Toxoplasma gondii, as one of the Apicomplexa phylum parasites, which repeatedly goes through a cycle of invasion, division and induction of host cell rupture, which is an obligatory process for proliferation inside warm-blooded animals. It is known that the biology of the parasite is controlled by a variety of factors ranging from genomic to epigenetic to transcriptional regulation. One of the mechanisms by which Toxoplasma gondii likely regulates responses to extracellular stimuli and life cycle transitions is through protein post-translational modifications (PTMs). PTMs, such as phosphorylation, ubiquitination, crotonylation, palmitoylation, succinylation, acetylation, methylation, glycosylation and 2-hydroxyisobutyrylation, regulate numerous metabolic processes and every aspect of T. gondii biology. PTMs can play a vital role in any time of their lifespan, which through alterations of target protein localization, structure, activity, protein-protein interactions, among other functions, dramatically increase proteome complexity and diversity. In this review, we highlight the advanced insights into PTMs in T. gondii and their crosstalk, laying a foundation for in-depth study on the biological characteristics of T. gondii.

Dermanyssus gallinae is one of the most serious ectoparasites in the poultry breeding industry. The most common mean of controlling Dermanyssus gallinaes is to use chemical drugs for prevention and control in poultry houses. With the development of healthy diet, more and more attention was paid to the problem of pesticide residues in eggs and chickens. Alternatives to chemical drugs have been explored, including vaccines, biological control methods, physical control methods and environment-friendly semiochemicals of Dermanyssus gallinaes. This article reviewed the current research status of semiochemicals such as aggregation pheromones, sex pheromones, alarm pheromones, kairomones and plant compounds, and provides theoretical basis to seek for green and pollution-free alternatives for the prevention and control of Dermanyssus gallinae.

Zinc finger(ZF) structure is a kind of common protein structure widely existing in animals, plants, microorganisms including many viruses. The characteristic group of amino acid residues in the peptide chain combines with Zn²⁺, thus forming a very short peptide space configuration which can self fold into "finger" shape. Zinc-finger proteins (ZNF) are proteins with zinc finger structure. It has been found that some viruses can encode zinc finger proteins, which can affect virus replication, packaging and release of mature virus particles, regulate and activate virus transcription and cleavage infection, and assist virus in innate immune escape. In this paper, the research progress of the role of ZNF in the process of virus infection is reviewed, which provides a reference for the further study of ZNF and the development of its targeting antiviral drugs.

Interferon-epsilon (IFN-ε) is a novel type I interferons (IFNs), which is also a type of cytokines secreted by varieties of cells. It is a glycoprotein, which can exert antiviral, immunoregulatory and anti-tumor biological activities. However, unlike other type I IFNs, IFN-ε is constitutively expressed by mucous epithelial cell and regulated by hormones without virus induction. IFN-ε plays an important role in mucosal immunity. At present, there are few reports on related studies. In this paper, we reviewed the research progress of IFN-ε origin of human and different animals, distribution in vivo, protection of reproductive system, regulation of nervous system and antiviral effect, to further understand and develop new IFNs reagent.

The purpose of this study was to estimate the heterosis of reciprocal crosses between Shanxia black pig (SX) and Lulai black pig (LL), and to screen the favorable intercross pattern of them, so as to improve the production efficiency of hogs with high-quality pork and meet the demand of people for high-quality pork. A total of 43 traits belonging to 4 categories were measured, including growth and fattening, body size and appearance(75-110 kg), reproduction, and carcass and meat quality(90-115 kg) in SX(164), LL(69) and the reciprocal crosses between them (6 SX ♂×25 LL♀ and 3 LL♂×35 SX♀), and the differences of these traits among the 4 populations were investigated and the heterosis of F₁ generation of the reciprocal crosses populations was estimated. The results showed that SX had favorable growth, fattening and body size traits, while LL had good reproductive and meat quality traits, and these traits of the reciprocal crosses populations were between them. The reproductive performance of SX×LL was better than that of LL×SX, but the growth and fertility performance was on the contrary. Because the sows of the 4 populations were pure breeds, there was no obvious heterosis in reproductive traits. Except fat thickness, thoracic vertebrae number, pH at 45 min and shearing force having no obvious heterosis and hybrid weakness, most carcass traits together with meat quality traits had obvious hybrid weaknesses. The heterosis of growth and fattening traits were not consistent in the reciprocal populations, the cross population of LL×SX showed significant heterosis, while the cross population of SX×LL had no obvious heterosis. In this study, the heterosis of reciprocal crosses between SX and LL were estimated, and the result would help to select the favorable intercross pattern between SX and LL.

The study aimed to explore the alternative splicing isoforms of pig C1QTNF3 gene and mechanism of miR-101 targeting C1QTNF3 expression in promoting porcine adipose-derived stromal vascular cells (ASVC) adipogenesis. The heart, liver, spleen, lung, kidney, stomach, biceps femoris, psoas magnus, abdominal fat and back fat were obtained from 3 30-day-old healthy Mashen male piglets. The full-length coding sequence (CDS) of C1QTNF3 transcripts were cloned by RT-PCR, the biological characteristics of C1QTNF3 protein were analyzed by bioinformatics methods. The expression patterns of C1QTNF3 transcripts in porcine tissues were detected by qRT-PCR. Bioinformatics prediction revealed that miR-101 was the upstream regulatory factor of C1QTNF3. The effect of miR-101 on C1QTNF3 was verified by dual luciferase reporter assay. The effect of overexpression of miR-101 on adipogenesis of porcine ASVC was detected by Oil Red O staining and qRT-PCR. Two transcripts of C1QTNF3 gene, C1QTNF3 and C1QTNF3-1, were successfully obtained in this study. C1QTNF3-1 was a newly identified transcript, which lacked 219 bp (73 amino acids) in the first exon compared with C1QTNF3. The phylogenetic tree analysis showed that the porcine C1QTNF3 and C1QTNF3-1 proteins sequence were highly related to Homo sapiens and Manis javanica. C1QTNF3 and C1QTNF3-1 were expressed in all porcine tissues, the expression level of C1QTNF3-1 was significantly higher than that of C1QTNF3 (P<0.01). Overexpression of miR-101 significantly reduced the relative luciferase activity of C1QTNF3 3'UTR(P<0.01) and mRNA expression of C1QTNF3(P<0.01); the mRNA expression of key genes for adipogenesis, PPARγ, C/EBPβ, SREBP-1c and FABP4 were significantly increased (P<0.01). In this study, two transcripts of pig C1QTNF3 gene were successfully cloned. The expression of C1QTNF3-1 was significantly higher than C1QTNF3 in different porcine tissues, suggesting it was the predominant transcript. In addition, miR-101 was identified as the upstream regulatory factor of C1QTNF3. By reducing C1QTNF3 gene expression, miR-101 promoted porcine ASVC adipogenesis, which enriched the biological role and regulatory network of C1QTNF3.

