The effects of different concentration (0, 1, 10 and 100 ng/mL) insulin-like growth factor-Ⅰ (IGF-Ⅰ) on the mRNA expression of genes related to milk protein and fat synthesis in bovine mammary epithelial cells cultured in vitro were evaluated by Real-time PCR. The results showed that the mRNA abundance of insulin like growth factor-Ⅰ receptor (IGFIR), insulin-like binding protein 3 (IGFBP3), α-s1-casein (CSN1S1) and κ-casein (CSN3) did not differ significantly (P>0.05) within different concentrations of IGF-Ⅰ. However, with the increasing concentrations of IGF-Ⅰ, the mRNA abundance of β-casein (CSN2), acetyl-CoA carboxylase (ACACA), fatty acid synthase (FASN) and fatty acid binding protein 3 (FABP3) increased significantly (P<0.05). These results indicated that IGF-Ⅰ, as an important cytokine, might involve in regulating the mRNA expression of genes related to milk protein and fat synthesis in bovine mammary epithelial cells cultured in vitro.

The Cap gene partially deleted nuclear localization signal sequence(pdCap) of porcine circovirus 2(PCV2)was cloned into pBACsurf-1 between the upstream gp64 signal sequence and downstresm gp64 mature domain to construct the recombinant plasmid pBACsurf-1-pdCap.The fusion gene containing pdCap and gp64 was subcloned into pFastBac^TM Dual to construct the recombinant plasmid pFastBac-pdCap.And then the plasmid was transformed into E.coli DH10Bac competent cells to obtain the recombinant shuttle vector Bacmid-pdCap.Eventually,the recombinant Bacmid-pdCap was transfected into Sf9 cells to produce the recombinant baculovirus BV-pdCap mediating by Lipofectamine^TM2000.Indirect immunofluorescent and Western blotting assay indicated that the recombinant baculovirus BV-pdCap could express pdCap protein with significant antigencity.

The paper was to study the effects of ENO1 overexpression with adenovirus vector on Bcl-2 in the granulose cell of Zi goose. Adenovirus vector with ENO1 were transfected to Zi goose primary cultured granulose cells, and Bcl-2 mRNA of granulose cells were detected by the method of Real-time fluorescence quantitative PCR. The results showed that ENO1 overexpression made the Bcl-2 mRNA expression in the granulose cells significantly increase (P<0.01). ENO1 overexpression made the primary culture follicle granulose cells in vitro improve Bcl-2 mRNA expression, which suggested that ENO1 might inhibit the apoptpsis of Zi goose granulose cell.

This study was aimed to detect the single nucleotide polymorphism (SNPs) of Holstein cows diacylglycerol acyltransferase 1 (DGAT1) gene in exon 8, and to predict the protein structure and function caused by the mutations and discuss the evolution relationship of DGAT1. PCR-SSCP combined with sequencing was used to detect the SNPs in exon 8 of DGAT1 gene. Bioinformatics approach was used to predict the physicochemical properties and structures of DGAT1 protein. The approach was also used to analyze the amino acid homology of DGAT1 between different species, and to build the evolutionary tree. The PCR-SSCP analysis results showed that a double mutations existed in exon 8 of DGAT1 gene, namely M694 and M695.The prediction of protein structure and function displayed that the double mutation had no influence on the physicochemical properties and structures of the protein, but led to the changes in the component of its protein function domain. The evolution analysis results indicated that amino acid sequence homology of DGAT1 between Holstein cows and sheep was the highest (99.1%), and the homology between Holstein cows and chimpanzees was the lowest (65.3%).

Primers were designed based on insert sequence IS1111 of Coxiella bumetii of Q fever, SYBR GreenⅠReal-time quantitative PCR assay was developed for indentification of Q fever. The recombinant plasmid containing the target sequence was constructed to detect the sensitivity and prepare the standard curve. The method could detect 10² of the plasmid copy numbers.Related coefficient was 0.991 of the standard curve,the amplification efficiency was 98%.The results of specific detection of nucleic acid sample for M.tuberculosis,Chlamydia,Brucella and bovine blood were negative. SYBR GreenⅠfluorescent quantitative PCR method developed in this study had high Brucella sensitivity and specificity,and could be used for clinical test.

In this study, we evaluated the stimulation of RAW264.7 macrophages with a purified expressed recombinant protein, catabolite control protein A (ccpA) of Streptococcus suis 2. The results showed that this protein could stimulate the secretion of NO and induce IL-6,TNF-α and IFN-γ in RAW264.7 macrophages if in a dose-dependent manner with specific pre-treatment of the cell cultures with iNOS, NF-κB, p38 and ERK 1/2 MAPK pathway inhibitors. This study might provide important proves for the development of regulatory factors on Streptococcus suis 2 infection.

