Single nucleotide polymorphism(SNP) are the important material of genetics research. In recent years, progress in the development of SNP genotyping methods provided the low-cost approach for researchers to obtain large volumes of genomic markers, and promoted the related research at genomic level. Genomic prediction constructs models from individuals with known phenotype and genotype data to predict genotype and phenotype of the individuals with unknown phenotype. It is of great importance in animal breeding. The combination of genomic-wide SNP genotyping and genomic prediction methods constitute the frontier of animal genomic selection. In this paper, we reviewed the advancements of the both methods, it will provide a reference for researchers working in molecular genetics, especially in complex traits.

The F-box and leucine rich repeat protein 3 (FBXL3) gene encodes the protein containing the F-box domain. It belongs to the F-box protein family and plays a role in the nucleus.The F-box and leucine rich repeat protein 21 (FBXL21) is located in the cytoplasm, and its function is antagonistic to the function of FBXL3 gene. FBXL3 and FBXL21 proteins both play important roles in the negative feedback cycle of molecular circadian rhythm in mammals. FBXL3 participates in the regulation of biological clock by forming SCF complex, which exercise E3 ubiquitin ligase function and promote the degradation of CRY protein through ubiquitination. FBXL21 makes CRY protein accumulation to participate in biological clock regulation through ubiquitination. The antagonism of FBXL3 and FBXL21 to CRY protein can stabilize the biological clock of mammals. In this review, the biological rhythm and the elements of biological clock were introduced firstly. Then, the regulation mechanism of FBXL family in circadian rhythm and the effects of FBXL family mutation on seasonal estrus traits in sheep were reviewed. It is helpful to explore the molecular mechanism of FBXL family mutation affecting sheep seasonal estrus deeply, and provide new ideas for changing sheep estrous season using modern biotechnology.

Zearalenone(ZEA) is a mycotoxin with estrogen activity,which is one of the most common mycotoxins in swine production.Zearalenone is widely found in cereal raw materials and compound feeds.Reproductive disorders may occur when sows are fed the feed contaminated by zearalenone,which can seriously affect the production performance and production potential of the sow.This paper reviewed the pollution status of zearalenone in pig feed, reproductive toxicity to sows, and toxic action pathways, which will provide a reference for further research.

As a class of polyphenolic compounds, tannins are widely distributed in various plants, and were generally classified as anti-nutritional factors in ruminants' feed due to their astringency taste which reduce the feed intake of animals. In recent years, studies have found that tannins can bind to proteins, which prevent proteins from being excessively degraded in the rumen, and increase the digestion of proteins in the hindgut, further improves the utilization of nitrogen (N) in ruminants. In addition, studies have shown that tannins play active roles in regulating rumen fermentation, inhibiting methane emissions, and possess certain antimicrobial capabilities. In this paper, the effects of tannins on ruminant production performance, rumen fermentation and microflora were reviewed in order to provide theoretical reference for its rational use in ruminant healthy farming.

The discovery of antibiotics has saved countless lives. However, the emergence of multidrug-resistant bacteria and ecological imbalance of normal microbial communities due to indiscriminate use of antibiotics has forced on finding effective substitutions of antibiotics. Antimicrobial peptides (AMPs) exist widely in animals and plants, which have effective activity against bacteria, tumor, virus etc., without inducing drug resistance. It has excellent application prospects in clinical and livestock industry. In order to avoid problems such as drug resistance and micro-ecological disorders, targeted antimicrobial peptides have aroused researchers' interests and gradually become a research hotspot, which have been expected as one of the effective substitutions of antibiotics. In this paper, the design ideas of targeted antimicrobial peptides in recent years and their application prospects in clinical and animal husbandry are summarized, in order to provide new ideas for the development of targeted antimicrobial peptides in the future.

