microRNA played an important role in the process of the animals skin development and their morphogenetic process in previous studies. The database imperfect was still the main reason for limiting its function studies in the study about the microRNA from the goat skin. Our research analyzed the 5.40% no annotation reads from the high-throughput sequencing total reads, reference the goat genomics gene, using the Mireap software to analyze the data, and analyzed the data including new microRNA length, expression, genomics location, predict the target gene and the KEGG pathway. Our research got 28 new microRNA, and their length were equal with the known microRNA in the goat and the cow. Their expression statistics showed that, there were 4 new microRNA their expression over the 100, the highest was the Novel-15, its expression got 555;There were 19 new microRNAs their expression between 10 to 100;There were 5 new microRNA their expression lower 10.We found the new microRNA evenly distributed over the goat chromosome by studying genomics location, exactly locate 2 microRNA on the number 5, 6, 7, 19 and X chromosome respectively. Predicted and analyzed the target gene data we got 6 519 target gene. Analyzed the KEGG pathway showed that, locate the 3 552 target gene on the 51 pathway, the three former pathway were Metabolic pathways, Pathways in cancer and MAPK signaling pathway. Our research found more new goat microRNA, had laid the foundation about the study of the goat skin development regulate.

In order to study differential expression in different tissues and period of MyoD Ⅰ gene in Guanling cattle. This study detected the expression of MyoD Ⅰ in longest back muscle, leg muscles, heart, liver, fat and small intestine tissues of Guanling cattle by Real-time quantitative PCR. Meanwhile, analyzed the expressing rules of longest back muscle in foetal, 18 and 30 months. The results showed that the MyoD Ⅰ was expression at high level in longest back muscle and in muscle tissues after birth had high expression, with the extension of born showed an upward trend. It was inferred that the reason of the high expression might be that longest back muscle was muscle enrichment place and increase of muscle fullness.

In order to provide the scientific basis for molecular epidemiology, genetic variation, comprehensive prevention and control of classical swine fever in Guangxi, the molecular characteristic of E0 and E2 genes of classical swine fever virus (CSFV) in Guangxi was analyzed. The E0 and E2 genes of CSFV from positive samples of swine was amplified using RT-PCR, cloned into the pMD18-T vector and sequenced. The sequences were analyzed using DNAStar software and the phylogenetic trees were drawn. The result indicated that the E0 and E2 genes of CSFV were amplified from positive samples, the homologies of nucleotide sequence and amino acid sequences of the GX2 strain to E0 gene of reference strains were 83.1% to 94 1% and 85.9% to 99.6%, respectively. The homologies of nucleotide sequence and amino acid sequences of the GX2 strain to E2 gene of reference strains were 81.7% to 93.7% and 89.0% to 97.0%. The E0 and E2 genes of CSFV belonged to group Ⅱ. No variation occurred at E0 protein amino acid sequences of two domains of RNase activity. No change was observed in 15 Cys sites of E2 protein of the GX2 strain. However, the variation of amino acids at sites S734R was observed. The variation trend of GX2 strain was similar to CSFV strains which were isolated in recent years in Guangxi. GX2 strain shared low homology with HCLV and Shimen strains and was not close related to HCLV and Shimen, but shared high homology with GXWZ02.

In order to study the prevalence of yak meat Shiga toxin-producing Escherichia coli (STEC) in Northwestern Sichuan and its stx2 subtype, five suspected colonies were picked from each enrichment culture of 204 collected yak meat samples (25 g). Double PCR assay was used to detect stx1 and stx2, stx2 was subtyped and its CDS region was sequenced. 8 STEC strains were isolated from 204 samples, in which the isolating rate of STEC was 3.9% (8/204). 4 different serotypes O detected were O38(4), O50(1), O74(2) and O150(1), respectively. 6 strains were found carry stx2, and 2 strains were stx2a and 4 strains were stx2c. The results revealed that yak-original isolated strains had a high degree of amino acid homology with human-original and cattle-original strains; Amino acid phylogenetic trees of stx2 A and B subunits showed that yak-original, human-original and cattle-original strains gathered into the same clade, indicating that their genetic distance was relatively near. Yak-original stx2 were distributed in their own little branch, compared with the human-original and cattle-original strains, although yak-original STEC stx2 genetic relations were closer with them, but still had some differences.

To discuss the mechanism how open reading frame 3 (ORF3) protein of genotype Ⅳ hepatitis E virus (HEV) affected the expression of matrix metalloproteinase (MMP) in target cells, Human Matrix Metalloproteinase Antibody Array was used to compare and analyze the expression levels of matrix metalloproteinase in stable cell lines that expressed or did not express HEV ORF3, screening the matrix metalloproteinases which were down-regulated or up-regulated. The results indicated that the expressions of TIMP-1, TIMP-2 and TIMP-4 in stable L02 cell line with expression of pEGFP-ORF3 were down-regulated, and which provided important information for explaining the molecular mechanism through which HEV pORF3 affected the expression of protein associated with matrix metalloproteinase in target cells.

The two dominant epitope sequences of pseudorabies virus (PRV) gB gene were joined together by overlap-extension PCR method with two pairs of primers. The products were inserted into pMD18-T vector. After properly sequenced, the multi-epitope sequence was linked with pET-32a(+) vector, and then transformed into BL21(DE3). By being induced with IPTG, the targeted protein was detected by SDS-PAGE and Western blotting. The results showed the fusion protein was successfully expressed with the size of 54 ku including 34 ku tandem protein and could be combined with monoclonal antibody against gB which showed that the expressed protein had good antigenicity.