Orf was an acute skin zoonosis caused by Orf virus (ORFV), which could affect sheep, goats and humans. This research used purified ORFV (OV/HLJ/4 strains) as experimental material, the macrophage cell line RAW264.7 as host, RAW264.7 were classically activated by LPS before ORFV inoculation in vitro, the effect for macrophage apoptosis upon ORFV infection was investigated simultaneously. Comparison of morphological features of activation in the cells treated with LPS and non-LPS, these resulted in significant morphological variation each other. The apoptosis quantity of macrophage cells treated with the LPS was bigger than the non-LPS group which was less of relative changes. Western blotting showed that the apoptosis signal Caspase-3 weakened at 24 h comparing with glyceraldehyde 3-phosphate dehydrogenase (GAPDH), such a poptosis signal that was significantly weakened, related to that macrophage apoptosis level declined at that time.

The study was aimed to establish a method for determining Irisflorentin in Shegan Dilong granules and inspection for the stability of the preparation. RP-HPLC method was adopted and the GL Sciences Inertsil ODS-3 (150 mm×4.6 mm, 5 μm) was used. The mobile phase was acetonitrile-0.2% phosphoric acid solution (39:61), with the flow rate of 1 mL/min. The detection wavelength was at 266 nm and the column temperature was 30℃. Under the high-light exposure experiment,accelerating experiment and room temperature storage observation, the results showed that the product was basically stable. The calibration curve of Irisflorentin was linear in the range of 0.3422 to 86.60 μg/mL, r was 0.9995 (n was 10), the average recovery rate was 99.80%, RSD was 1.50% (n was 9). The method was simple, sensitive and accurate with good repeatability, which could be used for the quality control of Shegan Dilong granules and the preparation had a good stability.

The study focused on the effects of peptides PrP106-126 on cerebellar granule neuron apoptosis-related genes expression. In this study, we detected expression changes of the cerebellar granule neuronal apoptosis gene affected by the peptide PrP106-126. The results showed that the expression of bax, caspase-3 and myc genes increased significantly after stimulated. On the contrary, the bcl-2 gene expression decreased significantly which, further confirmed that bax, caspase-3 and myc genes could induce cell apoptosis and bcl-2 gene could inhibit cell apoptosis.

In November 2011, an infectious disease characterized by systemic macular hemorrhage, diarrhea and obviously inguinal lymph node enlargement, broke out in a swine farm located in Beijing. The lymph node, kidney and spleen tissues were taken from infected pigs and grinded to obtain tissue suspension which was then filtered through 0.2 μm filter and inoculated into the PK15 cells. D-glucosamine was added to the infected cell which was then gone through 3 blind passages,and DNA was extracted from the harvested virus suspension. Identification were done by PCR method and immunofluorescent assay,and the complete genome was also amplified and sequenced,which was then analyzed by using DNAStar software. The results demonstrated that the isolated strain belonged to PCV2 (named BJ20111113 strain) with whole genome size of 1767 bp. At nucleotide sequence level, its homology could reach 94.5% to 99.8% compared with the domestic and foreign reference strains. Among them, the most closely related strain was HM038017 (BDH strain) with homology of 99.8%, and the far distantly related strain was EU148504 with homology of only 94.5%. The phylogenetic analysis result indicated that BJ20111113 strain belonged to genotype PCV2b.

To establish a rapid, specific and sensitive double PCR identification method of Streptococcus suis and Streptococcus equi subspecies pathogen,primers were designed based on gene conserved region of the M-like protein of Streptococcus equi subspecies and GDH protein of Streptococcus suis. The amount of M-like and GDH primers were both 1 μL (20 pmol/L). The optimal annealing temperature was 52.3℃. The sensitivity of double PCR were 10 CFU for Streptococcus suis and 100 CFU for Streptococcus equi subspecies. The specific trials had shown that five kinds of common pathogens had non-specific bands appeared in the double PCR system. 1 Streptococcus suis and 2 Streptococcus equi subspecies were detected by this method.

Effect of E.coli on intestinal mucosal immunity-associated cells of rabbits was studied, and the curative effect was compared by intestinal mucosal immunity, forty rabbits were randomly divided into four groups. At different times mucosal immunity-associated cells were observed after injection of E.coli by abdominal cavity. The results showed that the number of mucosal immunity-associated cells was increased after intraperitoneal injection of bacilli, intestinal mucosal immunity was stimulated. There were no differences among cidomycin, lactolin and control groups or cidomycin and lactolin groups, better treatment effect was made.

