterozygosity (H_E) (0.381 3 vs. 0.333 5-0.334 6), and genetic distance (0.305 7 vs. 0.279 8-0.280 6), estimated by array SNPs were higher than those by the SNPs of sequencing panels, and the phylogenetic tree constructed by array SNPs was of much difference from those by the SNPs of sequencing panels. These may be caused by the tendency of choosing higher minor allele frequency (MAF) SNPs in array design. 2) All sequencing panels had small impact on the analysis results of H_O (0.320 0-0.324 1), H_E (0.333 5-0.334 6), genetic distance (0.279 8-0.280 6) and phylogenetic tree, but had great impact on the number (784-106 547) and length (0.20-13.51 Mb) of runs of homozygosity (ROH) and genomic inbreeding coefficient (F_ROH) (0.127-0.263). Currently used 50 K genome-wide SNP arrays in the analysis of inbreeding coefficient of livestock and poultry is good at detection of large fragments of ROHs, but weak in identification small and medium ones. So the genomic inbreeding coefficient estimated by them may be lower than the actual value. In summary, different SNP genotyping techniques have a significant impact on the analysis results of genetic diversity, phylogenetic tree, and ROH. In the sequencing group, different SNP densities had a small impact on genetic diversity and phylogenetic tree analysis results, but had a significant impact on ROH and F_ROH analysis results.

This study aimed to reveal the genome-wide genetic variation of Wangyue black pigs, screen the SNP loci for molecular markers characteristic. Twenty-four Wangyue black pigs weighing 110 kg were randomly selected and re-sequenced at whole-genome level, and genome-wide variation types was detected and population genetic diversity parameters was analyzed; Combined with the public database information, the SNPs informations of Beijing black pigs, Duroc, Large White pigs, Lantang pigs, Min pigs and Shenxian black pigs were downloaded, and the SNPs of 133 individuals of 7 breeds were divided into two datasets by using the method of selecting the maximum classification ability, and the characteristic sites of Wanyue black pigs were selected, and the gene function of the screened SNPs were annotated. The variation information scanning results showed that a total of 315 343 84 SNPs, 43 673 SVs and 3 258 CNVRs were obtained. A feature database of black pigs in Anhui was constructed, and finally 33 specific sites of Wanyue black pigs were screened, and the genes involved in 33 SNPs were functionally enriched, and it was found that these genes were related to growth (SUSD4, NELL1, GPC6), reproduction (KHDRBS2, CRISP1), immunity (NECTIN1), neuromodulation and environmental adaptation (GRIK4); The results of the relevant parameters of the genetic structure of the population showed that the genetic distance between individuals was between 0.157 4-0.287 3, with an average of 0.243 5±0.022 2; the effective population size was 4.2, the expected heterozygosity was 0.320, the observed heterozygosity was 0.328, the polymorphism marker ratio was 0.788, the total individual ROH length was 14.6-178.0Mb, the average ROH length was (61±9.4) Mb, and the inbreeding coefficient of the population was 0.025±0.004, with a low inbreeding coefficient. In summary, the abundant genomic variation resources of Wanyue black pig indicate that Wanyue black pig retains rich genetic diversity in the breeding process, and most of the individuals have a long genetic distance, which is a characteristic and valuable germplasm resource. This study provides scientific reference for the further breeding of Wanyue black pig.

In the process of pig breeding, it was difficult to extract and recognize audio features of mixed pigs. A blind source signal separation method based on sparse theory was proposed for underdetermined pigs. The 4 months old and 150 kg sows in good health were selected, and the audio signals obtained by mixing their cries in different states according to different coefficients were used as observation signals. Short-time Fourier transform was used to convert the audio signals. Single source points in the signals were selected by grouping, and AP algorithm of adaptive damping coefficient combined with singular value decomposition was used. The single source points were clustered to estimate the mixing matrix, and the audio signal was reconstructed by optimizing the minimum l_p norm. Two groups of experiments were designed. One group explained the general process of the experiment, and the other group analyzed the performance of the whole separation algorithm by comparison. Similarity coefficient, signal-to-noise ratio and mean square error were used to measure the separation audio quality. The results showed that:1) In the underdetermined pig blind source separation of 3 source signals and 2 observation signals, the similarity coefficient, signal-to-noise ratio and mean square error of the separated audio signals and the corresponding source signals at different time ranged from 0.67-0.92, 7.9-9.7 dB and 0.005-0.08, respectively. From the view of waveform, the separation performance of the algorithm was independent of the length of time and the number of tests, and the results had a certain stability. 2) When the number of source signals and observed signals were 3 and 2, 4 and 2, 4 and 3, 5 and 2, 5 and 3, 5 and 4, the average similarity coefficient, signal-to-noise ratio and mean square error of reconstructed signals and source signals were 0.785-0.957, 7.468-10.347 dB and 0.019-0.092, respectively. After comparative analysis, this research method had certain reliability. 3) When the number of source signals was certain, the more the number of observed signals, the better the index measured, and the better the separated audio quality. In conclusion, this method can effectively separate each source signal component of mixed pig audio signal, which layed a foundation for feature extraction of mixed pig audio in real environment.

The aim of this study was to analyze the correlation between genetic variant loci of Wnt family member 3a (Wnt3a) gene and skin hair follicle density in Chongren partridge chicken based on gene polymorphism.Six hundred Chongren partridge chickens, 200 protospecies hens and 100 roosters and 200 B strain hens and 100 roosters, reared under the same environment and the same management level until date of listing (120 d), were randomly selected in good health and condition. After slaughter, hair follicle density was measured at the same locations on the skin of the back, chest and thighs, the morphological structure of hair follicles was observed by HE staining of skin tissue sections, and the diameter of hair follicles and skin thickness were measured. The genetic variant loci of Wnt3a gene were screened by PCR amplification followed by direct sequencing, and their correlation with the hair follicle density of chicken skin was analyzed. The expression of Wnt3a gene in different skin sites was detected by real-time fluorescence quantitative PCR technique. The results showed that:1) Comparing the same sex and skin area of protospecies and B strain, the hair follicle density on the back of protospecies hens was extremely significantly higher than that of B strain (P<0.01), the hair follicle density on the thighs was significantly higher than that of B strain (P<0.05), and the hair follicle diameter on the back and thighs was significantly lower than that of B strain (P<0.05). The density of skin follicles on the back of the protospecies cock was highly significant higher than that of B strain (P<0.01), the density of hair follicles on the chest was highly significant lower than that of B strain (P<0.01), and the diameter of hair follicles on the back and thighs was significantly lower than that of B strain (P<0.05).The thickness of the skin on the chest of males in B strain was signifi-cantly higher than that of protospecies (P<0.05), and the thickness of the skin on the thighs of both males and females in B strain was significantly higher than that of protospecies (P<0.05). 2) Three SNPs loci were screened for the Wnt3a gene and their correlation with the skin follicle density at each site was analyzed. g.2555812 T>C and g.2555377 T>C loci were found to have significantly higher skin follicle density on the back of individuals with CC genotype than that of individuals with other genotypes (P<0.05). 3) Real-time fluorescence quantitative PCR results showed that the Wnt3a gene in hair follicle tissues of all 3 skin sites of the protospecies and the B strain had relatively high expression, and the expression in the dorsal part of the protospecies was significantly higher than that in the dorsal part of the B strain (P<0.05). In conclusion, the skin pores of protospecies of Chongren partridge chicken were finer compared to the B strain, and the CC genotype of the Wnt3a gene at the g.2555812 T>C locus and the CC genotype at the g.2555377 T>C locus were favorable genotypes for the skin follicles of Chongren partridge chicken and could be used as molecular markers for the chicken skin follicle density trait. In this study, we investigated the genetic effects of Wnt3a as a candidate gene for hair follicle growth and develop-ment and the correlation between SNPs and skin hair follicle traits, in order to provide a reference basis for molecular marker screening and marker-assisted selection of quality chicken carcass packaging traits.

