Buffalo CTCF gene was cloned in the present study, the CTCF sequence was systemically analysised by bioinformatics techniques, and the expression patterns of CTCF in different tissues were also assayed with Real-time fluorescence quantitative-PCR(QRT-PCR). The results showed that, with RT-PCR a 2239 bp buffalo CTCF gene fragment was cloned and sequenced, including the whole CDS of 2184 bp (coded 727 amino acids). The molecular weight and isoelectric point of buffalo CTCF protein were predicted as 82.7 ku and 6.57 respectively. The sequence multialigned results showed that buffalo CTCF gene shared 99%, 96%, 96%, 94% and 92% of similar nucleotide sequence with Bos taurus, Equus caballus, Sus scrofa, Homo sapiens and Mus musclus,respectively. It was predicted that buffalo CTCF protein contained eleven ZnF_C2H2 domains, which meaned that it was a DNA binding protein. In addition, we also analyzed the tissue expression of CTCF gene through QRT-PCR. The results showed the mRNA of CTCF gene existed in all the six detected buffalo tissues with the most abundant expression in liver, followed by brain, muscle, spleen and ovary, the minimal expression in skin was observed.

To obtain recombinant ccpA of Streptococcus suis type 2 expressed in prokaryotic system and preliminaryly evaluate its antigenicity and specificity by Western blotting.ccpA gene was obtained by PCR and cloned into pET-28a vector,then transformed into Escherichia coli BL21 to induce,express and purify,the antigenicity of recombinant ccpA was identified by Western blotting. The double enzyme analysis and sequencing proved that recombinant plasmid pET-28a-ccpA was constructed correctly.The expressed recombinant protein,with a relative molecular mass of about 38000,was recognized by positive serum antibody.The ccpA of Streptococcus suis type 2 was successfully expressed in E.coli,which provided a material for development of serological diagnostic method for Streptococcus suis type 2.

In this study,we identified the metabolites of cyadox in vivo produced by chicken using ultra performance liquid chromatography/quadruple time of flight tandem mass spectrometry(UPLC-Q-TOF MS).Besides,it had been viewed that proposed metabolic pathways of cyadox in vivo of chicken. Animals were administered a single of cyadox (800 mg/kg) by oral gavage.We used UPLC-Q-TOF MS to analyze chicken feces, collected before and after the experiment, which were simple pretreatment. A total of fifteen metabolites were identified after the collected data processed by the software of Metabolynx^xs . This experimental results showed that cyadox occurred widely metabolism in vivo of chicken.

The male Small Tail Han sheep were inoculated with Brucella suis S2 vaccine strain, the suppression subtractive hybridization (SSH) library was constructed from buffy coat of sheep. After the recombinant plasmids were sequenced, the cDNA sequence of the conserved domain of lysozyme (LYZ) was recognized based on BLAST in GenBank, with the size of 770 bp, and continuously encoding 148 amino acid residuals, the encoded protein was a natural anti-infective substances.The sequence had been submitted to GenBank(JX263305).Quantitative Real-time PCR result showed that the LYZ gene was slightly up-regulated in buffy coat at 14 and 30 days post-challenge (dpc) without statistical significance (P>0.05), and then recovered to the normal level at 40 dpc.The lysozyme was further proved as an essential natural immune factor which laid the foundation for the effective prevention and control of brucellosis.

Signal transduction molecules AKT-1 and mTOR were essential for milk protein gene transcription and translation in mammary epithelial cells.Insulin,dexamethasone and prolactin composed of prolactin complex (DIP), the variety of lactation signal pathway activation was a necessary condition,including AKT-1/mTOR. In this study,the qRT-RCR and Western blotting were used to test above-mentioned two genes's expression level of mRNA and protein in different time points.The results showed that IGF-Ⅰ promoted significantly the expression levels of mRNA and protein for AKT-1 gene in the basis of DIP compared to the absence of IGF-Ⅰ(P<0.05), and it also promoted significantly the expression of mTOR gene in mRNA and protein levels(P<0.05).

The research was aimed to carry out the bioinformatics analysis on the coding region (CDS) of BPI gene among different species. By using the bioinformatics method, the genetic diversities of coding regions of BPI gene from the Homo sapiens, Mus musculus, Rattus norvegicus, Bos Taurus, Nomascus leucogenys, Gallus gallus, Oryctolagus cuniculus, Sus scrofa, Macaca mulatta, Equus caballus, Xenopus laevis, Pongo abelii and Canine were analyzed. The amino acid sequence, composition, signal peptide, hydrophobicity/hydrophilicity, trans-membrane structural domain, secondary structure and conservative structural domain of BPI protein were predicted and deduced. In 29 CDS sequences of BPI gene from 13 species, 532 polymorphic sites were detected and 15 haplotypes were generated. The coding region of BPI gene had the rich genetic diversity within and among species.BPI gene had strong codon bias.The theoretical isoelectric point of BPI protein was higher than 7. The protein was alkaline,and N terminal almost had the signal peptide. The peptide chain presented the hydrophilicity. There was a trans-membrane structural domain.The main secondary structure of BPI protein were α helix,β folding and random coil.There were two conservative structural domains named BPI1 and BPI2.

In this study, the bovine viral diarrhea virus (BVDV) strain Yak was used as material to sequence the complete genome.Nineteen pairs of primers were designed according to the complete genome sequence of BVDV-Ⅰ virus, the whole genome of BVDV strain Yak was cloned and sequenced. Sequence analysis showed that the genome was 12214 nucleotides (nt) in length, which contained a 288 nt 5'-untranslated region (UTR) and a 232 nt 3'-UTR. The complete genomic sequence of BVDV strain Yak had already submitted into GenBank (GenBank accession NO: JQ799141). Homologous analysis showed that homology of the whole genome sequence was low between BVDV strain Yak whole genome sequence and other 8 strains of BVDV whole genome sequences. Evolutionary tree results showed that the genetic evolution relationship was farther between BVDV strain Yak and other 8 strains of BVDV.