To set up a sensitive,specific,rapid and high throughput serological antibody detection method for porcine reproductive and respiratory syndrome virus(PRRSV),M protein were expressed using prokaryotic expression technology,an indirect ELISA antibody detection method for PRRSV were set up using the purified recombinant M protein as coating antigen. Based on published PRRSV M genome sequence,one pairs of specific primers were designed,amplified 435 bp M gene fragment by RT-PCR,the fragment was cloned into pET32a(+) expression vector,and induced by IPTG,the recombinant M protein was expressed in inclusion body forms,after recombinant protein purification,Western blotting showed the protein had good antigenicity and specificity.After optimization of reaction conditions,using the purified recombinant M protein as antigen established an indirect ELISA method for PRRSV,the established M-ELISA detected six kinds of swine disease (CSFV,JEV,TEDV,PRV,PPV,PCV2) positive sera were negative,the coefficient variation of the method intra-assay and inter-assay repetitive tests were less than 5% and 10%,compared with IDEXX ELISA kit, the coincidence rate of the method was 95.3%. The established M-ELISA detection method provided a simple,rapid,high throughput serological antibody detection method for the PRRS vaccinated pigs antibody level monitoring,and the wild PRRSV infection diagnosis and epidemiological investigation.

Using molecular biology software Vector NTI, the transferring-binding proteins-B of 15 Haemophilus parasuis(Hps) reference serovars and 12 isolates were analyzed with AluⅠ,AvaⅡ and RsaⅠ endonucleases. Reference serovars produced 11 different genotypes. GenotypeⅠ(AAA) included serovar 1,3,15 and genotypeⅡ(BBB) was represented by serovar 2, 9,11. Isolates had 10 genotypes, and 6 of those were new. The fact suggested heterogeneity of Hps isolates and PCR-RFLP was an useful method for studing the epidemiology of outbreaks of Glasser’s disease. Therefore, further validation with more field strains was needed.

The liver,spleen and kidney tissue of suspected Aeromonas hydrophila were isolated and identified by common nutrient agar,tryptic soy agar medium and biochemical reagent,three bacteria were aquired,then the genes sequence of three bacteria were tested by 16S rRNA common primer and compared with BLAST, regression test was used to determine pathogenicity of identified Aeromonas hydrophila. At the same time, the common antibiotics azithromycin,amikacin,streptomycin etc.ten kinds was used for drug sensitive test.The results showed that three bacteria i.e. Aeromonas hydrophila,Stenotrophomonas maltophilia,corruption watts bacteria were identified by separate culture,biochemical identify and 16S rRNA common primer,regression test confirmed Aeromonas hydrophila pathogenicity.The drug sensitive test showed that the bacteriostat diameter of azithromycin was 105 and 95 mm respectively for corruption watts bacteria and Stenotrophomonas maltophilia,the amikacin bacteriostat diameter was 105, 95 and 93 mm respectively for three bacteria. The grass carp tissue of suspecting Aeromonas hydrophila was isolated and identified three bacteria,identified Aeromonas hydrophila was proved its pathogenicity,amikacin was the most sensitive for the three bacteria,Stenotrophomonas maltophilia and corruption watts bacteria was sensitive for azithromycin.

To detect and identify two fimbriae genes (K88 and K99) of enterotoxigenic Escherichia coli (ETEC), a double PCR method was developed based on 2 pairs of primers for K88 and K99 genes amplification. 2 different virulence genes of the reference E.coli strain which passed was detected and the double PCR was proved to be specificity and sensitivity. The PCR amplification conditions ultimately were determined that dNTP concentration were 0.4 mmol/L, K88 and K99 primers concentration were both 25 μmol/L and the annealing temperature was 52℃. Furthermore, a total of 23 samples of E.coli were tested by the double PCR assay and the results showed 2 samples were K88, 3 samples were K99, and 5 samples were K88 and K99 positive. The double PCR assay provided a method to detect ETEC which caused diarrhea in piglets.

To develop a method to detect the antibody of rabbit hemorrhagic disease virus, an indirect ELISA was successfully developed to detect anti-RHDV antibody using recombinant VP60 protein as coating antigen. The method detected the positive sera of other three rabbit diseases(LaRV,SeV,Cl.welchii) were negative. Lowest antibody titer of positive control serum was 1:12800. Coefficient of variability percent (C.V%) of intro-batch and inter-batch duplicative tests were less than 4.9% and 6.9%, respectively. Therefore, the VP60-ELISA had good specificity, sensitivity and repeatability, which provided a technical means of rapid diagnosis for RHDV antibody detection and epidemiological investigation.

