The author performed the comparative analysis of carnitine palmitoyl transferase(CPT)gene in different tissues from sheep and goats by quantification PCR. The results showed that CPT1A mRNA expression in sheep liver and spleen was higher than CPT1B and CPT2 expression in sheep liver and spleen. Expression of CPT1B mRNA in the external oblique muscle of sheep was higher than CPT1A, CPT2 mRNA expression in the external oblique muscle of sheep. CPT1A mRNA expressed in goat spleen was higher than the expression of CPT1B and CPT2 mRNA in goat spleen. CPT1B mRNA in goats hind legs, biceps femoris and external oblique muscle were higher than CPT1A, CPT2 mRNA in goat hind shares two expression in the biceps. CPT1A mRNA expression in the liver of sheep was significantly higher than the expression of CPT1A mRNA in goats (P＜0.05). CPT1A, CPT1B, CPT2 gene expression in sheep heart, spleen, lumbar major muscle was significantly higher than the expression in goat(P＜0.05), the CPT1A, CPT1B, CPT2 gene expression in the hind legs in goat was significant higher than the amount of expression in sheep(P＜0.05). CPT family gene expression in the organization of sheep, goat differences might be associated with each gene expression patterns in different types of muscle tissue fiber content of fat deposition and other factors.

Constructed recombinant prokaryotic expression vector pET32a-VP73 was transformed into E.coli BL21, major antigenic epitope region of VP73 protein was induced stably and efficiently by IPTG, the result of SDS-PAGE showed, when final concentration of IPTG was 1.0 mmol/L, and pET32a-VP73 was induced 5 h, expression of major antigenic epitope region of VP73 protein was highest, the expressed protein was fusion protein, molecular weight was approximately 42 ku. The protein was identified as major antigenic epitope region of VP73 protein by SDS-PAGE and Western blotting. The purified major antigenic epitope region of VP73 protein was injected to New Zealand rabbits, serum was collected before and after injection. Antibody titer was determined by ELISA. Specificity of this protein was determined by ELISA whose primary antibody were ASF positive serum, rabbits serum before and after injection and positive serum of CSF, PRRS, pig pseudorabies. The result showed the titer of antiserum reached up to 1∶1024000, and gave rise to reactions with VP73 major antigenic region protein. The polyclonal antibody built the foundation for immunology research of VP73 and serological diagnosis of African swine fever.

The objective of this study was to detect the role of QRDR, acrR, marR and soxR in genetic characterization of highly fluoroquinolone-resistant clinical Escherichia coli strains. PCR method was used to analyse the mutations of QRDR, acrR, marR and soxR. RT-PCR method was used to detect the expression level of efflux pump and porin related genes. Mutations of QRDR mainly fell into GyrA(Ser83Leu and Asp87Asn) and ParC(Ser80Ile), and other mutations had also detected, such as ParC Glu84Gly, Glu84Lys, Glu84Val and Glu84Ala,ParE Ser458Ala,et al. There was an insertion of 777 bp fragment within acrR in ED28. 12 of 18 stains including ATCC25922 had double mutations (Gly103Ser and Tyr137His) in MarR and two (EP26 and EG42) of them had an insertion. ED40 had amino acid substitution (Thr38Ser and Gly74Arg) in SoxR. In multiple mutations strains, there were markedly increased expression of AcrAB and decrease expression of porin or porin loss.

Macrophages were acquired from the F.hepatica metacercariae infected IL-4 monoclonal antibody treated and wild type female BALB/c mice, respectively. All F.hepatica elicited macrophages and control macrophages were cultured in vitro. mRNAs from macrophages were extracted for the detection of Relmα, Ym1 and Arginase1 genes using Real-time polymerase chain reaction. Pictures and fluorescence intensity at different time points were acquired with a confocal microscopy and image software respectively after staining with serial purified antibodies. Results showed that levels of Relmα, Ym1 and Arginase1 in F.hepatica elicited macrophages(FeMΦ)and control macrophages (TeMΦ) from IL-4 monoclonal antibody treated mice were significantly decreased comparison to that in wild type mice.FABP2 was detected as spots in the cytoplasm. The fluorescence intensity of FABP2 increased from 12 to 24 h, but decreased afterward to 48 h. The fluorescence intensity of FABP2 in TeMΦ was similar to that in FeMΦ.

According to the published sequences of bovine Mycobacterium paratuberculosis in GenBank, a pair of primers were designed and synthesized for the C-2 chromosome(ISMav2)gene of Mycobacterium paratuberculosis, and a SYBR-GreenⅠfluorescent Real-time quantitative PCR assay was developed. The positive standard plasmid pMD-ISMav2 were used as quantitative template to make the standard curves by optimizing the reaction conditions, the correlation coefficient (R²) was 0.999. By using the positive standard as template, specificity and sensitivity were tested. According to the experiment, all negative controls such as Brucella, Escherichia coli, Salmonella and Streptococcus showed negative detection. By sensitivity analysis, the Real-time PCR indicated that a minimum of 1.96×10¹ copies of plasmid DNA was detected. As a result of the specificity and sensitivity of the assay with a relatively rapid and simple procedure, the Real-time PCR can be used as a routine assay for the diagnosis of Mycobacterium paratuberculosis.