A total of 80 7-week-old male BALB/c mice were randomly divided into four groups (Ⅰ,Ⅱ,Ⅲ and Ⅳ groups),and 20 mice in each group. Each group was fed with the diet containing 0.045,0.1,0.4,and 0.8 mg/kg Se,respectively. The results showed that the content Se of Ⅰ group in the liver reduced significantly (P<0.05). Glutathione peroxidase 1 (GPx1)mRNA levels of Ⅰ group was significant lower than Ⅱ and Ⅲ groups (P<0.05),while Ⅱ and Ⅲ groups were significant differences in the liver (P<0.05),and no significant differences in testis (P>0.05). Glutathione peroxidase 4 (GPx4) mRNA level of Ⅰand Ⅲ groups in liver had no significant difference (P>0.05),but were significant lower than group Ⅱ (P<0.05),while each group had no difference in the testis (P>0.05). Relative mRNA expression of thioredoxin reductase 2 (TrxR2) in liver and testis improved significantly (P<0.05) with the increasing of dietary Se supplemention. In conclusion,in liver and testis,deficient Se dietary could significantly reduce selenium deposition and relative mRNA expression of GPx1,GPx4 and TrxR2,however,excessive Se dietary could significantly reduce GPx1 mRNA levels (P<0.05) and significantly increase TrxR2 mRNA levels (P<0.05),it showed that there was difference for different organs.

This study was aimed to analyze antigenicity of E2 protein of classical swine fever virus (CSFV). The E2 was amplified by RT-PCR from the genomic RNA of CSFV C-strain and cloned into expression vector pET-30a-c(+) to obtain recombinant pET30a-E2. The truncated E2 protein was expressed with high level in transformed Rosetta (DE3) after induction with IPTG. The results of SDS-PAGE and Western blotting indicated that the gene cloned in downstream of pET30a-E2 was expressed in high level and the recombinant fusion protein had immunologically reactive.

To investigate the characteristics of novel duck flavivirus (DFV) epidemic strain in Guangxi province, a virus was isolated from sick laying berry valley ducks and designated as LG/01/11. Then, full length of viral envelope protein E gene was cloned and sequenced. Results showed that E gene of LG/01/11 was comprised of 1503 nucleotides, encoding 500 amino acids, and shared 99.3% to 99.6% nucleotide sequence homology with other duck flaviviruses isolated in China since 2010. No genetic deletion and insertion, but several scatter nucleotide substitutions were found in E gene of LG/01/11.

The aim of this paper was to amplify bovine Myf5 gene, and analyzed the protein biological characteristics. A pair of primers were designed according to bovine Myf5 gene sequences published in GenBank.Myf5 gene was amplified by RT-PCR from bovine back waist longest muscle, then linked to PUCm-T vector, restriction digested and sequenced. At the same time, we predicted the basic properties, the secondary structure and phosphorylated sites of Myf5 protein by the online tools. Having amplified coding areas sequence was 768 bp in length, encoded 255 amino acids. Protein biological characteristics analysis showed that Myf5 had the family genes typical basic helix-loop-helix domain. Serine(Ser) content was the highest in amino acid composition, and the phosphorylation site located in serine(Ser),threonine(Thr) and tyrosine(Tyr) residue.

The research aimed to carry out the cloning, identification and sequence analysis of genomic DNA of carp interferon γ-2β (IFNγ-2β). On the basis of the full-length cDNA of carp, we designed two pairs of primers in its bilateral untranslated regions and the exons to perform PCR that used the carp spleen genomic DNA as the template, the IFNγ-2β full-length genomic DNA sequence was obtained. The full-length genomic DNA was 2084 bp, the GenBank accession number was JX181981, the gene sequence analysis showed the evolution of conservative and contained 4 exons and 3 introns were like most of the other species, the number of bases of each exon closed to the mammalian exons correspondingly. The exon and intron splice sites were complied with GT-AG rules.

Based on the sequence of Torque-Teno canis virus registered on NCBI (Cf-TTV10, accession No.AB076002), we designed primers for nested PCR to detect the Torque-Teno canis virus in serum and fecal samples. It was showed that there were 14.6% Torque-Teno canis virus positive canine serum samples and 12.7% positive fecal samples, and it was verified that the gene type was same. There was no significant difference in different extraction samples(P>0.05). This research provided the basis for the further study of TTV characteristics and molecular epidemiology.

Yeast (Saccharomyces cerevisiae) CUP1 encodes a cysteine-rich protein with 61 amino acids in length and 6165 u in molecular weight. Yeast CUP1 is a copper metallothionein responsible for resistance to copper. Yeast is induced by copper, and the CUP1 expression is greatly enhanced at the mRNA lever driven by the strong combination of the CUP1 promoter and the factor ACE1.This article summarizes the progress on the yeast CUP1 gene. It mainly describes the structure, the role of binding metal-ion, and the analysis on the promoter of CUP1 gene. Also, it describes the application of the CUP1 gene, especially for the CUP1 promoter, for the transgenic organisms. It plays an essential role on the further research of the physiological mechanism of CUP1 gene, and provides theoretical assistance for the transgenic organisms, especially for the transgenic animals and plants.

Duck virus hepatitis (DVH) was one of the most important diseases to cause economic losses in the duck industry. The test confirmed the virus of dead duck for duck virus hepatitis type Ⅰby viral isolation, duckling regression test, the DHV ELD₅₀ determination, ducklings protection test, neutralization test in chicken embryos, innoculation test in duck embryo and electron microscopy observation and so on. There were some differences in pathological features of regression experiments of duck hepatitis virus type Ⅰ and standard virus that the determination and analysis of gene sequences needed to be further studied.

