Machine learning method is an important branch of genomic selection, and deep learning has become a new research hotspot in the field of machine learning in recent years. The principles and application development of machine learning and deep learning genomic selection were introduced in this paper, and the research progress of deep learning genomic breeding value estimation from the aspects of model framework, model parameters, feature selection were elaborated. Some problems in deep learning genomic selection research were explored and prospects in the future were discussed.

Fat content is one of the main factors affecting the quality of livestock meat. The distribution of fat in muscles is closely related to the color, palatability, flavor, tenderness and hydration of meat, and intramuscular fat is regarded as a key indicator for evaluating meat quality. Intestinal microbiota and mitochondria, as the largest microecosystem and cellular "power factories" in the body, play important bridging roles between lipid metabolism and cellular metabolism. At present, research on the gut microbiome and mitochondria is relatively extensive, but the impact of their interaction on fat deposition is not yet clear. This article reviews the research on gut microbiota and their metabolite functions and fatty acid metabolism, fat deposition and mitochondrial function, as well as the regulatory mechanisms of gut microbiota and their metabolite interactions with mitochondria on fat deposition, in order to provide references and ideas for improving animal meat quality through gut microbiota and mitochondria.

Meat quality improvement is one of the important goals of pig breeding. Meat quality traits are influenced by a variety of factors such as genetics, feed nutrition and management, and new research suggests that intestinal microorganism may also be a key factor. Intestinal microorganism is a complex system that is gradually formed and perfected as pigs come into contact with the external environment after birth, and it plays an important role in the growth and development of pigs. Earlier studies on porcine intestinal microorganism primarily focused on its impact on the host's nutrient digestion and absorption, as well as the immune system. In recent years, a new field of inquiry has emerged to improve pork quality through intestinal microorganism, focusing on three aspects including muscle fiber type transition, intramuscular fat content, and metabolism of skeletal muscle. Therefore, based on a brief introduction of muscle development and intramuscular fat deposition, as well as the composition and role of intestinal microorganism, this paper reviews the studies on the intestinal microorganism regulating muscle fiber type conversion, intramuscular fat deposition, and metabolism of skeletal muscle to improve the quality of pork, aiming to provide a scientific basis for the further exploration of the intestinal microorganism regulating the quality of pork.

With the improvement of milk production of dairy cows, China's dairy industry chain is also continuously optimized and upgraded, making progress towards modernization, standardization and internationalization. However, the reproductive performance of high producing dairy cows is gradually decreasing, which has become a common problem faced by dairy cows related researchers. Follicle development is closely related to reproductive efficiency, and normal follicle development is a prerequisite for reproductive activities such as estrus and ovulation. During follicle development, follicular granulosa cells can interact with oocytes and theca cells to produce a variety of hormones and factors, and act as receptors for a variety of hormones and factors to affect follicular development. Therefore, this paper summarized the research on the function of follicular granulosa cells and follicular development, and focused on the role of follicular granulosa cells in follicular development and the influence of granulosa cells on oocytes, in order to provide theoretical reference for future related studies.

Periparturient cows undergo intricate regulation influenced by multifaceted factors. Owing to alterations in physiology, diet, environment, and management practices, these cows become predisposed to negative energy balance and immunosuppression, potentially culminating in various diseases and a reduction in reproductive efficiency. Drawing upon recent research, this article systematically examines the nuanced alterations in energy dynamics, hormonal profiles, as well as vitamin and mineral levels, and cytokine patterns within periparturient cows. The investigation reveals that pivotal components such as blood glucose, insulin, fat, and amino acids undergo systematic and rhythmic changes during this critical period for dairy cows. Furthermore, reproductive hormones and other regulatory factors exhibit marked fluctuations. Additionally, there are noteworthy adjustments in the requirements for vitamins and minerals. Given the intricacy of these physiological transformations, it becomes imperative to offer well-founded recommendations for the dietary management of periparturient cows. This not only aids in averting and managing energy metabolism disorders but also contributes to the enhancement of milk quality and the amplification of overall production efficiency in dairy cows.

Endometrial fibrosis is caused by persistent damage stimulation of the endometrium leading to its impaired repair, characterized by excessive deposition of extracellular matrix, which may lead to endometrial structural damage and dysfunction or even failure of the endometrium, affecting the fertility and reproductive quality of domestic animals. Therefore, analyzing the formation mechanism of endometrial fibrosis is of significance for improving the fertility of domestic animals. This article elaborates on the trigger factors of endometrial fibrosis, the relationship between the cellular and molecular mechanisms of fibrosis formation, cellular metabolic regulation and extracellular matrix, further discusses the therapeutic strategies for fibrosis and future research directions aiming to provide a reference for the study of endometrial fibrosis in domestic animals.

Feline upper respiratory disease (FURTD) is widely spread in cats, characterized by upper respiratory, oral, and eyes symptoms. There are various pathogens caused FURTD. Except for common pathogens such as feline calicivirus (FCV), feline herpesvirus type 1 (FHV-1), Chlamydia felis (C.felis), Mycoplasma felis (M.felis), and Bordetella bronchiseptica (Bb), there are still many other sporadic infections in clinical practice. Clinically, there are often cases of co-infection by multiple pathogens, and the continuous evolution easily leads to the increased pathogenicity and changes of tissue tropism. The complexity of this etiology often makes the prevention and treatment of FURTD more difficult. Meanwhile, due to the similar symptoms caused by many pathogenic infections in cats, it is not possible to accurately determine the cause of the case solely based on clinical symptoms. To understand the characteristics of different pathogens and master detection methods specific for different pathogens are important to rapidly diagnose and provide efficient treatment strategies specific for FURTD. At present, lots of cases of FURTD caused by different pathogens are reported frequently. The diagnostic methods, treatments, and vaccine development for FURTD haven't been yet mature. Here, this article will systematically clarify the FURTD-associated emerging and remerging pathogens, diagnostic methods, treatment methods, and prevention and control measures, aiming to provide more ideas for the prevention and treatment of FURTD.

Hypoxia-inducible factor-1α (HIF-1α) is a central regulatory factor in cellular response to hypoxia. Stable HIF-1α has multiple functions and is involved in various biological processes, including angiogenesis, metabolic regulation, autophagy, and apoptosis. The stability of HIF-1α is regulated by multiple signals, including oxygen levels, pathogen infections, and metabolic intermediates. In recent years, increasing evidence has shown that HIF-1α plays an important role in infectious diseases. Therefore, this article discusses and summarizes the regulatory mechanisms of HIF-1α stability and its role in viral, bacterial, and other pathogen infection, as well as in the development of related diseases, aiming to provide references for further study on the role of HIF-1α in defending against various pathogen infections and its potential as a target for targeted therapy.

Japanese encephalitis (JE) is a mosquito-borne animal disease caused by Japanese encephalitis virus (JEV), although commercially vaccines are availabe, however, the vaccines has certain limitations and there is no effective drug treatment so far, and once the outbreak occurs, people's life and health as well as the public safety will be threatened, thus resulting in significant economic losses. JEV contains a large number of encoded proteins with immune escape function. The mechanism of these proteins escaping host immune function is diverse and complex, and the mechanism of which still remains unclear. At the same time, the complex immune escape mechanism of JEV may be the key factor hindering the immune protection effect of vaccine. Therefore, this paper mainly summarizes the mechanism of JEV escaping innate and adaptive immunity by inhibiting interferon response, regulating apoptosis, pyroptosis, autophagy and host miRNA to provide ideas and theoretical basis for the research of JEV pathogenesis and vaccine development.

With the implementation of the strategy of "reducing antimicrobials, banning antimicrobials, and making products antimicrobial-free" in China's aquaculture industry, the use of plant extracts as feed additives has become an important research direction. Glycyrrhiza polysaccharide is a kind of plant polysaccharide extracted from licorice, and the extraction methods include solvent extraction, ultrasound-assisted extraction, and microwave-assisted extraction, etc. As natural biomolecules, Glycyrrhiza polysaccharides consist of monosaccharides such as arabinose, glucose, galactose, rhamnose, mannose, xylose, etc., which have a variety of biological activities such as antioxidant, anti-inflammatory, immunomodulatory, anti-tumor and so on. It has a significant role in protecting the health of animals. The addition of Glycyrrhiza polysaccharide to animal diets can improve body weight growth rate, feed utilization, and meat quality, in addition reducing the negative effects on animals caused by intestinal diseases and stress. Due to its anti-inflammatory and antioxidant effects, Glycyrrhiza polysaccharide effectively reduces inflammation and damage induced by disease and stress in animals. In this paper, the extraction methods, biological activities, and mechanisms of action of Glycyrrhiza polysaccharides were reviewed, their applications in animal production were also discussed, in order to provide a scientific basis for the application of Glycyrrhiza polysaccharides in animal breeding.

The aim of this study was to estimate the genetic parameters of the reproductive traits of a sow farm in Beijing, evaluate their genetic progress, and provide a theoretical basis for developing a reasonable breeding scheme. In this study, a total of 29 269 original breeding records, 20 770 original birth records, and 146 871 original pedigree records were collected from a sow farm in Beijing from 2016 to 2022. The genetic parameters of reproductive traits of French Large White sows in the company were estimated. The best linear unbiased prediction (BLUP) method based on animal model was used to estimate heritability, genetic correlation, phenotypic correlation and breeding value by DMU software using single-trait and double-trait repeatability models. The results showed that the TNB was 12.67 heads in this study, the NBA was 10.90 heads, NSB for 1.35 heads, the MUM was 0.14 heads, the LWT was 13.99 kg, the LW 21 was 73.93 kg; The estimated heritability of reproductive traits ranged from 0.017 to 0.134, belongs to the low heritability; Genetic correlations among reproductive traits was between -0.368 and 0.856, phenotypic correlation was between -0.213 and 0.847; By mapping the phenotypic trends and genetic trends of reproductive traits, individuals showed increasing reproductive trait phenotypic values, breeding values rose first and then decreased. The results of this study show that the genetic improvement in this population is good, and the results can provide reference guidance and theoretical basis for the breeding and genetic improvement of French Large White pigs.