In order to understand the role of the wild birds in the transmission of avian influenza viruses (AIV), fresh stool from migratory birds and resident birds in Weining Caohai acquisition were collected and detected by RT-PCR. A flu virus positive sample was detected and the haemagglutinin (HA) gene was cloned and sequenced. Results showed that the AIV was subtyped into H3 and the HA fragment was 1794 bp belonging to the Eurasian lineage, which had one open reading frame and encoded 566 amino acids including 6 potential glycosylation sites. In addition, it had conserved amino acid sequences of influenza viruses of avian origin such as A, E, L, G, Q and G at position of 154, 206, 210, 241, 242 and 244, respectively, in the receptor binding site on the HA. The deduced amino acid sequences of the HA cleavage site had a typical low pathogenic motif (PEKQTR/GLF). The result showed that the low pathogenic H3 subtype AIV strain was circulating in wild birds of Guizhou province.

Porcine reproductive and respiratory syndrome virus (PRRSV) enters the host cells via a mechanism of receptor-mediated endocytosis. The entry mechanism is mediated by attachment to one or more cellular receptors and/or coreceptor on the cell surface and internalization. This article mainly describes several receptors for PRRSV. As far, it is reported five receptors of PRRSV which are independent and associated, heparin sulphate (HS), sialoadhesin (Sn),vimentin, CD163 and non-muscle myosinⅡA. It plays a significant role for the mechanism of PRRSV infection or disease prevention and treatment by studying the function of cellular receptors of PRRSV.

Mycoplasma suis (M.suis) is also called haemotrophic bacteria,causing anemia and jaundice disease, and is harmful for farming industry.The molecular biology of M.suis was summarized. It concluded the classify of M.suis based on the 16S rRNA sequencing,the complete genome sequence of strain Illinois and KI3806, the study of adhesion protein MSG1 and α-enolase,and diagnosed with PCR of M.suis.

Lipoprotein lipase (LPL) is a 54 kb glycoprotein, and its activity is affected by nutritional factors and hormone levels ect conciliation. Because the particularity of the yak living environment intended Tianjun yak the LPL-Exon7 gene polymorphism at the molecular level, and with the growth and development traits associated analysis for the yak nutritional regulation of species resources protection, genetic breeding scientific theory basis. Qinghai Haixi Tianjun yak anticoagulant 106 extracted blood samples were taken DNA, and polymorphism analysis of candidate genes LPL-Exon7 adopted the use of PCR technology and high-resolution melting curve analysis, and measured yak weight, body length, body height, bust, tube Wai growth traits and genetic effects. The results showed that the polymorphic yak and wild blood yak of the LPL-Exon7 gene were polymorphism, there were three genotypes, named AA, AB, BB, controlled by a pair of alleles A and B, A allele was the dominant allele. Loci balanced detection χ² test, in line with the law of Hardy-Weinberg, a state of equilibrium (P>0.05). The female and male of domestic yak and wild blood yak polymorphic information were 0.3588 and 0.2103, 0.3219 and 0.3343, polymorphic information content of domestic yak cows was PIC<0.25, at the low degree of polymorphism, the polymorphic information content of domestic yak bull wild blood yak public was in the 0.25<PIC<0.50, was a moderate polymorphism. LPL-Exon7 site polymorphism to yak weight,height and pipe circumference the existence of a significant effect (P<0.05).Yak body slanting length and heart girt were no significant effect (P>0.05). Therefore, the LPL-Exon7 was considered as one of yak genetic markers for marker-assisted selection.

To investigate the long-term effect of feeding flaxseed on the production performance and metabolism of laying hens.48 healthy, approximate weight, fifty-five weeks Hyline Brown hens were divided into four groups, corresponding to treatment 1, 2, 3 and 4 groups were supplemented with 0, 6%, 12% and 18% flaxseed for 10 weeks in the diets respectively. The test results showed that treatment 2 and 3 groups had no effect on weight gain (P>0.05), and treatment 4 group significantly decreased body weight (P<0.05). The excreta moisture content had no significant change with supplement of flaxseed (P>0.05). The treatment 2 and 3 groups could improve laying performance, but treatment 4 group could reduce laying performance. The treatment 2 group did not affect the apparent metabolizable energy and nutrient apparent metabolizable rate of diet, treatment 3 group could reduce the apparent metabolizable energy rate and P apparent metabolizable rate of diet (P<0.05), and treatment 4 group could reduce apparent metabolizable energy and apparent metabolic rate of nutrients. Therefore, supplemented with 6% flaxseed in diet could be used for long-term feeding of laying hens.

The experiment was conducted to study the effects of the level of protein in dietary on growth performances and blood biochemical indices in Hainan pigs. A total of 36 Hainan pigs with the weight of (15.88±1.82)kg were randomly allocated to three treatments, treatment one group (CP 16.5%), treatment two group (CP 18.5%) and treatment three group (CP 18.5%), each treatment group had three replicates and 4 pigs per replicate. The experiment lasted 40 days. The results showed that there were no significant effects of the level of protein in dietary on growth performances in Hainan pigs (P>0.05), but when the level of protein in diatary was 18.5%, the growth performances of Hainan pigs was the best. The concentrations of total protein and globulin in blood significantly influenced by the levels of protein in diets(P<0.05), as dietary protein level increased, Hainan pig blood total protein and globulin concentrations were elevated. In conclusion, Hainan pig’s optimal dietary protein level was 18.5%.