Rumen anaerobic fungi is a special flora besides in the rumen plays an important role,attracting more attention due to the application value of the secretion of high activity cellulase. Because of the limitation of the traditional culture,the use of molecular biological techniques to do research is in the ascendant. In this paper,the characteristics and diversities of anaerobic fungi,anaerobic fungal degradation of structural carbohydrates,starch and protein,and the ecological environment in the rumen had been analysed. Meanwhile,the molecular biology methods of ITS,DGGE,RFLP,RT-PCR and metagenomics were discussed,we expected to provide a reference for the anaerobic fungal research.

According to the published IBRV genome sequence, we designed and synthetized one pair of specific primers, amplied about 846 bp gD gene fragment. The fragment was cloned into prokaryotic expression vector pET-30a, after enzyme digestion and DNA sequencing were correct, the recomninated vector was transformed into BL21(DE3) bacteria and induced by IPTG, recombinant protein was expressed in partially soluble form. After purification of recombinant protein, Western blotting proved that it had good antigenicity and specificity. The present study using prokaryotic expression system had good antigenicity of gD protein, which laid a material foundation for IBRV further research and development of diagnostic reagents.

A pair of specific primers was designed according to the sequence of porcine cytomegalovirus (PCMV) gB gene(FJ870563.1) in GenBank. The gB gene was amplified from the final diagnosed PCMV samples by PCR,the amplified fragments was cloned into the pMD19-T vectors,and the positive recombinant T-PCMV-gB was sequenced. The results showed that the complete gene was consisted of 2580 bp.The results of homology analysis showed that gB gene shared 98.0% to 99.6% homology with reference gB sequence, and 42.9% to 43.6%,40.5% to 40.6% of amino acid homology with HHV-6 and HHV-7 respectively. The effects of isolates owing to these amino acid substitutions on biological characterization were unknown.

This paper was to research the effect of different proportions of fermented cottonseed meal included in diets on small intestinal mucosa morphology, T lymphocyte cell subset and intestinal microflora in Yellow chicken. Selected 14-day-old healthy broiler 320, were randomly divided into 4 groups, each group four replicates of 20 chickens per replicate. Ⅰ Group was the control group fed with the basal diet, Ⅱ, Ⅲ and Ⅳ groups were added 3%, 6% and 9% fermented cottonseed meal in diets,respectively. The aim was to study the effect of different content of fermented cottonseed meal on the early (14 to 28 days), medium (29 to 45 days), and later (46 to 65 days) periods on small intestinal mucosa morphology, T lymphocyte cell subset of medium and later periods and intestinal microflora of later periods in broilers.The results showed that the early, medium, the height of villus of duodenal of the wholes periods in Ⅲ group were significantly increased respectively by 31.90%,25.17% and 49.41% (P<0.01). The depth of crypt of duodenal in Ⅲ group were significantly reduced by 9.42%,11.31% and 10.83% (P<0.01).The V/C of duodenal in Ⅲ group were significantly increased respectively by 45.65%,41.19% and 67.84% (P<0.01). The height of villus of jejunum of the wholes periods in Ⅲ group were significantly increased respectively by 28.72%,28.38% and 27.61% (P<0.01). The depth of crypt of jejunum in Ⅲ group were significantly reduced by 6.65%(P<0.05),6.18% (P<0.01), 7.37%(P<0.01). The V/C of jejunum in Ⅲ group were significantly increased respectively by 37.84%,36.69% and 37.82% (P<0.01).The height of villus of ileum of the wholes periods in Ⅲ group were significantly increased respectively by 76.64%,40.36% and 67.20% (P<0.01). The depth of crypt of ileum in Ⅲ group were significantly reduced by 4.09%,6.07% and 6.02% (P<0.05).The V/C of ileum in Ⅲ group were significantly increased respectively by 83.79%,49.27% and 78.26% (P<0.01).The CD4⁺/CD8⁺ of broilers in Ⅳ group was significantly increased respectively by 42.86% and 34.95% (P<0.05) in the medium and later periods. With the increasing of the amount of fermentation of cottonseed meal, the number of broilers of the duodenum, jejunum, ileum of lactic acid bacteria increased gradually, excepte the difference of lactic acid bacteria of duodenum and jejunum of Ⅱ group was not significant, the number of the intestines of lactic acid bacteria of rest group increased gradually significantly. The number of the Escherichia coli and Salmonella reduced gradually, excepte the difference of the duodenal segment of Escherichia coli of Ⅱ group and the different of lactic acid bacteria of the duodenum, jejunum, ileum of the intestines. In a word, 6% fermented cottonseed meal in improving the villus heigh of the duodenum, jejunum, ileum, reducing its crypt depth, and improving the V/C was better than 3%, 9% fermented cottonseed meal group, while 9% fermentation cottonseed meal group in improving the CD4⁺/CD8⁺ and the number of lactic acid bacteria and reducing the number of the Escherichia coli and Ssalmonella was better than 3% and 6% fermented cottonseed meal group.