This study aimed to apply multilayer perceptron(MLP) to genome selection of sex-limited traits in sheep, and compare it with other classical genome selection methods in various situations. In this study, Qmsim software was used to simulate the phenotype data and genotype data of 2 sheep population(Popl and Pop2). Artificial neural network (ANN) was used in MLP, and residual maximum likelihood (REML) method was used in linear model to estimate genetic parameters of different populations. Using Python to write MLP model, using DMU software to achieve best linear unbiased prediction (BLUP), genomic BLUP (GBLUP) and single-step GBLUP (SSGBLUP). The differences in the estimation of heritability (h²) and breeding values under different conditions were evaluated. In all cases, MLP and SSGBLUP were significantly (P<0.05) better than GBLUP and BLUP. There is no significant difference between h² estimation of MLP and SSGBLUP in the 3 conditions:When h² was 0.05, the QTL number was 100 and the marker number was 10K in the Pop2 population; when h² was 0.2, the QTL number was 500 under the two marker number in Pop1, and the QTL number was 100 under the marker number of 50K in Pop2; when h² was 0.5 and QTL number was 100, the marker number of 10K in Pop1 and the marker number of 50K in Pop2. Except for the above, the h² estimates of MLP were significantly better (P<0.05) than SSGBLUP, GBLUP, and BLUP. Under different prior values of h², when QTL number, marker number changed, the difference between the estimated h² of MLP and that of contemporary population was smaller than that of SSGBLUP, GBLUP, and BLUP. The h² estimation results of SSGBLUP and GBLUP methods were very different under different marker number, and the MLP difference was small. In all cases, the prediction accuracy of GEBV by MLP was the highest. When the prior value of h² was 0.05, the GEBV accuracy of MLP at 10K was slightly higher than that of SSGBLUP at 50K. Under the same h², number of QTL and marker number, the EBV prediction accuracy of each method in the Pop2 population was improved compared with that in the Pop1 population. According to the above simulation results, MLP is superior to other classical genome selection methods in the genome selection of sex-limited traits in sheep.

This study aimed to analyze the genetic diversity, kindship, and population structure of Jining gray goat conservation population. The Illumina 70 K goat SNP chip was used to screen the whole genome SNPs in 40 Jining gray goats, the genetic structure, kinship, and inbreeding coefficient of the breeding Jining gray goats were analyzed by Plink software, GCTA tool and R language to reveal the genetic relationship between the individuals. The results showed that a total of 67 088 SNPs were detected, and the call rate was higher than 98%. There were 57 991 SNPs met the demand of quality control, and 85.5% of which were polymorphic sites. The average effective allele number was 1.697, the average polymorphism information content was 0.283, and the minor allele frequency was 0.293, which implied that the genetic diversity was abundant in Jining gray goats conservation population. The average observed heterozygosity was 0.409, greater than that of expected heterozygosity (0.418). The average IBS genetic distance was 0.333 4, and the IBS genetic distance of 23 breeding bucks was 0.330 3. A total of 347 runs of homozygous (ROH) were detected in 40 individuals. The inbreeding coefficient of whole population was 0.047 9 based on the ROH value, which indicated a low level of inbreeding and rich in population genetic diversity. Combined the results of IBS matrix, G matrix, and phylogenetic tree, 23 bucks were clustered into 14 families according to the coefficient of relationship was 0.1. In conclusion, the Jining gray goats have rich genetic diversity, low level of inbreeding, and with remarkable conservation effect. It is suggested that, the number and structure of families in Jining gray goat conservation population should be focused in the future work.

This study aimed to estimate whole-genome inbreeding levels of Chinese Holstein cattle from different herds in Henan province by using the runs of homozygosity (ROH), and identify ROH enriched regions and screen candidate genes associated with the traits of economic interest. A total of 900 Chinese Holstein cattle form 7 dairy herds in Henan province were used to detect genome-wide ROH by the GGP Bovine 150K Beadchip. The number, length and frequency of ROH in Holstein population was counted. The genome inbreeding coefficient(F_ROH) was calculated according to ROH, and the high frequency ROH regions were annotated. ROH was identified in all animals, 55 908 ROH were identified, with a mean length of 4.23 Mb. The estimated inbreeding coefficients of ROH in 7 herds ranged from 0.082 (H7) to 0.123 (H2), with an average F_ROH of 0.106 in all animals.Moreover, 79 genes related to the economic traits of dairy cows in the genomic region with high frequency ROH were identified. Among these genes, AKAP3, C5H12orf4, and FGF6 were related to the body size and height of cattle, CAPN3 was associated with carcass and reproductive traits, CHST14 was directly related to pregnancy maintenance and fetal growth, the traits of milk protein composition were affected by IL5RA, and FGF10 was involved in regulating fetal folliculogenesis. Notably, a high-frequency ROH region was detected on chromosome 14(22.78-23.38 Mb), where more than 80% of individuals carried ROH fragments. The genes TGS1, LYN and CHCHD7 related to growth and feed conversion were identified in this region. Evaluation of dairy cattle inbreeding based on ROH information could be a useful tool for selection and mating strategies. The candidate genes identified could be used for marker-assisted selection in dairy cattle breeding.

This experiment was conducted to compare the gene expression profile changes of porcine testicular sertoli cells (ST) exposed to environmental estrogen bisphenol A (BPA) for different times. The porcine ST cells were divided into group C and group B with 3 replicates per group. Group C was a blank control group, and group B cells were exposed to BPA at a concentration of 50 μmol·L^-1. At 6 and 24 h after BPA exposure, total RNA samples from C₆, B₆, C₂₄ and B₂₄ group cells were collected, and a sequencing library was constructed. High-throughput transcriptome sequencing was performed using a paired-end 150 bp sequencing method. The assembly data were used for functional annotation, differentially expressed gene (DEG) analysis, and GO and KEGG enrichment analysis. The expression of some key DEGs were validated through real-time PCR method. The number of DEGs (6 928) significantly increased at 24 h after BPA exposure, compared with 3 940 DEGs at 6 h. The expression of SPP1 gene encoding secretory phosphoprotein 1 increased at 6 h after BPA exposure, but decreased at 24 h after BPA exposure. The expression of lumen binding protein-encoding gene BIP, ubiquitin B-encoding gene UBB, ubiquitin C-encoding gene UBC, and ornithine decarboxylase 1-encoding gene ODC1 were significantly upregulated both at 6 and 24 h after BPA exposure. Enrichment analysis showed that the TNF signal pathway was the most significantly enriched at 6 h, and p53, IL-17 and MAPK signal pathways were also significantly enriched, indicating that ST cells exhibited proinflammatory response. At 24 h after BPA exposure, DEGs in pig ST cells were enriched in amino acids and sugar metabolism pathways, including arginine and proline, histidine and glycolysis metabolic pathways. Real-time PCR results showed that PTGS2 and other DEGs related to the inflammatory response pathway at 6 h after BPA exposure and ARG1 and other DEGs related to the amino acid metabolic pathways at 24 h after BPA exposure were significantly up-regulated. In conclusion, the damage mechanism of BPA on porcine sertoli cells shows obvious time effect. Porcine ST cells exhibit inflammatory responses characterized by activation of TNF and other signaling pathways at the early stage of BPA exposure, and the amino acid and glucose metabolism pathways are significantly activated in ST cells at the late stage of BPA exposure.