In this study, in order to provide a nuclear for somatic cell nuclear transfer and production of transgenic animals, cationic liposome-mediated transfection of homologous recombination FGF5 gene knockout vector to somatic cell. After positive and negative selection obtained 51 cell clones, which remaining 12 clones after propagation, and ultimately get 4 positive cell clones by the molecular indentification of DNA and RNA levels, namely established the FGF5-knockouted Inner Mongolia Arbas cashmere goat fetal skin fibroblast cell line. Compared with normal somatic cell, the four FGF5 -knockouted positive cell clones had occurred very significant changes in cell morphology, traits and growth characteristics: bigger cell volume;fuzzy boundaries, reduced stereoscopic;growth rate dramatic decline;trypsin digestion time required was significantly increased. This study showed that these changes might be due to FGF5 gene knockout, cell transfection, drug selection and other roles.

Internal transcribed spacer (ITS) and 5.8S of ribosomal DNA (rDNA) of 3 species (5 samples) of the genus Coronocyclus were amplified and sequenced using polymerase chain reaction (PCR) techniques. Sequence analysis demonstrated that the length of the ITS1-5.8S-ITS2 sequences ranged from 748 to 843 bp, and there were 119 variable sites (including gaps) and 18 parsimony informative sites. The length of the ITS1 sequences ranged from 367 to 370 bp, with 14 variable sites and 9 parsimony informative sites. The length of the ITS2 sequences ranged from 228 to 320 bp, with 105 variable sites and 9 parsimony informative sites. The 5.8S gene sequences of all 3 species were identical. For all 3 species, the G+C content was higher in the ITS1 (48.0% to 48.5%) than in the ITS2 (37.7% to 40.3%). Pairwise comparison in 5 sequences revealed inter specific differences ranged from 1.9% to 3.5% in the ITS1 and from 5.6% to 31.8% in the ITS2, however, intra specific variation was low (0 to 0.5% and 0 to 0.9%, respectively). Furthermore, the homology were 99.07% to 99.41% compared with the sequences in GenBank. The study demonstrated clearly that the internal transcribed spacer sequences provided genetic markers for the species identification of the genus Coronocyclus.

In order to establish a gene-chip detection method of equine virus, the highly conserved DNAs of African horse sickness virus (AHSV ) was obtained by artificial splicing, West Nile virus(WNV) and equine corona virus(ECV) were acquired by molecular cloning, and then spotted on the treated glass slides as the diagnostic gene-chip. And the cDNAs amplified by multiple asymmetric PCR with the template of splicing and cloned nucleic acid sequence were labeled with fluorescence as probes. Following specific hybridization of deposited gene chip and labeled probes, fluorescence signals were scanned by laser scanner and the obtained image was analyzed by Qiamt Array software with the digital computer. The results showed that the prepared gene chip could detect and distinguish the three equine viruses. And its sensitivity was about 10⁴ copies of AHSV and 10² of ECV and WNV. The hybridization specificity was confirmed by the presence of fluorescence signals on the corresponding sites with samples from the three relevant viruses DNA template and by the absence of positive signals with the specimens from irrelevant viruses was negative by gene chip. The evidence suggested that gene chip, which was quick, specific, sensitive, and reliable, could provide a practical alternative to screen and quarantine a large number of samples within a very short period of time.

Porcine reproductive and respiratory syndrome(PRRS)is a highly infectious disease of swine caused by porcine reproductive and respiratory syndrome virus(PRRSV). PRRS caused reproductive failure in sows,such as abortion,fetal losses, premature,parturition, mummies,stillbirths and neonatal death. E protein was encoded by ORF2b, a non-glycosylated structural protein, which size was 10 ku.It can affect viral replication, prompt the shelling of the virus particles, but do not work on the assembly of the virus, and have the ion channel protein activity. The characteristics, subcellular localization and topology,ion channel protein activity of E protein were summarized in this paper.

This paper developed a new method in rapid detection of canine distemper virus(CDV) based on loop-mediated isothermal amplification(LAMP)assay which overcame the difficulties found in other measures,such as efficiency and high cost. In order to improve the specificity and sensitivity, we designed 6 primers recognizing the 8 conservative sequences on N gene of CDV. The result could be easily confirmed by agarose gel electrophoresis, color reaction or turbidity comparison. In conclusion, The newly developed RT-LAMP detection method for CDV was more specific, sensitive, simple and rapid than the others, even more the proceeding reaction was visible by the naked eye, and that was also to say, it offered a simple solution in field veterinary diagnose for CDV control.