The study aimed to clone the angiotensin converting enzyme 2 (ACE2) gene, analyze its structural and expressing characteristics. In this experiment, the Xueshan chicken was selected as the research objects, the coding region of ACE2 gene was amplified by PCR, the structural characteristics were analyzed by the method of bioinformatics, the standard plasmid of ACE2 gene was constructed by TA cloning, and the expression characteristics of ACE2 gene was determined in the tissues of Xueshan chicken by absolute quantitative method. The results showed that CDS (2 427 bp) of ACE2 was cloned and encoded 808 amino acids, it was consistent with the ACE2 gene in white-feathered broilers, but a large number of mutations were found in yellow-feathered broilers which led to changes in amino acids. The homology analysis found that sequence of ACE2 gene of Xueshan chicken had 99% homology with red jungle fowl, followed by mallard duck 88%, and 73%-76% with other animals. The bioinformatics analysis showed that ACE2 belonged to type I transmembrane protein(secretory protein), and signal peptide was located in 1-17aa, contained 46 protein phosphorylation sites. The quantitative results found that the expression of ACE2 in the small intestine was significantly higher than that in other tissues. The expression of ACE2 in bursa, trachea, gallbladder and spleen was lower, and the expression in the spleen was the lowest. The expression of ACE2 in jejunum, cecum, spleen and lung of hens were significantly higher than those of roosters. The expression of ACE2 in kidney and heart of roosters were significantly higher than those of hens. The expression of ACE2 in the duodenum, jejunum, lung, spleen and ileum tissues of 90-day-old Xueshan chicken were significantly lower than those of 45-day-old Xueshan chicken, and the results showed an opposite trend in the bursa and kidney. The coding region of ACE2 gene was obtained in Xueshan chicken. Bioinformatics analysis showed that ACE2 gene was conservative in poultry. The expression level of ACE2 gene was higher in the intestinal tissues than that in other tissues, higher in hen than that in rooster, and the expression level decreased with time. The results laid a foundation for the study of function of ACE2 gene in the gut of yellow feather broilers.

Follistatin (FST) gene of Taihang chicken was analyzed by bioinformatics methods and its expression differences in different tissues and follicles at different developmental stages was explored, and the effect of FST on the laying performance of Taihang chickens was analyzed. Coding region of FST gene was cloned and sequenced for 43-week-old healthy Taihang chicken, functional structure was predicted by bioinformatics softwares. Quantitative PCR (Q-PCR) was used to detect the expression differences of FST gene in different tissues and follicles at different developmental stages of Taihang chickens. Three replicates were set in each group and three parallel experiments were carried out. The results showed that the coding region of FST gene of Taihang chicken was 1 032 bp, encoding 343 amino acids. Homology analysis indicated that FST protein had the highest homology with Gallus gallus (100.0%), and both had Sec signal peptide. The mature protein had two conserved domains: FOLN and KAZAL_FS. Physicochemical properties analysis of FST protein showed that it was hydrophilic protein, whose molecular formula was C₁₆₁₉H₂₅₆₇N₄₅₉O₅₂₀S₄₄, molecular weight was 38.192 6 ku, and theoretical isoelectric point was 5.59, the highest content was Cys (C, 10.50%). The secondary structure consisted of α-helix, extended chain, β-corner and random coil, accounting for 16.33%, 16.62%, 5.25% and 61.81%, respectively. qPCR revealed that FST gene was expressed in all tissues, the highest expression was found in liver. During follicle development, FST gene was expressed highest in the large white follicles (P<0.01), there was a 1.6-fold difference in expression before and after follicle selection. These results provided important basic data for studying FST gene in Taihang chicken, and provided a reference for the further mining candidate gene for egg-laying performance of hens.

The aim of this study was to investigate the effects of intravenous infusion of atrial natriuretic peptide (ANP) on blood lipid metabolism hormones and tail lipid transcriptome of sheep, and to provide reference for elucidating the mechanism of ANP in regulating lipid metabolism in sheep. Eight healthy Altay ewes with the age of 1.5-year-old, body weight (BW) of (45.6±6.5)kg were selected, and self control design was used. During the control period, 100 mL saline was infused intravenously in 45 min, and during the trial period, 100 mL of ANP saline solution with a dose of 1.125 μg·kg^-1BW was infused intravenously in 45 min. Intravenous infusion lasted for 4 days. At the end of each period (4th day), blood samples were collected at 0, 15, 30, 45, 60, 90 and 120 min after infusion, and the content of ANP, adiponectin, insulin, leptin and cyclic guanosine (cGMP) in plasma were deterimed. The sheep tail fat samples were collected for trans-criptome sequencing and biological information analysis, and qRT-PCR was used to detect the change of relative candidate genes expression. The results showed that: 1) After intravenous infusion of ANP, compared to the control period, the plasma ANP level at 30 and 90 min was significantly higher during the trial period(P<0.05); Adiponectin level at 30 min significantly increased (P<0.05), and at 90 min extremely significantly increased (P<0.01); The plasma cGMP level at 30 and 90 min extremely significantly increased(P<0.01), and at 45 and 60 min significantly increased (P<0.05); The plasma insulin at 0, 30 and 120 min extremely significantly increased(P<0.01), at 15, 45, 60 and 90 min significantly increased (P<0.05); The plasma leptin level at 0 and 15 min significantly increased(P<0.05), at 30 min extremely significantly increased(P<0.01). 2) Transcriptome analysis showed that there were 3 686 differentially expressed genes, of which 1 482 genes were up-regulated and 2 204 genes were down-regulated; GO function enrichment analysis showed that differentially expressed genes were significantly enriched into 118 biological functions, mainly related to mitochondrial respiratory chain and oxidative phosphorylation; KEGG pathways analysis revealed that differentially expressed genes were significantly enriched into 46 pathways, mainly involved in ribosome, thermogenesis, oxidative phosphorylation and regulating lipolysis in adipocytes and other signaling pathways; qRT-PCR analysis results showed that the expression trends of AQP7, FABP4, PLIN5, ADIPOR2, MGLL,IDE and ACSL1 were consistent with the RNA-seq results, which proved that the trans-criptome sequencing data was reliable. In conclusion, exogenous ANP promotes lipolysis by changing the level of lipid metabolism-related hormones and increasing the oxidative phosphorylation and thermogenesis of adipose tissue in sheep.