This study aimed to explore the changes of protein levels in chicken embryos during the middle and late developmental stages. One hundred and twenty fertilized eggs with (65±0.2)g were selected and randomly divided into 2 groups (E14 and H1), 3 replicates per group, and 20 eggs in each replicate. The liver tissues were collected at E14 and H1, respectively. Proteomics approach based on isobaric tags for relative and absolute quantitation (iTRAQ) was employed to screen the core differential proteins in liver. The results showed that 10 core differential proteins with different expression levels were identified in H1 compared with E14, these proteins mainly enhanced fatty acid degradation (upregulated ACOX1, ACSL1, ACSL5, CPT1A and ECI2),and gluconeogenesis (upregulated ALDH3A2, FBP1, FBP2, GPI and PGM2,downregulated LDHB and ALDH9A1). These results indicate that the main energy supply pathway for embryo development is fatty acid degradation, not glycolysis. In addition, increased gluconeogenesis promotes glycogen storage to cope with shelling and changes in the nutritional environment.

In order to better understand the genetic structure changes of Qingyu pig in the process of generation replacement, and better protect and utilize the genetic resources of Qingyu pig, the 50K SNP chip was used to measure SNP of 141 healthy adult individuals (26 boars, 115 sows) in Qingyu pig population. The pedigree of Qingyu pig population and each generation was corrected by various analysis softwares, and then the genetic diversity, genetic distance and genetic structure changes were analyzed. The results showed that the closed population was composed of 3 overlapping generations, the effective size of the population was 12, and the whole population could be divided into 6 families with boar and 1 family without boar. Among them, the effective size of the 3rd generation was the least, only three. The proportion of polymorphism markers decreased with the increase of generation. The average genetic distance of 141 Qingyu pigs was (0.260 4±0.025 2), and that of 26 boars was (0.263 3±0.023 7). With the increase of reproductive generations, the genetic distance of each generation population had a slight upward trend, and the genetic relationship and genetic distance between some breeding pigs were relatively closer; in 141 Qingyu pig population, 1 481 long homozygous segments (ROH) were detected, 78.01% of which were within 200 Mb. The inbreeding coefficient based on the ROH value showed that the average inbreeding coefficient of the whole population was 0.055, and the inbreeding coefficient of each generation continuously rose, reaching 0.075 by the 3rd generation. In conclusion, the study on the genetic structure of Qingyu pig population at the molecular level shows that there is a loss of population genetic diversity in the process of closed subsequent breeding, and it is necessary to strengthen the selection mating system or introduce external blood to ensure the long-term preservation of the genetic resources of Qingyu pig.

The purpose of this study was to screen the optimal DNA barcode for the rapid identification of hares in Xinjiang. A total of 4 hare breeds including 40 samples were collected, CO1, ND4, 16S rRNA and ITS2 genes were selected as candidate sequences. After PCR amplification and sequencing, the identification ability of candidate sequences at species level were analyzed and compared. The results showed that ITS2 fragments was first eliminated because no PCR amplification product was acquired for sequencing after repeated experiments. Compared with 16S rRNA, CO1 and ND4 showed stronger variation levels in sequence characteristics, variation characteristic parameters, and rank sum test results, their genetic frequency distribution map had wider gap, and the identification results of the systematic clustering tree had higher accuracy. Further, the characteristics of interspecies variation, interspecies rank sum test results and genetic frequency distribution map all indicated that ND4 was slightly better than CO1. By comprehensive analysis, the optimal DNA barcode obtained in this study was ND4, while CO1 could serve as the alternate one. These results would provide reference for the study of distribution and classification of hares in Xinjiang and their protection and utilization.