Ten gene fragments contains two strains of porcine reproductive and respiratory syndrome virus which were designated ZS-GD strain and ZH-GD strain complete genome were amplified respectively by RT-PCR, and cloned into pMD18-T vector for sequencing. Phylogenetic analysis of two strains of PRRSV complete genome sequence by bioinformatics software showed that the complete genome of ZS-GD and ZH-GD were 15320 bp, the genome of ZS-GD strain and ZH-GD strain shared 88.1% to 97.8% and 88.4% to 98.0% of nucleotide homology with American type strain, respectively; shared 60.6% and 60.4% of nucleotide homology with Europe type strain(Lelystad virus). There were 30aa depletion in Nsp2 protein of two stains. The results showed that these two strains were depletion and variant strain of American type.

The ORF2 gene of porcine circovirus type 2(PCV2) including complete open reading frame(ORF)was amplified by PCR,and it was cloned into the pFastBac1^TM vector of Bac-to-Bac baculovirus expression systems to construct the recombinant transfer vector pFast-ORF2.The transfer vector was transformed into DH10Bac competent cell which contained the BmNPV bacmid. A recombinant shuttle vector which contained ORF2 gene was constructed by site-specific transposition and the colonies containing recombinant bacmid were collected by white selection. The cultured Sf9 cells were transfected with recombinant bacmid DNA mediating with cellfectin reagent and the pure recombinant baculovirus Bacmid-ORF2 was obtained. After transfecting this transformant into Sf9 cells,Cap protein expression in Sf9 cells was detected by indirect immunofluorescent assay and Western blotting. The experimental results showed that the recombinant baculovirus was obtained and the Cap protein with a molecular weight of 31 ku was expressed in insect Sf9 cells. The recombinant protein could be recognized by PCV2 positive serum. This would lay a foundation for developing PCV2 subunit vaccine and diagnosis method.

Interferon products, with the broad-spectrum antiviral and immunomodulatory activity, have broad application prospects in the field of medicine and clinical veterinary medicine. In the actual practice, liquid dosage form of interferon preparations has the problems of poor thermal stability, difficult long-term preservation, long-distance transportation, et al. However, the shelf life and stability of the lyophilized interferon preparations could be increased. In this paper, the principle of freeze-drying technology, screening of lyoprotectant and freeze-dried process for interferon products are reviewed.

To analysis the OmpA and 16S rRNA gene of Riemerella anatipestifer isolated strains in Yunnan province and explore the correlationship between OmpA and 16S rRNA gene on phylogenetic trees, OmpA and 16S rRNA gene was amplified respectively by PCR to construct the phylogenetic trees. Results analysis showed that the OmpA gene of 18 Yunnan isolated RA strains were situated in two groups and their homology were 86% to 99.2% and 92.6% to 100% respectively, the 16S rRNA gene of 18 Yunnan isolated RA strains were situated in one group and their homology was 96.1% to 100%.It’s worth noting that the RA-1,RA-2,RA-1 and RA-39 isolated strains belonged to the same subgroup on both the OmpA phylogenetic tree and 16S rRNA phylogenetic tree with the higher homology 97.3% to 99.2% and 99.6% to 100% respectively, the two phylogenetic trees showed a significant correlation within RA-1,RA-2,RA-1 and RA-39 isolated strains and showed no correlation within the other 14 isolated strains.

Chromosome is the main carrier of genetic factors in cell. It affects the biological character on the cellular level and plays an important role in the speciation during evolution history. As research methodology of the cytogenics cytogenetics improves, the study of the chromosome of cervid goes further gradually. The paper reviewed the advances on the chromosome of cervid in China and gave some suggestions on existents problems and further research.

Trehalose is a non-reducing disaccharide whose function in some organisms is as a source of energy or a protectant against the stress. The exogenous trehalose is contributed to protecting the bioactivitie of some substances like cells and antibodies in vitro. This review summarizes the biological properties of trehalose, such as energy source, stress resistance, cell stabilizer and helping to stabilize antigens or antibodies in ELISA. The trehalose is also helpful for improving ELISA detection.

The gB and gE gene of bovine herpesvirus-1 Bartha Nu/67 strain were amplified by PCR.The gB and gE fragment were cloned into pGEM-T-easy vector. The gB and gE gene were subcloned into baculovirus transfer vector and the recombinant Baculovirus vector pFBHgB, pFBHgE were constructed successfully. Then they were transferred into E.coli DH10Bac and cultured in LB plate. The white bacterial colonies were positive recombinant bacmid named as BacmidgB, BacmidgE. Thus the result mentioned above laid down theoretical and practical foundations for the expression of gB and gE gene in insect cells.