The cobalt chloride (CoCl₂) was utilized in this study to simulate hypoxia in muscle cells and employed transcriptomic sequencing to analyze the effects of hypoxia on muscle fiber type and genes expression that related to muscle fiber types. Primary chicken myoblasts were treated with different concentrations of CoCl₂ (0, 50, 100, 200 and 400 μmol ·L^-1), with 3 repeat wells in each group. The CoCl₂ concentration was screened by detecting the expression of hypoxia marker genes and observing cell morphology to determine the most suitable concentration for modeling. Subsequently, the collected cell was used to detect the gene expression profile of chicken myoblast in hypoxia treatment and control treatment by Illumina technology and assess differential gene expression and enriched signaling pathways between treatments. The results indicated that 200 μmol ·L^-1 CoCl₂ was the most suitable concentration for constructing the hypoxic model in chicken embryonic myogenic cells. Transcriptomic results showed a total of 1 474 and 2 028 differentially expressed genes in the hypoxia treatment compared to the control group at 24 and 48 h, respectively. Interestingly, 959 genes showed differential expression in both time points.GO and KEGG analyses revealed significant alterations in key factors related to signaling pathways such as regulation of signaling receptor activity, integral component of plasma membrane, neuroactive ligand-receptor interaction, PI3K-AKT signaling pathway and muscle contraction in chicken muscle cells under hypoxic conditions. Consistent findings from sequencing and real-time quantitative polymerase chain reaction (qRT-PCR) demonstrated that slow muscle fiber-specific genes such as MYH7B and CSRP2 were significantly upregulated after hypoxia treatment, while fast muscle fiber-specific genes including MYH1F, MYH1D, TNNI2, and SOX6 were significantly downregulated, which indicated a transition of chicken muscle cells from fast to slow muscle fibers was induced by hypoxia. This study provides preliminary insights into the mechanisms underlying the hypoxia-induced transition of muscle fiber types in chicken myogenic cells and offers a valuable gene expression database for future research on poultry meat quality and hypoxia-related studies in mammals.

The purpose of this study was to preliminary investigate the function of the SOX18 gene in Hu sheep hair follicle dermal papilla cells. In this study, immunofluorescence staining was used to identify cells isolated from the hair follicles of Hu sheep. Subsequently, an overexpression vector of the SOX18 gene was conducted, and its overexpression efficacy was assessed using qRT-PCR and Western blot. The dermal papilla cells successfully identified and well-grown were divided into two groups. One group was transfected with SOX18 overexpression vector, while the other was transfected with empty plasmid. Each group had 3 replicates. The total RNA was extracted from 6 samples using the Trizol method, followed by the construction of a cDNA library. The Illumina Novaseq6000 platform was used for sequencing. Subsequently, sample correlation and differentially expressed genes were analyzed. The differentially expressed genes were categorized based on GO and annotated using the KEGG pathway analysis to identify candidate genes and pathways associated with the regulation of the SOX18 gene. To confirm the accuracy of the transcriptome data, 9 differentially expressed genes were randomly chosen for qRT-PCR analysis. The immunofluorescence results revealed dermal papilla-related genes had high expression in isolated cells, indicating a high purity of the isolated dermal papilla cells. qRT-PCR and Western blot results showed that overexpression of the SOX18 gene could increase the mRNA and protein levels of SOX18 in dermal papilla cells, indicating that the SOX18 overexpression vector could be used for subsequent experiments. Transcriptome analysis results showed a good correlation between samples. A total of 157 genes were differentially expressed between the SOX18 overexpression group and the control group, of which 103 genes were up-regulated after SOX18 overexpression and 54 genes were down-regulated. qRT-PCR analysis showed high accuracy of transcriptome data. GO functional analysis showed that some differential genes were enriched in cell progression and hair follicle development. KEGG enrichment analysis showed that some differentially expressed genes were enriched in signaling pathways related to cell proliferation and hair follicle development. GO functional analysis and KEGG enrichment analysis indicated that SOX18 played a role in the proliferation of dermal papilla cells and the development of hair follicles. In this study, transcriptome sequencing analysis found that SOX18 may affect the proliferation of dermal papilla cells and the development of hair follicles by affecting genes and signaling pathways related to cell proliferation and hair follicle development. The study results provide a theoretical basis for understanding the molecular regulatory mechanism of the SOX18 gene in Hu sheep dermal papilla cells and hair follicles.

The aim of this study was to explore the effects of PPARγ on the cell viability, apoptosis, cell migration rate and lipid accumulation of sheep trophoblast cells by interfering with the expression of PPARγ gene, so as to provide a basis for further study of the molecular mechanism of PPARγ in trophoblast cells.To design and synthesize siRNA interference fragment of PPARγ gene, the siRNA with the highest interference efficiency was screened by Western blot and RT-qPCR. Then, the siRNA was transfected into the sheep trophoblast cells. After interfering with the PPARγ gene, the accumulation of lipid droplets in trophoblast cells was measured by BODIPY staining, the amount of triglycerides by colorimetric method, and the transcript levels of genes involved in lipid metabolism was detected using RT-qPCR (real-time quantitative PCR). CCK-8 assay, wound healing test and flow cytometry were used to detect the effects of PPARγ interference on cell viability, migration rate and apoptosis of trophoblast cells. siPPARγ-1 fragment with the highest interference efficiency was screened and used to transfect trophoblast cells to interfere with the expression of PPARγ gene. Interference with PPARγ gene expression in trophoblast cells reduced the accumulation capacity of lipid droplets and cellular triglyceride content (P < 0.01), and the transcription levels of related genes CD36, FABP4 and PLIN2 were also inhibited (P < 0.01). At the same time, interference with PPARγ gene expression significantly reduced cell viability (P < 0.01) and migration rate (P < 0.05), while the cell apoptosis rate was increased significantly (P < 0.01).This study showed that PPARγ gene plays an important role in regulating the function of trophoblast cells and its specific molecular mechanism can be studied in depth with a view to using it in reproductive breeding practices.

The purpose of this study was to explore and evaluate 6 different methods for extracting genotype feature of single nucleotide polymorphisms (SNP). Six methods were analyzed and compared: principal component analysis (PCA), gene-principal component analysis (gene-PCA), SNP-Pearson correlation coefficient (SNP-PCC), linkage disequilibrium (LD), and genome-wide association study (GWAS) and random sampling (RS). The prediction accuracy of GEBV in 2 sets of data (Beijing duck, 542 samples, SNP loci 39 932; Duroc pig, 2 549 samples, SNP loci 230 884) and 3 sets of phenotypes (Beijing duck body length, Duroc pig backfat thickness and teat number) was evaluated. Results showed that SNP-PCC combined with 5 GS methods (GBLUP, BayesA, BayesB, BayesC, and Bayesian Lasso) achieved relatively reliable prediction accuracy for the Pecking duck body length phenotype and achieved the highest average prediction accuracy in pig backfat thickness and teat number phenotypes (increased by 5%, reaching 32.3%), and significantly improved computational efficiency (on average 5-7 times faster). In summary, this study found that selecting appropriate feature extraction methods can effectively improve the accuracy and computational efficiency of GS prediction, laying the foundation for in-depth research on the impact of different feature extraction methods on GS prediction accuracy, and providing reference for their application in breeding practice.

This study investigates the effect and mechanism of rhodiola sachalinensis polysaccaride (RSP) on the proliferation and apoptosis of procine leydig cells under hypoxia conditions. The appropriate hypoxia injury time, optimal RSP concentration and the pre-treattime duration determined by CCK-8 assay. The experiment was divided into 3 groups, normal group, hypoxia group, and hypoxia+RSP group, and cell cycle proliferation and apoptosis were detected by flow cytometry method, the cell proliferation and apoptosis-related genes (CCND1, CDK4, IL-6, TNF-a, Bcl-2, Bax and Caspase-3) mRNA expression levels was detect by RT-qPCR, Western blot was used to detect the expression of Bax, Bcl-2, p21, JNK, p-JNK and p53. The viability of porcine leydig cells was decreased under hypoxia, which could be alleviated after 18 hours of pretreatment with 0.012 5 mg·mL^-1 RSP. The Flow cytometry results showed that RSP could alleviate cell cycle arrest induced by hypoxia in porcine leydig cells. RSP significantly up-regulated the expression of Bcl-2, CCND1, CDK4(P < 0.05), and down-regulated the expression of TNF-α, IL-6, Bax, Caspase-3, and p21, p53, and pJNK/JNK (P < 0.05). RSP helps ensure the normal progression of leydig cells from S-phase to G2-phase under hypoxia conditions, promote cell proliferation and inhibit apoptosis. It is speculated that RSP regulates cell proliferation and apoptosis through p53/p21 and JNK pathways.