The purpose of this study was to explore the effects of different dry rearing systems on the reproductive system development of male geese. Taking 100 healthy male Sichuan White geese hatched from the same batch as the experimental subjects, at 120 days of age 60 male geese with similar body weights were selected and equally divided into two groups (30 individuals per group), which were reared under the cage-rearing system (CRS) and net-floor mixed rearing system (MRS), respectively, and were slaughtered at 270 days of age to collect the testes and external genitalia for subsequent histomorphological analysis. Morphological results showed that the weight and organ index of geese left-, right- and bilateral testes were extremely significantly higher in CRS than that in MRS (P<0.01), and the long diameter of geese right testis and short diameter of geese bilateral testis was significantly higher in CRS than that in MRS (P<0.05). The natural length and basal diameter of geese external genitalia were significantly higher in CRS than that in MRS (P<0.05), and the straightened length was extremely significantly higher in CRS than that in MRS (P<0.01). Histological results showed that the number of geese testicular spermatogonia was significantly higher in CRS than that in MRS (P<0.05), and the ratio of testicular parenchyma to interstitium, the thickness of seminiferous epithelium, the number of sertoli cells and the number of germ cells were extremely significantly higher in CRS than that in MRS (P<0.01). The thickness of keratinized epithelium of geese external genitalia was significantly lower in CRS than that in MRS (P<0.05). The results of correlation analysis showed that there were significant correlations between the morphological and histological parameters of either geese testes or external genitalia (P<0.05); moreover, there were significant correlations in the histomorphological indicators between geese testes and external genitalia under both rearing systems (P<0.05), but the correlations in the histomorphological indicators between geese testes and external genitalia in CRS were much stronger. Taken together, the developmental processes of geese testes and external genitalia are closely related under both dry rearing systems. Compared with MRS, the 150-day period of CRS starting from 120 days of age significantly promotes the reproductive system development of male geese and the testicular spermatogenic ability.

The aim of this study was to investigate the differential expression of FGFs (fibrolast growth factor)/FGFRs (fibrolast growth factor receptors) and genes in their mediated Ras and MAPK signaling pathways in undifferentiated spermatogonia of yak and cattleyak, in order to explore the causes of spermatogenesis arrest in cattleyak. In this study, testicular tissues of 3 healthy male Maiwa yaks and 3 F1 generation cattleyak aged 24 months were collected and divided into two sample groups, yak and cattleyak, with 3 replicates in each group. HE staining was used to detect the difference of spermatogenesis between yak and cattleyak. The difference of proliferation ability of undifferentiated spermatogonia of yak and cattleyak was detected by population doubling times, CCK-8 and EDU staining. The differential expressions of 6 FGF family members, 3 FGFRs, FGFs/FGFRs-mediated Ras and MAPK signaling pathway genes in undifferentiated spermatogonia of yak and cattleyak were detected by RT-qPCR. Compared with yak, cattleyak had abnormal seminiferous tubule development, single germ cell type, and the proliferation capacity of undifferentiated spermatogonia was significantly lower than that of yak (P<0.05). The expression of FGF2, FGF4, FGF5, FGF8, FGF9 and FGF21 in the testis of cattle-yak and the FGF receptors FGFR1, FGFR2 and FGFR3 in the undifferentiated spermatogonia of cattleyak were significantly lower than those of yak (P<0.01). In FGFs/FGFRs-mediated Ras signaling pathway genes, except for HRas and ARaf, the remaining genes (Raf1, BRaf, MEK1 and ERK) were all expressed at lower levels in cattleyak undifferentiated spermatogonia (P<0.01). p38, MEKK3, MEKK6 and ASK1 in the FGFs/FGFRs-mediated MAPK signaling pathway genes were lower expressed in the undifferentiated spermatogonia of cattleyak (P<0.01). Among the genes related to cell proliferation and regulated by the Ras pathway, only SHISA6, ID4 were highly expressed in cattleyak undifferentiated spermatogonia (P<0.01), while the remaining genes (TAF4B, ETV4, GFRα1 and Ret) were highly expressed in yak undifferentiated spermatogonia (P<0.01). Among the genes related to cell proliferation and co-regulated by the Ras and MAPK pathways, ETV5 and BCL6B were both highly expressed in undifferentiated spermatogonia of yak (P<0.01). Among the genes related to cell differentiation and regulated by the Ras pathway, except for PIWIL4, SOX3, NGN3 and stra8 were all highly expressed in yak undifferentiated spermatogonia (P<0.01). Therefore, the abnormal expression of FGFs/FGFRs and their mediated Ras and MAPK pathway genes in cattleyak reduces the proliferative activity of the undifferentiated spermatogonia, and leads to the insufficient number of undifferentiated spermatogonia entering the differentiation stage. This may be one of the important reasons for the stagnation of the reproduction of the cattleyak.

The aim of this experiment was to study the effect of rutin addition to the diet on rumen fermentation, rumen flora structure and antioxidant properties in perinatal Hu sheep. Thirty 24-month-old perinatal Hu sheep with similar perinatal period and similar body weight[(62.90±2.80) kg] were selected and randomly divided into three groups with 10 replicates in each group. The rutin additive dose was 0 mg·kg^-1 BW (Control group), 50 mg·kg^-1 BW (Group Ⅰ) and 100 mg·kg^-1 BW (Group II) to the basal diet, respectively. The pre-test period was 7 d and the formal test period was 56 d. At 28 days postpartum before morning feeding, five sheep in each group were randomly selected for slaughter, and rumen tissue morphology, rumen fermentation parameters, antioxidant properties and rumen flora structure were examined. The results showed that:1) The differences in daily roughage intake, daily dry matter intake and daily neutral detergent fiber intake of perinatal Hu sheep were not significant (P>0.05) among the groups; Postpartum body weight, FBW and ADG were higher in the 2 experimental groups of perinatal Hu sheep than in the Control group, but the differences were not significant (P>0.05), there was no significant change in the number of lambing and the weight of lambs in each group (P>0.05); 2) The rumen papilla length of perinatal Hu sheep in Group II was significantly increased compared with that in Control group (P<0.05); 3) GSH-Px and CAT activities in rumen fluid of Group I were significantly higher than those in Control group (P<0.05); 4) Compared with the Control Group, the concentration of bacterial protein, protozoan protein, acetic acid, propionic acid, valeric acid and total volatile fatty acids (TVFA) in the rumen fluid of perinatal Hu sheep in Group II were significantly higher (P<0.05), and the concentration of microbial protein was extremely significantly higher (P<0.01); 5) The first dominant bacterial phylum in the rumen of perinatal Hu sheep in the Control group and Group I was the Frimicutes, and the second dominant bacterial phylum was the Bacteroidetes; The first dominant bacterial phylum in the Group II was the Bacteroidetes, and the second bacterial dominant phylum was the Frimicutes. Compared with the Control group, the relative abundance of both the Frimicutes and the Spirochaetes were significantly lower in Group II (P<0.05), and the relative abundance of the Bacteroidetes was significantly higher in Group II (P<0.05). The dominant genus of rumen bacteria in all groups of perinatal Hu sheep was Prevotella. Compared with the Control group, the relative abundance of Prevotella was significantly higher in Group I (P<0.05); The relative abundance of Prevotella was highly significantly higher in Group II (P<0.01), and the relative abundance of Treponema was significantly lower in Group II (P<0.05). In summary, rutin has improved the morphology of rumen tissue in perinatal Hu sheep, and can improve the antioxidant capacity of rumen, change the structure of rumen flora, and effectively improve the fermentation function of rumen in perinatal Hu sheep.