To explore the effect of two kinds of nutrition additives on fattening and slaughter performance of Holstein bulls, 60 Holstein bulls were selected with similar body weight and month-old, and were randomly divided into 3 groups. The control group was fed the basal diet, the groups 1 and 2 added 100 g/d additives Ⅰ and Ⅱ in the basal diet. The adjustment period lasted for 14 days,and the experimental period lasted for 31 days.The results showed that the average daily gain in the groups 1 and 2 increased 0.32 and 0.37 kg (P<0.05) compared with the control group. Compared with the control group, carcass meat percentage, meat-bone ratio, thirteen cut meat weight of the group 2 showed significant differences (P<0.05),and dressing percentage, meat percentage and percentage of thirteen cut meat to meat weight in the groups 1 and 2 were not significant differences (P>0.05), but respectively increased by 2.96%,3.05%,1.47% and 1.89%, 2.82%, 2.39%. Compared with the control group, the profit of the groups 1 and 2 increased 1233.80 and 1370.20 yuan in the experimental period. The results suggested that fattening effect of the two kinds of nutrition additives on Holstein bulls were better and could produce better economic benefits.

By the comparative study on the beef quality of growing Holstein dairy bull,Simmental cattle, Xinjiang Brown calf and Xinjiang local cattle,the assay was aimed to explore the internal beef quality of the dairy bull. Four different breeds of 18 month age cattle were slaughter in the same nutrition model. Carcass were taken back up the right muscle as meat sample. Edible quality,normal composition of nutrition and amino acids were detected and analysed. The results showed that marbling, beef color, water losing rate,water retention capacity and cooking percentage of Holstein dairy bull were better than Xinjiang Brown calf and Xinjiang local cattle, but weaker than Simmental cattle. The tenderness of Holstein dairy bull was better than Xinjiang local cattle. The contents of crude protein and crude ash were 20.14% and 1.11%,there were no significant differences during each breeds (P>0.05). The content of dry matter was 26.30%, it was significantly higher than Xinjiang Brown calf and Xinjiang local cattle (P<0.05), lower than Simmental cattle (P>0.05). The content of crude fat was 10.04%, which was significantly higher than other breeds (P<0.05). Moreover,Holstein dairy bull contained every amino acids which were needed by human body. Among them,the contents of methionine,glutamate,glycine,histidine and taurine were rich, and the proportion of amino acid was also well.

To study the effect of a kind of new acidifier on the growth performance and gut health of broiler chickens, 126 1-day-old healthy male Abor Acre broiler chickens were allotted to two groups, the control group was fed with the basal diet, and trial group was fed the supplementation of acidifier in the basal diet. The whole trial period was 49 days, and the early, medium term, and laster stage were 1 to 21, 22 to 42 and 43 to 49 days. The results showed that during 1 to 21 days, the addition of acidifier in diet had the trend to decrease feed intake, increase body weight gain and the body weight at 21 day, and to decrease the feed conversion ratio. No significant difference was found for the broiler growth performance during 22 to 49 days. The addition of acidifier in diet also had the trend to rise the volatile fatty acid and reduce the total volatile basic nitrogen content in ileal chime at day 49. It was concluded that acidifier could improve the gut health of broilers and increase the growth performance, and 1 to 21 days had a better feeding effect than during 22 to 49 days.

In present experiment,meat quality was analysed for randomly selected 120 day-old 10 chickens(♀:5,♂:5) from the same grazing conditions of Sizhou chicken and to provide scientific information for its exploitation. The results indicated that Sizhou chicken had excellent muscle tenderness,high dry matter,rich calcium,lower selenium and iron,moderate zinc. Oleic acid,linoleic acid,palmitic acid and stearic acid accounted for 92.20% in all of 11 fatty acids in chest muscle. Unsaturated fatty acids and essential fatty acids was 69.35% and 15.31%,respectively. Glutamic acid,aspartic acid,lysine and leucine accounted for 44.22% in all amino acids. Essential amino acids,palatable amino acids,sweet amino acids,aromatic amino acids,branched chain amino acids and antioxidant amino acid contents were 39.40%,26.18%,20.09%,7.06%,18.08% and 11.61%,respectively. Therefore,Sizhou chicken had excellent meat quality and good flavour.

It was found that trimeric autotransporter adhesin (TAA) was the important virulence factor for Actinobacillus pleuropneumoniae(APP) adhere to host in the recent years. In this research, we compared the culture characteristic and the pathogenicity to piglet of △adh and 5b wild strain. The results showed, the growth rate of △adh was higher than 5b obviously; the △adh showed weaker aggregation compared with 5b in liquid culture condition; the piglet showed typical symptoms after the infection, the observation of HE showed pathogenicity of the △adh was weaker than 5b wild strain. The result of the comparisons conformed the importance of adh in the progress of APP attacking to host and set basics in the pathogenic research of APP for the next step.

To investigate the effect of porcine circovirus type2 (PCV2) infection on immune function of porcine alveolar macrophages (PAM), forty-day-old healthy piglets infected with PCV2 were used to detect the kinetics of pattern recognition receptors(PRR) mRNA of PAM. Results showed that the trends of TRL2, TRL4 and CD16 mRNA were similar. Compared to the control, these molecules from the infected groups increased at 3 and 7 d, and decreased at 14 d, with significant differences (P<0.01) of TLR4 and CD16 at 3d and a significant difference (P<0.05) of TLR2 at 14 d. The transcription of CD18 mRNA lowered with significant difference PRR (P<0.05) at 3 d, and went up with significant difference (P<0.05) at 7 d. All these suggested that the recognition and phagocytosis functions of PAM on foreign object post-infected with PCV2 and thereafter its signal transduction function could be disorders, which influence the abilities of phagocytosis and clear microorganisms of PAM.