Six strains of virus isolated from several areas of poultry occurred Newcastle disease, through HA,HI,neutralization test, and molecular biology experiments were identified as Newcastle disease virus. According to F gene sequences published in GenBank, a pair of specific primers was designed and synthesized, length of approximately 610 bp was amplified, alignment and analysis of nucleotide and amino acid sequences were conducted. The results showed that there were 3 lines with virulent Newcastle disease virus strain characteristics, also had the Newcastle Ⅶ genotype features. According to MDT, ICPI and IVPI index determination result, we finally determined that the 3 strains were virulent strains of NDV and belonged to Ⅶ genotype. The homology of nucleic acid and amino acid sequence between remaining 3 strains of virus and La Sota were both 99%.

A fragment of about 972 bp long was amplified by RT-PCR technique with a pair of specific primers based on published Japanese encephalitis virus(JEV) genome sequence. Then the target fragment was directionally cloned into pET30a vector. After identifying with enzyme cutting and sequencing, the recombinant plasmid was transformed into E.coli BL21(DE3). The recombinant protein E was expressed in inclusion body form in E.coli after induction with IPTG. After denaturation, purification and renaturation, the purified protein was analyzed by Western blotting, the results showed that the purified recombinant protein retained better antigenicity and specificity.

According to the GenBank login duck reovirus genome sequence, using biology software to design and synthesis two pairs of primers, established a nest RT-PCR detection method for the detection of Muscovy duck reovirus(MDRV), and used the detection method of the isolated virus and other poultry virus for application testing. The results showed that the method could amplify specific target fragment from MDRV, sensitivity to 0.1 pg virus RNA, while other viruses of avian reovirus (ARV), chicken infectious bursal disease virus (IBDV), Muscovy duck parvovirus(MDPV), goose parvovirus (GPV), duck hepatitis virus (DHV) and sample amplification results were negative. Therefore, the research provided a reference for MDRV disease detection and diagnose.

An isolate of avian influenza virus,designated WUDI-H9N2,was isolated from Shandong area of China in Auguste 2011 and identified for pathogenicity to chicken embryo by hemagglutination test, hemagglutination inhibition test and RT-PCR. A pair of primers was designed to amplify the HA strain based on the reported gene sequences in GenBank by Oligo 6.0 and for WUDI-H9N2 HA gene amplifying and sequence analysis. The separated AIV H9 strain and other serotype were respectively analysed by nucleotide and amino acid phylogeny tree. The results of sequencing showed that the HA gene was 1708 bp in legth, no nucleotide insertion and deletion, the cDNA contains whole open reading frame(ORF) of HA gene was 1683 bp and encoded a 560 amino acid protein. Compared with reference strains, the nucleotide sequence similarity of HA was 92.9% to 98.1%. On the basis of phylogenetic analysis of HA gene, we concluded that the HA shared 98.1% similarity of structure protein encoding sequence with A/chicken/China/AH-10-012010(H9N2) strain.

To amplify pig interferon-γ(pIFN-γ) cDNA from ConA-activated peripheral-blood lymphocyte by RT-PCR, then was cloned into pMD18-T and sequenced, and recombined with pET32a(+) vector, the target protein His×6 IFN-γ was induced by IPTG;the sequence analysis showed that the nucleotide sequences of the IFN-γ in the study shared 98.6% to 100.0% homology with that of pig published in GenBank, and the identity of amino acid was 96.4% to 99.4%.This recombinant protein, a molecular weight of about 27 ku and mainly presenting in inclusion bodies, was approved to have immunological activity by Western blotting analysis and SDS-PAGE. It laid a foundation for further research of IFN-γ.