This experiment was conducted to evaluate the effects of dietary crude protein levels on growth performance, nitrogen metabolism, nutrient apparent metabolic rate, serum biochemical indices, muscle chemical composition and slaughter performance of female pheasant aged 11 to 17 weeks, and to study the optimal dietary crude protein level for female pheasant aged from 11 to 17 weeks. A total of 200 11-week-old healthy female pheasants with an initial body weight of (539.43±11.97) g were randomly divided into 5 groups with 5 replicates per group and 8 chickens per replicate. A single-factor experimental design was adopted. Dietary crude protein levels were 15.00%, 16.50%, 18.00%, 19.50% and 21.00%, respectively. The pre-feeding period was 7 days, and the formal period was 49 days. The results showed that:1) Dietary crude protein level significantly affected the average daily gain and feed to gain ratio of pheasant(P<0.05), and the average daily gain in CP 15.00% group was significantly lower than that in the other 4 groups (P<0.05), and the feed to gain ratio in CP 15.00% group was significantly higher than that in the other groups(P<0.05). 2) Dietary crude protein level had significant effects on nitrogen intake, fecal nitrogen, nitrogen retention and nitrogen utilization rate of pheasant (P<0.05), The content of nitrogen intake and fecal nitrogen in 19.50% and 21.00% CP groups were significantly higher than those in the other three groups (P<0.05), and the content of deposited nitrogen in 15.00% and 16.50% CP groups was significantly lower than that in the other three groups (P<0.05). The nitrogen utilization rate of 15.00% CP group was significantly lower than that of 18.00% and 19.50% CP groups (P<0.05).3) There were no significant differences in dry matter metabolic rate, ether extract metabolic rate and gross energy metabolic rate among all groups (P>0.05). 4) There were no significant differences in high-density lipoprotein cholesterol contents among all groups (P>0.05). The serum total protein content in 15.00% CP group was significantly lower than that in 18.00%, 19.50% and 21.00% CP groups (P<0.05), and the serum albumin content in 15.00% CP group was significantly lower than that in 18.00% and 19.50% CP groups (P<0.05). The serum urea content in 15.00% CP group was significantly lower than that in 19.50% and 21.00% groups (P<0.05). The serum triglyceride content of 21.00% CP group was significantly lower than that of 15.00% and 16.50% groups (P<0.05), and the serum low density lipoprotein cholesterol content of 21.00% CP group was significantly lower than that of 15.00%, 16.50% and 18.00% groups (P<0.05). The serum total cholesterol content of 21.00% CP group was significantly lower than that of the other four groups (P<0.05). 5) There were no significant differences in the contents of crude protein and ether extract in leg muscle among all groups (P>0.05), but the content of crude protein in breast muscle of CP 15.00% group was significantly lower than in 18.00%, 19.50% and 21.00% groups(P<0.05). The ether extract content of pectoral muscle in 15.00% CP group was significantly higher than that in 18.00%, 19.50% and 21.00% groups(P<0.05). 6) There were no significant differences in dressing performance(except percentage of abdominal fat) and organ index among all groups (P>0.05).and the percentage of abdominal fat in 15.00% group was significantly higher than that in 18.00%, 19.50% and 21.00% groups (P<0.05). In conclusion, pheasants with 18.00% and 19.5% crude protein levels can obtain higher average daily gain, lower feed/gain, and better nitrogen utilization. When CP is greater than 15.00%, serum lipid indexes are better, serum protein and urea contents are lower, and the protein content in pectoralis is higher and the fat content is less. Therefore, it is recommended that the crude protein level of female pheasants aged 11 to 17 weeks should be 18.00%-19.50%.

This study aimed to establish a flow cytometry (FCM) method for sorting secretory immunoglobulin A (sIgA) coated bacteria from the digestive tract contents of ruminants. Four healthy adult meat sheep in good physical condition were randomly selected and their ileal contents were collected to prepare the bacterial solution. The loading concentration of the bacterial solution, the working concentration of the bacterial dye 4', 6-diamidino-2-phenylindole (DAPI) and the anti-IgA antibody were optimized. The results showed that the optimal loading concentration of the bacterial solution of the contents was from 2.5 to 5 mg·mL^-1, the best staining concentration of DAPI was 5 μg·mL^-1, and the optimum amount of anti-IgA antibody was 5 μg. The reproducibility test results showed that the relative bias of the proposed method is less than 5% and that it is stable and reproducible. The sorting method of sIgA-coated bacteria established in this study is high selectivity, less sample consumption and easy operation and can provide technical support for related studies of the composition and function of sIgA-coated bacteria in ruminant digestive tracts.

Viruses, mainly bacteriophage, are significant components of the ruminal microbiota in ruminants. All the ruminal viruses are called ruminal virome, and could be studied by metagenomics. Understanding the components and functions of goat ruminal viruses is expected to provide a more comprehensive understanding of rumen microecology. Accurate profiling of complex rumen virome requires DNA extraction methods that provide adequate quality and quantity, as well as sufficient coverage of the original community. In this study, four procedures to extract viruses-like particles (VLPs) from rumen fluid were compared:(P1) centrifugation + dilution, (P2) centrifugation + dilution + filtration, (P3) centrifugation + dilution + filtration + polyethylene glycol (PEG) precipitation, (P4) centrifugation + dilution + filtration + PEG precipitation + chloroform treatment. The nucleic acid in four procedures was extracted and DNA concentration, purity and integrity were compared. Then genomic DNA sequencing was performed using Illumina Novaseq 6000 and virus population structure was analyzed. The results showed that there were positive effects of filtration and PEG precipitation on viral nucleic acid quality. The chloroform treatment, severely reducing virus-like particles (VLPs) yield, should be used with caution when performing high-throughput sequencing. The viral communities in the rumen were dominated by bacteriophages belonging to the viral order Caudovirales (i.e., Myoviridae, Podoviridae, and Siphoviridae). The results provide the basis for future investigations regarding the ecological importance of viruses in rumen microbial ecosystems.