This study was aimed to express viral protein genome-linked (VPg) of porcine sapovirus, using the RNA of the CH430 strain of Northwestern China as a template, the VPg gene was amplified by RT-PCR and cloned into pMD19-T Simple Vector, and then recombinant plasmid was confirmed by double-endonuclease digestion and DNA sequencing. VPg gene was subcloned into a prokaryotic expression vector pET-30a to obtain the prokaryotically expressed plasmid pET30a-VPg. The positive recombinant plasmid was transformed into E. coli BL21 (DE3) cells to induce expression. The fusion purified protein was injected into rabbits to produce hyper-immune serum, the specificity and titer of the hyper-immune serum were determined by Western blotting and ELISA. The results indicated that the full-length of VPg gene was 339 bp, encoding 113 amino acids. The expression of recombination protein reached a maximum level when the recombinant bacteria at the condition of 37 ℃, final concentration of IPTG at 1.0mmol/L and the time of induction at 6 h. The purpose protein at size of 22 ku was consistent with expectation and expressed in the form of inclusion bodies analyzed by SDS-PAGE. Western blotting exhibited that expression products had good reactogenicity with the hyper-immune serum. ELISA result showed hyper-immune serum titer was 1:12 800 and had good specificity. The hyper-immune serum laid the foundation for further investigation on the structure and function of non-structural protein VPg of porcine sapovirus.

The study was aimed to select the acid sensitive sites of foot-and-mouth disease virus (FMDV) type O capsid. Using the bovine FMDV type O ON strain as the research object, six acid resistance monoclonal strains were obtained after treatment in pH 6.0acidic environment for serial passages. The P1 region which encoding their capsid proteins of the mutagenic viruses was sequenced and compared with the parental virus, then seven mutation sites were found, of which the mutation sites A1334G(Y445C), A1643G(Q548R), A1822G/A1823C (N608A) of P1 region found in six strains were sense mutation, while the mutation site G1371A was nonsense mutation. In addition the mutation site C1849T(P617S) of P1 region found in m2 was sense mutation and the mutation site G468T of P1 region found in m1 was nonsense mutation. This research laid a good foundation for the reveal of the acid sensitive mechanism and the acid sensitive enhance of FMDV type O capsid.

In this study, the CDS sequence of buffalo BDNF gene was cloned, and then the homology, evolutive tree of BDNF amino acid sequence and the physicochemical characteristics, secondary and tertiary structures of BDNF protein were analyzed and predicted by bioinformatics techniques, also, the expression pattern of BDNF mRNA in different tissues of fetal and adult buffalo was investigated by QRT-PCR. The results showed that with RT-PCR a 800 bp buffalo BDNF gene fragment was cloned and sequenced, including the whole CDS of 753 bp (coded 250 amino acid). The multiple sequence alignment result showed that, the BDNF gene in buffalo shared 99%, 94%, 93%, 90%, 90% and 89% similar nucleotide sequence with that of Bos taurus, Sus scrofa, Canis lupus familiaris, Homo sapiens, Equus caballus and Mus musculus, respectively. The evolutive tree analysis showed that BDNF gene was highly conservative in evolution. The molecular weight and isoelectric point of BDNF protein were predicted as 28 173.36 u and 9.12, respectively. The secondary structure of BDNF protein was composed of several α-helix, β-sheet, T-turn and randon coil structures, and the tertiary structure of BDNF protein was composed of several α-helix and 3 pairs of antiparallel β-sheet structures and so on, which configuring an active center aslo called the NGF functional domain. The QRT-PCR result showed that BDNF mRNA completely expressed in heart, lung, kidney, brain, muscle, ovary and testis in both of fetal and adult buffalo, but the mRNA expression in adult buffalo was higher than that of fetal buffalo.

To investigate the prevalence and genetic diversity of canine distemper virus (CDV) in the main breeding areas of fur animal in China in recent years, the tissue samples of the minks, foxes and raccoon dogs with symptoms of canine distemper (CD) were collected from Shandong, Hebei and Liaoning provinces in the years 2012 to 2014.The signal peptide of fusion protein (Fsp) genes of 15 CDV wild-type strains were cloned and sequenced after the samples were identified to be positive of CD by one step CDV rapid test kit and RT-PCR. The results showed that the identities of nucleotide and amino acid sequences of the 15 CDV wild-type strains were 93.6% to 100.0%, while the identities of nucleotide and amino acid sequences of the isolates and vaccines were 80.7% to 81.7%. The phylogenetic analysis of Fsp genes revealed the 15 CDV wild-type strains were characterized as an Asia-1 genotype that were circulating currently in China. The alignment analysis of amino acid sequences indicated there were obviously diversity at the amino acid sites 3 to 14, 16 to 37, 47 to 67.The sequences analysis of Fsp gene of the currently circulating CDV wild-type strains suggested this Fsp gene region had high divergence and could be used to identify the CDV genotypes that would provide foundation for the prevention and control of CD in the fur animal.