The experiment was aimed at the effects of different structural starters on growth performance and diarrhea rate in calf. Three days of health Holstein cows were choosed as a test sample, then divided into four groups according to the principle of similar weight randomly, and each group contained 6 calves. The four groups were fed different grain material respectively. The results showed that compared with the control group, at the age of 30 days, calves weight in 2 group were significantly higher (P<0.05); at the age of 60 days, calves weight in 2 and 3 groups were significantly higher (P<0.05), the body height and chest circumference in 2 group were significantly higher (P<0.05). During the whole trial period, the feed intake of calf in 2 and 3 groups were significantly higher than the test group and control group (P<0.05). In the rate of diarrhea, the control group was minimum, but 1 group was the highest. From the growth performance and body measurements, the results showed that the calf starters with ratio of calf feed pellets, steam flaking corn to steam flaking wheat being 7:2:1, and adding 5% molasses spray were better than other groups, and had the highest feed utilization rate.

With the development of the living standard of people, the consumers made more and more attention to the safe of animal production, and the quantity of transforming into mass. So the research of meat quality evaluation and quality regulating became one of the focus in the study of nutrition scholars. In this paper the author introduced the meat quality evaluation indices, and the effect factors of meat flavor. At the same time, the paper reviewed the two aspects of genetic and feedstuff to discuss the influence factors of meat quality, for the next step to improve meat quality to provide the theory basis and the reference material.

To establish a method of isolation, cultivation and purification of dendritic cells from mouse bone marrow in vitro. Mononuclear cells isolated form mouse bone marrow were induced into immature dendritic cells by being cultured with rGM-CSF and then examined from aspects of morphology, phenotype and function of T lymphocyte proliferation. The result showed that a large number of typical dendritic cells were observed after culturing for 9 days, and high expressed CD11c 81.09%, middle exproessed MHCⅡ, low expressed CD40, CD80, CD86. A method to generate large number of DCs from mouse bone marrow in vitro had been established.

The aim of the study was to investigate the effects on plup peroxidase (MPO),milk catalase (LP) and immune globulin-M (IgM), immune globulin-A (IgA), immune globulin-G (IgG) levels in serum of experimental mastitis that transplanted rat bone marrow mesenchymal stem cells (BMSCs) through the breast base.5 rats selected from 70 normal femal SD rats were slaughted after 72 h of postpartum, namely test -1 d, at the same time the rest of 65 rats were perfused 0.05 mL (50 μg) endotoxin through the nipple tube as the reference to domestic common way, 5 rats that the data obtained from these rats would be equally compared and contrasted with data from other groups later were randomly selected and slaughted after 1 d namely test 0 d that established experimental mastitis model successfully, at the same time the rest of the 60 rats were randomly divided into three groups. The control group (injection of saline in breast base, 50 μL), antibiotic group (injection of antibiotic in breast base ,50 μL, (5000 U/side)) and BMSCs transplantion group (injection of BMSCs in breast base μL (1×10⁶)), and slaughted 5 rats each group at 1, 3, 5, 7 d respectively, cardiac blood was collected from the dead bodies and preserved for detecting serum MPO, LP, IgM, IgA and IgG. The results showed that compared with control group, the MPO level of BMSCs transplantion group was significantly lower at any time points after treatment (P<0.05), and LP level at 1, 7 d (P<0.05). IgA, IgG and IgM level at 1, 5 d and IgG, IgM level at 7 d were significantly higher than control group (P<0.05), IgG level at 1 and 7 d, IgA level at 3 and 5 d, IgM level at 5 d were significantly differences compared with antibiotic group (P<0.05).So the level MPO and LP of antibiotic and BMSCs groups were lower, and improved the levels of IgM, IgA and IgG in mastitis serum of rats, and BMSCs transplantion group was more efficient than antibiotic group overall.

This assay was aimed to study the differences of different doses of streptozocin(STZ) such as 65,70 and 75 mg/kg,and different genders in establishing diabetic cataract models of rats.Single intraperitoneally injection of different doses of STZ to establish male and female diabetic cataract.We measured blood sugar after 36 h of injection, examined diabetic cataract after three weeks, measured food and drink consumption every day and weight changes every week, etc. The results showed that 70 mg/kg STZ was faster than 65 mg/kg STZ in establishing diabetic cataract,and had lower motality compared with 75 mg/kg STZ.We found that 70 mg/kg STZ was more superiority than the other two doses in the time required for the appearance of cataract, model stability and other aspects. Besides,male rats was more suitable in establishing diabetic cataract models.