The assay was aimed to explore the early cytokine profiles in hepatic mononuclear cell from mice infected with Fasciola hepatica. Mice were classed into 2 groups, infected and non-infected. In each group, mice were treated or untreated with IL-4R α antibodies, respectively. Hepatic mononuclear cells were harvested and cultured. The supernatant was used to measure the cytokine levels and the livers were used to measure mRNA of IL-4 and IL-12P40 by Real-time PCR. Results demonstrated that the profiles of IL-4 and IL-5 were similar. Both were quite low without infection of F.hepatica whatever the antibodies were treated or not. However, when mice were infected, both cytokines in mice without antibody were significantly higher than mice treated with antibody (P<0.05). The IL-13 showed different profile to IL-4 with the high level in mice treated with antibody. IFN-γ increased when mice were infected, but no differences were found when mice were treated or untreated with antibody (P>0.05). The mRNA level of IL-4 in mice treated with antibody but uninfected with F.hepatica was significantly lower than wild type mice (P<0.05). However, it was extremly significantly higher in mice infected but untreated with antibody than mice that infected and treated with antibody (P<0.01). mRNA level of IL-12P40 showed no differences between groups when mice were infected with F.hepatica. Our data showed that IL-4Rα was crucial to the expressions of IL-4 and IL-5, but not to the IL-13 and IFN-γ during the acute phase of F.hepatica infection.

The aim of the study was to detect the distribution of FecTT mutation of growth differentiation factor 9(GDF9) gene in Jining Grey, Guizhou White, Dazu Black, Shannan White, Gulin Ma, Matou, Boer, Chengdu Ma, Saanen Dairy, Wendeng Dairy, Banjiao, Jintang Black, Guanzhong Dairy, Chuandong White, Angora, Liaoning Cashmere and Inner Mongolia Cashmere goats by PCR-RFLP. The results indicated that the FecTT mutation of GDF9 gene was not detected in seventeen goat breeds tested. The FecTT mutation of GDF9 gene that had significant effect on fecundity of Thoka sheep might be have no significant effect on fecundity of the above seventeen goat breeds.

In order to understand the endocrine regulation process in a morphological view and reveal luteotropic hormone(LH) mechanism action in the female mammalian ovary, the distribution and localization of luteotropic hormone receptors(LHR) in ovaries of Duolang sheep and Karakul sheep were determined by immunohistochemical SP method. The results showed that 5 positive continuous sections were selected respectively. LHR-immunoreactive substance was mainly distributed over granulose cells, theca cells, oocyte, and ovary blood vessel surrounding interstitium. LHR was mainly loaded by the granulose cells and theca cells. Receptors regulation was showed during the earlier follicle period. With the development of follicles, the quantities and the staining intensity of the LHR-immunoreactive cells presented an increasing trend.

In order to explore the expression of prostaglandin E₂ and F_2α synthase (PGES and PGFS) mRNA in bovine oviductal epithelial cells by estrogen,investigation the function of physiology in bovine. We used bovine regenerating epithelial cells in vitro training system,different times(0,2,4,8,16,24 and 48 h)and different concentrations of estrogen (0,10^-12,10^-11,10^-10 and 10^-9 mol/L) were added, and measured PGES and PGFS mRNA expression by using Real-time PCR. The results showed that different concentrations of E₂ or different stimulation time in the same concentration of E₂ could increase the expression of prostaglandin E₂ synthase (PGES), but in addition of E₂ 4 h and the concentration was 10^-10 mol/L compared with the control was extremly significant different (P<0.01), while PGFS in addition of E₂ 24 h and the concentration was 10^-12 mol/L compared with the control was extremly significant different (P<0.01).Experimental results showed that estrogen could promote the expression of PGES and PGFS mRNA in oviduct epithelial cells of bovine, and regulate the mRNA expression of prostaglandin synthase PGES and PGFS.

The physicochemical properties and proliferative characteristics in BHK-21 cells of Japanese encephalitis virus(JEV) LS strain were studied in the test. The results indicated that JEV LS strain was sensitive for temperature (56 ℃), acid(below pH 5.0), diethyl ether and trypsin. The viral titer of LS strain remained at the same level undergoing a repeated freeze-thaw test. The TCID₅₀ and hemagglutination titer of LS strain kept at the high level (TCID₅₀=10^7.75/mL,2⁸) after 20 serial passages in BHK-21 cells. According to the proliferation dynamic curve of JEV LS strain in BHK-21 cells, the optimum time to harvest JEV was 28 to 40 h after virus innoculation. This research laid a foundation for the further research on biological characteristics of JEV LS strain.

The germ cells come from the primordial germ cells(PGCs), in vivo differentiation mechanism of which has been basically understood. With the development of germ cell studies, in vitro induction of germ cells has become the reality. Embryonic stem cells(ESCs) can be induced to differentiate into epiblast-like cells, then into primordial germ-like or primordial germ cells. When transferred into fetal gland they have the abilities to undergo meiosis and create sperm; when transferred together with ovum into gonad of newly born mouse lacking of seminiferous tubule, they can engender healthy offspring. Therefore, induction of germ cells in vitro further elucidates the mechanism of the ESCs differentiating into germ cells, and lays the foundation for more effective utilization of stem cells to benefit mankind.

Bone metabolism included bone formation and bone absorption. Parathyroid hormone(PTH) was a factor and played an important role in regulating bone remodeling and formation,in where PHT through the G protein coupled receptors,parathyroid hormone/parathyroid hormone related peptide receptor activity regulating bone metabolism. Recent years, PTH had now been approved for clinical diagnose in the treatment of bone loss in osteoporosis as bone anabolic drugs and opened up a new age. The author summarized the mechanism of PTH on the bone remodeling and function of related regulatory genes in bone metabolism.

The assay was aimed to study the relationship of ROS and MAPK family protein in pseudolaric acid B (PAB)-induced MCF-7 cell apoptosis. Contrast phase microscope was used to observe cellular morphologic alternation; MTT analysis detected the inhibitory ratio; flow cytometric analysis of DCFH-DA staining determined the level of ROS, and Western blotting detected the protein expression. We found that PAB induced MCF-7 cell apoptosis through increasing ROS and increased the phosphoration of JNK and decrease the phosphoration of ERK.