In order to construction porcine bactericidal/permeability-increasing protein (BPI) eukaryotic expression vector and obtain porcine BPI protein recombinant vector. In this study, BPI coding sequence was cloned into vector pUC57 according to GenScript's CloneEZ^® PCR Cloning Kit. After correct identification with digestion and sequencing, the recombinant plasmid pUC57-BPI was transfected to 293-6E cells, then the expression of the recombinant protein was detected by SDS-PAGE and Western blotting. The results showed that we successfully constructed the eukaryotic expression vector pUC57-BPI. Furthermore, porcine BPI recombinant protein was expressed in cultural supernatant of 293-6E cells in eukaryotic expression system, and the establishment of porcine BPI recombinant protein expression system in vitro provided the basis for the deep study of porcine BPI biological function and the preparation of porcine antibacterial protein.

One strain pathogenic bacteria named PMSD was isolated from the infected mink in Liaoning province, and identified as Proteus mirabilis by morphological observation and VITEK 2 Compact System from bioMerieux. Antibiotic susceptibility test of PMSD showed that it was highly sensitive to aminoglycosides and quinolones while resistant to beta-lactam and sulfonamides. The 16S rRNA gene of 1 504 bp was amplified by PCR from PMSD strain, which was deposited in GenBank (accession No. KM229530). BLAST analysis showed that the gene shared more than 99% homology with the sequences of Proteus mirabilis. Furthermore, the phylogenetic tree analysis of 16S rRNA gene indicated that PMSD 16S rRNA gene shared higher homology with other 20 Proteus mirabilis strains available in GenBank. The homologies of nucleotide was from 98.9% to 99.7%. The highest homology was 99.7% with BB2000 strain, HI4320 strain, B1 strain and FCC141 strain. This study provided a foundation for the prevention and control of mink diseases caused by Proteus mirabilis.

The aim of this study was to develop a Real-time PCR method for detection of chicken infectious anemia virus (CIAV). According to the CIAV genome conservative area, a pair of specific primers which could amplify about 180 bp fragment was designed. By building the pGM-T-CIAV recombinant plasmid, and preparing positive standard, the SYBR GreenⅠReal-time PCR standard curve was established. At the same time, sensitivity test, specificity test and repeatability test were determined. The results indicated that CIAV Ct threshold had good linear relationship with the standard concentration. The correlation rate and slope were 0.998 and -3.443, respectively. The Tm was about 86 ℃. No cross-reactions were found between REV, ALV J subtype, MDV and IBDV. The sensibility was 5.33 coppies/μL, which was 1 000 times than the traditional PCR. The coefficient of variation value was less than 3%. The CIAV SYBR Green Ⅰ Real-time PCR detection method could be realized on the early diagnosis of the disease and infection degree quantitative analysis of the test.

A duplex PCR assay for detection of pseudorabies virus (PRV) gE-deleted vaccine and PRV was established using the primers designed based on the characteristics of PRV possessing surface antigen gD and gE virulence genes, and PRV gE-deleted vaccine only owing gD gene but no gE gene. The specificity, sensitivity and repetition tests were conducted, moreover, samples which were taken from clinic suspicious PRV infected pigs had been testified by the established duplex PCR assay. The results showed that a duplex PCR assay for detection of PRV gE-deleted vaccine and PRV was successfully established. The sensitivity and specificity of the duplex PCR assay revealed that the threshold of the duplex PCR was 100 copies/μL of gD or gE gene, and gD/gE gene could be amplified specifically from wild virus or gD gene from PRV gE-deleted vaccine but no products were amplified from the nucleic acid of PK-15 cell or the other 8 kinds of pathogenic viral or bacterial microorganism. The four repetition tests indicated that the duplex PCR was repeatable. Total of 267 gD and gE positive samples and 28 gD positive samples were detected from 835 clinic suspicious PRV infected pigs. Selecting 26 PRV gD and gE positive samples detected by the duplex PCR assay for pathogen cell culture, the coincidence rate between the two methods was 96.1%. The study suggested that the established duplex PCR method was highly specific and sensitive, and was suitable for clinic rapid differential diagnosis of PRV gE-deleted vaccine and PRV.

This study was aimed to express and purify the flagellins, and contribute to research and gain high efficient adjuvants. The fljB, fljB' and fliC genes were amplified from Salmonella Typhimurium genomic DNA by PCR, and the amplified PCR products were cloned into pMD19-T Simple Vector to construct cloned plasmid pMD19-fljB, pMD19-fljB'and pMD19-fljC. The cloned plasmids were digested by double enzymes, and purified genes were subcloned into pET-30a vector to construct prokaryotic expression vector pET30a-fljB, pET30a-fljB'fliC and pET30a-fliC. Transformed colonies were screened by PCR, restriction enzyme analysis and sequencing. Then the positive expression vectors were transformed into E. coli BL21(DE3) strain. After induced by IPTG, the expressed proteins were extracted from E. coli BL21(DE3) strain using Ultrusonic Cell Disrupter System. The fljB, fljB' fliC and fliC were purified by Ni-NTA Columns. The recombinant proteins were further confirmed by Western blotting analysis using anti-flagellin serum from Guinea pig. Through optimizing the expression conditions, flagellins were expressed in dissoluble form. The purified flagellins would lay the foundation for researching their adjuvant properties later.