In order to explore the effect of inactivated Salmonella induced maggot antimicrobial peptides, maggot were cultivated with inactivated of Salmonella and Salmonella, extracting antimicrobial peptides from maggot. The concentration of maggot antimicrobial peptides and antibacterial property were determined at different time. Meanwhile bacterial content of maggot in vivo was counted by colony forming units (CFU). The results showed that the concentration of induced antimicrobial peptides was significant higher than the non-induced group (P<0.05), and the biggest concentration of antimicrobial peptides was showed at 24 h;inhibition zone diameter of induction group was significant bigger than non-induced group for Salmonella (P<0.05), but less than antibiotic group (P<0.05), and the value of MIC was 0.5 mg/mL;the bacterial content of maggot in vivo was 1.6×10⁵ CFU/g, was lower than the national standard of the number of bacteria in the feed 2×10⁶ CFU/g. The experiments indicated that the expression of maggot antimicrobial peptides could be enhanced by induction of inactivated Salmonella, and the antimicrobial peptides showed a strong anti-bacterial property and security.

To compare the effects of saturated ammonium sulfate method and caprylic acid-ammonium sulfate method for purifying the rabbit anti-AR73 IgG. The IgG were tested in protein content,relative molecular weight, the antibody titer by BCA method,sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and indirect enzyme linked immunosorbent assay (ELISA).The results indicated that the protein content of IgG by saturated ammonium sulfate method was 36.44 mg/mL and the antibody titer was 1.6×10⁵, the protein content of IgG by caprylic acid-ammonium sulfate method was 15.79 mg/mL, and the antibody titer was 3.2×10⁵. The purity of purified antibody by caprylic acid-ammonium sulfate method was higher than that by saturated ammonium sulfate method.

The assay was aimed to screen the polymorphisms of AMPD1 gene promoter and analyze the effect of SNPs on function elements of promoter. Three goat breeds (Small-xiang goat, Qianbei ma goat, Nubian goat) were selected to construct DNA pools, SNPs sites were screened by direct sequencing subsequently and analyzed by BLAST. The results showed that 5 single nucleotide polymorphisms (SNPs) were found in the promoter region which included T-614A, G-326A, G-309A, T-287C and T-165C. Furthermore, bioinformatics tools were used to predict the core region of the promoter and identify various transcription factors binding sites. It demonstrated that 7 new transcription factors binding sites emerged while 9 previous transcription factors binding sites disappeared based on the SNPs found in this study, the remarkable change of secondary structure of RNA were also detected but the range of CpG islands were not predicted by the analysis of various softwares.

The polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to detect the polymorphism of keratin associated protein 7 (KAP7) in Shanbei White cashmere goat,Taihang Black goat and Nanjiang White cashmere goat. The results showed that the KAP7 presented AA and AB genetypes in these three breeds,AA genetype was the dominant genetypes. The PCR production of different genotype of KAP7 were purified and sequnenced. One mutation,129 bp(C→G) was found in KAP7. This mutation site was in 5' regulatory region.

Factors on culturing of embryonic gem cells (EG)in pigs were studied in order to establish porcine EG lines. In order to analysis genetic background influencing the culturing of the porcine embryonic germ cells, fetuses were collected from Wuzhishan pigs (WZSP) and White Minitype pigs respectively. Under the same culturing conditions, when the first time of EG colonies coming forth (T1),the first time of large quantities of EG colonies coming forth (T2),the first time of passage (T3),average passage intervals (T4),passages (P) were considered. The results showed that there was no notable difference in EG culturing between the two types of pigs(P>0.05). That was WZSP and White Minitype pigs were all suitable materials to establish porcine EG lines; In order to analysis fetal stages influencing the culturing of porcine EG, fetuses at 26 d, 27 d and 28 d were collected respectively, the length of the fetuses and the length of the fetal mesonephros were measured, the 5 indexes above mentioned were still considered, the results showed that under the same conditions, there was no notable difference in EG culturing using fetuses collected on 26 d, 27 d and 28 d respectively(P>0.05).