In this study, the skinfold thickness over the neck (STN), rib (STR) and udder (STU) of Holstein cows (referred to as neck, ribs and breast skinfold thickness, respectively) and body condition score (BCS) were measured in a large population. The purpose was to explore the population characteristics of skinfold thickness in different body positions and BCS, and to estimate the genetic parameters of skinfold thickness in different body positions and BCS of Holstein cows by modeling. The STN, STR, STU and BCS of 10 915 lactating Holstein cows from 12 dairy farms were measured in summer from 2017 to 2020. The single-trait and four-trait animal models were employed to perform genetic analysis, and heritabilities and genetic correlations for STN, STR, STU and BCS were obtained. Furthermore, the approximate genetic correlations between skinfold thickness, BCS and important functional traits (longevity and reproduction traits) were calculated. The results showed that the STU, STN, STR and BCS in Holstein cows were (7.49±1.45) mm, (7.27±1.34) mm, (11.74±1.90) mm and (2.94±0.79), respectively. There were moderate heritabilities for STN (0.15) and STU (0.11), and moderate to high heritabilities for STR (0.28) and BCS (0.22). The moderate to high genetic correlations among the skinfold thickness over the different body positions were found, ranging from 0.21 (STN and STU) to 0.69 (STN and STR). There was a large variation for genetic correlations between BCS and skinfold thickness over the different body positions, ranging from -0.25 (BCS and STU) to 0.28 (BCS and STN). The moderate genetic correlations between skinfold thickness, BCS and some functional traits were found. In this study, the genetic analysis on skinfold thickness and BCS traits were performed, and the heritabilities and genetic correlations for skinfold thickness over the different body positions and BCS were estimated. The results from the current study could help us understand the genetic foundation for fat deposition in dairy cattle, and was beneficial for balance breeding in Chinese Holstein population.

The purpose of this study was to explore the biological characteristics of serum proteins in horses and donkeys, and to provide a theoretical basis for the comparison of the physiological characteristics among equine species and health protection. In this study, 9 Mongolian horses and 9 Liaoxi donkeys, female, aged 4-10 years old, were selected from an intensive farm in Dalian city, Liaoning province. They were all in the interestrus of normal estrous cycle. They were healthy, disease-free, mental state and eating well, and provided the same feeding conditions. Nine serum samples of horses and donkeys were collected and randomly divided into 3 groups, respectively. Three serum samples in each group were evenly mixed into one biological repeat, and 3 serum biological repeats were obtained in each group. In this study, Label-free proteomics and bioinformatics methods were used to compare the serum protein components of horses and donkeys,serum proteins were extracted, and then the proteins were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The expression of proteins in horse and donkey serum samples was analyzed by database search. The different key regulatory proteins in serum between horses and donkeys were screened by bioinformatics method.The results showed that a total of 361 proteins were identified, of which 288 proteins were identified in horse serum with molecular weight from 1.52 to 511.24 ku, and 244 proteins were identified in donkey serum with molecular weight from 1.53 to 611.47 ku, and 231 differentially expressed proteins were obtained (fold change≥1.5, P≤0.05). These proteins were mainly involved in protein activation, complement activation, immune response, regulation of coagulation and lipoprotein oxidation and other biological processes. The significantly enriched KEGG pathways included the complement and coagulation cascade, phagosome, protein processing in endoplasmic reticulum, antigen processing and presentation, metabolism of glycine, serine and threonine, proteoglycans in cancer, and so on. The interaction analysis of differentially expressed proteins showed that the most significant function modules enriched by differentially expressed proteins were metabolic pathway, complement and coagulation cascades and phagosome. HSP90AA1, HSPA8, APOD, APOM, SERPING1, MASP1, CALR, TUBA1B and TUBB were important nodes in network. There are some differences in serum protein composition between horses and donkeys under physiological conditions. This study provides a foundation for further revealing the differences of physiological characteristics among equine species and effective health protection on them.

This study aimed to propose a new genomic relationship matrix and validate its simulated application efficacy for an across-breeds population. The bovine phenotypic and genotypic data were simulated by QMsim software; General G matrix was constructed by Gmatrix software; The new G matrix was constructed by R language. Compared with the general G matrix, the new G matrix took into account the Hardy-Weinberg disequilibrium loci in combined population; The ssGBLUP method in DMU software was used to calculate the estimated genomic breeding values; The GEBV accuracies predicted by two G matrixes were compared for different situations. The results indicated that under different heritabilities and QTL numbers, the GEBV accuracy of the general G matrix using A₂₂ matrix weighting could be achieved without using the A₂₂ matrix weighting for the new G matrix. When the partial pedigree information missed, the GEBV accuracy of new G matrix without weighting outperformed general G matrix with weighting. In conclusion, new G matrix could perform better on GEBV prediction for across-breeds population when the partial pedigree information missed.

This study aimed to select candidate biomarkers for different pregnancy status of cows on 17 days after artificial insemination. Healthy Holstein dairy cow with a weight of (550±50)kg and similar body condition scores in a dairy farm in Ningxia was used as the test object. After synchronization of estrus, blood was collected from tail vein before morning feeding on the 17th day after artificial insemination. Pregnancy status of cows was diagnosed by pedometer and B-ultrasound instrument at the later stage. According to the diagnosis results, cows were divided into pregnant group (group A, n=12) and non-pregnant return to estrus group (group B, n=24). Blood samples from the two groups were collected for analyzing metabolic profile and metabolites change. Principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) showed that the plasma metabolic profile of group A and B were significantly changed. A total of 8 differential metabolites with area under the ROC curve (AUC)> 0.8 were selected. Ala-Leu, Glycine, L-Proline, L-Norleucine, DL-Phenylalanine, Sarcosine, Pyrrole-2-carboxylic acid and Val-Met were expected to be the different metabolites for the identification of pregnancy cows (group A) and non-pregnant return to estrus cows (group B). In conclusion, the 8 metabolites in plasma could be used as potential biomarkers in the pregnancy recognition period, which could provide new ideas for early pregnancy diagnosis.

The aim of this study was to clone the establishment of sister chromatid cohesion N-acetyltransferase 2(ESCO2) gene, analyze its expression and localization in the testis at different developmental stages in cattle-yak and yak, and provide a theoretical basis for further study the role mechanism of ESCO2 in the male meiosis process. Healthy male cattle-yaks and yaks were used as experimental animals in this study, they were divided into fetal cattle group (5-6 months old), juvenile group (1-2 years old) and adult group (3-4 years old), with 3 animals in each group. The CDS region of ESCO2 gene of cattle-yak and yak were cloned by RT-PCR and its sequence characteristics were analyzed by bioinformatics softwares. qRT-PCR was used to detect the expression patterns of ESCO2 gene in different tissues of cattle-yak. The expression of ESCO2 mRNA in testes at different developmental stages of cattle-yak and yak were detected by qRT-PCR, the differences of cell location and expression of ESCO2 protein were detected by immunohistochemistry (IHC) staining. The results showed that the CDS region of ESCO2 genes in cattle-yak was 1 833 bp(MW198470), it encoded 610 amino acids. Compared with yak, the ESCO2 protein sequence of cattle-yak had more 19 amino acids at position 301-319 aa and there were 3 amino acid mutations. The cattle-yak and cattle had higher homology than other mammals based on the protein sequence of ESCO2. Interaction network showed that ESCO2 mainly interacted with SMC3, SMC1A, PDS5A, PDS5B, STAG2 and these proteins were related to sister chromatid condensation, meiotic cell cycle, DNA repair, cell division and chromosome remodeling. ESCO2 was expressed in all tissues of the cattle-yak, but its relative expression level in the testis was significantly higher than those in other tissues (P<0.05). In addition, the spatial expression of ESCO2 in testes of cattle-yak showed an upward trend with age, and the relative expression abundance of ESCO2 in cattle-yak at juvenile and adult stages was significantly lower than that of yak(P<0.05). IHC staining showed that meiosis of male cattle-yak was blocked in primary spermatocytes and ESCO2 protein was differentially expressed in primary spermatocytes between cattle-yak and yak. The research suggested that there was obvious difference in the sequence and expression pattern of ESCO2 between cattle-yak and yak, which may be one of reasons caused male sterility in cattle-yak, but the specific mechanism need further study.