The study was aimed to express and purify the dairy cattle early pregnancy factor (EPF) protein and prepare EPF polyclonal antibody. The recombinant plasmid pET32a-EPF was expressed in E.coli BL21 (DE3). Polyclonal antibodies were developed by immunizing BALB/c mice with the SDS-PAGE gel extraction purified fusion protein. The recombinant protein and the polyclonal antibody were separately detected by Western blotting and ELISA. The results showed that the recombinant protein with a molecular weight of 37 ku was expressed successfully in E.coli and its content was approximately 48.6% of total bacteria proteins. The purity of target protein could reach 93% after purification. The recombinant protein was recognized by the prepared polyclonal antibody in Western blotting analysis. The antibody titer was 1:12 800.These results indicated that we had successfully obtained and purified the dairy cattle EPF protein, which laid a foundation for the further study of dairy cattle pregnancy diagnosis.

The aim of this study was to identify the most stable gene to be used as reference genes for qRT-PCR analysis in Sika deer antler tissues. The expression patterns of 6 housekeeping genes were analyzed, including glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta-2-microglobulin (B2M), NADH dehydrogenase (NADH), 60S ribosomal protein L40 (RPL40), glutathione peroxidase 7 (GPx) and beta actin (ACTB), in Sika deer antler at different stages (10, 20, 40 and 60 d). The stability of housekeeping gene expression was analyzed with use of 2 software packages, including geNorm and NormFinder. The results showed that GAPDH, ACTB, RPL40 were suitable reference genes for efficient normalization of qRT-PCR data, whereas NADH and GPx were not suitable for analysis of Sika deer antler. These findings were confirmed by comparative profiling of 5 antler genes associated with antler rapid growth (ANXA5, HSP27, PRD2, CRABP1 and LGALS1), while it was observed that the 5 genes were significanly expressed in antler aged 10 d. These results will provide a necessary basis for the further research on rapid growth of antler and ossification genes.

This experiment was to study the effect of cold stress on the immune system and heat stress proteins 70 (Hsp70) in Hu sheep. Using Real-time fluorescence quantitative PCR and ELISA, the expression quantity of Hsp70 mRNA in Hu sheep's liver, lung, spleen, lymph node tissue and concentrations of IL-4, IL-2 in serum were detected respectively before and after cold stress. The results showed that the Hsp70 mRNA expression quantity in liver, lung and spleen tissues of Hu sheep was extremely significantly increased after cold stress compared with before (P<0.01), and the expression of the liver was particularly significant. After cold stress, the IL-2, IL-4 concentrations in Hu sheep serum showed a decreasing trend, especially IL-4 concentration decreased extremely significantly (P<0.01). The results indicated that enhanced expression of Hsp70 mRNA after cold stress in Hu sheep tissues could improve the animal's function of self-protection and enhance resistance to adverse external stimulate, but the decreases in cytokines IL-2 and IL-4 concentrations indicated that cold stress suppressed the body's immune system.

This test mainly studied the effects of supplemental different levels of amino acids on the performance, plasma antioxidant capacity and other metabolic index of Yanqi horse. 11 Yanqi horses were randomly divided into three groups according to performance, body weight, body measurement, age. The groups were referred to as the control group (3 horses), groupⅠ(4 horses) and group Ⅱ(4 horses). The control group was fed basal diet. Horses in the groups Ⅰ and Ⅱ were fed with diet supplemented with different levels of amino acids. All horses were participated in 20 km racing, on the 20th day. Temperature, heart rate, respiratory rate of all horses were measured, 30 min after the racing. Simultaneously, blood samples were collected from the jugular vein. The result showed that the heart rate value of 0 min after the racing, groups Ⅰ and Ⅱ were significantly lower than control group (P<0.05), the heart rate of 20 min after the racing and respiratory rate of 5 min after the racing, groupsⅠ and Ⅱ were highly significantly lower than the control group (P<0.01);The plasma GSH-Px, T-SOD activity and GLU concentration of groups Ⅰ and Ⅱ compared with control group respectively increased by 34.41% (P>0.05), 37.02% (P<0.01), 37.41%(P<0.01) and 32.06%(P>0.05), 35.46%(P<0.01), 26.98%(P<0.05);The plasma MDA, LA concentration of groups Ⅰ and Ⅱ compared with control group respectively decreased by 38.46%(P<0.05), 17.56%(P<0.05) and 38.26%(P<0.05), 13.04%(P>0.05). The results showed that the performance and antioxidant capacity of Yanqi horse could be improved by the basic ration added different levels of amino acids. The physiological function of Yanqi horse could be fast recovery after the racing. The symptoms of fatigue of Yanqi horse were mitigated during the race. Compared, added 0.25% lysine feed supplement was especially effective.

In order to study the effect of defeathering on feather performance, growth performance and serum biochemical indexes of Zhedong White geese. A total of 120 120-day-old Zhedong White geese were randomly divided into control group and experimental group. The control group was defeathered only at the end of the experiment (45 d), while the experimental group were defeathered at the beginning (1 d) and the end of the experiment (45 d), respectively. The trail lasted for 45 days. The results showed that defeathering could promote the growth of feather and improve the feed conversion efficiency. Meanwhile, it could bring stresses syndrome to geese in a short time. There were significant differences in the serum level of TP, ALB, GLB, GLU, Ca and P 4 d after defeathering compared with 1 d before defeathering (P<0.05). There was no significant difference in the serum activity of AST 4 d after defeathering (P>0.05). Serum level of of BUN and serum activity of transpeptidase, ALT and ALP were not significantly changed before and after defeathering (P>0.05). In conclusion, it's feasible to defeather in 120-day Zhengdong White geese. At the same time more attention should be paid to the feeds and managements during the first week after defeathering.