The associations between SNPs in the PLTP gene and production performance of pigs were reported in this study.The results showed alleles detected in the sequence were allele A and a with frequencies of 0.4172 and 0.5828,0.4155 and 0.5845,0.4309 and 0.5691 in F4, F5, F6 generation,respectively. There were 4 mutations of 3 for conversion and 1 for transversion,4 SNPs (1085,1192,1286 and 1305 bp, respectively) were found by the sampling sequence. Associations between the founded Hae Ⅲ RFLP in intron 4 and birth weight, body weight at 45 days, body weight at 4 months, body weight at 6 months, body height at 6 months, body length at 6 months,cannon circumference at 6 months and back-fat thickness at 6 months were tested by least square analysis. In F4 generation,no significant difference between HaeⅢ RFLP and all growth traits(P>0.05). In F5 generation, the birth weight and back-fat thickness at 6 months with Aa genotype were higher than that with AA genotype (P<0.05). In F6 generation,the body weight at 4 months(P<0.01)and body height at 6 months (P<0.05) with aa genotype were higher than that with AA genotype. The cannon circumference at 6 months (P<0.01) and the birth weight (P<0.05) with Aa genotype were higher than that with AA genotype. In addition, the cannon circumference were the highest significant than that with AA genotype male (P<0.01). The birth weight and back-fat thickness at 6 months were higher than that with AA genotype female (P<0.05). These results suggested that the Aa genotype of PLTP gene had some effects on the growth traits of porcine. These SNP might be considered as a potential genetic marker for selection.

The experiment was a preliminary study on fine-wool sheep hair follicle stem cells, which were isolated and cultured, in order to establish the fine-wool sheep hair follicle stem cells culturing system in vitro. It separated and cultivated the fine wool sheep hair follicle stem cells by three methods, one was the two-step enzyme digestion, another one was mechanical separation, and the third was the combination of two-step enzyme digestion and mechanical separation. By measuring the number, cloning efficiency and proliferation ability of hair follicle stem cells, the experiment intended to evaluate the advantages and disadvantages of the three methods synthetically, for not only obtaining a large number of hair follicle stem cells conveniently, but also seeking out ideal culture conditions of reducing their differentiation. Under the inverted microscope, the cells obtained by three cultural methods were paving stone-like, by which expressed keratin K19 and integrin-β1, indicating successful acquirement of sheep hair follicle stem cells and establishment of in vitro culture system. Finally, the experiment showed that, it was the optimal method to obtain and culture hair follicle stem cells by the combination of two methods of two-step enzymatic digestion and mechanical separation.

It is very important to assess sperm quality in animal for artificial insemination and in vitro fertilization. At present, in the human reproductive clinical and animal husbandry production, semen quality evaluation methods are mostly several target union examination, making the semen quality evaluation results to be more scientific and to be accurate.This paper reviewd the recent animal semen quality testing of new technology, new methods and new progress, so as to the exactly reflect sperm quality and fertility.

In this test we collected goat’s lesions of lung tissue in Inner Mongolia which was probably infected with pleuropneumonia,and inoculated it on the mycoplasma selection medium under sterile conditions to isolate Mycoplasma.Through the cultivated characters,morphology and culture characterization and biochemical reactions,the isolated strain was preliminarily identified as Mycoplasma ovipneumoniae(MO).Then we extracted the isolated strain’s genomic DNA and amplified its 16S rRNA by the commom primers,33 reference strains were loaded down from GenBank,the result of sequence analysis indicated that the isolated strain exhibited 99% homology with reference strain Y-98.The results indicated that the isolated strain was MO.

Knowing the research advance in the resistance mechanism of bacteria and the dissemination of antimicrobial resistance gene elements will help to guide the use of antimicrobials, and reduce the prevalence of antimicrobial resistance as well as provide a firm support on the development and use of new antimicrobials.

To find out the situations about infection of Salmonella and its drug resistance somewhere in Jilin province,the bacteria were isolated from 150 tissue samples of sick broiler of 7 farms in this city,and hemagglutinin test, pathogenity test and drug sensitive test were carried out. 26 strains were isolated, and 19 strains were identified positive by the HA,of which five were strongly positive and killed broiler, drug sensitive test results showed that the 5 strains of Salmonella were more sensitive to cefotaxime, and very high resistance to ampicillin,tilmicosin,lincomycin,azithromycin and cephradine,resistance rates as high as 100%. Norfloxacin,enrofloxacin,ciprofloxacin,doxycycline and chloramphenicol didn’t have synergistic or additive effect with mequindox which was used in clinical. These results suggested that Salmonella was one of the main pathogens harmful to the broiler of farms, and resistance was very serious.

Foot-and-mouth disease (FMD) is a highly contagious and economically devastating viral disease. Vaccination is the main measures to prevent and control the disease. With the development of scientific technology, the research of foot-and-mouth disease vaccines are improving and developing, this paper reviewed the development of foot-and-mouth disease vaccines.