The aim of this research was to clone the spermatogenesis associated 3 (SPATA3) gene in yak, identify its expression pattern in various tissues of yak, and in different developmental stages in yak and cattle-yak testis, respectively, explore the effects of SPATA3 on testicular development and sperm maturity, and provide reliable basis data for studying the mechanism of SPATA3 in spermatogenesis of yak. The samples of yak heart, liver, spleen, lung, kidney, small intestine, brain, ovary, uterus and testis were collected after slaughtering. The total RNAs in different samples were extracted. The coding sequence of SPATA3 gene was cloned and the expression of SPATA3 in different tissues was detected by RT-PCR. The structure and function of SPATA3 were analyzed by bioinformatics softwares. The mRNA expression of SPATA3 gene in testicular different development stages of yak and cattle-yak was compared by quantitative real-time PCR (RT-qPCR). Then, the localization and expression of SPATA3 in yak and cattle-yak testis were detected by immunohistochemistry. The results showed that the CDS region of SPATA3 gene was 693 bp, encoding 230 amino acids; It had high homology with SPATA3 sequence of cattle, wild yak and buffalo. SPATA3 gene was specifically expressed in testis tissue, and no expression was observed in other tissues of yak. The results of RT-qPCR showed that SPATA3 gene was expressed during testicular development of yak, and the expression level in testis of adulthood yak(4-5 years old) was extremely significantly higher than that of fetal period yak(5-6 months old, P<0.01). And its expression in testis in different developmental stages of yak were all extremely significantly higher than that of cattle-yak (P<0.01). The results of immunohistochemistry showed that SPATA3 was expressed in cytoplasm of round spermatid and elongating spermatid, and its expression level in testis of adulthood yak was extremely significantly higher than that in testis of adulthood cattle-yak (P<0.01). The above results show that SPATA3 is highly conserved during evolution, and it is differentially expressed in testis tissues of yak and cattle-yak. Its low expression may cause sperm formation disorder and participate in sperm maturation process, but the specific function should be researched further.

The objective of this study was to investigate the effect of vitrification/thawing on developmental competence of immature oocytes and cumulus-oocyte-complexes (COCs) transcriptome of yaks (Bos grunniens), in order to provide theoretical foundation to improve vitrification techniques of yak COCs. Vitrified/thawed yak immature COCs were divided into two groups. Group A:COCs were in vitro matured (IVM) and in vitro fertilized (IVF) with cattle sperms, then in vitro cultured (IVC) in G-1 for 72 h followed by IVC in G-2 for 96 h; Group B:after IVF, zygotes were IVC in G-1 for 120 h followed by IVC in G-2 for 48 h. Fresh immature yak COCs were used as the control (Group C):after IVF, zygotes were IVC in G-1 for 72 h followed by IVC in G-2 for 96 h. Yak fresh immature COCs (n=3) and vitrified/thawed immature COCs (n=3) were used for amplification, library preparation and RNA-seq analysis. The results showed that cleavage rate and blastocyst rate in Group B were significantly higher than those in Group A (P<0.05), but cleavage rate and blastocyst rate in both Group A and B were significantly lower than those in Group C (P<0.05). When|log₂(fold change)|≥ 2 and Q-value <0.05 were set as thresholds for identifying deferentially expressed genes (DEGs), a total of 851 DEGs were detected, of which, 846 were up-regulated and 5 were down-regulated in virtrified/thawed COCs compared to fresh COCs. GO analysis showed DEGs were classified into 3 categories:biological processes, cellular components and molecular functions. KEGG annotation showed that there were 258 pathways, of which, 16 were significantly enriched (P<0.05). In conclusion, the results showed that IVC in G-1 for 120 h after IVF could increase subsequent developmental competence of vitrified yak oocytes. Vitrification affected transcriptome of yak COCs, which reduced developmental potential of yak vitrified oocytes. The result provided a theoretical basis for improving vitrification techniques of yak COCs.