The aim of this study was to explore the impact of breed factors on the efficiency of commercial rapid propagation of OPU-IVEP. This study used healthy 8-month-old to 3-year-old black headed Suffolk, black headed Dorper, and Texel meat breeding ewes identified by breed identification as donors. OPU technology was used to obtain oocytes at Inner Mongolia Saino Sheep Technology Co., Ltd. for in vitro embryo factory production or in vivo embryos obtained through laparoscopic insemination and surgical flushing. Embryo transfer was performed, and relevant data of oocyte recovery, embryo development, and transfer pregnancy were recorded for statistical analysis. The differences in oocyte recovery efficiency, oocyte in vitro fertilization and embryo development ability, embryo transfer pregnancy and lambing efficiency among different breeds in large-scale in vitro embryo production was compared; At the same time, the application effects of in vivo and in vitro embryo production technologies of different breeds were also compared. The results showed that: 1) there was a significant difference in oocyte recovery rates among the 3 breeds (P < 0.01), with the highest recovery efficiency observed in the black headed Dorper; 2) There was no significant difference in the in vitro fertilization and cleavage rates among the 3 breeds (P > 0.05), but the blastocyst development rate of Texel sheep was significantly higher than the other 2 breeds (P < 0.01); 3) There was no significant difference (P > 0.05) in the conception rate and average number of lambs produced by embryo transfer between different breeds, whether in vivo or in vitro, and there was no significant difference (P > 0.05) in the average conception rate and lambing rate of in vivo and in vitro embryo transfer in different breeds. In summary, in the process of OPU-IVEP industrial and large-scale application in sheep in vitro embryo production, there are differences in the efficiency among different breeds, such as follicle development, number of retrieved oocytes, and blastocyst development rate. Texel sheep, which has the lowest average number of follicles and average number of retrieved oocytes, has the highest blastocyst development rate; The conception and lambing situation after transplantation, which directly affects the efficiency of rapid reproduction, did not show significant differences among different varieties, and there was no significant difference in the number of offspring obtained from single production of in vivo and in vitro embryos. However, the donor utilization rate of OPU-IVEP is higher than that of MOET.

The aim of this study was to investigate the effects of heat stress on epigenetic modifications and developmental capacity of bovine oocytes and their embryos. In this study, oocytes were cultured under in vitro heat stress conditions (41 ℃ for 12 h+38.5 ℃ for 12 h) for in vitro maturation (IVM) and in vitro fertilization (IVF). The developmental ability of oocytes and embryos and the modification levels of epigenetic modifications histone H1, histone H2, histone H4, and DNA methylation were examined in control (38.5 ℃ for 24 h) and heat-stressed cattle. In this study, we also examined oocyte reactive oxygen species (ROS) levels, mitochondrial membrane potential (ΔΨm) levels, and expression levels of genes associated with epigenetic modifications and developmental competence. The results showed that the maturation rate ((59.21±4.29)%), oocyte cleavage rate ((57.78±4.58)%) and blastocyst rate ((22.31±1.67)%) of the heat stress treatment group were significantly lower than those of the control group ((85.10±6.75)%, (78.64±2.46)%, and (42.64±1.38)%, P < 0.05); The levels of epigenetic modifications histone H1, histone H2, histone H4, and DNA methylation of bovine oocytes and embryos at all stages in the heat stress group were significantly lower than those of the control group (P < 0.05); the level of ΔΨm of bovine oocytes in the heat stress group was significantly lower than that of the control group (P < 0.05); the level of ROS in bovine oocytes of heat stress group was significantly higher than that of the control group (P < 0.05); the mRNA expression levels of genes related to epigenetic modification of bovine oocytes DNMT1, DNMT3A, DNMT3B, Histone H2A, SMYD3, and IGF-2R were significantly lower in the heat stress group than in the control group (P < 0.05); the mRNA expression levels of the genes C-MOS, GDF-9 and POU5F1 related to oocyte developmental competence were significantly lower in the heat stress group than in the control group (P < 0.05). The results showed that heat stress significantly reduced the levels of epigenetic modifications histone H1, histone H2, histone H4, and DNA methylation, significantly decreased the levels of ΔΨm and mRNA expression of genes related to epigenetic modifications and developmental competence, and significantly increased the levels of ROS and reduced the quality of bovine oocytes and embryos.

The aim of this study was to investigate the effects of combination of insulin-like growth factor 1 (IGF1), coenzyme Q10 (CoQ10), and melatonin (MT) on development of bovine oocytes, embryos and heat stressed blastocysts. In this study, bovine ovaries were collected from the abattoir and cumulus-oocyte complexes (COCs) were extracted in the laboratory. IGF1, CoQ10 and MT were added to bovine oocyte in vitro maturation (IVM) solution and bovine embryo in vitro culture (IVC) solution. The developmental ability, level of reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) of bovine oocytes from each treatment were detected. By applying 41 ℃ heat stress at the blastocyst stage, the effects of heat stress and combined supplementation on the developmental ability and apoptosis level of bovine IVF blastocysts were determined, and the mRNA expression levels of embryo quality related genes were detected by qRT-PCR. Each experimental group was repeated 3 times. The results showed that the combination of IGF1, CoQ10 and MT (ICM group) significantly increased oocyte maturation rate ((90.40±2.06)% vs. (65.41±0.63)%), cleavage rate ((93.33±1.96)% vs. (59.77±2.93)%) and blastocyst rate ((51.43±5.34)% vs. (26.92±3.24)%)) (P < 0.05) compared with the non added group (CT-0 group). In the ICM group, the ROS level of bovine oocytes was significantly decreased and the ΔΨm of bovine oocytes was significantly increased (P < 0.05). Compared with the heat stress group (HS group), the combination of IGF1, CoQ10 and MT (ICM+HS group) significantly increased expanding rate ((62.00±2.97)% vs. (30.77±8.66)%, P < 0.05) of heat stressed blastocysts, inhibited apoptosis of heat stressed blastocyst cells, and increased mRNA expression levels of IGFBP3, ATP1A1, DSC2, and IFNT2 (P < 0.05) of heat stressed blastocysts. In summary, the combination of IGF1, CoQ10 and MT can effectively enhance the developmental ability, reduce the ROS level and increase the ΔΨm of bovine oocytes. Heat stress reduced the expanding rate of bovine blastocysts, promoted apoptosis of bovine blastocyst cells and damaged the quality of blastocysts, while the combination of IGF1, CoQ10 and MT alleviated the damage from heat stress.

The study aimed to reveal the mechanism of melatonin in alleviating testicular damage caused by busulfan, improve the efficiency of busulfan in preparing spermatogonial stem cell transplantation recipients, and explore its therapeutic effect on male spermatogenic function caused by busulfan's toxicity. In this study, ICR male mice at 6-7 weeks were used as model animals and divided into control group, busulfan treatment group, and busulfan+melatonin treatment group. Each group had 8 replicates and samples were taken on the 7th day after injection. The levels of inflammatory cytokines in testicular tissue was detected using ELISA, and perform HE staining on the testes and epididymis. In addition, immunohistochemistry or immunofluorescence were used to detect the expression of blood testis barrier related proteins to analyze the effect of melatonin on BTB. The results showed that after injection of busulfan into mice, combined with melatonin treatment, the trend of decreased testicular weight and testicular body rate was significantly alleviated, and the rate of decrease in sperm count was slowed down; HE staining of testicular tissue and detection of inflammatory cytokines showed that melatonin significantly slowed down the upward trend of nuclear transportation factor κB, p-NF-κB, interleukin-1β, tumor necrosis factor-α and myeloperoxidase, thereby reducing the infiltration of inflammatory cells in the tissue; immunohistochemical or immunofluorescence detection showed that melatonin slowed down the decrease in the levels of blood testis barrier constituent proteins (Occludin, connexin-43, N-cadherin) and cytoskeletal proteins (F-actin and Vimentin) caused by busulfan treatment. Melatonin alleviates the mouse orchitis response induced by busulfan by down regulating the levels of inflammatory cytokines in testicular tissue; and by slowing down the decreasing trend of BTB related proteins content, the damage of busulfan to BTB and sertoli cells is alleviated, thereby alleviating the damage to mouse spermatogenic functionn

This experiment was conducted to investigate the effects of dietary rumen-protected lysine (RPLys) and rumen-protected methionine (RPMet) supplemented in low-protein diet on growth performance, nutrient apparent digestibility, serum biochemistry and meat quality of Hulun Buir sheep. Sixteen 2.5-month-old weaned healthy lambs were randomly divided into two groups evenly: control group (fed low protein diet with crude protein content of 7.84%) and treatment group (supplemented with 10 g·d^-1 RPMet and 15 g·d^-1 RPLys on the basis of low protein diet). The pretrial period lasted for 7 d and the experimental period lasted for 56 d. The results showed that: 1)The average daily feed intake of treatment group was significantly lower (P < 0.05), but its average daily height increase was significantly higher (P < 0.05), and feed to gain ratio of treatment group was lower than that of control group. 2) There was no significant difference in slaughter performance or nutrient apparent digestibility between two groups (P > 0.05). 3) The contents of total protein (TP), albumin (ALB), creatinine (CREA), lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and low density lipoprotein (LDL) in the serum of treatment group were significantly lower than those of the control group (P < 0.05), and the content of free fatty acid (NEFA) was significantly higher (P < 0.05) than that of the control group. 4) There was no significant difference in meat color, conventional nutrients, the composition and content of fatty acid between the two groups (P > 0.05). The content of threonine (Thr) in muscle of treatment group was significantly higher (P < 0.05), and the contents of isoleucine (Ile), leucine (Leu), tyrosine (Tyr) were significantly lower than those of the control group (P < 0.05). The proportion of flavor amino acids and essential amino acids in total amino acids (DAA/TAA, EAA/TAA) in muscle of treatment group were significantly higher (P < 0.05), and the ratio of essential amino acids to non-essential amino acids (EAA/NAA) was significantly higher than those of the control group (P < 0.05). In conclusion, under the condition of the current experiment, adding 10 g·d^-1 RPMet and 15 g·d^-1 RPLys into the low protein diet has a positive effect on the growth and development of Hulun Buir sheep, and without affecting its nutrient digestibility and slaughter performance. It can also increase the proportion of delicious amino acids and essential amino acids in muscle and improve muscle quality.