This experiment was to identify the pathogen of calves which was suspected of infecting bovine respiratory syncytial virus and Pasteurella mutocida simultaneously. Conventional test of virus and bacterium, PCR methods were all used to identify viral and bacterial respectively. The results indicated that the virus could grow well, propagate and induce typical cytopathic effect with syncytium form ation in BT cells. The isolate could not agglutinate and adsorb erythrocytes of human, some animals and chicken. The virus could be neutralized by bovine respiratory syncytial virus (BRSV) standard positive serum. The RNA extracted from the cell culture of the isolated virus was subjected to RT-PCR analysis. According to the colony morphology, bacterial staining characteristics and biochemical characteristics, the isolated bacterium was Pasteurella multocida. The calves were infected by bovine respiratory syncytial virus and Pasteurella mutocida simultaneously.

A strain of goat pox viruses was isolated from suspected goat pox cases in Inner Mongolia province, and was named as NM strain. Samples from skin varioliform and blisters were collected and inoculated BHK-21 cells and 10-day-old SPF chicken. Typical cytopathic effect was observed in BHK-21 cells, significant variola was observed in vaccination site. The NM strain cytotoxic and embryo virus could be inhibited by goat pox virus positive serum. Using goat pox virus specific detection primer amplified of the NM strains by PCR, obtained consistent with the expected size of 445 bp fragment. Then the amplification product was cloned into pMD18-T vector, after identifying with double-enzyme cutting. The sequencing of positive clones showed that the NM strain fragment had a high degree of homology with published goat pox virus, the homology was 99.2% to 100%.

Morphological and molecular identification were done with the high protease-producing strains screened out from 245 marine organism and environmental samples,and their biological characteristics and compatibility effects between different strains were investigated. The results showed that, both the isolates Pe and Pc were identified as Bacillus subtilis, Ps was identified as Bacillus licheniformis. All the three isolates entered the growth platform phase after 16 h cultivation. The optimum pH and temperature for the growth of Pc and Ps strains were 7.0 and 35℃, respectively. And the optimum pH and temperature for the growth of Pe strain was 6.5 and 30℃, respectively. The best enzyme production occurred when the proportion of Pe and Ps was 1:1.

The cattle pica allotriphagia was a kind of common disease, leading to malnutrition, growth retardation and decreased production performance, and had brought huge economic losses to cattle industry. The author analyzed the causes of cattle pica allotriphagia, put forward treatment measures, and discussed its clinical symptoms and disease prevention, in order to improve the breeding benefit.

Ceratozdes arborescens was a considerable amount of forage resource, but the understand level of feeding value had rest on a traditional standard. So the feeding value experiment was developed in order to fully understand and utilize Ceratozdes arborescens. The experiment was conducted to feed sheep on particle diet combined by Ceratozdes arborescens in different ratio of concentrate to roughage. Then the weight gain caused by Ceratozdes arborescens diet, alfalfa diet and millet straw diet on sheep were analyzed comparatively. The results showed that the largest weight gain appeared at the particle diet which made from powder of Ceratozdes arborescens, alfalfa and millet straw mixed with 50% and 30% concentrated feed were the best, the daily gain was 127 g/d. In desert steppe, the primary cause of limiting alfalfa popularization was high price. In the level of 30% concentrated feed, the Ceratozdes arborescens and millet straw had the same weight gain effect as alfalfa, and could increase net income by 80 yuan per sheep.

This study was to find the difference of nutrient component and egg quality between the ordinary ducks and Chansi ducks. A total of 50 ordinary eggs and Chansi eggs respectively produced from 300-hundred-day adult ducks were randomly selected. Egg traits such as weight, eggshell thickness and strength, eggshell color, albumen height, egg shape index, yolk color, Haugh unit etc were determined, and crude protein, crude fat calcium, phosphorus, iron, zinc, and other nutrients content were measured as well. The results indicated that as with egg weight, eggshell thickness, eggshell color, strength of eggshell and egg shape index, the differences between Chansi eggs and ordinary eggs were not significant (P>0.05), as for albumen height, Haugh unit and yolk color were very significantly higher than ordinary eggs (P<0.01), there were also more protein, calcium, zinc and iron, and less fat and phosphorus in Chansi eggs than in ordinary eggs. Therefore, the quality of Chansi eggs was better, and they contained more nutrition than the ordinary ones.

Antibacterial peptide was a kind of bioactive micromolecule polypeptide, inducing from living organisms. There were included Defensions and Cathelkicidins, antibacterial peptide protegrin belonged to Cathelkicidins. Antibacterial peptide protegrin had strong antibacterial and rapid sterilization activity. This article mainly reviewed structure,gene expression, the mechanism and effect factors of gene expression and application prospect of antibacterial peptide protegrin.

Rumination time (RT) was an important indicator to evaluate health in dairy cattle. Recent researches showed that daily RT at estrus significantly reduced compared with that on nonestrus days. Based on this results, RT could be acted as a new method of heat detection in dairy cattle. In the article, the relation between daily RT and estrus, as well as the application prospects of the potential heat detection method, were reviewed in order to improve the production efficiency of dairy cattle.