This study aimed to investigate the effects of Hermetia illucens larvae meal on the number of main microbes and metabolites in the cecal digesta of finishing pigs. Seventy-two 130 days of age healthy (Duroc×Landrace×Large White) crossbred pigs with similar body weight of (76.0±0.56) kg were randomly allocated to 3 different dietary treatments with 8 replicates for each treatment and 3 pigs for each replicate. Pigs in the control group were fed basal diets, pigs in the experimental groups were fed diets with 4% (4% group) and 8% (8% group) Hermetia illucens larvae meal to attain the same nitrogen and energy level, respectively by adjusting diet formula. After 42 days formal feeding, one pig from each replicate in each group was randomly selected and slaughtered, and the cecal digesta were collected for correlated parameters analysis. The results showed that:1) Compared with control group, dietary with 4% Hermetia illucens larvae meal significantly increased the number of Bifidobacterium, Lactobacillus, Prevotella, Clostridium cluster IV, and Roseburia in the cecal digesta of pigs (P<0.05), while significantly decreased the number of Escherichia coli (P<0.05); Meanwhile, dietary with 8% Hermetia illucens larvae meal significantly increased the number of Prevotella and Bifidobacterium in the cecal digesta of pigs (P<0.05). 2) Compared with control group, dietary with 4% and 8% Hermetia illucens larvae meal significantly increased the concentrations of lactate, total short-chain fatty acid, and acetate in the cecal digesta of pigs (P<0.05), and dietary with 8% Hermetia illucens larvae meal significantly decreased the concentration of valerate (P<0.05). For nitrogen metabolites, compared with control group, dietary with 4% Hermetia illucens larvae meal significantly decreased the concentrations of total amines, cadaverine, putrescine, spermine, p-cresol, and phenol in the cecal digesta of pigs (P<0.05), and dietary with 8% Hermetia illucens larvae meal also significantly decreased the cadaverine and phenol (P<0.05). In conclusion, under the conditions of this trail, dietary with 4% Hermetia illucens larvae meal increased the number of beneficial microbes and the concentrations of beneficial metabolites in the cecal digesta of finishing pigs, while decreased the number of potential pathogenic microbes and the concentrations of nitrogen metabolites. These findings indicated that appropriate proportion of Hermetia illucens larvae meal could be used as high quality protein feedstuff in pig production.

The purpose of this experiment was to study the effects of diets with different calcium (Ca) and phosphorus (P) levels on colostrum composition, fatty acid composition of colostrum fat, the plasma physiological and biochemical indexes including plasma Ca and P and hormones related with Ca and P metabolism, growth and reproduction in Yili mares after delivery, and to provide reference for defining the appropriate Ca and P requirement of Yili mares during the late gestation period. Twenty-five healthy Yili mares without kinship, and with the age of 12-13 years old, body weight of (380±32) kg, parity of 4-5 births and about in the 10 th gestational month were selected and randomly divided into 5 groups with 5 replicates in each group. Ca feeding levels of the 5 groups were 36.00, 39.00, 42.00, 45.00 and 48.00 g·d^-1, respectively and P feeding levels were 26.30, 28.30, 30.30, 32.30 and 34.30 g·d^-1, respectively. The pre-test period was 10 days, and the experimental period lasted from the 11th day of the trial to the 2nd day after delivery. Samples of colostrum and fasting blood were collected within 12 hours after delivery to determine the milk composition, fatty acids content in milk fat, and plasma concentration of Ca, P, parathyroid hormone (PTH), calcitonin (CT), osteocalcin (OC), placental prolactin (PL), pituitary prolactin (PRL), estrone (E₁), estradiol (E₂), progesterone (PROG), thyroxine (T₄), triiodothyronine (T₃), growth hormone (GH), and insulin-like growth factor-Ⅰ (IGF-1). The results showed that:1) Dietary Ca and P levels had no significant effect on the Ca, P, milk fat percentage and lactose percentage in colostrum, but there were significant differences in milk protein percentage, total solids, somatic cell counts, total saturated fatty acids (SFA) and unsaturated fatty acids (UFA) percentages in milk fat among groups (P<0.05). With the increase of dietary Ca and P levels, the concentration of P, milk protein percentage and UFA percentage in colostrum significantly increased linearly (P<0.05), total solids significantly increased quadraticly (P<0.05), and lactose, SFA and myristic acid percentage significantly decreased linearly (P<0.05). 2) Plasma concentrations of OC and PROG were significantly affected by dietary Ca and P levels (P<0.05). With the increase of dietary Ca and P levels, the concentration of plasma ionic Ca, CT and OC significantly decreased linearly (P<0.05), while the concentration of PTH and PROG significantly increased linearly (P<0.05). Dietary Ca and P levels had no significant effect on the concentration of reproduction hormones such as PL, PRL, E₁, E₂, and growth and metabolism hormones such as T₃, T₄, GH and IGF-1. In this experiment, it can be concluded that dietary Ca and P levels affect colostrum compositions by increasing the milk protein percentage and total solids content, changing the saturated fatty acid and unsaturated fatty acid content in milk fat, affect Ca and P metabolism by reducing OC concentration, and affect fertility status by increasing PROG concentration.