The aim of this experiment was to clarify the changes of rumen metabolites and their metabolic pathways of dairy cows before and after parturition. Ten healthy Holstein cows with similar body condition and parity were selected to collect fasting rumen fluid before and after parturition. The changes of rumen fermentation products and their metabolic pathways were analyzed by using UPLC-MS/MS metabonomics technology and pathway analysis method with MetaboAnanlyst 5.0. The results showed that the concentration of 57 rumen metabolites in dairy cows changed significantly before and after delivery, of which 34 metabolites were down-regulated and 23 were up-regulated after delivery. And 9 metabolic pathways (taurine and hypotaurine metabolism, arginine and proline metabolism, arginine biosynthesis, D-glutamine and D-glutamate metabolism, alanine, aspartic acid and glutamate metabolism, starch and sucrose metabolism, galactose metabolism, purine metabolism and vitamin B₆ metabolism) consequently underwent strikingly variation. These results indicated that the composition of rumen microflora related to amino acids, sugars, nucleotides and vitamins occured responsive change in dairy cows at 7~10 days before delivery.

This study aims to establish a high-efficiency, rapid, high-sensitivity and high-specificity TaqMan double real-time fluorescence quantification for duck hepatitis virus type 1 (DHAV-1) and duck hepatitis virus type 3 (DHAV-3) PCR (q-PCR) detection method and applied to the detection of clinical suspected samples. According to the conservative regions of the VP1 gene of DHAV-1 and DHAV-3, two pairs of specific primers and probes were designed and synthesized respectively. On this basis, the TaqMan dual real-time fluorescent quantitative PCR method was established and optimized. Results were as follows: The correlation coefficient (R²) of the optimized standard curve is above 0.999, the amplification efficiency (E) is between 105% and 110%, and the coefficient of variation (i-CV) and the coefficient of variation between groups (C-CV) are all below 0.77%. The double q-PCR method was used to detect mixed plasmids and viral nucleic acids of different dilutions. The results showed that the established TaqMan double q-PCR has high specificity; the sensitivity of simultaneous detection of DHAV-1 and DHAV-3 can reach 10 copies, Which are 1 000 times and 100 times higher than conventional PCR methods. Forty tissue samples of duck suspected of duck hepatitis virus infection from central Jiangsu and northern Jiangsu were tested. The results showed that 36 of them were positive by q-PCR method, with a detection rate of 90%; while conventional PCR methods failed to detect samples with high Ct values, and the detection rate was 72.5% (29 positive samples).The establishment of the TaqMan dual q-PCR detection method provides an efficient, sensitive, and specific tool for the detection of clinical samples of DHAV-1 and DHAV-3, and promotes clinical molecular epidemiological investigation and viral quantitative analysis.

The study focused on analysis of the molecular characteristics and genetic evolution of the antigen gene E^rns of different subgenotypes of bovine viral diarrhea virus (BVDV) in northwest China from 2013 to 2019. Total RNA was extracted from 150 EDTA-anticoagulated blood samples sent for BVDV detection, which were collected from cattle suspected of BVDV infection on large-scale farms in Gansu, Qinghai provinces and Ningxia Hui Autonomous Region. The RNA samples were submitted for RT-PCR with primers targeting at the E^rns-E1 region of the viral genome, followed by cloning, sequencing, and genetic evolution analysis by constructing phylogenetic trees. The virus of individual BVDV subgenotype identified in the samples was isolated with Madin-Darby bovine kidney (MDBK) cells, followed by biotype determination. Results were as follows: The overall BVDV positive rate was 37.33% by RT-PCR, with positive rate of 37.68%, 35.71% and 40.00% in Gansu Province, Qinghai Province and Ningxia Hui Autonomous Region respectively. E^rns-E1 DNA was obtained by RT-PCR from 56 blood samples and 33 different E^rns coding sequences with 681 bp in length were harvested by cloning and sequencing. Sequence analysis showed that the epidemic strains belonged to 10 BVDV genotypes: two within BVDV-1a, five within BVDV-1b, one within BVDV-1c, three within BVDV-1d, 11 within BVDV-1m, one within BVDV-1o, four within BVDV-1p, four within BVDV-1q, one within BVDV-1v, and one within BVDV-2a. One non-cytopathogenic BVDV isolate of individual subgenotype of BVDV-1a, BVDV-1b, BVDV-1v, BVDV-2a and two isolates of BVDV-1d susgenotype were obtained. Higher nucleotide similarity of E^rns gene among different strains within individual subgenotype was observed in the classical subgenotype BVDV-1a to 1d (79.8%-85.9%) or in BVDV-1m to 1q and the novel subgenotype 1v (81.0%-87.3%), with the highest nucleotide similarity shared by strains within BVDV-1m and BVDV-1p (87.3%). The RNase active sites and the dsRNA-interacting motif (₁₃₉KKGK₁₄₂) were conserved among strains within individual subgenotype while the N-glycosylation site at position N26 (26 NRSL) was translocated (24 NVSR) in strains within the 1m to 1q and 1v subgenotype. For the first time, a phylogenetic tree based on the nucleotide sequences of E^rns gene was constructed, illustrating the close genetic relatedness in the evolution among strains within subgenotypes 1m to 1q and 1v. For the first time, the E^rns gene was selected as a target for homology and phylogenetic analysis of BVDV of bovine origin from Gansu, Qinghai provinces and Ningxia Hui Autonomous Region in northwest China. Epidemic strains within 10 subgenotypes were detected, with the BVDV-1m subgenotype being the most prevalent and close genetic relationship shared by BVDV strains within BVDV-1m to 1q and 1v subgenotypes.