This study aimed to investigate the effects of lactoferrin supplementation on gut microbial diversity in weaned piglets. Twelve weaned DianTai piglets with similar initial body weight of (6.12±0.54) kg were randomly divided into Control group, Bacitracin group and Lactoferrin group. Piglets in Control group were fed with basal diet, Bacitracin group was added with 0.5 g·kg^-1 bacitracin premix in the basal diet, while Lactoferrin group was supplemented with 150 mg·kg^-1 lactoferrin in the basal diet. The experimental period was 28 days. Fecal samples were collected on days 7, 21 and 28, and fecal microbial diversity was detected by high-throughput sequencing of bacterial 16S rRNA gene sequencing. The results showed that there was no significant difference in intestinal microbial diversity of weaned piglets (P>0.05). At the phylum level, Firmicutes and Bacteroidetes were dominant phylum on days 7, 21 and 28. At 7th day, there was no significant difference in the relative abundance of Firmicutes or Bacteroidetes among all groups (P>0.05). Compared with the control group, the relative abundance of Firmicutes in lactoferrin group decreased by 1.43% and that of Bacteroidetes increased by 16.4%. On day 21, compared with Control and Bacitracin groups, there was no significant difference in the relative abundance of Firmicutes and Bacteroidetes in Lactoferrin group (P>0.05). Compared with Control group, the relative abundance of Firmicutes decreased by 16.4% and that of Bacteroidetes increased by 34.1% in Lactoferrin group. On the 28th day of the experiment, compared with the control group, the relative abundance of Firmicutes in Lactoferrin group was significantly decreased (P<0.05). At the genus level, Lactobacillus was the dominant genus on days 7, 21 and 28, and there was no significant difference in the relative abundance of Lactobacillus among all groups (P>0.05). On day 7 of the experiment, the relative abundance of Lactobacillus in Lactoferrin group was reduced by 50.21% compared with the Control group. On day 21, the relative abundance of Lactobacillus was 8.42% lower than that of the Control group. On day 28, the relative abundance of Lactobacillus in the Lactoferrin group was 16.09% lower than that in the Control group.In conclusion, in this study, it was found that dietary lactoferrin supplementation did not significantly affect the intestinal microbial richness and diversity of weaned DianTai piglets. The dominant phylum in intestinal microbial flora of piglets at 7, 21 and 28 d after weaning were Firmicutes and Bacteroides, and the dominant genus was Lactobacillus.

Peste des petits ruminants virus (PPRV) is an important pathogen affecting the global livestock industry. The purpose of this research is to establish a stable and reliable PPRV reverse genetic manipulation platform, to provide an effective technical platform for the analysis of PPRV pathogenic mechanism, immune escape mechanism, virus replication mechanism and other basic theoretical research, at the same time, it will lay the necessary preliminary foundation for the development of safer and more effective new vaccines. The RNA of the PPRV Clone9 strain was extracted, and the PPRV antigenome cDNA was amplified by RT-PCR in six sections. The full-length antigenome of the PPRV Clone9 strain was digested and ligated to obtain pB-PPRV, and the PPRV nucleoprotein (N), three helper plasmids for phosphoprotein (P) and large protein (L). pB-PPRV and helper plasmids were co-transfected into BHK-T7 cells mediated by liposomes. Three days after transfection, Vero cells were infected by freezing and thawing three times. Significant cytopathic changes (CPE) were observed in the second generation. The results of indirect immunofluorescence, Western blot, RT-PCR and sequence determination showed that the infectious virus was rescued. The rescue virus (rPPRV-Clone9) was stably passaged on Vero cells, and the proliferation dynamics were similar to the parental virus. In this study, the reverse genetics system of PPRV Clone9 strain was successfully established.

This study aims to preliminarily screen the regulatory genes and virulence genes that interact with the host during African swine fever virus (ASFV) infection, so as to provide a molecular theoretical basis for the development of specific drugs and vaccines against ASFV. We re-analyzed the differentially expressed genes in the host during high and low virulent ASFV strains infection using GSE145954 and literature mining. The Metascape database was used for gene function enrichment analysis and the protein interaction network was established through the STRING database. The key genes in the regulatory network of viral infection host were obtained through network topology screening, and then the ASFV virulence genes regulating the key genes were screened at the cell level. The study found that ECE1, CCL2, CTSB, SCARB2 and CD14 were significantly up-regulated in ASFV infected mononuclear macrophages and whole blood of animals, while HMBS, DYNLL1, UBB, EZH2 and SERPINE1 were significantly down-regulated. PPI analysis showed that UBB, CCL2, CTBB, EZH2, SERPINE1, CD14 and DYNLL1 in host cells are key genes in the regulatory network of host infected with ASFV, in which UBB played a central role. B119L, I215L and MGF360-13L of ASFV can inhibit the expression of UBB. The results suggest that UBB, CCL2, CTSB, EZH2, SERPINE1, CD14 and DYNLL1 are the key genes in the regulatory network, and UBB is the hub gene during ASFV infection. B119L, I215L and MGF360-13L of ASFV inhibit UBB expression, which are important virulent genes of ASFV.

The aim of this study was to monitor the prevalence of pseudorabies virus (PRV) in pig slaughterhouses in Hubei Province. A total of 1 795 blood samples and 2 081 lung samples (lung and hilar lymph nodes) were collected from slaughterhouses of different sizes in nine cities in four regions (East Hubei, North Hubei, West Hubei, Central Hubei) of Hubei Province from 2020 to 2022, and antibodies in serum and pathogens in lung samples were detected by ELISA and PCR targeting the PRV gE gene. Virus isolation and identification, whole genome sequencing, and phylogenetic analysis were performed on positive tissue samples. Serological detection results showed that 104 serum samples were positive, with an average positive detection rate of 5.79%. The results of pathogen detection showed that 82 lung samples were positive, with an average positive detection rate of 3.94%. According to different factors, The highest detection rates of antibody and pathogen were 18.57% and 9.74% respectively in spring. The antibody detection rate of eastern Hubei was the highest (10.77%), and the pathogen detection rate of central Hubei was the highest (4.93%). The detection rates of pathogens and antibodies in grade A small slaughterhouses were significantly higher than those in the other three types, reaching 16.28% and 6.01%, respectively. A total of 17 PRV strains were isolated from PRV positive samples. The results of genetic evolution analysis of whole genome and gB, gC and gE genes showed that the isolates belonged to genotype Ⅱ variants, which were in different genetic evolution branches from early foreign isolates such as Bartha, NIA-3 and Becker. HBXG is suspected to be a recombinant strain of genotype Ⅰ and genotype Ⅱ, and is in the same clade2.2 genetic branch as the earlier reported recombinant HuB1/CHN2017. In conclusion, mutant strains were mainly prevalent in pig farms associated with slaughterhouses in Hubei Province. At the same time, recombination and mutation of vaccine strains and wild viruses were still occurring in vaccinated pig farms. Therefore, this study is of great significance for the continuous monitoring of PRV and development of novel vaccines.