This paper was to research the effect of different proportions of fermented cottonseed meal included in diets on the index of blood biochemistry and immune performance in Yellow-feathered chicken. Selected 320 14-day-old healthy broilers, were randomly divided into 4 groups, each group four replicates of 20 chickens. GroupⅠ was the control group fed with the basal diet, group Ⅱ, group Ⅲ and group Ⅳ 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 of the index of blood biochemistry and immune performance in broilers. The results showed that compared with group Ⅰ, the levels of total protein of the early, medium and later period in group Ⅲ significantly increased respectively by 41.96%,52.57% and 28.53%(P＜0.05). The levels of albumin of the early, medium and later period in group Ⅲ significantly increased by 21.31%(P＜0.01), 18.10%(P＜0.05) and 19.00%(P＜0.05).The levels of albumin of serum Ca of the medium and later period in group Ⅲ were significantly increased respectively by 18.03%(P＜0.01) and 51.23%(P＜0.05).The levels of albumin of serum P of the early and medium period in group Ⅲ significantly increased respectively by 34.55%(P＜0.05),31.16%(P＜0.05), and 27.65%(P＜0.01). The levels of cholesterol of the later period in group Ⅳ significantly reduced 21.22%(P＜0.05).The levels of immunoglobulin A (IgA) of the later period in group Ⅱ, group Ⅲ and group Ⅳ significantly increased respectively by 11.07%(P＜0.05), 24.41%(P＜0.01) and 13.07% (P＜0.01).The levels of immunoglobulin G (IgG) of the later period in group Ⅱ, group Ⅲ and group Ⅳ significantly increased respectively by 15.27%(P＜0.05), 28.09%(P＜0.01) and 16.63%(P＜0.01). The levels of immunoglobulin M (IgM) of the later period in group Ⅲ, group Ⅱ and group Ⅳ significantly increased respectively by 26.02%(P＜0.01), 11.13%(P＜0.05), 10.26%(P＜0.05).The index of spleen of the early period in group Ⅲ and group Ⅳ significantly increased respectively by 55.86%(P＜0.05) and 75.17%(P＜0.01). The index of spleen of the medium and later period in group Ⅳ were significantly increased respectively by 30.66%, 44.68%(P＜0.05).The index of bursal of the early period in group Ⅱ significantly increased respectively by 28.08% (P＜0.05). The index of bursal of the early and medium period in group Ⅲ and Ⅳ significantly increased respectively by 69.23%, 46.54%, 39.74%, 37.18%(P＜0.01).The index of bursal of the later period in group Ⅲ was significantly increased by 64.71%(P＜0.05).Therefore, adding 6% fermented cottonseed meal in broilers diet could improve the levels of total protein,albumin, Ca, P, IgA, IgG, IgM and the index of bursal were better than 3%, 9% fermented cottonseed meal added to diets, but 9% fermented cottonseed meal in broilers diet, in reducing the levels of cholesterol and increasing the index of spleen were better than 3%, 6% fermented cottonseed meal added to diets.

This study was investigated the effects of administering beef cattle antimicrobials at subtherapeutic concentrations on the abundance and persistence of tetracycline resistance genes within the whole microbial community of fecal waste.Cattlwere administered chlortetracycline, chlortetracycline plus sulfamethazine and no antimicrobials. Model fecal deposits (n=3) were prepared by mixing fresh feces from pens into a single composite sample. Real-time PCR was used to measure concentrations of tetracycline resistance genes tet(B), tet(C), tet(L), tet(M), tet(W) and 16S rRNA in DNA extracted from composite feces after 7, 14, 28, 42, 56, 70, 84, 98, 112, 126 and 175 days in the field. The concentrations of 16S rRNA in feces were similar across treatments and increased by day 56. Generally, initial concentrations of tetracycline resistance genes were greater (P＜0.05) in fecal pats from animals fed chlortetracycline. For all fecal treatments, tet(B) and tet(C) increased 1 to 2 log units by 56 d, and then decreased to the initial 7 d levels by 175 d (P＞0.05). The concentrations of tet(M) and tet(W) were greater than other tetracycline resistance determinants. Tetracycline resistance genes could persist in fecal waste from cattle beyond 175 days and the initial load of some genes might underestimate concentrations at later time points. Temporal changes in the concentrations of resistance genes were likely due to shifts in microbial populations.

Bioactive peptides belong to a multifunctional factor.It receives more and more attention by people with discovering its biological activities and physiological functions. The author reviewed different physiological functions according to the different sources of bioactive peptides in order to provide reference for the research and development of biological active peptide.

The objective of this study was to investigate the effects of supplementing different ratios of short-medium chain fatty acids (SMCFA) and long chain fatty acids (LCFA) on the immune function in mid-lactating dairy cows. Seventy-two Chinese Holstein cows were blocked by the milk yield, lactation day age and parity, and then assigned to six groups. The animals were fed 0 g/d, 400 g/d LCFA, 80 g/d SMCFA, 320 g/d LCFA, 400 g/d butter fat, 240 g/d SMCFA and 160 g/d LCFA and 400 g/d SMCFA respectively on the basis of ordinary diets for the control, group A and group B,group C, group D and group E. The experiment lasted 63 days and on the last day, blood samples were collected to determine the concentrations of serum immunoglobulin IgA, IgG, IgM, prostaglandin E₂ (PGE₂ ) and cytokines. The results showed that among all the groups, IgM and PGE₂ were not affected by supplementing fatty acids (P＞0.05). The concentrations of IgA in group A was higher than that in groups B and E and the concentrations of IgG in groups B and D were higher than the other four groups(P＜0.05). Interleukin-4 and 10 increased after supplementing fatty acids, especially in group D(P＜0.01). In conclusion, supplementing fatty acids which consisted of 240 g/d SMCFA and 160 g/d LCFA could improve the immune functions in mid-lactating dairy cows well.