In order to obtain and analyze the complete genome sequences of enzootic nasal tumor virus 2 (ENTV-2) in goat intranasal tumor tissues obtained from a Fujian farm, six specific fragments of the ENTV-2 were amplified from these tumor tissues by RT-PCR. The products were then sequenced and assembled using biological software DNAStar to obtain the whole genome sequences. We found that whole length of the viral genome was 7 443 bp, and the virus was named as ENTV-2-FJ. The structure of ENTV-2-FJ genome is similar to other retroviruses, which has a 5'-U5-gag-pro-pol-env-U3-3' canonical structure with four open reading frames, flanked by non-coding regions and terminal repeats sequence. In order to prepare rabbit anti-ENTV-2 gag protein polyclonal antibody, a pair of specific primers was designed and used to clone the gag gene that was then ligated to pET-21a (+) vector. The recombinant plasmid pET-21a-gag was constructed, transferred into BL21 (DE3) competent cells, and the recombinant fusion protein with a molecular weight of about 70 ku was successfully induced by IPTG. Western blot analysis showed that the prepared rabbit anti-gag protein antibody specifically detected the gag protein in the tumor tissues in diseased goats and cells expressing gag gene. Furthermore, we generated K562 cell lines overexpressing the gag gene and conducted tumorigenic experiments on nude mice. The results showed that overexpression of gag gene promoted tumor growth through activation of JAK2-STAT5 signaling pathway. Taken together, this study provides the complete genomic sequences of ENTV-2-FJ and rabbit polyclonal antibodies against viral gag protein, and reveals an important role of the gag protein in tumorigenesis mediated by ENTV-2-FJ.

This study intends to evaluate the interference of porcine reproductive and respiratory syndrome virus modified live attenuated virus vaccine (PRRSV MLV-vaccine) on the immune effect of PCV2 and CSFV vaccine, and analyze the effects of different immunization methods on the growth of piglets. It is expected to provide data for exploring the combined immunization of PRRSV MLV-vaccine and PCV2 and CSF vaccine. In this study, 100 piglets were selected and randomly divided into A, B, C, and D 4 groups. Group A was immunized with PCV2 vaccine 7 days after the PRRSV MLV-vaccine; group B was simultaneously immunized with PRRSV MLV-vaccine and PCV2 vaccine; group C was immunized with PCV2 vaccine and group D was immunized with PRRSV MLV-vaccine. Then another 100 piglets were selected and randomly divided into E, F, G, and H 4 groups. Group E was injected with CSFV vaccine 12 days after the PRRSV MLV-vaccine; group F was simultaneously immunized with PRRSV MLV-vaccine and CSFV vaccine; group G was immunized with CSFV vaccine and group H was immunized with PRRSV MLV-vaccine. After 4 weeks of immunization, the level of antibodies in the serum of the piglets was measured. The weight of piglets in each group was measured before and after the experiment, and the average daily gain (ADG) of piglets in different groups was calculated. In groups A-D, group A and B produced high level of antibody responses against PRRSV and PCV2. Group C only produced high level of antibody responses against PCV2, while group D produced high level of antibody responses against PRRS. Among the 4 groups of E-H, the group E and F produced high level of antibody responses against PRRSV and CSFV. Group G only produced high level of antibody responses against CSFV, while group H only produced high level of antibody responses against PRRSV. The ADG of group B among the A, B, C, and D was the highest; while the ADG of group F among the of E, F, G, and H was significantly higher than that of the other three groups. Both simultaneous immunization with PRRSV MLV-vaccine and PCV2 or CSFV vaccine, and subsequent immunization with PCV2 or CSFV vaccine after vaccination with PRRSV MLV-vaccine, could induce high levels of antibody production; in terms of humoral immunity, neither immunization nor non-immunization of the PRRSV MLV-vaccine showed significant immune interference to the other two vaccines. Therefore, “simultaneous immunization with PRRSV MLV-vaccine and PCV2 vaccine at 28 days of age, followed by immunization with CSFV vaccine 12 days later” not only induced high levels of antibody responses, but also resulted in a higher ADG.

Mycoplasma hyorhinis is ubiquitous in the pig population and could cause systemic diseases such as polyserositis and polyarthritis. At present, there is no standard challenge model for M. hyorhinis, as well as standard virulent strain. In this study, a M. hyorhinis strain was isolated from the synovial fluid of a pig with typical clinical symptoms. It was identified to be M. hyorhinis by analysis of PCR, colony and bacterial morphology identification and multi locus sequence typing (MLST). To evaluate the pathogenicity of the strain, 1-month-old snatch-farrowed, porcine-colostrum-deprived (SF-pCD) pigs were inoculated with the isolated strain. The infected piglets showed obvious joint swelling and lameness. The daily weight gains of the infected piglets were significantly lower than those of the pigs in the control group. Three of the infected piglets died during the observation period. At necropsy, the infected pigs showed extensive pleurisy, peritonitis, pericarditis and arthritis. The histopathological analysis of the lung tissues showed thickened alveolar septa but no shrimp-like lesions. The challenge strain can be re-isolated from the tonsils, lungs, hearts and joint effusions of the infected pigs. To sum up, a strain of M. hyorhinis was isolated in this study, and the pigs artificially infected which showed typical symptoms. The establishment of the challenge model laid an important foundation for the study of M. hyorhinis pathogenic mechanisms and vaccine development.

This study uses the IPEC-J2 cell line, which is widely used in studying the mechanism of intestinal infection, to explore the role of poCRIP2 in porcine gastrointestinal inflammation. The full-length sequence of poCRIP2 gene was identified and the relevant characteristics and its tissue expression pattern were analyzed. An effective three-dimensional structure model of poCRIP2 was established to explain its function, and to understand the immune function of poCRIP2 in the process of bacterial infection and its relationship with NF-κB pathway. Using reverse transcription polymerase chain reaction (RT-PCR) technology to obtain the full-length sequence of poCRIP2 from pig heart; using Primer Premier5.0 and NCBI Primer BLAST program to design a specific primer pair, using the protection gene β-actin normalizes the results to test the tissue distribution of the CRIP2 gene in pigs; using bioinformatics methods to analyze the characteristics of poCRIP2 to construct the poCRIP2 protein model; using the ROBETTA server and the high homology mouse LIM-homeodomain protein islet 1 (Isl1) (PDB: 4 JCJ) to simulate the three-dimensional structure of poCRIP2; the model was analyzed by PROCHECK and ProSA; the stereochemical and topological analysis of protein folding was performed by Procheck, and applied ProSA to evaluate protein folding; PyMOL program was used to evaluate the modeling results and analyzes the conserved domains of poCRIP2; designs an intestinal infection test to analyze the function of poCRIP2 in intestinal immunity. The results showed that the full-length sequence of poCRIP2 cDNA obtained from porcine heart was 1 118 bp. The homology of poCRIP2 protein to human and mouse were 94.23% and 93.75%, respectively. It also contained two conserved regions (LIM-TLP and LIM-CRP). Tissue expression pattern analysis of poCRIP2 showed that poCRIP2 was expressed in all tissues, but less in the small intestine, lung, stomach, spleen and muscle and other tissues. The expression level of poCRIP2 under the action of Gram-negative bacteria is 1~2 times higher than that of the control group, but the NF-κB pathway is activated under the same action without inhibition. PoCRIP2 may not interact with the NF-κB pathway, but may have combined effect with other pathways on intestinal immunity. This study has successfully obtained the full-length sequence of the pig CRIP2 gene. The tissue expression pattern of poCRIP2 shows that poCRIP2 is similar to human CRIP2. A reliable 3D structural model has been established. Based on this model, it is speculated that the functions contained in poCRIP2 are mainly composed of β folding and α-helical structure are realized, and poCRIP2 can be significantly upregulated after Gram-negative bacteria infect the intestine. PoCRIP2 may not depend on the NF-κB pathway and play other roles in intestinal immunity.