The purpose of this study was to investigate the effects of red clover extract on rumen microbial composition and urea metabolism in vitro. The experiment was divided into control group (no red clover extract added) and supplement with red clover extract treatment group. Rumen fluid collected from 3 healthy cows were mixed for in vitro fermentation. The fermentation fluid samples were collected at 0, 0.5, 1, 2, 4, 6 and 24 h of fermentation, respectively, and the contents of urea, ammonia nitrogen (NH₃-N), microbial crude protein and total amino acids were determined. 16S rRNA sequencing was used to reveal the changes of microbial diversity, community composition and function in rumen. The results showed that in the red clover extract treatment group, urea decomposition rate was significantly reduced within 4 h of fermentation, ammonia nitrogen content was significantly reduced after 2 h of fermentation, rumen microbial protein content was significantly increased after 6 h of fermentation, and total free amino acid content was not significantly affected. 16S rRNA sequencing results showed that supplemented with red clover extract significantly increased the Chao 1 and Shannon indexes of microbial α diversity and changed the rumen microbial community structure in the in vitro fermentation. The abundance of Prevotella (ASV 35 and s_unidentified_rumen_bacterium_RF18) and Ruminobacter were changed, Bilophil were decreased. In summary, the addition of red clover extract can reduce the rate of urea hydrolyzing by rumen bacteria, and improving the ability of rumen bacteria to convert ammonia nitrogen into microbial crude protein.

This study aimed to investigate the effects of tannic acid added to the diet of weaned piglets on liver index, inflammatory response, antioxidant function, and related gene expression in weaned piglets under lipopolysaccharide stress. The trial was divided into two parts. In the first part, a number of 72 28-day-old ternary hybrid weaned piglets (Duroc×Landrace×Yorkshire) with similar weight and good health condition were randomly divided into 2 groups with 6 replicates each and 6 piglets per replicate. The basal diet feeding group was fed a basal diet, and the tannin feeding group was fed the basal diet supplemented with 1 000 mg·kg^-1 tannic acid for 28 days. At the end of the first part of the experiment, twelve 56-day-old piglets with the body weight close to average weight were selected from the basal diet feeding group and the tannin feeding group, respectively, and then randomly divided into 4 groups of 6 piglets in each group. The piglets in control group (CON) and LPS stress group (LPS) were from the basal diet-feeding group of the first part of trial; The piglets in tannin group (TA) and the tannin+LPS group (TA+LPS) were from the tannin feeding group of the first part of trial. At 8 am of the day when the piglets are 56-day-old, the piglets in the LPS group and TA+LPS group were injected intraperitoneally with 1.5 mL of LPS at a concentration of 1 mg·mL^-1, and piglets in CON and TA groups were injected intraperitoneally with 1.5 mL of normal saline. After 1.5 h of injection, piglets were slaughtered and sampled. The results showed that, LPS stress increased the liver index, enhanced serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels, decreased catalase (CAT) and total superoxide dismutase (T-SOD) activities in liver, and enhanced the mRNA levels of Kelch-1ike ECH-associated protein-1(Keap1), Toll-like receptor(TLR4), Myeloid differentiation factor 88(MyD88), Nod-like receptor protein 3(NLRP3), Interleukin-6(IL-6) and Tumor necrosis factor-α(TNF-α), reduced the mRNA levels of Nuclear factor erythroid2-related factor 2(Nrf2), NADH Quinone Oxidoreductase 1(NQO1), Manganese superoxide dismutase(Mn-SOD) and Catalase(CAT) of liver in piglets (P < 0.05). The addition of TA to the feed reduced the levels of AST and ALT in serum, increased the levels of CAT and T-SOD of liver, and significantly affect the contents of IL-6, IL-10 and TNF-α in liver, and down-regulated the mRNA levels of Keap1, TLR4, MyD88, NLRP3, IL-6 and TNF-α in liver, and up-regulated the mRNA levels of Nrf2、NQO1、Cu/Zn-SOD、Mn-SOD and CAT in liver of piglets (P < 0.05). Compared with the CON group, the decreased level of AST was displayed in serum of piglets in TA+LPS group (P < 0.05). Compared with the LPS group, the serum AST and ALT levels decreased in TA+LPS group, the contents of IL-6 and TNF-α reduced, the expression levels of genes Keap1, TLR4, IL-6 and TNF-α decreased, and the expression levels of Mn-SOD and CAT increased in liver of piglets (P < 0.05). In conclusion, under the circumstances of the current study, tannic acid can alleviate the liver stress damage state caused by LPS stress through improving the liver index, enhancing liver antioxidant capacity, inhibiting the secretion of liver inflammatory factors, and regulating the expression of genes related to Nrf2 signaling, TLR4 signaling and NLRP3 inflammasome signaling.

This experiment was conducted to study the effects of Lactobacillus plantarum and Lactobacillus plantarum postbiotics on growth performance, immune status and intestinal health of growing female minks. A total of 40 gray female minks at 12 weeks of age were selected and divided into 4 groups with 10 replicates per group. The minks were fed basal diet, basal diet supplemented with 0.1% Lactobacillus plantarum (viable bacteria number > 10⁶ CFU·mL^-1), basal diet supplemented with 0.3% postbiotics of Lactobacillus plantarum, basal diet supplemented with 0.1% Lactobacillus plantarum and 0.3% postbiotics of Lactobacillus plantarum, respectively. The pre-experimental period lasted for 1 week, and the experimental period lasted for 8 weeks. During the experiment, bodyweight and feed intake were recorded to determine the growth performance. At the end of the experiment, blood, jejunum tissue, jejunal content were collected to determine the immune indexes and microflora. The results were showed as follows: 1) Compared to the minks in group without Lactobacillus plantarum, minks in Lactobacillus plantarum group had greater body weight at week 4 and week 8 (P < 0.05), average daily gain and average daily feed intake for the first 4 weeks and the entire 8 weeks (P < 0.05), sIgA content in jejunal mucosa (P < 0.05), and had less IgG content in serum and TNF-α content in jejunal mucosa (P < 0.05). 2) Minks supplemented with Lactobacillus plantarum postbiotics had greater IgG content in serum and IFN-γ content in jejunal mucosa (P < 0.05), and less IL-2, IL-12 and TNF-α content in jejunal mucosa (P < 0.05). 3) Neither Lactobacillus plantarum nor postbiotics had significant effects on Alpha diversity index of intestinal flora for growing female minks (P>0.05). Minks in Lactobacillus plantarum group had less proportion of Sporosarcina, Aminobacter and Agathobacter, and greater proportion of Fastidiosipila in rectal flora than minks in group without Lactobacillus plantarum (P < 0.05). Minks in the postbiotics groups had greater proportion of Staphylococcus, Weissella, Brevibacterium, Dietzia, Brachybacterium, Carnobacterium, Aerococcus and Sphingomonas, and less proportion of Kocuria, Plesiomonas, unclassified_f_Lachnospiraceae, Sanguibacter, Microbacterium, Glutamicibacter and Paracoccus in rectal flora than minks in group without postbiotics (P < 0.05). 4) Interactions between Lactobacillus plantarum and Lactobacillus plantarum postbiotics were significant for serum IgA and IgM contents, jejunum IL-2, IL-10, TNF-α, IL-8 levels (P < 0.05). In conclusion, dietary Lactobacillus plantarum supplementation could improve the growth performance of female minks, both Lactobacillus plantarum and Lactobacillus plantarum postbiotics improved the immune function of females and increase the relative abundance of beneficial bacteria.

This study was conducted to investigate the effect and mechanism of heat shock protein 27 (HSP27) on the proliferation of vesicular stomatitis virus (VSV) in vitro and the retinoid acid-inducible gene-I-like receptor (RLR) signaling pathway mediated by VSV infection. Real-time quantitative PCR, Western blotting, Co-immunoprecipitation, TCID₅₀ assay, and immunofluorescence methods were employed to reveal the effects of VSV infection on the mRNA and protein expression of HSP27, and then the effects of overexpression and interference with HSP27 on the proliferation of VSV, and further explore the effects of HSP27 on the RLR signaling pathway mediated by VSV infection. Finally, the interaction and co-localization between HSP27 and the target molecules in RLR signaling pathway were analyzed. The results showed that VSV infection could increase the gene transcription and protein expression of HSP27, both stable and transient overexpression of HSP27 significantly inhibited the proliferation of VSV in vitro. Interfering with HSP27 expression in host cells promoted the proliferation of VSV. Further studies showed that HSP27 enhanced both VSV and retinoic acid-inducible gene I protein (RIG-I) mediated IFN-β production and the protein expression of RIG-I, and that HSP27 interacted with RIG-I and co-localized in the cytoplasm. This study for the revealed the molecular mechanism by which HSP27 targets RIG-I to upregulate its expression, enhance the transduction of RLR signaling pathways, and then negatively regulate the proliferation of VSV in vitro, laying a foundation for further revealing the mechanism of host factor HSP27 in viral infection.