α-galactosidase can catalyze α-galactosis bond hydrolyzing,and decompose anti-nutrients α-galactose galactosides contained in feed and leguminous food,and improve nutrients. Some α-galactosidases have a transglycosylation action at high substrates concentrations,and it's proved to be used for the oligosaccharides synthesis and cyclodextrin modification based on this feature. Moreover,α-galactosidases also have applications in the pharmaceutical,thickening agent treatment and paper industry. The author reviewed the study and application situation of α-galactosidases,and the research direction and development trends of α-galactosidases were also summarized and evaluated.

The intestinal barrier is the largest interface between animal and the external environment,which includes physical barrier,chemical barrier,immune barrier and microorganism barrier.The integrity of intestinal barrier has an important role in preserving health.Many kinds of stress can influence the integrity. The author reviewed the effect of prolonged exercise,restraint stress and heat stress on the intestinal barrier,and the measures to solve it.

Antimicrobial peptides have important effects on the innate resistance system of human and livestock due to their activity against bacteria, viruses and fungi. They also are expected to be used as preventive and therapeutic medicine. Therefore they are called 'new generation antibiotics'. Antimicrobial peptides are cationic peptides, and the most numerous group of them is a family of defensins. Among them the most common are peptides with a beta-sheet structure containing three intra-molecular disulphide bonds, called defensins, comprising three classes: alpha-, beta- and theta-defensin. The family of beta-defensins is the largest one, and their transcripts have been found in many tissues of human and livestock. The aim of this paper is to present the current knowledge about antimicrobial peptides from the defensins family in farm animals, their expression, polymorphism, as well as the potential of their use as genetic markers of health and production traits.

Embryo diapause was an importance factors which affecting mink farming, researchers had made a great deal of studies. In this presentation we addressed the characteristics of the embryo in diapause, analyzed the factors of embryo diapause and focused on the mechaminsms of regulation of this phenomenon, including the environmental stimuli that induced and terminated this condition, the hormonal regulatory pathways, and the phenomenon of cell cycle arrest and reactivation, overcome the embryonic diapause and shorten pregnancy period. On these bases, the existing problems and further research directions were pointed out and provided scientific ideas for further research on the mink embryo implantation in the future.

Oxidative stress is involved in the defect of embryo development, reactive oxygen species (ROS) can originate from embryo metabolism or embryo surroundings. Some exogenous factors can improve the production of ROS and cause embryo developmental arrest, it seriously limits embryos related studies. There are many mechanisms to protect embryos against ROS, therefore, it is essential to combine with ROS resources and anti-oxidative mechanisms at different developmental phase, in order to improve the inner anti-oxidative efforts, control culture conditions and in practical operation but it is a complex problem to defend oxidative damage during embryo culture, and complete culture system, decrease the induction effects of outer factors on ROS, produce in vitro embryos effectively.

The Mycoplasma ovipneumoniae (MO) SC01 isolates were cultured in the modified Thiaucourt's medium and collected in different times. The growth curve of MO was determined by the color change unit (CCU), nephelornetry and Real-time PCR. The results of CCU showed 0 to 4 h was the lag phase, 4 to 16 h was logarithmic phase and the most of lg CCU/mL was 10.7000?0.4700, 16 to 20 h was the stationary phase, and 20 h later was decline phase of SC01 isolates. The results of nephelornetry method showed that D_{420 nm} of five folds concentration of SC01 cultures gradually increased with the culture time, following the 0.0028?0.0002 in 2 h, slowly increased in 4 to 12 h, fast increased in 12 to 24 h, the 0.8609?0.0045 of highest level in 24 h and then stabilizaed. The results of Real-time PCR showed that the copy of SC01 isolates steadily increased in 2 to 30 h that the lg copy/mL was 4.5889?0.0048 in 2 h and the most of lg copy/mL was 7.6165?0.1089 in 30 h. Furthermore, the correlation analysis exhibited that, in lag phase and logarithmic phase, the CCU had a high correlation with the Real-time PCR and nephelornetry, and Real-time PCR also existed a high correlation with nephelornetry, following that the correlation coefficients were 0.967,0.720 and 0.849, respectively. During 2 to 30 h,there was a high correlation between the Real-time PCR method and nephelornetry and the correlation coefficients was 0.912. Based on the above results of the time and accuracy of three detection methods, the study indicated that the Real-time PCR method was fast, accurate and easy to standardization and could replace for the CCU and nephelornetry methods.

The aim of present study was to investigate the antimicrobial resistance, and the prevalence of methicillin-resistant Staphylococcus aureus (S.aureus) isolated from bovine mastitis in Gansu province, to provide credible theory evidence for prevention and treatment on bovine mastitis. Eight commonly used antimicrobial agents were used for determining antimicrobial susceptibility of 17 total S.aureus strains by disk diffusion method. Agar screen method was used for determining the oxacillin and vancomycin minimum inhibitory concentration value as well. Disk diffusion method using the cephamycin antibiotics cefoxitin and detection of mecA gene by PCR assay were performed to detect the presence of MRSA. Most of strains showed a high resistance for penicillin and sulfafurazole, yet keeping complete sensitivity for ciprofloxacin, cefazolin, vancomycin and oxacillin. None MRSA isolate was identified by the phenotypic detection method, but eight MRSA isolates with the MIC of oxacillin lower than 2 μg/mL were found to carry mecA gene via PCR approach. This study suggested that the antimicrobial resistance of S.aureus of bovine origin was very serious. High frequency of mecA-positive oxacillin-susceptible S.aureus (OS-MRSA) were present in our herds.