To scientifically select and use brucellosis antibody detection method, and promote the standardization of brucellosis diagnostic reagents. In this study, the sensitivities of four methods, which included Brucella fluorescence polarisation assay (FPA) antibody test kit, animal brucellosis competitive ELISA (cELISA) antibody test kit, bovine brucellosis indirect ELISA (iELISA) antibody test kit and the improved micro complement fixation test (mCFT) developed by the National/OIE Reference Laboratory for Brucellosis, were determined with the brucellosis positive serum standard. The sensitivity and specificity of the four methods were assessed by the detection serum samples with known background. At the same time, the clinical bovine serum samples were detected by the above four methods, and the coincidence rates were compared. The results showed that the sensitivities of the four methods were the same. The detection results of brucellosis positive serum standard diluted 1∶20 (50 IU·mL^-1) was positive, and the detection results of brucellosis positive serum standard diluted 1∶40 (25 IU·mL^-1) was negative. The sensitivity of FPA, cELISA, iELISA and mCFT were 97.14%, 100.00%, 100.00%, 98.57%, respectively. The specificity of the four methods were 96.34%, 95.12%, 97.56%, 100.00%, respectively. By detecting the 315 clinical samples, the result showed that the coincidence rates of different methods were higher than 90.00%. According to the confirm result of mCFT, the coincidence rates of iELISA, FPA and cELISA were 97.14%, 96.83% and 92.70%, respectively. Compared with iELISA, the coincidence rates of FPA and cELISA were 95.24% and 93.65%, respectively. Compared with cELISA, the coincidence rate of FPA was 91.43%. The identical degree of iELISA, FPA and mCFT was the highest, while the identical degree of cELISA and the other three methods was slightly lower.

NMB/NMBR had been involved in anti-influenza A virus (IAV/H1N1/PR8) infection by regulating the expression of cytokines. To explore the signaling pathway of NMB/NMBR against IAV/H1N1 infection, the MLE-12 cells and mice were infected with IAV/H1N1 subtype PR8 or WSN strain respectively, and the cells were treated with NF-κB inhibitor BAY11-7028 alone or in combination with NMB, meanwhile, the mice were injected with NMB or NMBRA. The expression profiles of NMB, NMBR, IL-6, IFN-α and NP genes in transcript levels were analyzed by the methods of RT-PCR and qRT-PCR, and the expression profiles of NMB, NMBR, P65/p-P65, IκBα and NP in protein levels were analyzed by Western blot. The results showed that the decreased expression levels of NMB, NMBR, IL-6 and IFN-α, and increased NP levels in MLE-12 cells after PR8 or WSN infection were induced by BAY11-7028 treatment. Meanwhile, the significantly decreased expression levels of NMB, NMBR and p-P65 proteins, and increased expression levels of IκBα and NP proteins were also induced by BAY11-7028. However, NMB combined with BAY11-7028 could induce a significant decrease in IL-6 and NP, and a significant increase in IFN-α in MLE-12 cells after PR8 or WSN infection. NMB could inhibit the expression of p-P65 and NP proteins, and promoted the expression level of IκBα protein in lung tissues of PR8 and WSN infected mice. NMBR inhibitor NMBRA combined with NMB could counteract the regulatory effects of NMB on the expression of these proteins after PR8 or WSN infection. These results suggest that NMB could regulate the phosphorylation of P65 protein and the expression of IκBα involved in NF-κB signaling pathway in PR8 or WSN infected MLE-12 cells and mice, and affect the expressions of downstream cytokines, IL-6 and IFN-α, thereby play the innate immune response against IAV/H1N1 infection.

This study was conducted to investigate the histological structure and effects of TGF-β2 and HIF-1α on growth and development of hair follicles in young yaks. Ten healthy young yaks, Skin tissues from their neck, back, breast, abdomen, crus, axilla and scrotum were collected. Paraffin sections were prepared from tissues, stained with HE and Sacpic, and then observed under optical microscope after which representative photomicrographs were taken for analysis. Hair follicles were counted in each part of the skin tissue and categorized as either hairy or less hairy skin. Then qRT-PCR, Western blot, and immunohistochemistry techniques were used to preliminary study TGF-β2 and HIF-1α expression and localization in the hairy or less hairy skin of young yaks. The results showed that hair follicles in the skin of young yaks were distributed in the dermis, often accompanied by sweat glands and sebaceous glands. The structure of hair medulla and inner root sheath was incomplete. Sacpic staining showed the inner root sheath of hair follicles as red, the outer root sheath as pale green, and connective tissue sheath as blue-green. The numbers of hair follicles in the abdominal skin and scrotal skin were (2 085±15)·cm^-2 and (158±15)·cm^-2 respectively. Immunohistochemistry results showed that TGF-β2 and HIF-1α were mainly distributed in the epidermis, outer root sheath of hair follicle, sebaceous glands, and sweat glands. The mRNA and protein levels of TGF-β2 in scrotum and axillary were significantly higher than those in abdomen and back. The mRNA and protein levels of HIF-1α in the abdomen were significantly higher than those in the other three sites. In conclusion, young yaks’ hair follicles were in the catagen phase. The abdomen had the largest number of hair follicles, followed by the back, and the scrotum, the least. The expression levels of TGF-β2 and HIF-1α were significantly varied in different parts of the skin, which provided a theoretical basis for further study on the effects of TGF-β2 and HIF-1α on hair follicle growth and development in yak skin.

Haemophilus parasuis disease is a systemic infection of swine caused by Haemophilus parasuis. It mainly affects piglets at the nursery stage and is extremely harmful to pig farms. β-lactam drug is a kind of antibiotics used more clinically, and has the advantages of strong bactericidal activity, low toxicity, wide indications and good clinical curative effect, also often used to treat Haemophilus parasuis. To understand the drug resistance of Haemophilus parasuis to β-lactam antibiotics, and guide clinical drug use and new drug development more scientifically, this study referred to the method of establishing the epidemiological cut-off values in CLSI, to summarize the susceptibility results of strains from different regions to β-lactam drugs (determined by broth dilution method)， and to establish ECOFFs for β-lactam drugs. The ECOFFs of cefaclor，cefepime, cefotaxime, cefoquinme, ceftiofur, cefalexin, amoxicillin-clavulanic acid, amoxicillin, clavulanic acid, ampicillin, penicillin, oxacillin, imipenem and meropenem were recommended to be 16, 0.5, 0.125, 0.031 25, 0.5, 32， 0.25， 1， 0.5， 1， 2， 8， 0.25， 0.062 5 μg·mL^-1. In the absence of CLSI and EUCAST sensitivity criteria, the appearance of non-wild-type strains can be recognized directly, which is beneficial to the development of drug resistance monitoring and has certain reference value for the treatment and control of Haemophilus parasuis.