Bovine viral diarrhea virus (BVDV) is an important pathogen causing bovine viral diarrhea/mucosal disease, and it is also the common source of contaminant in bovine serum and related biological products. In this study, a cytopathogenic (CP) type BVDV (GS2018 strain) was isolated from commercial FBS and identified by transmission electron microscopy (TEM), molecular identification, immunofluorescence assay, complete genome sequencing and genetic evolution analysis respectively. The results showed that MDBK cells presented obvious cytopathic effect (CPE) when inoculated with the BVDV GS2018 strain, and the virus titer was up to 10^6.2·0.1 mL^-1. The purified virus was detected by transmission electron microscope (TEM), round-shaped virions were observed with the diameter ranging from 50 to 60 nm, and specific positive fluorescence signal was found clearly by immunofluorescence assay. Meanwhile, the complete genome of GS2018 was sequenced as 12 235 nucleotides (nt), and BLAST result of 5'UTR region revealed that the strain had high homology with the reference sequences of BVDV-2. The phylogenetic evolutionary analysis of 5'UTR, N^pro and E2 genes showed that the GS2018 strain was classified into BVDV-2a subtype obviously, sharing the highest homology (up to 98%) with BVDV-2a representative strain USMARC-60764. Moreover, an interesting phenomenon was found that no nucleic acid fragment insertion was present in the NS2/3 region with further analysis of the genome, unlike most of CP type BVDV-2. The diversity of BVDV2 genome suggested that the mechanism of BVDV induced cytopathic effect is complicated and further research of genomic variation will contribute to answering this question.

An indirect ELISA was developed for detecting antibodies to Mycoplasma mycoides subsp. mycoides SC (MmmSC), the agent of contagious bovine pleuropneumonia (CBPP), and used for CBPP surveillance. Based on bioinformatics assay of the whole genome sequence of MmmSC isolates Ben-1, lipoprotein rP0308 was selected as coating antigen. An indirect ELISA was established after optimization of a series of conditions, and its performance was evaluated. As a result, the diagnostic sensitivity and specificity of the indirect ELISA were 92% and 96%, respectively. The ELISA did not react with other antibodies against Mycoplasma bovis, Mycoplasma bovirhinis, Mycoplasma agalactiae, foot and mouth disease, bovine infectious rhinotracheitis, Mycobacterium tuberculosis. Reproducibility analysis revealed that the coefficients of variation of samples within and between runs were 2.41%-6.03%, and 2.94%-6.59%, respectively. One thousand six hundred and forty-eight serum samples from our lab were tested using this ELISA system and a commercial kit, a total of 1 405/1 576 (89.1%) of the sera found positive in the commercial kit were also positive in the ELISA, and 57/72 (79.2%) of the sera found negative in the both system. The overall coincidence was 88.7%. The above results suggested that the established ELISA demonstrated high specificity, sensitivity and reproducibility, and could be a promising tool applied in the field.

Francisella tularensis (F. tularensis) is an important zoonotic pathogen, but its virulence factors and molecular pathogenesis are still unclear. This study aimed to analyze the distribution of unknown functional protein FTN_0109 in F. tularensis, and to explore its influence on the virulence of F.tularensis. The biological function of FTN_0109 was predicted by bioinformatics softwares and its subcellular localization was detected by hypervelocity centrifugation. The FTN_0109 gene deleted strain and its trans-complemented strain were constructed by homologous recombination method. The invasion and intracellular proliferation ability of the wild-type strain and its derivates in three macrophagocytes from mice were detected, and their pathogenicity and proliferation dynamics in BALB/c mice were analyzed. The results showed that FTN_0109 was predicted to be a kind of lipoprotein, mainly distributed in the inner membrane of F. tularensis. FTN_0109 mutant strain could not proliferate in mouse macrophages and its LD₅₀ in BALB/c mice increased more than 10⁴ CFU compared with wild-type strain. The bacterial load of FTN_0109 mutant strain in infected mice showed a decreasing trend and was significantly lower than that of wild-type strain(P<0.05). The results suggested that FTN_0109 was an inner membrane lipoprotein, and its deletion could seriously reduce the survival ability of F. tularensis in macrophages and mice, and thus reduced its pathogenicity in mice.