20 strains of E.coli from meat, chickens and mutton collected from product market and supermarket of Hebei Jidong areas were to be identified with serotype identification and K88 fimbriae gene of different serotypes was detected. O serotype of the strains of E.coli were identified with conventional methods and PCR. These isolates distributed in 7 serogroups (O38,O78,O88,O11,O107,O91,O9) and O38 and O78 were the dominant serotypes, the positive rates were both 25%(5/20). Detection rate of K88 fimbriae gene from food was 30%(6/20).The serotypes of O78 and O38 in Jidong areas from food were commom, and the detection rate of K88 fimbriae gene from food was 30%.

To better understand, we investigated the SO (H1N1) IV in 40 pig farms from 9 cities in Shandong province. Serum samples (263) and nasal swabs(235)from pigs with flu-like symptoms were collected, respectively. The positive rates of nasal swabs and blood samples were measured using Real-time RT-PCR and HI-test, respectively. Results demonstrated that, the positive rates of serum samples were 23.07% to 63.33%, accounting for 30.63%; the positive rates of nasal swabs were 10.0% to 64.28%, accounting for 30.63%. This result provided the basis to understand the SO (H1N1) IV distribution in Shandong province.

Clostridium perfringen isolate was isolated from rabbit's farm in Shandong. The isolate was identified by colonial morphology, culture characteristic, biochemistry reaction, pathogenicity test on mouse and rabbit. The results indicated that the pathogen designated as SD12 strain,was anaerobic and identified A genotype Clostridium perfringen with high pathogenicity on mouse and rabbit.

In order to survey the status quo of fish pathogens resistant to fluoroquinolones sourced from Beijing area, and provide the basis to regulate the fluoroquinolones used in fish farming,there were 16 pathogens isolated from diseased fish organs or focus with typical symtom in Beijing, and then the resistance phenotype of fish pathogens on seven kinds of fluoroquinolones were assayed by Kirby-Bauer disk diffusion method according to CLSI and the status of which carried gyrA,qnrA and qnrS genes were detected by PCR method. The data results showed that,on one hand,the rate of fish pathogens detected quinolones resistance gene-positive was higher than that of resistance phenotype,and the reason might be that the resistance genes detection were more exact than resistance phenotype tested. On the other hand, half of the fish pathogens resistant to a kind of fluoroquinolones at least, while 87.5% of the pathogens carried a quinolones resistance gene at any rate.Thereby,it was serious that the fish pathogens sourced from Beijing resistant to fluoroquinolones at genetic level, and it was necessary to strictly control these drugs used in fish farming.

The canine parvovirus disease was an acute infectious disease caused by CPV. The major features of this disease included severe vomiting, hemorrhagic enteritis, significant reduction in white blood cells and myocarditis. The disease with high incidence rate, highly infectious and high mortality was one of the serious infectious diseases threatening dog raising in our country. In this research ,260 cases of canine parvovirus case from Taizhou Aite pet clinic during 2010 January to 2011 March were analyzed.This article disclosed the epidemiology of CPV in Taizhou area of Jiangsu province, aimed to provide useful guide for the clinical treatment of CPV in the future.

In order to screen efficient Chinese herbal medicine against Newcastle disease virus, 8 kinds of compound Chinese herbal medicine against Newcastle disease virus were determined by chick embryo test. The result showed that some compound Chinese herbal medicine had better inhibition effect, including jingfang baidu powder,yinqiao powder,baitouweng decoction and shuanghuanlian oral liquid.

This paper investigated the advantages and disadvantages of three methods to detect classical swine fever virus(CSFV).We used three kinds of methods, including reverse transcription-multiplex nested polymerase chain reaction(RT-nPCR),fluorescence antibody method and colloidal gold immunochromatographic assay(CGIA),to detect 30 CSFV samples.The results showed that 11 samples of detected 30 samples were positive by RT-nPCR,13 samples were positive by fluorescence antibody method, 8 samples were positive by CGIA,and 8 samples were positive and 17 samples were negative by the three methods.Although fluorescence antibody method sowed time, it needed a seasoned staff to judge results. What's more, there were many false positive results arising by the method, and its specificity was rather poor.Gold dipstick method's biggest advantage was simple and easy to operate,and it was also appropriate for primary level,while the disadvantage was that it had a low sensibility.RT-nPCR's advantage was that it had a high-speed and sensitive operation, but it needed certain equipment and mature technology tests. In additionto this, it could differentiate testing samples whether they were vaccine strain or wild strain, and this was the point which other two methods couldn't match.

Infectious hematopoietic necrosis(IHN) as the national second-class animal disease was a serious viral disease of cold-water fish. The following aspects of IHN were discussed and reviewed in this papers in order to provide the data for the prevention and control of the disease, such as biological characteristics, clinical symptoms and popular feature, the viral genome and structural protein characteristics,detection and diagnosis technology, the prevetion and control measures.

In order to investigate E.coli isolates collected from different growth stages pigs in a pig farm of Xinjiang to commonly used antibiotics. Fecal samples were taken from the fattening pigs, pregnant pigs, nursery pigs and lactating pigs. Minimal inhibitory concentrations of the antibiotics to these E.coli isolates which were confirmed were determined by the broth micro-dilution method. Results showed that isolation rate of E.coli isolated from the fattening pigs, pregnant pigs, nursery pigs and lactating pigs were 85.2%, 83.0%, 81.0% and 83.0%, respectively. E. coli isolates from pigs in different growth stages were highly resistant to ampicillin and amoxicillin/clavulanic acid, while were more sensitive to ceftiofur. Resistant condition of E. coli isolates was serious in the nursery pigs and lactating pigs, and most of the isolates were resistant to 5 kinds of antibiotics. E.coli isolates from pregnant pigs were sensitive to all antibiotics measured. E.coli isolates from fattening pigs were mainly resistant to 3 kinds of antibiotics. Antibiotic resistant of E.coli isolates from different growth periods were different.