Various unprocessed corn, steam-flaked corn (thickness 1.5 mm, SFC 1.5), and steam-flaked corn (thickness 2.0 mm, SFC 2.0) was used in the present study to compare the chemical compositions, starch gelatinization and ruminal fermentation characteristics in vitro of corn grains as affected by various processing based on chemical analysis, in vitro ruminal fermentation and gas production method. The results showed that the contents of crude protein and ether extract decreased (P<0.05), but the starch gelatinization increased (P<0.05);Gas production rate and starch digestibility of SFC were higher than unprocessed corn (P<0.05), but NH₃-N concentrations were lower (P<0.05);The starch gelatinization of SFC 1.5 (88.76%) increased by 17% than that of SFC 2.0 (71.60%);The gas production rate, the digestibility of dry matter and starch of SFC 1.5 in 4 h significantly increased than that of SFC 2.0 (P<0.05). No differences on 48 h gas production, 12 and 24 h pH, total VFA production, the molar proportion of acetate, propionate, acetate/propionate, isovalerate, isobutyrate and valerate (P>0.05). It was concluded that the various thickness steam-flaked corn grains could increase starch digestibility and degradation rate, slow ruminal ammonia release, but would not affect 48 h gas production and other ruminal fermentation traits.

A series of experiments were conducted to analyze moisture, crude protein, crude fat, crude ash, parts of vitamins and minerals, amino acid and fatty acid composition in the powder of Zophobas morio larvae by methods of the national standards (GB), and to investigate the storage characteristics of the powder. The results showed that the moisture, crude fat, crude protein and ash in the fresh young larvae were 58.13%, 16.68%, 18.71% and 1.52%, as well as 52.65%, 18.03%, 16.84% and 1.89% in that of the old larvae, respectively. The crude fat in the powder of young and old larvae (YOL) was 30.02% and 33.80%. The ratio of the unsaturated fatty acids to that of total fatty acids were 58.12% and 58.68%, containing high amount of oleic (C18:1) and linoleic acid (C18:2). Whereas, the ratios of the saturated fatty acids to that of total fatty acids were 41.88% and 38.12%, containing high amount of palmitic acid (30 63% and 28.80%, respectively). The crude protein in the powder of YOL was 49.12% and 45.70%. They were all comprised of 17 kinds of amino acids (Try not measured), and the contents of the total of amino acids were 42.66 and 39.36 g/100 g. The essential amino acids in YOL were high, accounting for 44.82% and 44.41% of the total amino acids. Moreover, the contents of vitamins and mineral matters were abundant in the powder of YOL, and the ratio of VE, VC and VB₂, as well as K, P, Zn and Fe were high in the vitamins and mineral matters, respectively. Rancidity of the powder of the young larvae was changed quickly under the conditions of the higher temperature, higher relative humidity and higher moisture content, with the storage time prolonged. Therefore, the proper storage conditions of the powder of the young larvae were at temperature below 20 ℃, relative humidity below 60%, with the moisture of the powder controlled less than 10.0%. So the powder of Zophobas morio larvae was developed as a new feed resource used in animal husbandry, with extremely high nutritional values.

This study investigated the effect of Bacillus licheniformis on growth performance, digestive enzymes activity of pancreas, disaccharidase of intestinal mucosa and the number of intestinal main microbial flora of weaned piglets raised in fermentation bed. One hundred and eight 35-day-old weaned piglets with similar average body weight were randomly divided into three groups with three replicates in each group and twelve piglets in each replicate. The weaned piglets in three groups were fed a basal diet (control group, CT), basal diet+40 mg/kg bacitracin zinc+20 mg/kg colistin sulphate (antibiotic group, ABT) and basal diet+300 mg/kg Bacillus licheniformis (probiotics group, PBT). The pre-test period lasted for 7 days and the trial period lasted for 52 days. The results showed as follows:①Compared with the control group, the supplementation of Bacillus licheniformis could increase ADG, ADFI and decrease F/G to some extent, but there were no significant differences in growth performance among all groups (P>0.05). ②Compared with the control group, the supplementation of Bacillus licheniformis extremely significantly increased the activitiy of amylase in pancreas, the activity of maltase in duodenum mucosa and the activity of lactase in jejunum mucosa (P<0.01), and significantly increased the activity of sucrase in duodenum and jejunum mucosa (P<0.05). ③Compared with the control group, the supplementation of Bacillus licheniformis significantly increased the number of Bacillus in caecum (P<0.05), and the number of Lactobacillus was increased by 4.09%, the number of Escherichia coli was decreased by 4.86%, but the differences were not significant (P>0.05). In conclusion, supplementation of 300 mg/kg Bacillus licheniformis could improve the activity of digestive enzymes in pancreas and intestinal mucosa of piglets, increase the number of Bacillus in caecum, and improve the growth performance to some extent.