The motility of E. coli is mainly driven by flagella, which is one of the important virulence factors of pathogenic E. coli. In this paper, the influences of Fur and its negatively regulated non-coding RNA-RyhB on the motility of avian pathogenic E. coli (APEC) AE17 strain were investigated to provide scientific basis for exploring potential targets for prevention and control of avian E. coli. The Red homologous recombination method was used to construct AE17△Fur and AE17△Fur/RyhB, then the motility characteristics of the absent and original strains were compared, and the effects of Fur and RyhB on the formation of APEC flagellum and biofilm in combination were explored with transcriptome data. The results showed that the AE17△Fur and AE17△Fur/RyhB were constructed successfully. Transcriptome results showed that the absence of Fur basically down-regulated all flagella-related genes. The absence of Fur significantly weakened the movement of APEC, and RyhB had no significant influence on the movement of APEC. △Fur/RyhB has increased its movement ability compared with △Fur. Biofilm formation ability of △Fur and △Fur/RyhB were significantly enhanced, and that of △RyhB was weakened, which were consistent with the trend of movement. Fur plays a very important role in flagellum assembly of pathogenic E. coli, and its negatively regulated RyhB can inhibit the effect of this mechanism to some extent, and further affect the formation of biofilm, so as to provide a possibility for finding relevant drug targets.

To explore the expression and chitin-binding manner of chondroitin proteoglycans 1 of Toxocara canis (TcCPG1) and to provide a basis for the functional study of this proteins in T. canis. In the present work, the chondroitin proteoglycans 1 gene(Tc-cpg-1)of T. canis was molecularly cloned and expressed in a prokaryotic expression system for recombinant protein TcCPG1, which was then purified using Ni-NTA affinity chromatography and tested in a chitin-binding assay. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to determine the transcriptional differences of Tc-cpg-1 between sexes and among tissues of T. canis. The results showed that the coding sequence of Tc-cpg-1 was 1 251 bp in length, encoding 417 amino acids. SDS-PAGE analysis showed that the recombinant protein TcCPG1 was expressed in inclusion bodies, and the molecular weight was estimated at~70 ku. Optimized conditions (induction by 0.8 mmol·L^-1 IPTG for 14 h and incubation at 16℃) facilitated an efficient production of the recombinant protein TcCPG1. Following Ni-NTA affinity purification, the recombinant protein bound chitin, suggesting a chitin-binding activity of TcCPG1. In addition, differential analysis showed a high transcriptional level of Tc-cpg-1 in female adult worm, particularly in the reproductive tissues, indicating potential roles of Tc-cpg-1 in reproductive process (embryonic and reproductive development) of female worms, which might be achieved by interacting with the chitin in egg shell.

This study aimed to investigate the effect of autophagy on the stemness of canine bone marrow mesenchymal stem cells (cBMSCs). The isolated cBMSCs were cultured with ordinary medium, rapamycin-containing medium, 3-MA-containing medium. Cells were collected after 12, 24 and 48 hours' treatment, then autophagy microtubule-associated protein 1 light chain 3 Ⅱ (LC 3Ⅱ) were detected by indirect immunofluorescence; autophagy and stemness related genes, e.g. LC 3, Beclin 1, autophagy-related 7 (Atg 7), SRY-related high-monility-group (HMG)-box protein-2 (Sox 2), special AT-rich sequence-binding protein 2 (Satb 2), were detected by fluorescence quantitative PCR. The cBMSCs differentiation abilities were detected by the dyeing experiment after adipogenic (Oil red O staining) and osteoplastic (alizarin red staining) differentiation. Results were as follows:The LC 3Ⅱ protein expression and gene expression level at the rapamycin group was increased, while which was decreased in the 3-MA group. The results showed that there was a significant increase at the stemness and autophagy related mRNA expression levels of the rapamycin group, while which was decreased in the 3-MA group. The cBMSCs in the rapamycin group showed morphological alterations obviously and the larger mineralized area, while the cBMSCs in the 3-MA group showed the smaller mineralized area. There were fewer lipid droplets at the rapamycin group, while the 3-MA group was more. These results indicated that autophagy could enhance the stemness and osteogenic differentiation of cBMSCs, it was providing theoretical and technical evidence for cBMSCs in the treatment of diseases in dogs.