Canine adenovirus type 2(CAV-2) is an important respiratory pathogen in Canine. The aim of this study was to investigate the prevalence and pathogenicity of CAV-2 in parts of Sichuan Province and Chongqing Municipality. The nasal swabs of 121 canines with respiratory tract diseases were collected from 2021 to 2022 for detecting CAV-2 by specific PCR primers, and the epidemic strain was isolated for further study, including its genome characteristics and pathogenicity. The results showed that the positive rate of CAV-2 in canine was 29.8%. Notably, 69.4% of the positive samples had identical 9 consecutive nucleotide deletions (1 035-1 043 nt) in the E3 genes, which resulting in 4 amino acid deletions in the E3 genes (345-348 aa). One strain with natural E3 deletion was successfully isolated, and the half infection value of MDCK cells was 10^-9.46TCID₅₀·0.1 mL^-1 after plaque purification. The puppies inoculated with the isolation showed typical respiratory symptoms, such as sneezing, runny nosing and coughing. The genome of the isolate was 31 786 bp, and shared the homology of 98.8%-98.9% in nt with the CAV-2 genomes in GenBank. Compared with the known CAV-2 genomes, besides the unique mutations of the E3 gene, the isolate in this study also had unique amino acid mutations in Fiber, Hexon, and E1A, which made this strain distinguish from the known CAV-2 strain in the genomic evolutionary tree. This study obtained the genome sequence of the Chinese CAV-2 strain for the first time, and found the strain with natural deletion in E3 genes, which is helpful to understand the prevalence and genetic evolution of CAV-2.

The purpose of this study was to analyze the biological function of 0580 gene of Mycoplasma bovis (Mb) PG45 strain. In the previous stage, we found that PG45、08M and 07801 strain of M. bovis can lyse mouse red blood cells. So, according to the amino acid sequence of the putative hemolysin related gene 0580 in PG45 strain, gene MBOVPG45_0580 presumed to be related to hemolysin was screened. Due to the protein encoded by this gene is predicted to have four transmembrane regions and is difficult to express, we constructed the prokaryotic expression vector Pcold III-0580-C terminal fragment without transmembrane region to induce expression and purification, The recombinant protein 0580-C was immunized in BALB/c to prepare polyclonal antibody. At last, we explored whether the 0580-C recombinant protein has the ability of lysing RBCs and influences the ability of the strain to adhere to cell; Besides, the obtained polyclonal antibody specificity and titer were detected. The putative hemolysin protein 0580 in strain PG45 was 100% homologous to M. bovis, and the C-terminal truncated form of protein 0580 which removed four transmembrane regions could be successfully expressed in E. coli. The relative molecular weight of the purified recombinant 0580-C protein was about 37 ku. The hemolytic test of mouse RBCs found that the recombinant 0580-C protein has no hemolytic activity, but the cell adhesion test show that the protein contributes to improve the adhesion ability of Mb to EBL cell, polyclonal antibody of 0580-C protein could effectively block the adhesion process. The titer of the polyclonal antibody against 0580-C protein was as high as 2¹¹. The polyclonal antibody could recognize the 0580 protein expressed by three strains of M. bovis (PG45, 08M, 07801), which indicated that the polyclonal antibody against 0580-C protein was specific. This study found that the C-terminus of protein 0580 may be a novel adhesive-related molecule, which provides new insights into the biological function of putative hemolysin associated protein and the pathogenic mechanism of Mycoplasma bovis.

One to three week old pigeons from a farm in Chongqing had large pieces of cheese-like substances in their throats, causing pigeons' throats to be blocked, resulting in inability to eat and death in succession. The purpose of this study was to identify the pathogens that caused the disease of a large number of young pigeons. The pathogen was isolated from the throat, crop, glandular stomach and intestines of diseased pigeons. The samples were collected from the throat, crop, glandular stomach and intestines of diseased pigeons, and inoculated on PDA plates containing 1% gentamicin to isolated pathogens. Pathogens were identified by gram staining, cotton blue staining, germ tube test, and PCR amplification. Animal regression test and antifungal susceptibility were carried out. Eight gram-positive yeast-like cells with oval, black or purple pink color were isolated from 5 diseased pigeons. The results of cotton blue staining showed that mycelia and spores grew from the isolates. These isolates were milky white smooth colonies with neat edges on PDA plates containing 1% gentamicin. When incubated in fetal bovine serum, they can grow into germ tubes. Transcriptional spacer sequence of Candida albicans specific rDNA genes were amplified from the isolates. The results of animal regression test showed that the isolated strains causes swelling of crops, and the degeneration, necrosis and abscission of crops' squamous epithelial cells. The antifungal susceptibility testing showed that the isolates were susceptible to amphotericin B, nystatin and clotrimazole. In this study, Candida albicans was identified as the pathogen of pigeons at a pigeon farm in Chongqing, and the Candida albicans isolates were susceptible to some antifungal drugs in vitro.

Enterococcus faecium, as an opportunistic pathogen in the human and animal intestines, could cause endocarditis and sepsis in the host under certain conditions. Recently, multiple studies indicated that Enterococcus faecium, antimicrobial resistance and pathogenicity of Enterococcus faecium in food animal and wild animal are increasing, but there are fewer studies about AMR in the wild animal. In order to explore the cause of the sudden death of a red-bellied squirrel in Beijing Daxing Wildlife Park, gross autopsy, pathological tissue section, pathogenic bacteria isolation, antibiotic susceptibility testing and pathogenicity analysis were performed on the dead red-bellied squirrel. Results showed that blue flora with short chains of circular or oval shape was observed in lung and kidney sections, and bacteria pathogen were isolated from lung, liver, spleen and abdominal effusion. Antibiotic susceptibility tests have shown that the isolated strains are highly resistant to sulfisoxazole and telomectin; The results of whole genome sequencing showed that the isolated strains all belonged to Enterococcus faecium ST-324, and all carried the virulence gene efaAfm and antibiotic resistance genes tetM and msrC; the core genome single nucleotide polymorphisms (SNPs) analysis showed no significant differences. Animal pathogenicity tests suggested that isolated strains have pathogenicity. The results suggested that the cause of death of the squirrel was sepsis caused by Enterococcus faecium infection.

To investigate the infection of intestinal parasites (especially Clonorchis sinensis) in pet cats in Henan Province and analyze related disease factors and clinical manifestations, a total of 898 fecal samples from 11 regions of Henan Province were examined by centrifugal sedimentation, Lugol's solution staining, and sugar-flotation technique. The results showed that the total infection rate of intestinal parasites was 10.58% (95/898). Among the 95 positive samples, the dominant species was coccidia (49.47%), followed by Clonorchis sinensis (29.47%), hookworm (16.84%), Blastocystis (7.37%), Giardia (3.16%) and Ascaris (3.16%). In general, the prevalence rate of intestinal parasites infection was higher in pet cats in the Jiaozuo region, 6 months of age and younger, not dewormed, not immunized and diarrhea. Except for gender (P>0.05), risk factors including age, deworming, and immunity affected the infection rate of C. sinensis in pet cats. There was also a significant correlation between the diarrhea of pet cats and the infection of C. sinensis. Clinical symptom analysis showed that pet cats infected with C. sinensis were more emaciated than those infected with other parasites (P<0.001). In summary, pet cat owners in Henan Province should pay more attention to scientific feeding, regular immunization and deworming of young cats to reduce the harm of intestinal parasites and the risk of transmission of zoonotic clonorchiasis.