The experiment was conducted to study the effects of Enterococcus faecium T013 complex preparation of the diets on the growth performance, fecal microbial, environmental indicators and serum indicators in growing pigs.120 crossbred (Duroc× Landrace× Large White) weaned piglets with average body weight of (31.75±3.78)kg were randomly assigned to three dietary treatments: the control(no supplementation), test Ⅰ(3% supplementation), test Ⅱ(6% supplementation)) with four replicates per treatment and ten pigs per replicate. The feeding trial lasted for 28 days. The results showed that Enterococcus faecium T013 complex preparation could increase the daily feed intake and average dairy gain, lower feed conversion, but the difference was not significant(P＞0.05), and could significantly reduce diarrhea rates(P＜0.05),experimental group Ⅱ could significantly reduce fecal pH and the number of Escherichia coli, increase the number of lactic acid bacteria(P＜0.01), experimental groups could significantly reduce emission of NH₃ (P＜0.01), experimental group Ⅱ could significantly increase the serum IgG levels and reduce blood urea nitrogen levels (P＜0.05);TP,ALB,A/G and GLU showed not effect significantly(P＞0.05).Therefore, Enterococcus faecium T013 complex preparation could reduce diarrhea of the growing pigs,improve the intestinal microbial environment and reduce emissions of NH₃, and enhance immunity of pigs.

Beef is a nutritional value of food which has high protein, low fat and low cholesterol. With the level of health and health awareness increased, more and more people were conscious of health and diet, the consumers just paid attention to evaluate the value of beef nutrition and asked to produce functional beef to improve the health and prevent the occurrence of diseases. So it had been the hotspot that all kinds of beef production contained functional constituents. In this article, recent researches on manipulation about beef abundance of functional fat acid production technology were reviewed.

Milk protein is an important component of milk, containing almost all the essential amino acids which are required by humans. However, many factors affect the synthesis of milk protein, such as the diet composition, the genetic factors, and the rearing environment and so on, but the specific mechanism is not clear. Therefore, it has become a hot and emphasis research on how to improve the content of milk protein by studying the milk protein synthesis mechanism combined with nutritional control measures. This article reviews the nutrition levels and mechanisms which affect milk protein synthesis.

Nutritional value of feedstuff has a direct impact on the animal performance and the quality of animal products. In vitro methods for laboratory estimations of feed degradation are important tools for ruminant nutritionist. These methods are measured either substrate disappearance by quantifying incubation residues or they record fermentation products such as microbial biomass, volatile fatty acid or gas production. In vitro gas production technology, which is based on the relationship between rumen fermentation and gas production, is a in vitro rumen simulation technique. It is simple, economic, quick and low cost, and wildly used to evaluate the nutritional value of feedstuff in ruminant production. The aim of this study is to describe the application and research advances of in vitro GWgas production technology on animal nutrition and classic mathematics models in this area.

The purpose of this project was to study the anti-oxidant activity in vivo of total flavonoids from Oxytropis glabra DC. Ultrasonic-assisted extraction was adopted to extract total flavonoids from Oxytropis glabra DC. Mice model of carbon tetrachloride (CCl₄) liver injury was used to assay the antioxidant ability in vivo of total flavonoids. In mice serum and liver, total flavnoids from Oxytropis glabra DC could markedly increase the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and decrease the content of maleic dialdehyde (MDA) and lipofuscim(LP). Therefore, total flavnoids from Oxytropis glabra DC had antioxidant activity in vivo to some extant, which provided for its further developing and researching.

N-carbamylglutamate (NCG) is a kind of new pig feed additive, which can improve the reproductive performance including increasing the total piglets born per litter and the birth weight of all piglets born of sow, enhancing the fecundity of boar and promoting the growth and development of piglets through promoting the endogenous synthesis of arginine. The use of NCG in pig was reviewed to provide examples for hoggeries to improve their efficiency of pig production.

The purpose of this study is to compare the growth of buffalo ovarian granulosa cells (GCs) cultured with different culture medium, and research the proliferation, karyotype and apoptosis of GCs to know well the growth characteristic of GCs in vitro, so that to establish a culture system of buffalo ovarian GCs in vitro. The results showed: the viability of GCs was about 60% after isolation. The growth speed and density of GCs were more dominant as culturing in DMEM medium than that of TCM-199 and DMEM/F12 medium. The GCs began to proliferate after cultured for 24 h, and reached a proliferation peak during cultured for 3 to 5 d. Karyotype analysis displayed that beyond 85% of GCs had normal karyotype in the 1, 3, 5, 7 passage, which did not show that significant difference among these passages. The apoptosis ratio of the 5 and 7 passages of GCs was higher as comparing with the 1 passage (P<0.05, P<0.01). These results indicated that the DMEM medium was more suitable for GCs culturing in vitro, and the GCs could be passaged and keep relatively stable karyotype, but the apoptosis ratio would be elevate with the increase of the passage in vitro culture.