The aim of this study was to investigate the effects of the high level of copper on mitophagy in cardiomyocytes of rats. Thirty-two eight-month-old healthy rats were randomly divided into 4 groups with 8 rats in each group. The rats were fed diet with different content of copper (control group:15 mg·kg^-1, high-copper group Ⅰ:30 mg·kg^-1, high-copper group Ⅱ:60 mg·kg^-1, high-copper group Ⅲ:120 mg·kg^-1). After feeding for 6 months, heart tissues were collected from anesthetized rats. The content of copper and the pathological change of myocardial tissues were measured. The mRNA levels of LC3A, LC3B, Parkin, PINK1 and p62 were detected by real-time quantitative PCR. Additionally, the protein expression of Parkin, PINK1 and LC3B/LC3A were measured by Western blot and the expression and localization of LC3B was detected by immunohistochemistry and immunofluorescence. The results showed that content of copper in the heart of the high-copper groups was higher than that of the control group, the myocardial fibers were loose and irregular in the high-copper groups. Additionally, compared with the control group, the mRNA expression of LC3A and LC3B in the high-copper groups increased. The p62 mRNA expression in the high-copper group Ⅱ and Ⅲ was lower than that of control group. Furthermore, compared with the control group, the expression of PINK1 mRNA in the high-copper group was significantly increased, and the expression of Parkin mRNA was increased but not significantly. Meanwhile, with the increase of copper content in the diet, the protein expressions of PINK1, Parkin and LC3B/LC3A were significantly up-regulated. Moreover, LC3B was located in the cytoplasm, and the LC3B expression in high-coppergroups was significantly increased compared with the control group. These results suggested that high level of copper could induce mitophagy of rat cardiomyocytes.

This study was conducted to investigate the clinical efficacy of BQJGHJ, a traditional Chinese medicine compound pharmaceutics, in the treatment of chickens artificially infected with Infectious bronchitis virus (IBV). One hundred and twenty chicks were divided into blank group, model group (IBV infection), positive drug group (IBV infection & Gan Dan oral liquid), and BQJGHJ high, middle and low dose groups (IBV infection & BQJGHJ), 20 chicks in each group. Except for the blank group,the other groups were modeled by infecting IBV-M41 strain, 4 days after modeling, the medicines were administrated through drinking water for 5 days. The clinical symptoms of each group were observed every day, the scores of clinical symptoms were recorded, the effective rate was counted, and the cure rate was used as the evaluation index. On the 3rd, 5th and 7th day post the first administration, the blood samples were collected from the wing vein and the serum were separated. The contents of IFN-γ, IL-6 and IL-10 in chicks serum were detected by ELISA. The results indicated that the cure rates of BQJGHJ high, middle and low dose groups were 75.0%, 80.0% and 65.0%, and the effective rates were 90.0%, 95.0% and 80.0%, respectively. The content of IL-10 in serum of chicks in model group decreased and the content of IL-6 increased, which was significantly different from that in blank group (P<0.05 or P<0.01). On day 3 and 5 of administration, the contents of IFN-γ (P<0.01 at day 5 in high dose group,P>0.05 for others) and IL-10(P<0.01 at day 5 in eath BQJGHJ gruop and positive drug group) in serum of chicks in BQJGHJ high, middle and low dose groups and positive drug groups increased, while the contents of IL-6 in serum of chicks decreased, which were significantly different from those in model group (P<0.05). On the 2nd day after withdrawal, the contents of IFN-γ and IL-10 in serum of chicks in BQJGHJ high, middle and low dose groups and positive drug groups increased, while the contents of IL-6 in serum decreased, which were significantly different from those in model group (P<0.05 or P<0.01); There was no significant difference in the contents of IFN-γ and IL-6, IL-10 in serum of chicks between each BQJGHJ group and the positive drug group (P>0.05). BQJGHJ can increase the content of IFN-γ,IL-10 in serum of chicks with infectious bronchitis, reduce the content of IL-6 in serum of chicks with infectious bronchitis, so as to achieve the effect of treating infectious bronchitis in chicks.