The purpose of the study was to evaluate therapeutic effects of dry suspension of andrographolide against diarrhea in broiler challenged with pathogenic Escherichia coli, and thus to provide experimental evidence supporting its clinical application. Three hundred healthy broilers at 12 days old were randomly divided into 5 groups with 4 replicates in each group and 15 broilers per replicate. Including negative control (no challenge), positive control (E. coli ETECO101 challenge only) and 3 treatment groups (ETECO101 challenge plus 100 or 200 mg·L^-1 of dry suspension of andrographolide, or ETECO101 challenge plus 60 mg·L^-1 of colistin). Starting from day 12 of age, broilers in the 3 treatment groups received colistin or dry suspension of andrographolide for 6 days in drinking water consecutively. Amount of 700 mL drinking water containing colistin or andrographolide were supplied twice a day for each replication. On day 15 of age, broilers in the 3 treatment groups and the positive control group were challenged with 0.2 mL saline containing 5×10⁸ cfu·mL^-1 pathogenic E. coli ETECO101 via intraperitoneal injection, while the negative control was intraperitoneally injected with 0.2 mL of normal saline. Among 4 replicates, one of the replicates in each group (15 broilers) was used for blood and organs sampling at 24 h and on day 5 post challenge (6 broilers for each time point). Liver histopathology, small intestine microstructure, immune organ index, and serum biochemical parameters were determined. The other 3 replicates in each group (45 broilers) were used to evaluate the efficacy of the drugs, including survival rate, fecal score, average daily gain(ADG), and average daily feed intake(ADFI) until day 7 post challenge. The results showed that, compared with the positive control, treating with 100 or 200 mg·L^-1 of dry suspension of andrographolide resulted in an increase in broiler survival rate by 13.3 and 17.8 percentage point, respectively (P<0.05), alleviated diarrhea caused by E. coli ETECO101 challenge, and significantly improved broiler ADG (P<0.01) and ADFI (P<0.01). In comparison with the positive control, treating with 200 mg·L^-1 of dry suspension of andrographolide significantly increased the spleen and bursa indexes (P<0.01) of broilers at 24 h post challenge, decreased levels of serum GLB and ALB on day 5 post challenge (P<0.05), effectively suppressed the increase of MDA content in serum caused by E. coli challenge, and enhanced activities of serum SOD (P<0.01) and GSH-Px (P<0.01). Meanwhile, the height of jejunum villi (P<0.05) and the ratio of villi height to crypt depth in the group of 200 mg·L^-1 of dry suspension of andrographolide were significantly higher than those in the positive control. In summary, supplementation of dry suspension of andrographolide in drinking water could reduce broiler death rate caused by E. coli ETECO101 infection, improve broiler growth performance, alleviate serum oxidative stress, improve intestinal morphology, increase bursa index, and thereby show remarkable therapeutic effects against broiler diarrhea caused by E. coli ETECO101 infection.

To investigate the intervention effect of Dahuang Mudan decoction on acute pancreatitis model rats and its possible molecular mechanism. Ninty-six rats were selected and randomly divided into 6 groups: sham operation group, model group, positive drug control group and Dahuang Mudan decoction high, medium and low dose groups (n=16). The acute pancreatitis model was established by retrograde injection of sodium taurocholate into the pancreatic duct. The model group and sham operation group were given normal saline by gavage; The positive drug control group was subcutaneously injected with octreotide; the with high, medium and low dose experimental group were given Dahuang Mudan decoction for 6 days. Blood was taken from the heart and pancreatic tissue was dissected by laparotomy. The expression levels of HMGB1, RAGE, p-NF-κBp65 and p-IκBα in pancreatic tissue were detected by IHC and Western blot methods. The expression levels of HMGB1 and RAGE in pancreatic tissue were detected by RT-PCR. The contents of IL-1β, TNF-α and IL-6 in pancreatic tissue were detected by ELISA. The results showed that:1) Compared with the sham operation group, the rats in the model group had a relatively poor general survival condition, and a significant swelling and hyperemia of the pancreatic tissue under the microscope. The protein expression of HMGB1 and RAGE, the phosphorylation of NF-κBp65 and IκBα, and the contents of TNF-α, IL-1β and IL-6 in pancreatic tissue were significantly increased (P<0.05); 2) Compared with the model group, the general living conditions of the rats in the treatment groups were improved in different degrees, interstitial edema and necrosis were significantly improved under the microscope. The protein and gene expression of HMGB1 and RAGE, the phosphorylation of NF-κB p65 and IκBα, and the contents of TNF-α, IL-1 β and IL-6 in pancreatic tissue were significantly decreased, especially in the large dose group of Dahuang Mudan decoction (P<0.05). This study indicate that Dahuang Mudan decoction can significantly improve acute pancreatic lesions in rats, and its mechanism is that it can inhibit the activation of HMGB1 /RAGE/ NF-κB pathway and block the inflammatory cascade.

The purpose of this study was to isolate the polysaccharide from Lagotis brevituba Maxim (LMP) and to investigate the structural features and its immunomodulatory activity in vitro. LMP was obtained by water extraction and ethanol precipitation, and purified sequentially by DEAE-52 column chromatography. Subsequently, high performance gel permeation chromatography (HPGPC), fourier transform infrared spectroscopy (FT-IR), ultraviolet spectroscopy (UV) were employed to characterize the structural properties of LMP. The immunomodulatory activities of LMP were studied by MTT, FCM and ELISA, and using murine bone marrow derived dendritic cells (BMDCs) as the target cells. The results showed that LMP was obtained from Lagotis brevituba Maxim and contained no nucleic acid and protein. The yield and total carbohydrate content of LMP were 18.5%±1.7% and 89.7%±1.9%, respectively, and the average molecular weight of LMP was 3.18 ku. In addition, the average particle size and Zeta potentials of LMP were 1 483.89 nm and-14.81 mV, respectively. SEM and AFM images showed that LMP was a sheet-like appearance with smooth surface and uniformly dispersed in height of 5.4 nm. At 3.13-50 μg·mL^-1, LMP promoted the cell proliferation of BMDCs, and significantly increased the expression of surface markers (CD80, CD86, MHC-Ⅰ and MHC-Ⅱ) (P<0.05) and the secretion content of cytokines (TNF-α and IL-12) (P<0.05). Moreover, LMP decreased the phagocytosis of BMDCs (P<0.05). LMP showed better immune activity and it could induce the maturation of the phenotype and functions of BMDCs, which would be potentially developed as an effective immunomodulatory agent.