Infectious bursal disease is a highly contagious, immunosuppressive disease of young chickens caused by infectious bursal disease virus (IBDV). In recent years, recombinant IBDV mainly threats to the poultry industry. In our study, we characterized whole-gene sequence analysis of three recombinant strains of infectious bursal disease virus and assessed its pathogenicity in specific pathogen-free chickens. Three strains of virus were identified and amplified by chicken embryo isolation. The nucleotide homology and amino acid evolution tree were compared with the reference strain by Mega 11 and MegAlign software. The pathogen was infected with SPF chickens to determine the pathogenicity, and the viral copies in organs were detected by real-time fluorescence quantitative PCR. Phylogenetic analysis revealed that GD-1 strain belonged to type A2dB3, and showed the characteristics of new variant A, very virulent B; Strain GD-2 and strain GD-3 belong to type A3B1, showed the characteristics of very virulent A, new variant B and very virulent A, early variant B. Pathogenicity assays showed that none of the three strains caused the death of chicks, GD-1 strain had no obvious clinical symptoms, and GD-2 strain and GD-3 strain showed depression and lying on the ground 7 days after the attack. It caused significant atrophy of bursa of Fabricius and enlargement of spleen. The load of toxin in bursa of Fabricius was higher than that in spleen. On the day 14 after challenge, the bursa of bursa of GD-1 strain was significantly higher than that of GD-2 strain and GD-3 strain. Three groups of experimental chickens bursa of fabricius follicles are shrinking, and GD-2 group and GD-3 pathological changes is more serious, the spleen is a GD-1 set of pathological changes is more serious. In this study, the sequence of the three strains was determined, the evolutionary tree was drawn, and the pathogenicity was determined. It may help to study the pathogenic mechanism and the prevention of infectious bursal virus in poultry.

Feline panleukopenia virus (FPV) is a highly pathogenic threat to cats. In recent years, an FPV variant harboring an amino acid mutation at the VP2 91 site (A to S) has emerged prominently in China, however, information regarding on its pathogenicity is lacking. The aim of this study was to investigate the infection and molecular characteristics of the FPV A91S variant. F81 cells were used to isolate the FPV A91S variant, and the hemagglutination properties, pathogenicity and genomic characteristics were determined. An FPV A91S variant was successfully isolated, with a virus titer of 10^4.6 TCID₅₀·mL^-1 after viral purification. This isolate could effectively agglutinate porcine erythrocytes at pH 6.0-6.5 and temperatures of 4 ℃ and 37 ℃. When orally administered to kittens, the isolate caused a spectrum of clinical manifestations including vomiting, diarrhea, severe leukopenia, purulent discharge from the eyes, and cat death within 5 days. Gross and histopathological examinations unveiled severe hemorrhaging in the intestines and lungs. The nearly full genome sequence of this isolate was obtained, and the key amino acid residues were consistent with those of the typical FPV. In addition to the A91S mutation in VP2, three distinctive amino acid mutations (D23N, I443V and H596Q) were observed in the NS1 protein. These amino acid mutations caused the FPV A91S strains to form a discrete branch on all the evolutionary trees. This study, for the first time, investigated the hemagglutination properties and pathogenicity of the FPV A91S variant, which contribute valuable insights into the biological characteristics and genetic variations of FPV.

This study aims to use metagenomic sequencing analysis technology to quickly diagnose the possible pathogens causing the death of an Asian elephant, and then combine with conventional diagnostic techniques to analyze and confirm the etiological cause of elephant diseases, so as to provide reference for rapid diagnosis and prevention of elephant-related diseases. Autopsies were performed on dead elephant, and organs such as the heart, liver, spleen, lungs, kidneys, lymph nodes, muscles, stomach, duodenum, jejunum, colon, cecum, and pancreas were collected and sectioned for HE staining and pathological analysis; Metagenomic sequencing analysis was performed on the inguinal lymph nodes; species-specific PCR were used to amplify and sequence the characteristic genes, and an evolutionary tree was drawn to analyze its phylogenetic relationship with other strains. Data showed that necropsy and case biopsy results of dead elephant are consistent with the clinical characteristics of endothelial herpesvirus infection in elephants. Metagenomic analysis detected a large number of elephant betaherpesvirus type 1 (i.e., parent endothelial herpes virus) in the inguinal lymph nodes of elephants. The death of the elephant was probably caused by endothelial herpesvirus infection. Metagenomic technology offers valuable application prospects for the rapid diagnosis of endothelial herpesvirus infection in elephants as well as other endangered animal clinical infectious cases.

The aim of this experiment was to construct a double-deletion strain of antitranscriptional terminator (Lm4b_02325) and unknown protein (Lm4b_02326) in Listeria pathogenicity island 4 (LIPI-4) of foodborne Listeria monocytogenes (LM). In order to study the effect of Lm4b_02325/26 gene on partial biological characteristics of LM. LM928ΔLm4b_02325/26 double-deletion strain was constructed by homologous recombination technology. The D_{600 nm} of LM928 and LM928ΔLm4b_02325/26 double-deletion strains were detected under BHI, different EtOH and NaCl concentrations, different pH and different temperatures, and the growth curves were plotted. The transcription level of some virulence factors of LM in BHI culture were detected by RT-qPCR. The adhesion, invasion and intracellular proliferation of LM-infected murine macrophages RAW264.7 were measured by plate count. And the bacterial load in liver, spleen and brain of C57BL/6 mice infected with LM intraperitoneal injection were detected. The results showed that, a genetically stable double-deletion strain LM928Δ Lm4b_02325/26 was successfully constructed. In the medium containing 0.5%~10% NaCl or 3%~4.5% EtOH, there was no significant difference between the growth of LM928ΔLm4b_02325/26 and LM928. The growth rate of LM928ΔLm4b_02325/26 strain was significantly higher than that of LM928 under the condition of pH5, and the growth rate of LM928Δ Lm4b_02325/26 strain was significantly lower than that of LM928 under the condition of pH9.In BHI medium at different temperatures (4 ℃, 37 ℃, 42 ℃), there was no significant difference between the growth of double-deficient strains and wild strains. The hly, inlC and PlcA genes of LM928Δ Lm4b_02325/26 strain in BHI were significantly upregulated (P < 0.01), while mpl, inlB, actA, inlP, PlcB, prfA, SigB, iap, and inlA genes were significantly downregulated (P < 0.01). The adhesion rate (14.86%) and invasion rate (2.23%) of LM928Δ Lm4b_02325/26 strain to RAW264.7 cells were significantly lower than those of wild strains (with adhesion rate of 22.93% and invasion rate of 4.28%) (P < 0.01). The number of bacteria in RAW264.7 cells infected by LM928Δ Lm4b_02325/26 strain was significantly lower than that of LM928 strain at 3, 6 and 12 h after infection (P < 0.01). In animal experiment, after the infection of mice, there was no significant difference in bacterial load between the two strains in the liver or spleen of mice, but in the brain tissue, the bacterial load of LM928Δ Lm4b_02325/26 strain was 10⁴, which was higher than that of the LM928 10^3.07, and the difference was extremely significant (P < 0.01). In summary, the Lm4b_02325 and Lm4b_02326 genes of LIPI-4 are involved in the expression of LM virulence factors and the colonization ability of brain tissues, and are related to adaptability under weak acid and strong base conditions and the adhesion, invasion and intracellular proliferation of LM to RAW264.7 cells. This study lays a base for the functional study of LIPI-4.

The aim of this study was to evaluate the immunoprotective effect of recombinant Eimeria intestinalis microneme protein 2 (rEiMIC2) in rabbits and to provide a reference for the study of subunit recombinant vaccine against Eimeria intestinalis. EiMIC2 gene was identified in E. intestinalis transcriptome data, then cloning, prokaryotic expression and protein purification were conducted. The immunoreactivity of the recombinant protein was also detected using immunoblotting. Thirty-two 35-day-old coccidia-free New Zealand young rabbits were randomly divided into 4 groups (unchallenged group, challenged group, pET-32a(+) vector group and rEiMIC2 immunization group), with 8 rabbits in each group. In the unchallenged and challenged groups, 1 mL sterile PBS was subcutaneously injected into the neck of each rabbit. While in the pET-32a(+) vector group and rEiMIC2 immunization group, each rabbits were inoculated with 100 μg pET-32a(+) protein or rEiMIC2. A second immunization was conducted after 2 weeks of the first immunization. Fourteen days after the second immunization, excluding the unchallenged group, each of the remaining three groups of New Zealand rabbits was orally inoculated with 5×10⁴ sporulated oocysts of Eimeria intestinalis; All New Zealand rabbits were euthanized 14 days post-infection. After immunizing rabbits with recombinant protein, the immune protection effect of rEiMIC2 was comprehensively evaluated by observing clinical symptoms, evaluating intestinal lesions, and measuring relative weight gain rate, oocyst reduction rate, anticoccidial index (ACI), feed-to-meat ratio, serum IgG, IL-2, IL-4, IL-10, IFN-γ. The results showed that, the open reading frame length of EiMIC2 gene was 1 605 bp, encoded a protein with a molecular mass of approximately 57.68 ku. Western blot showed that rEiMIC2 could be recognized by positive sera, indicating its good immunoreactivity. The immune protection experiment showed that after the challenge, the symptoms of depression and diarrhea appeared in each group, and one rabbit died in each challenged group and the rEiMIC2 immunization group; the relative weight gain rate of the rEiMIC2 immunization group was 73.97%, and the oocyst reduction rate was 69.98%, the ACI was 136.97, and the feed-to-meat ratio was 3.98∶1, which were significantly different from the challenged group (P < 0.05). Compared with the intestinal lesions in the challenged group, the intestinal lesions in the rEiMIC2 immunization group were milder, and the level of the IgG in serum was significantly higher than that in the challenged group (P < 0.05). In summary, after immunizing rabbits with rEiMIC2, it can effectively reduce the excretion of oocysts and the loss of weight gain, and reduce intestinal damage, which providing a certain protective effect of the rabbits.