To obtain the transgenic swine of liver specific expression Cre recombinase, we constructed porcine fibroblast line of liver specific expression Cre recombinase. The promoter of alpha1-human antitrypsin and the BGHpolyA were amplified respectively from the homo-blood genome and the vector of pcDNA3.1(+) through PCR . Cre recombinase gene of porcine origian was obtained by the method of overlap PCR. The FRT2neo cassette was digested with XhoⅠ and SalⅠ from the vector pGC-FRT2neo, which was awarded by professor Stefano Casola of Italy. We combined the above four fragments through SOE-PCR splicing and restriction enzyme ligation, and then linked the four fragmens to the eukaryotic expression vertor pC1-neo. The recombinant was transfected into porcine fibroblast by Lipofectamine^TM2000. Successfully constructed the Cre recombinase expression vector and integrated to the genome of the porcine fibroblast and obtained the fibroblast of liver specific expression Cre recombinase. The obtaining of the porcine fibroblast of liver specific expression Cre recombinase should be of great value to the construction of the liver specific expression Cre recombinase transgenic swine.

Plateau environment is characterized by low oxygen partial pressure, plateau nation and animals settled in highlands at different time and levels height, subject to different degrees and different times of hypoxia selection. Physiological characteristic of blood is an important performance of hypoxia adaptation, and blood physiological parameters can directly reflect organism’s adaptability to hypoxic environment. At the same time, genes related hypoxia adaptation play very important roles in the regulation process of body hypoxia adaptation. The researchs on hypoxia adaptation of plateau nation and animals are concentrated in blood physiology and its molecular mechanism, more and more genes that involve in regulation pathways of hypoxia adaptation are found with developing of molecular biology techniques. In this paper, we review the progress of research on blood physiological characteristics and related genes of hypoxia adaptation.

The skin sample of cashmere goat was incubated in 0.02% trypsin at 4 ℃ overnight and the epidermis was separated,then they were placed in asterile tube containing 0.25% trypsin at 37 ℃ for 30 minutes and the dissociated cells were adhered with collagen type Ⅳ to culture dishes for 10 minutes. 10 minutes later, the unattached cells were removed and the rapidly adherent epidermal cells were cultured in stem cell growth medium(80% DMEM-F12 + 20% FBS + 25 μg/mL hydrocortisone + 100 IU/mL penicillin + 100 μg/mL streptomycin + 15 μg/mL insulin + transferring + 20 μg/mL EGF) for 24 h. And then dissociated cells were cultured on layer cells which the adult cashmere goat fibroblast were treated with 20 μg/mL mitomycin C for 4 h. After 2 weeks, these cells could form different shapes of clones. Dyed with AKP, it showed deep purple and cells were judged as positive.

In order to research the effects of swimming stress on lipid peroxidation of kidneys, lungs and brain tissues in mice. 50 Kunming mice were conducted the swimming experiment, 10 mice selected randomly were sacrificed before swimming and at 10, 20, 30, 50 min after swimming respectively, kidneys, lungs and brain tissues of mice were collected, maleic dialdehyde (MDA) contents, superoxide dismutase (SOD) activities and glutathione peroxidase (GSH-Px) activities were determined by spectrophotometry. This results showed that MDA contents of kidneys, lungs and brain tissues of mice at 10 or 20 min after swimming were increased significantly (P<0.05), and then decreased in a different extent; SOD activities of kidneys during the swimming process continued to rise, and 30 and 50 min groups significantly increased (P<0.05), SOD activities of lungs at 20 min after swimming reduced markedly (P<0.05) and increased slightly at 30 min after swimming, a little decrease was found at 50 min after swimming again, SOD activities of brain tissues were no significant difference during the whole swimming progress; GSH-Px activities of kidneys at 20 and 30 min after swimming significantly increased (P<0.05) and decreased markedly at 50 min after swimming(P<0.05), GSH-Px activities of lungs during the first 40 min swimming process continued to decrease and significantly reduced at 30 min swimming group (P<0.05), GSH-Px activities of brain tissues at 10 and 20 min after swimming significantly decreased (P<0.05) and then fluctuant changes occured. These results indicated that lipid peroxidation of kidneys, lungs and brain tissues in mice was affected by swimming stress and speculated that swimming stress may cause lipid peroxidation reaction of the whole body of mice.

To study the skin pharmacokinetics of interferon α1b spray, the pharmacokinetics of interferon α1b spray on the skin was carried out in the integral, permeable and injured skin of the minipigs. 1, 3, 5, 25 μg/mL of recombinant human interferon α1b were consperged onto animal’s skin. Skin tissues were shearing and homogenated at 0,1,2,4 h after consperge. The biological activity of interferon α1b was measured to show the drug’s antiviral effect. The interferon α1b was detected in integral skin and achieved to therapeutic concentration in permeable and injured skin of minipigs within 4 h after consperging; the concentrations of interferon α1b in tissues were correlated with dosage and time, those differences between groups were significant (P<0.05). 5 μg/mL interferon α1b spray showed antiviral effect on skin region when sprayed every 4 h.