Steam-flaked technology originated in the United States, the initial application in ruminant production. Steam-flaked technology is a method of grain processing, which grain has been made into flake after a series of processing such as steam hardening and tempering, roller press rolling and cooling. By a variety of effect of physical and chemical in the process of processing, the digestibility of starch and other chemical composition was improved, the contact area of nutrients and digestive tract digestive juices was increased, and the digestive site was changed to a certain extend. Thus, it could increase the animal digestion and absorption rate of grain. As a consequence, the steam-flaked technology could improve feeding value of grain, increase the efficiency of animals used for grain and induce feed waste and environmental pollution. In the article, the technological process of steam-flaked, the influence of steam-flaked on grain nutrients and its application in livestock and poultry production have been reviewed.

The effect of sodium humate with different concentration in the MRS liquid medium on the growth, metabolism and morphology of Lactobacillus plantarum were investigated, to probe the sterilization mechanism of sodium humate. The result showed that in the MRS liquid medium sodium humate (1% to 5%) made the viable count of Lactobacillus plantarum reach the highest point of growth 6 h earlier than control, while it could make the number of Lactobacillus plantarum sightly decreased. Adding 3% sodium humate changed the metabolism of organic acid and made the colony of Lactobacillus plantarum smaller and darker. The result of transmission electron micro-scope showed that the boundaries blur between cell wall and cell membrane. So it followed that in the liquid medium sodium humate inhibited the growth of Lactobacillus plantarum by changing their metabolism and morphology.

In order to select the best dose of certain purified Chinese herbal medicinal compound polysaccharide (cCHMPS), which had been proven to significantly affect the immunity of mice, three hundred one-day-old male Liangfeng Green-feet Ma chickens were randomly divided into six groups, namely control group, levamisole hydrochloride (LM) group, Astragalus polysaccharide (APS) group, high, medium and low-dose purified cCHMPS group, and with 50 chickens for each group. Eight-day-old chickens were vaccined with physiologic saline, LM, APS, high, medium and low-dose purified cCHMPS, once a day for 7 consecutive days. On days 8, 14, 21, 28, 35 and 42 after the first vaccination, 5 chickens' blood for each group were sampled for determination of T, B lymphocyte proliferation activities and NDV antibody levels. The results showed that APS and purified cCHMPS at different doses could significantly or extremely significantly promote the proliferation of peripheral T, B lymphocytes (P<0.05;P<0.01), increase the concentrations of NDV antibody, enhance cellular immunity and humoral immunity, and the medium-dose purified cCHMPS was the best.

The study was aimed to produce recombinant ricin A chain (r-RTA) with high biological activity and compare its toxicity with native ricin (n-RT). We synthesized the published RTA gene sequence on NCBI and cloned it into the pET-28a vector, and then prokaryoticly expressed by the E. coli. After purification with Ni²⁺-NTA resin column, the r-RTA was eluted by 500 mmol/L of imidazole solutions. The purified protein was identified by SDS-PAGE and Western blotting, and the immunogenicity was determined by ELISA. Animal experiments and cell toxicity analysis were conducted to compare the toxicity between the r-RTA and n-RT. The rate of recombinant expression was 31.2%. About 20 mg fusion proteins were obtained from 1 000 mL cultures with the protein purity of ≥90%. The results of ELISA showed that the immunogenicity of r-RTA were about 1.27 times of that of n-RT. The half inhibitory concentration (IC₅₀) of n-RT to RAW264.7 cells was 0.01 μg/mL and the median lethal dose (LD₅₀) to mouse was (3.27±0.44) μg/kg. We used the same dose of toxin to challenge the Raw264.7 cells or mouse, and found that the virulence of n-RT was 2 700 times of that of the r-RTA. The mortality rate of n-RT was 40%, while r-RTA was all survive. Our results suggested that single RTA, without the assistance of RTB, had a significantly reduced toxicity, which provided vital data and theoretical supports for the development of RTA-based vaccine.

The assay was aimed to establish the method to simultaneously determine praziquantel (PZQ) and eprinomectin (EPR) contents of compound praziquantel pour-on preparation by HPLC in single wavelengths. The chromatographic conditions were as follows, C18 column (250 mm×4.6 mm, 5 μm) with a column temperature of 30 ℃, the mobile phase was consisted of acetonitrile-water-trifluoroacetic acid (70:30:0.01) with a flow rate of 1.0 mL/min, and the detection wavelength was set at 245 nm. The results showed that the linear relation of PZQ was good in the linear range between 0.7506 and 7.5063 mg/mL, the linear relation of EPR was good in the linear range between 31.266 and 312.660 μg/mL, related coefficients were both 0.9999, and the average recovery was 99.0% with RSD 1.02% and 98.1% with RSD 0.32%, respectively. The established HPLC method was quick, simple, accurate and reliable, and could be applied to determine the PZQ and EPR contents of compound praziquantel pour-on preparation.

This study was designed to investigate the effect of epicatechin (EC) on mitochondrial DNA (mtDNA) copy number and developmental competence of rat oocytes following maturation and culture in vitro. Cumulus-oocyte complexes were respectively cultured in the maturation medium supplemented with different concentrations of EC (0, 5, 10, 15 and 20 μmol/L) for 16 h. And then the mtDNA copy number in matured oocytes was detected with the method of Real-time PCR. Moreover, the subsequent developmental competence of embryos from parthenogenetic activation (PA) was examined. The PCR result showed that the mtDNA copy number in each EC treatment group were increased than that of control group, and there was remarkably increase in the both groups of 10 and 15 μmol/L compared with the control group (P<0.05), but the number of 10 μmol/L group was more closed to that of the nature ovulation cycle oocytes (P>0.05). Furthermore, the result of in vitro culture showed that adding with 10 μmol/L EC into the maturation medium, there were no significant effect in the maturation rate of oocytes (P>0.05), but the developmental rate of blastocyst after PA was significantly improved than that of control group (P<0.05). The datas above showed that adding 10 μmol/L EC into the maturation medium could increase the mtDNA copy number of oocytes, and contribute to improve its subsequent developmental competence.