This paper aimed to discuss the mechanism of the Scutellaria baicalensis water extraction on the damage and repair of jejunum in the model of enteritis mice. Thirty mice were randomly divided into control group, model group, low dose group, medium dose group and high dose group. Each mouse in the model group were fed with 0.4 mL·d^-1 enterotoxigenic Escherichia coli (3×10⁸ CFU·mL^-1) by the method of intragastric administration for 3 days to establish the model of enteritis. Each mouse in the low (0.3 g·d^-1), medium(0.6 g·d^-1) and high dose (1.2 g·d^-1) groups were pre-administered with the Scutellaria baicalensis water extraction for 6 days before establishing the enteritis model. Results showed that, compared with the control group, the mRNA expression of IL-6 in the jejunum of model group was extremely significantly increased (P<0.01), and the villous structure of jejunum was seriously damaged. However, the mRNA expression of MAPK in the model group that could repair the intestine damage did not change significantly. Compared with the model group, the mRNA expression of IL-6 in the jejunum of Scutellaria baicalensis water extraction groups were all extremely significantly decreased (P<0.01), as well as the mRNA expression of JNK/ERK (P<0.05). Moreover, the villous structures of mice in the Scutellaria baicalensis water extraction groups were all improved. In conclusion, Scutellaria baicalensis water extraction could repair the damage of jejunum by weakening intestinal inflammation and decreasing the mRNA expression of JNK and ERK in the model of enteritis mice.

To explore the biological characteristics of Clostridium cadaveris isolated from goat, physiological and biochemical analysis, 16S rDNA gene amplification, animal infection test, drug-resistant phenotype and genetic testing for drug resistance of LJH-1 strain were carried out. The results showed that strain LJH-1 was Gram-positive bacteria. A fragment of 1 023 bp was obtained by PCR amplification. LJH-1 and Clostridium cadaveris were clustered, so it was identified as Clostridium cadaveris. All mice in the experimental group died. There were many red blood cells and inflammatory cells in the central hepatic lobular vein of the mice. Blood vessels in the renal stroma were congested. Intestinal mucosa epithelial cells shed. LJH-1 was sensitive to cefotaxime and amoxicillin, and resistant to cefradine and amoxicillin. Ant(3″)-Ia, aac(3)-IIa, aph(3')-IIa, Sul2, Sul3 and TEM were detected. This provided scientific data for the detection, prevention and treatment of the bacterial diseases.

In order to establish a SYBR Green Ⅰ fluorescence quantitative RT-PCR method for the rapid detection of Porcine astrovirus 4 (PAstV4), a pair of specific primer was designed based on the conservative ORF1a gene. The amplified fragment was cloned into pMD19-T vector. The acquared recombinant plasmid, as a standard, was detected by SYBR Green Ⅰ fluorescence quantitative RT-PCR assay. And the sensitivity, specificity and repeatability were verified. The results showed that the minimum detectable amount of the method was 50.3 copies·μL^-1, which was 100 times more sensitive than the traditional PCR, and there was no cross reaction with CSFV, PRRSV, PRV, PEDV and PDCoV. The established standard curve showed a good linear relationship with R²=1.00. The established method was used to detect 43 clinical samples collected from 2018 to 2019, with a positive rate of 18.6%. These results indicated that a convenient, specific and sensitive qPCR assay was successfully established for the detection of PAstV4.