Different types of adjuvants are different on the alloantigen immune adjuvant effect. To screen the ideal adjuvant of the inactivated vaccine of Mycoplasma ovipneumoniae (MO), the inactivated vaccines using 4 different adjuvants were respectively immunized adult rabbits and the determination of serum antibody titers were detected using the indirect hemagglutination kit after immunization from 1 to 9 weeks. The results showed that the 4 different vaccines could quickly produce antibodies after immunization. Furthermore, the rabbits antibodies using Buonavoglial's and ISA-760 adjuvant vaccines peaked at 3 weeks after vaccination ((5.67±0.70) log2,6.00 log2), and maintained the peak until in 9 weeks (6.00 log2,(6.00±1.00) log2). In addition to vaccines with MO ISA-206 and white oil adjuvant, the rabbit antibodies peaked in 4 weeks after immunization ((4.33±0.57) log2,5.00 log2) and decreased a titer until 9 weeks. According to the statistical analysis, the results showed that the immune enhancement effect of ISA-760 and Buonavoglial's adjuvant for the MO was significantly better than that of ISA-206 and white oil adjuvant (P<0.05). Therefore, the above results indicated that ISA-760 and Buonavoglial's were an ideal candidate for MO inactivated vaccine adjuvant and provided a scientific basis for studing the efficient of the inactivated vaccine for MO.

A HPLC method for the determination of icariin in different particle sizes of Epimedium was developed. Chromatography was performed on a Shimadzu VP-ODS column(150 mm×4.6 mm,5 μm). An acetonitrile-water (V:V=25:75) as the mobile phase was used on the wavelength of 270 nm. The results showed that the calibration curve was linear in the range from 0. 08 to 0.40 mg/mL with r=0.9999. The relative standard deviation was 1.62% and the recovery was from 97.11% to 101.08%. The method was accurate and rapid for the determination of icariin in different particle sizes of Epimedium. And the dissolution amount of icariin was increased with the diminution of particle size. But when the size surpassed 100 meshes, the dissolution amount remained constant.

Tetracycline with the broad-spectrum antibacterial ability,has been largely used in animal husbandry industry. However,illegal administration will lead to residues in the animal food.Traditional detection methods have the disadvantages such as complex program,time and energy consuming,costly equipment,and difficulty to implementon-line,real-time and mass detection,but the immunoassay can make up these shortcomings.In this paper,tetracycline residues,damage and immunoassay are reviewed.

The experiment was aimed to establish the mice model of subacute kidney injury induced by gentamycin (GM), and study the effects of saccharum alhagi which was Uighur medicine on nephrotoxicity caused by gentamicin. Twenty eight mice were divided into 7 groups, group A(125 mg/kg GM), group B(100 mg/kg GM), group C(80 mg/kg GM), group D(125 mg/kg GM+saccharum alhagi), group E(100 mg/kg GM+saccharum alhagi),group F(80 mg/kg GM+saccharum alhagi) and group G(saline), respectively, intraperitoneally injected for 7 d. Mice were sacrificed at 8 d after blood collection. The blood was measured for concentration of non-protein nitrogen (NPN) and the weight of livers, spleens and kidneys. The results showed that compared with group A and C respectively, concentration of NPN and kidney coefficients of group D and group F had significant difference(P<0.05); compared with group B, concentration of NPN of group E had no significant difference(P>0.05), but kidney coefficients had significant difference(P<0.05). Kidney damage was found in the groups A, B and C. Saccharum alhagi had palyed an extremly role in the protection of kidney injury induced by 125 and 80 mg/kg GM, but didn't play a significant role in the protection of kidney injury induced by 100 mg/kg GM.

Coating technology has a wide range of feed additives. It plays an important role to improve the stability of the product, cover up the bad smell, control the release of the product in the digestive tract and improve the bioavailability. Film coating technology in animal husbandry is introduced and the development prospects for film coating technology is proposed finally.

Acathopanas senticosus polysaccharides (ASPS) is a biologically active polysaccharide, not only has strong anti-tumor effect and antioxidant activity, at the same time is also an ideal immune enhancer and helps to regulate the gut microflora balance. So ASPS as a new green feed additives would have good prospects for development in animal production. This review summarizes physiological function of ASPS and its application in animal production.

This study established a competitive enzyme-linked immunosorbent assay (ELISA) method for detecting zilpaterol in animal derived food. Zilpaterol polyclonal antibody was obtained by immunizing New Zealand rabbit with zilpaterol-KLH conjugates. Checkerboard titration method was used to determine the optimum concentration of antibody and enzyme-labelled antigen. Coating condition, reaction time and the color developing time of substrate were also detected. The results showed that an ELISA kit with a linear sensitivity ranged from 0.15～10 ng/mL was successfully established. Half inhibitory concentration (IC₅₀) ranged between 0.43～0.79 ng/mL. The recovery rates for spiked zilpaterol in urine, feed, milk powder, milk and tissue were 65%～90% , with less than 15% coefficient variations (CV). The cross-reaction for other similar medicines was less than 0.1%. The ELISA kit established in current study for detecting zilpaterol in animal derived food and showed the good reproducibility, stability and specificity, hence it could be used to detect the zilpaterol residues in animal derived food.

The Hippochase rhamnoides is a new high-value multi-nutrient plants addiction, which contains a variety of active chemical ingredients.The Hippochase rhamnoides can affect production performance, immune function, antioxidant properties of poultry. This paper reviews physicochemical properties of Hippochase rhamnoides and the impact on poultry production performance and its prospects for development in the feed industry.