To explore the epigenetics mechanism for the slow growth in Xiang pig, 60 piglets in 20 days were divided into two groups with high or low body weight. The methylation modification at cytosine in the genomic DNA from blood and liver was investigated by methylation-sensitive amplifying polymorphism (MSAP) technique. A total of 5 977 sites were counted based on MSAP results. Compared to the low-weight group, the total DNA methylation level in high-weight group was lower. Although the half DNA methylation levels in blood samples were higher than that in liver samples, the both/total DNA methylation levels in blood were lower than that in liver. It showed that the total DNA methylation level tended to decrease as the increasing of body weight. Five specific fragments from A1 to A5 were sequenced which were differently methylated between two groups. Both of A1 and A4 detected from high-weight group while the others got from low-weight group. Fragment A1 and A2 detected from blood genomic DNA and A3 to A5 from liver samples. A3 was apart far from two genes without coding region for RNA. The others were located in genes related with metabolism and immune. The results indicated that genomic DNA methylation level was different among individual piglet and related with the body weight of Xiang piglet in 20 days. The four specific methylation fragments would be useful for research on the mechanism of growth in Xiang pig breed.

In this study, the genetic polymorphism of 5'-flank region of follicle-stimulating hormone receptor (FSHR) gene in yaks and cattle-yak were detected in order to offer the materials and theoretical foundation for solving the problem of lower yak's reproduction rate and screening the molecular markers for reproductive traits. The polymorphism of the partial 5'-flank region of the FSHR gene in 110 individuals from Maiwa yaks, Jiulong yaks, Datong yaks and cattle-yak was analyzed by PCR-SSCP. The parameters of alleles frequency, genotype frequency, linkage equilibrium of Hardy-Weinberg, heterozygosity (H), effective numbers of alleles (Ne) and polymorphism information content (PIC) were calculated by statistical methods. The results showed that the 5'-flank region of FSHR gene in Maiwa yak, Datong yak and Jiulong yak had polymorphism, but no polymorphism found in cattle-yak. There were three genotypes (AA, AB and BB) in Maiwa yak, two genotypes (AA and AB) in Jiulong yak and Datong yak, AB genotype was favorable genotype in three yak breeds, and A allele was favorable allele. The polymorphism information content were 0.3693, 0.3565 and 0.3705 in Maiwa yak, Jiulong yak and Datong yak, respectively, which reached a moderate polymorphism (0.25<PIC<0.5). It indicated that there was greater genetic variation in all yak breeds.

In order to construct the assessment model of general disease resistance in Yorkshire, eight important cytokines levels (IL-1β, IL-4, IL-6, IL-8, IL-10, TGF-β, TNF-α and IFN-γ) were detected in 30-day-age Yorkshire piglets, respectively, and then the principal component analysis was carried out on the immune traits. The results showed that fore 5 character values were selected as the principal components (occupied 87.721% of accumulative contribution rate), and these five components contained nearly all the original information. The combined scores of diarrheal and health weaning piglets in 30-day-age were compared by the assessment model in different genotype population, and the results showed that the combined score of weaning piglets of health group was 0.080, and weaning piglets of diarrheal group was -0.009.This study initially identified the main components which affected the general disease resistance of Yorkshire, and made the preliminary verification of it. The result provided some guidance and theoretical basis for the early disease resistance breeding of Yorkshire in the future.

During the last decade of the 20th century, after high-speed development, decelerating of the rate of goose production of China. Apart from the limit of land use in animal husbandry, the harm of animal epidemics and the rising cost of manpower, the main reason is the lack of technology innovation and breakthrough. Goose artificial insemination technology(AI) is an advanced breeding technology. Improvement and application of this technology, not only can increase the gander breeding efficiency, but also can speed up the genetic progress of goose breeding. If the semen cryopreservation success, goose artificial insemination can also be used for conservation breeding and overcome the limits of time and space. Goose breeding backwardness and low reproductive rate is the bottleneck restricting the development of goose industry, the application of artificial insemination technique will push to great action of the goose industry development. Thus, this paper review the AI techniques developed over the years, including semen collection, ejaculate quality, semen storage and cryopreservation, and artificial insemination in goose.

Age at the first egg (AFE) remained to be an important economic implication because the earlier onset of sexual maturity meaned the more number of egg production. There were many articles concerning the genetic basis on AFE, based on inheritance, heritability and genetic markers with sibling design, GWAS and QTL mapping. However, few review summarized the previous study. Most studies have concluded that medium heritability, ranged between 0.24 and 0.55, had exists for effective selection on AFE. High CV in AFE meaned uniformity would be selected in the future. It had been reported that polymorphisms of candidate genes, such as genes involved in hypothalamus-pituitary-ovary axis regulation, were associated with AFE. However, many of these results had needed further confirmations. Based on the results from genome-wide linkage analyses on different subject populations, 11 loci located on 9 chromosomes were reported to harbor AFE genes in chicken. It's expected that the dissections in genetic architecture of AFE would provide a basis on selection and genetic improvement on local breed.