The aim of this study was to explore the molecular mechanism of Sihuang Zhili Granule in the treatment of piglet diarrhea based on the functional module, and to explain its compatibility law. TCMSP, TCMID and HERB databases were used to screen the active ingredients of Coptidis Rhizoma, Phellodendri Chinensis Cortex, Rhei Radix et Rhizoma, Scutellariae Radix, Isatidis Radix, Glycyrrhizae Radix et Rhizoma, and collect the related targets. Disease targets of piglet diarrhea were selected from GeneCards、NCBI、OMIM database. Then, the intersection of drug targets and the disease targets were collected by Venny platform, and uploaded to the STRING database for protein interaction analysis. Protein-protein interaction (PPI) network was constructed by Cytoscape software, and its functional module was identified with MCODE plug-in. KEGG pathway enrichment analysis was performed with David database. The results were showed as follows:1) Network analysis showed that a total of 180 active ingredients were obtained, mainly included alkaloids, flavonoids, anthraquinones and glycosides; A total of 88 drug targets and 470 disease targets were screened, and there were 35 intersection targets. 2) The PPI network analysis showed that module 1 composed of 17 target proteins was the functional module of Sihuang Zhili granules. 3) The KEGG enrichment analysis determined that the pathways mainly involved in module 1 include 17 signaling pathways such as TNF, IL-17 and NF-κB. 4) The analysis of meridians showed that Sihuang Zhili granule could act on 11 collateral channels of internal organs of animal body, and mainly on spleen and stomach meridians. In conclusion, alkaloids, flavonoids, anthraquinone and glycosides are the material basis of Sihuang Zhili granules. They mainly regulate the signaling pathways such as TNF, IL-17 and NF-κB by acting on the key targets included in functional module 1, regulate multiple meridians of the body such as spleen and stomach, and play the role of treating piglet diarrhea from multiple levels such as protein targets, signaling pathways and meridians.

To investigate the drug resistance and virulence gene carrying of Klebsiella pneumoniae isolates, sampling investigation and drug resistance analysis were carried out in a slaughterhouse in Guangzhou. Klebsiella pneumoniae was isolated and identified from pig lung samples, and six common capsular serotypes (K1, K2, K5, K20, K54, K57) were identified, and the minimum inhibitory concentration (MIC) of 13 commonly used antibiotics was detected by the agar dilution method. The resistance genes (bla_KPC-2, bla_NDM-1, bla_NDM-5, bla_CTX-M1, tetA, tetB, sul1, sul2, oqxA, qnrB, aac(6')-Ib-cr) and virulence genes (rmpA, entB, iucA, iroN, ycfM, fimH, mrkD, wabG) were detected by PCR. The results showed that 116 strains of Klebsiella pneumoniae were successfully isolated and identified from 150 pig lung samples, the isolation rate was 77.3%. The bacteria showed red short rod shape by Gram staining microscopy, and serotype of capsule was mainly K57 (50.9%). The results of drug sensitivity test showed that most of the isolates showed multi-drug resistance phenotype, and were highly resistant to tetracycline, β-lactam and sulfonamides, and sensitive to meropenem, colistin and enrofloxacin. The detection of drug resistance genes showed that the highest detection rate was sul2 gene (60.3%) and oqxA gene (59.5%), and the lowest detection rates were bla_NDM-1 and bla_NDM-5 genes (both 0.9%). The virulence gene detection showed that the highest positive rate was ycfM gene (87.1%), and the lowest positive rate was iroN gene (1.7%). The results showed that the detection rate of Klebsiella pneumoniae isolated from slaughterhouse was high, the situation of multiple drug resistance was severe, and the resistance genes and virulence genes were abundant, providing reference for the prevention and control of swine Klebsiella pneumoniae.

This experiment was to investigate the role of glutamine in the regulation of apoptosis in Bacille Calmette-Guerin vaccine (BCG)-infected macrophages. In this study, a cellular model of apoptosis caused by BCG infection was established using mouse mononuclear macrophages RAW264.7, and the optimal time of BCG infection and the number of infection replicates were clarified. Then, a glutamine deprivation model was constructed by glutamine-free medium and combined with BCG infection, and the Key metabolic markers of glutamine were detected by ELISA, immunoblotting, immunofluorescence and flow cytometry. The results showed that BCG infection significantly upregulated the expression of Caspase 3 and PARP in mouse macrophages RAW264.7 (P<0.001) and promoted the catabolism of glutamine. Glutamine deprivation significantly reduced glutamate (P<0.001) and glutathione (P<0.01) levels in BCG-infected macrophages and increased intracellular ROS content (P<0.001). Meanwhile, glutamine deprivation highly significantly up-regulated the expression of Caspase 3, PARP, Cytc and Bax (P<0.001) and highly significantly down-regulated the expression of the apoptosis suppressor protein Bcl-2 (P<0.001). These results indicated that glutamine deprivation in BCG-infected RAW264.7 cells promotes apoptosis by reducing intracellular glutathione levels, causing ROS accumulation and thus promoting apoptosis through an endogenous pathway.

The study aimed to clarify the SLA genetic background of experimental Rongchang pigs and further study SLA-1 molecule-related antigen presentation and immune response mechanism. In this study, the anticoagulant bloods were collected from 27 Rongchang pigs, and peripheral blood lymphocytes were isolated. Total RNA were extracted and SLA-1 genes were amplified using specific primers with RT-PCR method, cloned and sequenced. Molecular genetic characte-ristics of the obtained sequences were further analyzed. The results showed that a total of 11 SLA-1 alleles were obtained in Rongchang pigs, of which nine were novel alleles. All alleles obtained GenBank accession numbers and official names were assigned by the ISAG SLA Nomenclature Committee. The frequency of SLA-1^*24:01 allele was the highest in this population. The full-length of the coding region of SLA-1 gene was 1 086 bp, with 127 nucleotide polymorphic sites. The number of non-synonymous SNPs was higher than that of synonymous SNPs. The nucleotide diversity was 0.044 9, the haplotype diversity was 1, the average nucleotide difference number was 48.782, and the G+C content was 64.9%. There were 75 amino acid variation sites among 361 amino acids encoded by SLA-1 gene. The coding region of exon 2 and exon 3 had the highest polymorphism, and the degree of amino acid variation in exon 2 region was higher than that in exon 3 region. Among the 33 key amino acid sites that make up the six pockets of the peptide-binding groove, 11 of them were highly conserved between human and Rongchang pig, and 10 of the 19 key amino acid sites were consistent with β2-microglobulin. Only two sites, 225 (Thr/Ser) and 228 (Thr/Met), were different among the key amino acid sites that CD8 molecules bind to MHC. The analysis of homology and phylogenetic tree showed that SLA-1^*10:03 and SLA-1^*18:03 had the highest homology with human HLA-A^*02:01 and HLA-A^*11:01 alleles, respectively. Rongchang pig was closely related to Asian boar, Bama miniature pig, Rongshui miniature pig and other Asian pig breeds. The SLA-1 alleles of Chinese Rongchang pigs were successfully identified, and it was found to have highly rich polymorphism. The results of this study provided a genetic foundation for revealing the SLA genetic background of Rongchang pigs and conducting xenotransplantation research.