In order to identify the species of nematodes isolated from dolphin fecal samples, the morphology of the isolated nematodes is first identified. The parasite's body length was observed to be 120-140 mm long, with a cylindrical shape and slender ends. Its appearance was milky white and translucent, and its internal structure was observed to be yellowish under the microscope. It can be seen that the front end of the esophagus is thin and gradually expands backwards. We extracted the genomic DNA of the isolated worm and amplified the 28S rRNA D2-D3 gene and ITS gene fragments by PCR for molecular identification. The PCR results showed that the amplification products of the 28S rRNA D2-D3 gene and ITS gene were 797 and 840 bp, respectively. BLAST alignment results showed that the sequence similarity between the 28S rRNA D2-D3 gene (GenBank accession number OR345165) and the Anisakis typical (GenBank accession number HF911524.1) was 95.36%. The sequence similarity between the ITS gene (GenBank accession number OR345164) and the Anisakis typical (GenBank accession number MF668919.1) was 97.11%, with a 91.90% sequence identity between the ITS-1 region and the Anisakis typical (GenBank accession number MF668919.1). There is a 94.97% sequence identity between the ITS-2 region and the Anisakis typical (GenBank accession number JN968938.1) and a 100.00% sequence identity between the 5.8S rRNA region and the Anisakis typical (GenBank accession number OQ244221.1). Phylogenetic analyses based on the ITS gene and the 28S rRNA D2-D3 gene showed that the parasite samples isolated in this study were genetically closest to the Anisakis typical, and on the same branch.In summary, the isolated parasite samples obtained from this study should be the larvae of Anisakis typical based on microscopic examination and molecular analysis.

This study aims to investigate the immunoreactivity and secretion characteristics of the Echinococcus granulosus (E. granulosus) annexin B5, B15, and B25 in the intermediate host tissues, laying the foundation for further research on the interaction between annexin and hosts. In this study, total RNA was extracted from the protoscolex of E. granulosus, and then reverse transcribed into cDNA. The full-length coding sequences of EgANXB5, EgANXB15, and EgANXB25 genes were amplified by PCR using the cDNA as a template. The recombinant plasmids pET32a-EgANXB5, pET32a-EgANXB15, and pET32a-EgANXB25 were constructed using homologous recombination method, and the recombinant proteins were expressed in Escherichia coli. The immunoreactivity of these recombinant proteins was analyzed using Western blotting, and their secretion in the intermediate host tissues was examined by immunofluorescence staining. The recombinant EgANXB5, EgANXB15, and EgANXB25 were successfully cloned and expressed. The coding sequence (CDS) of EgANXB25 was corrected (GenBank: OR245515.1). These three proteins could be specifically recognized by positive canine sera for E. granulosus and positive mouse sera for E. granulosus, indicating that they had strong immunoreactivity. Moreover, immunofluorescence staining revealed that EgANXB5, EgANXB15, and EgANXB25 were primarily distributed in the liver parenchyma near the E. granulosus cyst. Recombinant EgANXB5, EgANXB15, and EgANXB25 exhibite strong immunoreactivity and these proteins have the potential to secrete into the hepatic parenchymal tissue surrounding the cyst, possibly further participating in the interaction between E. granulosus and their host.

The purpose of this study was to determine the effects of knockdown of Echinococcus multilocularis emu-let-7-5p on mouse peritoneal macrophages polarization and worm growth. A recombinant adeno-associated virus vector expressing emu-let-7-5p sponge sequence of E. multilocularis was constructed, and its inhibitory effect and specificity were validated in 293T cells. Seventy clean-grade BALB/c mice were injected intravenously (tail vein) with AAV8-si-let-7-5p, AAV8-control, and PBS, respectively. After two weeks, two mice in each group were randomly selected, and the expression of GFP, emu-let-7-5p and its target C/EBP δ in liver were detected by Western blot and qRT-PCR. At 3 months after intraperitoneal inoculation of 1 000 protoscolexs in each mouse, the transcriptional levels of emu-let-7-5p, C/EBP δ, M1 type (IL-1β, iNOS and IL-6) and M2 type (Arg-1, IL-4 and IL-10) related molecules in peritoneal macrophages were detected by qRT-PCR, and cysts weight and protoscolexs number of E. multilocularis in each group were analyzed. The results showed that, on the 15th day after injection, AAV8-si-let-7-5p successfully targeted the mouse liver and caused knockdown of emu-let-7-5p and up-regulation of C/EBP δ. At 3 months after infection, the expressions of emu-let-7-5p and M2 related molecule (Arg-1) in peritoneal macrophages were significantly down-regulated in AAV8-si-let-7-5p group, while C/EBP δ and M1 related molecules (IL-1β and iNOS) were significantly up-regulated. In addition, in AAV8-si-let-7-5p group, the expressions of IL-4 and IL-6 were down-regulated, while the expression of IL-10 was up-regulated, and cysts weight and protoscolexs number in AAV8-si-let-7-5p group were significantly lower than those in the control group. The above results suggested that knockdown of emu-let-7-5p in mice infected with E. multilocularis can promote the expression of C/EPB δ, and thus inhibit M2 macrophage polarization and worm growth.

In order to understand the infection status of chicken coccidia in some regions of Zhejiang province, and to identify the new species, Eimeria latan, Eimeria nagambie and Eimeria zaria, 349 fresh chicken feces were collected from 7 farms in Hangzhou, Lishui and Jiaxing. The number of coccidia oocysts per gram of feces was examined and counted by McMaster counting method. The species of coccidia in positive samples were determined by microscopic observation and identified by PCR. Subsequently, the infection of chicken coccidia was analyzed according to regional sources, breeding methods, uses and age. The result showed that the total infection rate of chicken coccidia in the investigated area was 72.78% (254/349), and seven species of Eimeria and Eimeria nagambie were detected, among which the dominant species was Eimeria acervulina, mixed infection were found in all farms. The PCR and sequence analysis results showed that four of the seven farms were infected with suspected Eimeria nagambie, the Eimeria nagambie of farm A had the closest homology with the COI gene of the Australian strain LR877717.1, the homology is 99.99%, and clustered into a branch. The results showed that the overall infection rate of chicken coccidia in Zhejiang province was high, and the field infection of E. acervulina was particularly serious. Some chicken farms in some areas of Zhejiang Province have been infected with suspected new species of coccidiosis Eimeria nagambie

This study aimed to investigate the effect of guanylate binding protein 2b (GBP2b) on M.bovis infection of RAW264.7 cells and its signal pathways in polarization of M1 and M2 during M.bovis infection. Downregulate the expression of GBP2b in RAW264.7 macrophages through small interfering RNA, and detect the expression of M1 and M2 macrophage surface markers and key proteins related to the MyD88 signaling pathway using qRT-PCR, Western blot, and flow cytometry, respectively. Dual luciferase reporter assay to examine the effect of GBP2b on NF-κB transcriptional regulation. The results showed that GBP2b was highly expressed in macrophages infected with M. bovis. After downregulating the expression of GBP2b in RAW264.7 macrophages and then being infected with M. bovis for 24 h, the expression of surface markers in M1 macrophages was downregulated, while there was no significant effect on the expression of surface markers in M2 macrophages. The transcription initiation of NF-κB is inhibited, and the downregulation of MyD88 signaling pathway related protein expression were due to the downregulation of GBP2b. The results indicate that GBP2b mainly promotes the polarization and inflammatory response of RAW264.7 macrophage infected with M. bovis towards the M1 phenotype by relying on the MyD88 signaling pathway, thereby helping macrophages clear intracellular M. bovis.

The aim is to study the pathogenic mechanism of porcine epidemic diarrhea virus (PEDV) by transcriptome analysis of the jejunum of two-day-old piglets infected with PEDV. Three two-day-old piglets were infected with the PEDV HB strain (PEDV-HB), which belongs to the currently prevalent G2b subtype. At 18 h post-infection, jejunum tissues were collected from piglets for pathology, PEDV detection, RNA-seq and transcriptome analysis. Finally; the transcriptome results were verified by quantitative real-time PCR (RT-qPCR). The result showed that the jejunal villi of piglets infected PEDV-HB were significantly shed and shortened; and was mainly distributed on the surface of intestinal villi. GO and KEGG enrichment analysis showed that differentially expressed genes (DEGs) were mainly related to immune response and inflammatory response, and were enriched in several classical pathways, such as cytokine-cytokine receptor interaction, Toll-like receptor signaling pathway, fat and glucose absorption and metabolism. PEDV infection of the jejunum of piglets activates the host immune response and associated antiviral signaling pathways and induces a cytokine storm as well as dysfunctions in fat and glucose absorption and metabolism. In all, this research can lay the foundation for the study of pathogenesis mechanism of PED.