Large White pig is a kind of excellent local pig breed, with the expansion of large scale of raising pigs, some strains decline sharply, even on the verge of extinction. This paper reviews the current situation and characteristics of‘Plum pig’, puts forward the importance of the protection of high-quality local livestock taking Plum pig as an example, focuses on the research progress on the present preservation method of pig genetic resources, the development and application of pig somatic cell cloning technology, the exploration of operation, we also discusses the direction of the development and utilization of Plum pig.

This research was aimed to investigate the Chinese material medica serum of Holly Bark combined with antimicrobials to induce in vitro antibacterial activity of bacteria. The Chinese material medica serum of Holly Bark was prepared. The minimal inhibition concentration (MIC) of antimicrobials were tested through twice micro-dilution method. Extended spectrum β-lactamases (ESBLs)-producing bacteria were induced and subcultured by the Chinese material medica serum of Holly Bark combined with antimicrobials. The Chinese material medica serum of Holly Bark combined with antimicrobials to induce bacteria passage for drug-resistant E. coli had antibacterial activity to a different extent. The effects of β-lactam antibiotics, aminoglycosides, quinolones and kaba oxygen against drug-resistant E. coli were obviously strengthened when combined with the Chinese material medica serum of Holly Bark in vitro.

Cysticercus pisiformis which is larval of Taenia pisiformis, infects the lagomorpha animals (eg. rabbit). As one of common parasite diseases in rabbits, cysticercosis is caused by Cysticercus pisiformis without obvious clinical symptoms and does not get enough attention by farmers. Thus, cysticercosis is treated by lack of standardization in rabbit, and negatively impacts on rabbit production. According to research data in recent years, this paper deals with the morphlogy, molecular biology, epidemiology, pathogenicity, diagnosis and preventive measures of cysticercosis in rabbit, which will provide a foundation for control of rabbit cysticercosis.

In order to obtain the lowest irradiation dose on effectively killing Ascosphaera apis, our study took Ascosphaera apis as the research object, and explored the killing effect of different irradiation dose on the Ascosphaera apis. Also, we hoped to provide a reference basis for Co60γ irradiating bee feed (especially for bee pollen). During the study, we had used bacteria-purification technology to obtaine Ascosphaera apis from cretaceous disease (bee chalkbrood diseases) larvae, and combined with morphology, lactic acid phenol medan dyeing and 5.8S rDNA sequence analysis technology to identificate this Ascosphaera apis. In addition, we preparated different levels of Ascosphaera apis spore suspension, and added them to 3 d larvae feed to test the half lethal dose (LC₅₀). Behind, we added LC₅₀ level of spore suspension into larvae diet, and then adopted different irradiation dose irradiation diet, finally fed larvae with that treated diets. The results showed that the isolated and purified fungal pathogen, which got from bee chalkbrood disease larvae, were bee balloon bacteria, identified by morphological and molecular biology. Furthermore, under the condition of indoor artificial breeding, the LC₅₀ of bacteria Ascosphaera apis on larvae was 9.5×10⁴/mL, and the effectively killing dose was 7.0 kGy. There was significant difference between larvae prevalence and the irradiation group (P<0.05). From this we could be sure that the minimum effective radiation dose of bee feed, which was added LC₅₀ balloon fungus spores was 7.0 kGy.

Foot-and-mouth disease (FMD) is an acute, febrile, highly contagious animal disease caused by foot-and-mouth disease virus (FMDV). FMDV has a variety of mechanisms resisting host innate immune response, leader protease (L^pro) has played a key role in the process. L^pro can cut off the host cell cap-dependent translation, inhibiting the synthesis of interferon. L^pro through destroying the integrity of NF-κB or reducing the expression of IRF3/7, inhibits the production of interferons mRNA. Besides, L^pro can take part in the deubiquitination of reninoic-acid-inducible gene Ⅰ (RIG-Ⅰ), TANK binding kinase 1 (TBK1), TNF receptor-associate factor 3 (TRAF3) and TRAF6, thus affecting the activation of type Ⅰ interferon signaling pathway.

In this study, we compared the effects of Fuzheng Jiedu super micropowder (FZSM) and Fuzheng Jiedu san (FZ) on anti-IBDV in vitro. Method of chicken embryo, 10-day-old SPF chicken embryos were inoculated with FZSM and FZ in three safe concentrations by two pathways of adding decoction before inoculation of virus and adding decoction after inoculation of virus, and then the effects of two decoctions on ELD₅₀ of IBDV to 10-day-old SPF chicken embryos were measured. Method of cell, CEF was inoculated with FZSM and FZ in three safe concentrations by two pathways of adding decoction before inoculation of virus and adding decoction after inoculation of virus, and then the effects of two decoctions on CPE of IBDV to CEF were observed. The results showed that the virus titer and inhibition rate of CPE of IBDV were reduced by FZSM and FZ, the effect of high concentration was better than its low concentration, at the same concentration, the anti-IBDV effect of FZSM was superior to that of FZ, and preventive effect was better than therapeutic action. These results indicated that FZSM and FZ had the function of anti-IBDV in vitro and the effect of FZSM was superior to FZ.