This study was conducted to study the effect of Huiyangjiuzhen technique on postoperative recovery quality and hemorheology index of dog. Eight healthy dogs were randomly divided into control group and acupuncture group and sterilized, with four dogs in each group. The control group was anesthetized and sterilized according to the normal operation procedure, while the acupuncture group was based on the treatment of the control group by acupuncture at Huiyangjiuzhen acupoints before waking up after the operation. The recovery time, circulatory and respiratory indexes, body temperature and hemorheology indexes and other indicators of the two groups were collected, expressed as mean±standard deviation, and analyzed by t test.The results showed that the recovery of swallowing reflex and the first head up time in the acupuncture group were significantly lower than that in the control group (P<0.05); the HR of the acupuncture group was significantly higher than that of the control group at the first head up time (P<0.05); after 90 minutes of anesthesia the hemorheological index of the acupuncture group were Hηb 200·s^-1, Hηb 150·s^-1, Mηb 50·s^-1, Mηb 30·s^-1, Lηb 10·s^-1, Lηb 1·s^-1, EDI, ET, high shear resistance, medium shear resistance， Low shear resistance, QwX, and the difference are significantly different from the control group (P<0.05), Hηr 200·s^-1, Mηr 30·s^-1, Lηr 1·s^-1, BR, Br, EAI, VAI, ERI have highly significant differences (P<0.05). It shows that acupuncture at the Huiyangjiuzhen acupoints can shorten the recovery time of the surgical dog, improve the heart rate during recovery, reduce the blood viscosity after anesthesia, and alleviate the rise of blood viscosity etc, so as to improve the recovery quality of dogs.

The study aimed to identify and map the genetic markers and functional genes which associated with milk production traits through genome-wide association analysis (GWAS) of milk production traits in Dairy Meade(DM) sheep. In this study, 135 DM sheep were collected and the whole genome resequencing technology was used. The single nucleotide polymorphisms (SNPs) were identified through SAMTOOLS software. The quality control (QC) was performed by Plink v1.90 software. The genome-wide association analysis of milk production traits in DM sheep was performed based on the QC results using the mixed linear model of GEMMA v 0.98.1. The results showed that there was one SNP which was significantly correlated and 8 SNPs were potentially significantly correlated with D90 average daily milk yield trait at genome-wide; The associated candidate genes included TRNAQ-CUG-2, LOC114117240, ACADL, MYL1, CHD6 and SLCO3A1. There were 2 SNPs which were potentially significantly correlated with D150 average daily milk yield trait; The associated candidate genes included PRMT6 and RNF180. There were 2 SNPs which were potentially significantly correlated with lactation period trait; The associated candidate genes included PRMT6, TRNAW-CCA-68 and TRNAS-GGA-61. Further gene functional analysis suggested that ACADL and SLCO3A1 were likely candidate genes which affect milk production traits of DM sheep. This study provides a certain basis for the exploration of milk production traits molecular mechanism in dairy sheep. The result could provide a certain theoretical reference for new dairy sheep breeding in China.

This experiment was conducted to investigate the effects of vitamin K₃ on coagulation time and vitamin K-dependent proteins of 14-day-old Pekin ducks. In order to screen sensitive indicators reflecting the nutritional status of vitamin K in meat ducks, and determine the vitamin K₃ requirements of Pekin ducks. An one-factor completely randomize design with 6 vitamin K₃ supplemental levels (0, 0.5, 1, 2, 3 and 4 mg·kg^-1) was used in this experiment. The diets were formulated by adding 6 different levels of menadione sodium bisulfite to the corn-soybean isolate basal diets. A total of 180 one-day-old male Pekin ducks were randomly divided into 6 groups with 5 replicates each group and 6 ducks per replicate. The experiment lasted for 14 days. The results showed as follows: Different vitamin K₃supplemental levels had no significant effect on the average daily gain, average daily feed intake and ratio of feed to gain of Pekin ducks from 1 to 14 days of age (P>0.05). The prothrombin time of the 0 and 0.5 mg·kg^-1 vitamin K₃ supplemental level groups were significantly higher than those of 2, 3, and 4 mg·kg^-1 vitamin K₃ supplemental level groups (P<0.05). The level of serum PIVKA-Ⅱ content of the 0 mg·kg^-1 vitamin K₃ supplemental level group was significantly higher than those of the 0.5, 1, 2, 3, 4 mg·kg^-1 vitamin K₃ supplemental level groups (P<0.05); The level of serum undercarboxylated osteocalcin content of the 0 mg·kg^-1 vitamin K₃ supplemental group was significantly higher than those of the 2, 3, 4 mg·kg^-1 vitamin K₃ supplemental level groups (P<0.05). Taking prothrombin time, serum PIVKA-Ⅱ and undercarboxylated osteocalcin as evaluation indicators, vitamin K₃ requirements of Pekin ducks from 1 to 14 days of age was determined as 2.00, 0.56 and 1.27 mg·kg^-1 by broken-line regression models. In conclusion, the supplemental of vitamin K₃ to the diet could enhance coagulation performance, decrease the levels of serum vitamin K-dependent proteins such as the PIVKA-Ⅱ and undercarboxylated osteocalcin. Under the condition of this experiment, according to prothrombin time, the evaluation of vitamin K₃ requirements of Pekin ducks from 1 to 14 days of age was 2.00 mg·kg^-1 by broken-line regression models.

This study was conducted to investigate the biological characteristics of the clpV2 gene in the type VI secretion system 2 (T6SS2) from avian pathogenic Escherichia coli (APEC). The clpV2 deleted strain was constructed by the Red recombination system. In addition, the clpV2 gene was cloned into the expression vectors of pBR322 to construct the complementary strain. The mutant strains were proved to be genetically stable. Comparison of biological characteristics was performed. The data demonstrated that, the mutant strains showed no differences in growth curve and sensitivity to multiple antibiotics. But compared with the wild type strain, the motility and biofilm formation of the deleted strain were decreased significantly. This work proved that the clpV2 gene had partial impact on the biological characteristics of TW-XM. Also, this work provided further basis to investigate the function of the clpV2 gene which might also help in understanding the pathogenesis of APEC.

In November 2020, the imported cattle in Binzhou and Dongying of Shandong Province showed circumscribed skin nodules, skin wounds and scab, which were suspected of lumpy skin disease (LSD). In order to confirm the cause of the disease in the two places and understand its genetic evolution relationship, fluorescent quantitative PCR(qPCR) was used for etiological diagnosis. GCPR gene was amplified by PCR and the alignment analysis and phylogenetic analysis were carried out. Lumpy skin disease virus (LSDV) was detected with qPCR in the collected samples. The disease occurred in the two cities were finally confirmed as LSD. The alignment analysis showed that there were 12 nucleotides insertion in the GCPR gene of China/SDBinzhou/2020 and China/SDDongying/2020, which was same as the insertion sequence in GCPR gene of vaccine strains, Neethling vaccine LW 1959 and Neethling-LSD vaccine-OBP, and vaccine-like strain, Saratov. Phylogenetic analysis showed that GCPR of China/SDBinzhou/2020 and China/SDDongying/2020 were closely related to LSDV strains found in Xinjiang. At the same time, LSDV strain found in China were allocated in a large branch. In conclusion, the disease occurred in Binzhou and Dongying were confirmed as LSD, which was the first confirmation of the imported case of LSD in Shandong province.