This experiment was conducted to establish a technical platform for metabolite analysis based on chromatography-mass spectrometry (UPLC-Q-TOF-MS/MS), three-week-old chickens were feed with equal dose of the Chinese veterinary drug andrographolide (AG) and the broad-spectrum antibiotic oxytetracycline (OTC) respectively, to analyze the metabolite changes in the excretion of chickens after intervention. Through statistical analysis of the data, differential metabolites were obtained, and the intervention mechanism of AG and OTC on chicken gut microbiota metabolism were also explored. The excretory samples of yellow-finned chickens in each group were detected by UPLC-Q-TOF-MS/MS, and the metabolic pathways were enriched and analyzed by MetaboAnalyst platform. The differential metabolites were screened by using QI software. The results showed that compared with the blank group, 12 and 10 differential compounds were found in OTC and AG group, respectively. Fifteen metabolic pathways were involved in the OTC group, of which 2 most relevant metabolic pathways were linoleic acid and glycerophospholipid metabolism; 17 metabolic pathways were involved in the AG group, of which 5 most relevant metabolic pathways were glycerophospholipid metabolism, sphingolipid metabolism, cysteine and methionine metabolism, unsaturated fatty acid biosynthesis and glycine, serine and threonine metabolic pathways. It can be concluded that the mechanism of action of OTC and AG affecting chicken metabolism is very similar, both can affect the normal metabolic function of chickens through changing phospholipid metabolism, fatty acid metabolism and amino acid metabolism, especially the distribution pattern of OTC group is significantly different from the control group.

The purpose of this study was to compare the effects of different isolation and culture methods on the culture of feline limbal stem cells (FLSCs) in vitro, and to establish the best isolation and culture system for the continuous and stable proliferation, normal structure and function of FLSCs. The limbal tissues of cats were collected under surgical microscope. Three different digestion methods which were the tissue adhesion method (T method), the mixed enzyme digestion method (N method) and the mixed enzyme digestion methods combined with tissue adhesion method (NT method) were used to separate FLSCs, to identify the expression of LSCs marker protein ABCG2, in order to screen the most suitable method for separation. Cells were cultured through three kinds of complete media which were BC, DLM, and SCM, the cell count and the cell morphology was observed every day for 7 days, and the positive markers of LSCs p63, vimentin and epithelial differentiation markers CK3, CK12 were selected to qualitatively analyze the 3rd, 4th, and 5th generation cells. The results showed that the expression of ABCG2 from cells separated by three methods were positive and that of NT method expressed the most positive, indicating that three methods can all be used to separate FLSCs, and the separation effect of NT method was the best. FLSCs can be cultured in all three media, with no difference in cell morphology. Cells proliferation speed were the fastest in group DLM. Compared with the other two groups, time of cell reaching the peak cell growth was shorter in DLM, and the difference was significant (P<0.05). The growth curve of FLSCs was showed a typical "S" type. The first 72 h of LSCs growth was in the lag phase, the 72-216 h was in the logarithmic growth phase, the 216-264 h was basically in the plateau phase, and the cells entered the decay phase at the 264th hour. The population doubling time of FLSCs was 38.9 h, and the average maximum proliferation concentration was 5.66×10⁵ cells·mL^-1. The 3rd, 4th and 5th generation FLSCs showed strong positive expression of vimentin and p63, but CK3 and CK12 didn't express. FLSCs can be isolated by NT method and cultured in DLM to establish a stable proliferation system of FLSCs in vitro.

Canine intestinal lymphadenopathy is a rare protein-losing enteropathy that is very rare in veterinary practice. In this report, a case of an 8-year-old female Teddy is described with clinical signs of decreased appetite, occasional vomiting, persistent diarrhea, coarse and disheveled coat, abdominal enlargement and panting. Ultrasonographic examination revealed a well-defined stratification of the jejunal wall, moderate thickening of the mucosal layer, echogenic enhancement within the mucosal layer with unclear borders, non-uniformly distributed bright spot structures, and a small amount of ascites in the posterior part of the abdominal cavity. The dog was diagnosed with canine intestinal lymphadenopathy based on a comprehensive analysis of its medical history, clinical signs, hematological examination and imaging findings. Based on the conventional symptomatic therapeutic measures, a combination of improved recipes and medication was developed according to the actual prognosis. After more than two years of long-term maintenance treatment and prognostic follow-up, the dog showed significant weight gain, normalization of biochemical parameters, and significant improvement of intestinal symptoms on ultrasound imaging. This is the first report of a clinical case and treatment plan for canine intestinal lymphatic duct dilatation in China, which has reference value and significance for the diagnosis, treatment and research of related diseases in veterinary clinic.

The fluoride widely present in water and food of livestock and poultry seriously threatens the liver health of livestock and poultry and the safety of animal food. To elucidate the internal mechanism of fluoride-induced liver injury, and clarify the regulatory role of IL-17A in fluoride-induced liver inflammation and hepatocyte apoptosis, this study randomly divided 24 wild-type C57 mice and 12 IL-17A knockout mice into control, NaF, and KO+NaF groups. In addition, HE staining, ELISA, flow cytometry, and immunohistochemistry were used to detect the changes of morphology, inflammatory cells, inflammatory factors, and apoptosis in the liver. The results showed that fluoride exposure induced liver morphology damage, increased the content of inflammatory factors (TNF-α, IL-17A, INF-γ, IL-23, TGF-β) and the levels of M2 macrophages and dendritic cells, decreased the levels of IL-1β, natural killer cells, γδT cells, CD4⁺T cells and the ratio of CD4⁺T cells/CD8⁺T cells in the liver. In addition, the results of apoptosis detection showed that fluoride exposure increased the apoptotic cells and the protein expression levels of key apoptosis genes Cyt-c and Caspase3 in the liver. However, compared with the NaF group, the liver injury was alleviated, the contents of inflammatory factors (INF-γ, TNF-α, TGF-β, IL-23, IL-17A) and dendritic cells were significantly reduced, and the number of apoptotic cells and the protein expression levels of Cyt-c and Caspase3 were significantly decreased in the liver of KO+NaF group. In summary, IL-17A knockout alleviated fluoride-induced inflammatory response and hepatocyte apoptosis. This study provides theoretical basis and new ideas for the research and the scientific prevention/treatment of fluorotoxic liver injury.

Neuromedin B(NMB) and its receptor(NMBR) could inhibit the infection of influenza A virus(IAV) H1N1 subtype through activating the pathway of NF-κB signaling. However, the regulation of NMB and NMBR on the expression of ubiquitination ligases related with NF-κB signaling pathway remains unclear. To explore the effects of NMB and NMBR on regulating the ubiquitination ligases involved in NF-κB signaling pathway during IAV/H9N2 infection, the expression levels of E3 ubiquitin ligase mind bomb-2(MIB2) and ring finger protein 8(RNF8), deubiquitin enzyme cylindromatosis(CYLD) involved in NF-κB signaling pathway were analyzed using the methods of RT-PCR, qRT-PCR, and Western blot(WB), based on the sh-NMBR cells and NMB stimulating A549 cells during IAV/H9N2 infection. The results showed that the increased expression levels of RNF8 and CYLD, and the decreased levels of MIB2 and P65 phosphorylation were observed in sh-NMBR cells during H9N2 infection, while the decreased levels of RNF8 and CYLD, and the increased levels of MIB2 and P65 phosphorylation were confirmed in NMB stimulating A549 cells. These results indicated that NMB and NMBR could affect the function of NF-κB signaling pathway by regulating the expression levels of ubiquitination ligases and P65 phosphorylation during IAV/H9N2 infection, which could provide a theoretical basis for further studying the mechanism of NMB and NMBR on inhibiting influenza A virus.