As an important surface molecule of NK cells, CD56 plays an important role in the development, subgroup typing and function of NK cells. Therefore, in order to facilitate the screening of different subpopulations of porcine NK cells and explore their biological functions, this study prepared porcine CD56 monoclonal antibodies and preliminically identified their epitopes. Firstly, the amino acid sequences of the dominant epitope of porcine CD56 protein extracellular region (50-499aa) was predicted and selected by bioinformatics software. Then the selected region of CD56 was expressed by prokaryotic expression system, and the protein was purified and renaturated. After that, the recombinant protein was immunized to BALB/c mice, and a hybridoma cell line with stable secretion of porcine CD56 monoclonal antibody was obtained through cell fusion and subcloning experiments, which was named 1G8. Western blot results showed that the prepared porcine CD56 monoclonal antibody could bind specifically to the total protein of porcine brain and spleen. To further explore the specificity of CD56 monoclonal antibody, NK cells were isolated from porcine peripheral blood, then indirect immunofluorescence assay (IFA) was performed by using the prepared CD56 monoclonal antibody as the primary antibody. The results showed that the prepared monoclonal antibody could bind specifically to NK cell membrane. Finally, in order to further investigate the epitopes recognized by monoclonal antibodies, we further truncated the selected porcine CD56 gene into four truncations (50aa-192aa, 193aa-294aa, 295aa-397aa, 398aa-499aa), which were insert into eukaryotic expression plasmids. And then these eukaryotic expression recombinant plasmids were transfected into 293T cells, respectively. Western blotting and IFA results showed that the epitopes recognized by the prepared CD56 monoclonal antibody were 193aa-294aa of porcine CD56 protein. In this study, porcine CD56 monoclonal antibody was successfully prepared, and its specificity and epitopes were preliminically identified, which is of great significance for screening porcine NK cell subsets and exploring the biological function of porcine NK cells

Hypothalamus is an important brain area involved in the regulation of food intake, which has a complex circuit regulation mechanism. However, it is not clear whether there are nuclei that also play a role in weight regulation outside the hypothalamus. Therefore, in this study, the neural circuit tracer technique was used to identify the distribution and projection pattern of neurons in the locus coeruleus (LC) projecting to the interscapular brown adipose tissue (IBAT), and to explore the morphological basis of central neurons and neural circuits regulating energy homeostasis. The reporter gene and retrograde tracing were used to identify the localization of vesicular glutamate transporter 2 (VGlut2) and extracellular regulated protein kinase (ERK) neurons in the brain, and the anterograde tracing experiment expanded the brain area where VGlut2 neurons sent innervation. The body weight and feeding regulation of VGlut2 immunoreactive neurons were verified by chemical genetics. The results showed that LC^{VGlut2:: ERK} neurons sent dense innervation signals to IBAT. ERK immunoreactive neurons are enriched and expressed in LC and can be used as specific markers of LC. LC^VGlut2 neurons project to the striatum (CPu), secondary motor cortex (M2), ventromedial hypothalamic nucleus (VMH) and dorsal motor nucleus of the vagus (DMV). Chemical genetic activation of LC^VGlut2 neurons significantly decreased body weight (P=0.016 5) and feed intake (P=0.029 0). These results suggesting that LC^VGlut2 neurons were involved in the process of body weight regulation in mice. Identification of glutamatergic neurons other than paraventricular nucleus of hypothalamus (PVN) and downstream neural circuits regulating body weight will further understand the feeding mechanism and provide some inspiration for the development of obesity drugs.

Foot pad dermatitis (FPD) is a contact dermatitis affecting the claw and toe surfaces of poultry. FPD is one of the most common paw-toe health problems, with negative effects on the welfare and health of poultry. This study aimed to explore the effects of FPD on production performance, egg quality, behavioral responses, and immune levels of laying hens in the furnished cages. In this study, 60 Hyline Brown laying hens with 48 weeks of ages (WOA) were randomly selected. After four-point scoring, that score 0 for no lesions on the foot pads, score 1 for excessive keratosis, a small lesion of the foot pad epithelium with redness or discoloration, score 2 for foot pad dermatitis with epidermis discoloration, cutaneous lesions slight to erosion; score 3 for bumble foot with swelling of the foot pad visible from dorsal view. Bird was raised in furnished cage until 55 WOA. Performance and egg quality of laying hens were recorded and video-recorded for 2 consecutive days at 51 and 55 WOA. Then, the tonic immobility (TI) tests and open field tests (OFT) were carried out. Finally, the expression of immune-related factors in the skin, spleens, livers, small intestines, and serum of hens were detected. We found that FPD had no significant effect on the production performance (P>0.05), but improved the body weight compared with the score 0 group (P < 0.05), and the protein height and Haugh Units were significantly reduced for hens at 51 WOA (P < 0.05). Behavioral observations showed that FPD reduced the expression of preening behaviors in laying hens (P < 0.05). However, there was no significant effect on other event behaviors (object pecking, toe pecking, pacing, comfort behaviors) (P>0.05). The results of behavioral tests showed that FPD had no significant effect on responses of fear and anxiety in laying hens (P>0.05). The relative mRNA expressions of immune cytokines (IL-2, IL-4, IL-6, IL-8, IL-10, IL-17, IL-1β, IFN-γ, and TNF-α) and immunoglobulin IgA and IgG in foot, liver, and spleen tissues were significantly increased in hens with FPD (P < 0.05). The concentrations of IL-4, IL-6, IFN-γ, and TNF-α were also increased in the serum (P < 0.05). FPD had no significant effect on the production performance of laying hens. FPD decreased the expression of preening behavior and increased the expression of immune-related factors in the skin of the paw toe, which might lead to the chronic inflammation of laying hens.

This experiment was conducted to analyze the pathogenic microorganisms in the intestines of dead white feather broilers, clarify the cause of death of white feather broilers, and provide new ideas for scientific disease prevention and treatment of white feather broilers. Intestinal contents of died white feather broilers were collected from a chicken farm, then metagenomic sequencing, sequence splicing, box splitting technology, evolutionary tree analysis, and species annotation were used to identify pathogenic microorganisms in the intestine of white feather broilers. The pathogenic and drug resistance mechanisms of pathogenic microorganisms were elucidated through annotation of genome virulence genes and drug resistance genes. The results showed that the pathogenic microorganism detected in the intestinal contents of diseased white feather broiler chickens was avian pathogenic Escherichia coli. The entire genome sequence of the pathogen was obtained through sequence splicing and box splitting techniques, with a size of 4 998 208 bp and an integrity of 99.23%. This bacterium contains 192 virulence genes, mainly encoding flagella, cilia synthesis related proteins, colonization related proteins, and bacteriocin synthesis related proteins. It contains 88 resistance genes, mainly involving 28 types of antibiotic resistance, indicating that the above types of antibiotics have no therapeutic effect on white feather broilers infected with pathogenic Escherichia coli in poultry, ultimately leading to the death of white jade broilers. This study utilizes macro gene sequencing technology to effectively analyze the pathogenic microorganisms, pathogenesis, and drug resistance mechanisms of white feather broilers. It can help breeders understand the etiology in the early stages of white feather broiler disease, and provide targeted guidance for scientific medication to reduce economic losses.

Mycoplasma synoviae (MS) is one of the most detrimental mycoplasmas in the large-scale poultry industry. Meanwhile, the development of antimicrobial resistance to MS is serious currently. In order to understand the current situation of antimicrobial resistance to MS, the research was conducted in reference to the method of establishing epidemiological/wildtype cut-off values (ECOFFs/CO_WT) in CLSI and EUCAST by collecting and summarizing MIC from different regions and sources in recent 5 years after searching relevant literature reports at home and abroad, and then the ECOFFs of various antimicrobial agents to MS were obtained through comprehensive analysis using visual estimation, ECOFFFinder software analysis and Normalized Drug Resistance Interpretation (NRI). Subsequently, the corresponding drug resistance rates were calculated. It showed that the ECOFFs of MS to tylosin, tilmicosin, tylvalosin, doxycycline, oxytetracycline, chlortetracycline, tiamulin, lincomycin, florfenicol and spectinomycin were 1, 0.5, 0.5, 2, 4, 2, 2, 2, 8 and 4 μg·mL^-1 respectively. In absence of sensitive breakpoint of MS in CLSI and ECUAST, the theoretical basis and scientific evidence are provided by this research for the rational use of drugs in clinical practice, development of antimicrobial resistance work for monitoring and reducing it, and also beneficial for the treatment and control of MS.

To explore the effect of doxorubicin on lncRNAs in canine breast tumor cells and find potential biomarkers and therapeutic targets for canine mammary tumor, RNA-Seq technology was used to perform transcriptome sequencing analysis on canine breast tumor cells treated with doxorubicin, and differentially expressed lncRNAs were screened. GO enrichment and KEGG enrichment analysis were used to annotate gene functions and pathways and verify their expression. The results showed that there were 277 differentially expressed lncRNAs in canine breast tumor cells after doxorubicin treatment, of which 147 were up-regulated and 130 were down-regulated, mainly involving biological processes such as cell metabolism, transcriptional program disorder, ECM receptor interaction and cell necroptosis. Three differentially expressed lncRNAs were randomly detected by real-time quantitative PCR, and their expression trends were consistent with RNA-Seq, indicating the accuracy of the RNA-Seq sequencing results. Based on transcriptomics technology, this study comprehensively analyzed the overall characteristics of the expression changes of lncRNAs related to doxorubicin against canine breast tumor cells, revealed its possible new regulatory mechanisms, and provided a reference for the targeted therapy of canine mammary tumor and the development and utilization of potential biomarkers.

The polymorphisms of forkhead box protein P3 (FOXP3), follicle stimulating hormone receptor (FSHR) and fragile X mental retardation 1 (FMR1) were studied, and the effects of gene polymorphisms on infertility and hormone levels in cows were discussed. The sequence of the gene mutation site was amplified by PCR, and the PCR results were identified by enzyme digestion and sequencing peak analysis, and the genotypes of the three genes were counted. The intergroup differences in the mutations of the three genes were analyzed. The concentrations of estrogen (E), follicle-stimulating hormone (FSH), prolactin (PRL), testosterone (T) and luteinizing hormone (LH) in the two groups were detected by ELISA. The correlation between gene polymorphism and hormone level was analyzed. Three genotypes of FOXP3 gene were detected: AA, AG and GG; three genotypes of FSHR gene were detected: AA, AC and CC; T base insertion was detected at 302 bp of 5'UTR of FMR1 gene. The results of chi-square adaptability test showed that the 92 377 635 A>G loci of FOXP3 gene and the 31 450 279 A>C loci of FSHR gene were lower than Hardy-Weinberg equilibrium (P < 0.05). Correlation analysis showed that FOXP3 gene G allele and FSHR gene C allele were significantly positively correlated with repeated infertility in cows. There were significant differences in E, FSH, PRL and T hormones between the infertile group and the healthy mother group (P < 0.05). The E hormone concentration of FOXP3 AA and GG genotypes was significantly different from that of AG genotype (P < 0.05). FSHR gene AA, AC genotype E hormone concentration was significantly different from CC genotype (P < 0.05). (concentration) was significantly different from AG genotype (P < 0.05); FSHR gene 31 450 279 A>C locus AA, AC genotype E hormone content (concentration) FOXP3 gene and FSHR gene are polymorphic in Simmental, Brown and Angus populations, which indicates that the gene may be a potential genetic factor affecting the infertility of cows. The G allele of FOXP3 gene was significantly different between the healthy group and the multiple infertility group (P < 0.05), and the C allele of FSHR gene was significantly different between the healthy group and the multiple infertility group (P < 0.05). The G allele of g.92 377 635 A>G locus of FOXP3 gene was significantly positively correlated with multiple infertility in cows (the C allele of PC locus was significantly positively correlated with multiple infertility in cows (P < 0.05, r=0.14).The polymorphism sites of FOXP3 gene and FSHR gene may be used as the marker identification sites of repeated mating infertility in cows, which can be used to screen the potential candidate genetic markers of repeated mating infertility cows, which needs further proof. The G allele of FOXP3 gene was weakly positively correlated with E concentration (P < 0.05, r=0.16), and the T base insertion of FMR1 gene was weakly positively correlated with E concentration (P < 0.05, r=0.187). However, how the variation of FOXP3 gene and FSHR gene regulates the reproductive ability of cows needs further study.

The present study aimed to investigate the regulation and its possible mechanism of glycyrrhiza polysaccharide (GPS) on LPS-induced orchitis in mice. Sixty 8-week-old male C57BL/6J mice were randomly divided into 6 groups (10 mice per group): Blank control group, model group (LPS group), positive control group (dexamethasone group) and GPS low, medium, high dose (100, 200, 400 mg·kg^-1) group. On the first 20 days, GPS low, medium, high dose group was given the corresponding dose of GPS by intragastric, and other groups were given the same dose of physiological saline by intragastric. On the 21st day, model group was intraperitoneally injected with 5 mg·kg^-1 LPS, and other groups were intraperitoneally injected with the same amount of physiological saline, 2 h later, the positive control group was given 5 mg·kg^-1 dexamethasone by intragastric, 12 h later, the mice were sacrificed according to animal ethical standards, blood and testicular tissues were collected. The morphological changes of testicular tissue were observed by HE staining, the content of testosterone (T) and the level of lactate dehydrogenase (LDH) in serum were detected by enzyme-linked immunosorbent assay (ELISA), and the changes of oxidative damage indexes (T-SOD, CAT, GSH) were detected by Nanjing Jiancheng Kit. Real-time fluorescence quantitative PCR was used to detect mRNA expression changes of tight junction proteins (Occludin, ZO-1), inflammatory factors (IL-18, IL-1β, IL-6, TNF-α) and key cytokines of pyroptosis (caspase-1, NLRP3, GSDMD) in testis, western blot was used to detect the protein expression of IL-18, IL-1β, NLRP3, GSDMD. The results showed that dexamethasone group and GPS medium, high dose groups could significantly relieve the LPS-induced testicular damage, increase the serum testosterone content, reduce oxidative damage, decrease the mRNA expressions of pro-inflammatory cytokines, and inhibit the occurrence of pyroptosis. In summary, GPS could effectively alleviate LPS-induced orchitis injury, which may related to reducing oxidative damage, inhibiting pyroptosis and preventing inflammatory response.

The present study aimed to assess the effect of antimicrobial use in the perinatal period on the performance of sows and piglets in PRRSV-positive farms. A total of 280 near-term pregnant sows were selected from a large-scale PRRSV-positive pig farm in Guangxi and divided into five groups randomly. During the perinatal period, the sows in each group were administered veterinary antibiotics such as tilmicosin, tiamulin, doxycycline, and tylvalosin commonly used in veterinary medicine, while the control group received no antibiotic treatment. The anterior vena cava blood of weaned piglets in each group was collected for PRRSV antigen and antibody detection to assess the effectiveness of the antibiotics in preventing PRRSV infection in piglets. The number and ratio of diarrhea cases were recorded for each group of weaned piglets to investigate the preventive effect of common antibiotics on piglet diarrhea. The weaning interval of weaned sows and the estrus mating rate within 7 and 10 days after weaning were counted in each group, and the production indexes such as the number of weaning per litter, weaning survival rate and weaning weight of piglets in each group were counted to evaluate the impact of perinatal health care with commonly used antibiotics on the production performance of sows and piglets and to calculate the economic benefits of using drugs. The weaned piglets in the tylvalosin and tilmicosin groups showed a PRRSV nucleic acid positivity rate of 0%, and their PRRS antibody levels were significantly higher than those in the control group. The tylvalosin group exhibited a lower diarrhea rate in piglets compared to the other treatment groups and the control group. Statistical analysis of relevant production indicators showed that compared to other groups, the weaning survival rate and weaning weight of piglets in the tylvalosin groups were the highest, the weaning interval of sows was the shortest, and the average litter loss was lower. In summary, this study assessed the impact of perinatal health care with antibiotics commonly used in veterinary clinic on the production performance of sows and piglets in PRRSV-infected pig farms. It was found that using tylvalosin can inhibit the vertical transmission of PRRSV, enhance piglet antibody levels, and significantly reduce the weaned piglet diarrhea rate. In addition, the use of tylvalosin significantly improved weaning survival rate and weaning weight of piglets, shortened the separation interval of sows, and reduced the loss of litter assets. The initiation of this study provides crucial experimental data and theoretical support for effective antimicrobial use in large-scale PRRSV-positive pig farms, and provided necessary supplementary measures for realizing cost reduction and efficiency increase in large-scale pig farms under the background of PRRSV pandemic.

This study assessed the mitigation effects of Viola yedoensis Makino (VYM) on the growth performance, meat quality, and gut microbiota of AA broilers under heat stress. Sixty 1-day-old male AA broilers were randomly allotted into 6 treatments. The NT group was fed with a standard diet at normal temperature. The NT+VYM-H group was fed the diet supplemented with 4.5% VYM at normal temperature; The HS group was fed with a standard diet under heat stress; The HS+VYM-L, HS+VYM-M, and HS+VYM-H groups were fed diets containing 0.5%, 1.5%, and 4.5% VYM under heat stress. All broilers were slaughtered at 42 days old. Growth performance, serrum biochemistry, meat quality, duodenum morphorlogy, related genes and composition of gut microbe were analyzed. Broilers fed with 1.5% VYM (w/w) had higher average daily weight gain and a lower feed intake/body weight gain (F/G) ratio under heat stress (P < 0.05). Under heat stress, VYM supplementation reduced serum HSP70 levels (P < 0.05), while the supplementation of 0.5% VYM increased duodenal lipase activity (P < 0.05), and upregulated myogenic regulatory-related mRNA (Pax3, Pax7, Myog, Myod, Myf5) expression in the breast muscle of broilers, and upregulated digestive-related mRNA (Ppara, Fatp1, B⁰at1, Pept1, Cat1, Eaat3) expression in the duodenum (P < 0.05). Under heat stress, 1.5% VYM supplementation decreased shearing stress and the color of meat (P < 0.05). In the cecum, VYM remarkably improved the richness of intestinal microflora (P < 0.05) and increased the abundance of Bacteroides, which was significantly positively related to meat pH (45 min). It also lowered the abundance of Subdoligranulum, which was significantly negatively correlated with duodenal amylase activity. In conclusion, under the conditions of this study, the degradation of growth performance and meat quality caused by heat stress can be reduced and the balance of intestinal flora of broilers can be regulated by the supplementation of 1.5% and 4.5% Viola yedoensis Makino in diet effectively. This work provides a valuable reference for the positive benefits of applying VYM in poultry production, giving a viable solution for coping with the high-temperature climate of poultry farming.