Defensins are a group of small molecular antimicrobial peptides and distribute mostly in animals and plants. The paneth cells defensins are kinds of antimicrobial peptides which have existed in the paneth cells of some mammal’s small intestinal crypts and have important roles in natural immunity of intestinal tract. The classification, distribution, molecular characteristicsm, antimicrobial activities and immune function of paneth cells defensins at home and abroad were summaried.

The polymorphisms of MspⅠ site at exton 4 of RBP4 gene in Yorkshire, Landrace and Duroc populations were detected with PCR-RFLP method, meanwhile the total number born (TNB), number born alive (NBA), number of alive weaning litter (NW), body weight of litter (BWL), weaning weight of litter (WWL) and weaning survive rate of different genotypes for the 2nd and 3rd parity were measured, and then the relationship between reproductive performance and different genotypes was analyzed in this study. The results showed that the polymorphic sites of RBP4 genes were found in all three breeds, the results of χ² test indicated that this site of Yorkshire didn’t fit Hardy-Weinberg equilibrium(P<0.05), while this site of Landrace and Duroc fitted Hardy-Weinberg equilibrium (P>0.05) . In addition, the dominant genotype in Yorkshire was AA genotype,however in Landrace and Duroc were BB genotype. For the reproductive performance,BB genotype of Landrace in 2nd parity produced 0.97 TNB, 1.28 NBA, 0.92 NW, 1.74 kg BWL and 4.42 kg WWL more than AA genotype, and produced 0.92 TNB, 1.02 NBA, 0.98 NW, 1.43 kg BWL and 4.98 kg WWL more than AA genotype. On the whole, BB genotype was also favourable genotype in 3rd parity.

To investigate the germ-carrying condition, molecular evolution and transitional condition of stx in 5 Escherichia coli O157 strains which were separated and identified from Henan and Gansu province during 2009 to 2010, the stx gene of the isolated strains was amplified and sequenced, and phylogenetic analysis had been completed. The results revealed that all of the isolated strains had stx1 and stx2 gene: the homology of nucleotide and amino acid of five isolated strains were high with each other; the stx1 gene of the five isolated strains showed that a high degree of omology with those referenced Escherichia coli O157 strains deprived from goat and food and their genetic distance of the cladogram are near; the stx2 gene of the isolated strains showed that a high degree of homology with those referenced Escherichia coli O157 strains deprived from cattle and human,though the isolated strains were in one main branch according to phylogenic trees of stx2 gene, the genetic distance of the isolated strain 27 was the farthest with other strains and only the strain 27 was in one secondary branch; the isolated strain L37 and W were in one secondary branch with the referenced strains deprived from cattle; the isolated strains 12 and 50 were in one secondary branch with the referenced strains deprived from human. We could infer from these that stx1 of the five isolated strains might be Escherichia coli O157 strains deprived from food or goat; the stx2 of L37 and W might deprive from cattle, 12 and 50 might from human while 27 was not apparent. The research revealed that all the five isolated strains had stx1 and stx2 gene, but the origin of the two genes were different.

The expression products of insulin-like growth factor-Ⅰ (IGF-Ⅰ) gene play an important role in muscle growth of pigs. In this research, IGF-Ⅰ gene was chosen as candidate genes for growth traits of Saba pigs. The HhaⅠ polymorphism of the gene was investigated by using PCR-RFLP technology, and the relationship between the polymorphism and 12 growth traits was analyzed. The results showed that there was a higher degree of polymorphisms on IGF-Ⅰ gene in the population. AA, AB, BB genotypes of IGF-Ⅰ gene were detected in Saba pigs, which frequency were 0.3473, 0.5149, 0.1378, respectively. The frequency of A and B were 0.6048 and 0.3952, respectively. The gene loci was in Hardy-Weinberg equilibration. The genetic variation of IGF-Ⅰ gene of Saba pigs had significant influence on growth traits (P<0.05 or P<0.01). The favorable genotype of the body weight and the daily gains at different growth stages was AB genotype,and the favorable genotype of the body size traits at 6 months was BB genotype except partial traits was AB genotype. Combining with the previous results, it could be primarily deduced that IGF-Ⅰ gene was probably a major gene or a QTL linked gene that associated with growth traits in pigs.

Reproductive traits of sheep has became an important research area in sheep industry. Genetic improvement of sheep fecundity is the only way to improve sheep production levels in China.The annual ewe lambing ability is mainly affected by the impact of seasonal estrus and the litter size of single births. In this view, the author gives a brief review on ovulation and seasonal estrus related genes and miRNAs, and prospects the genome-wide comparison used in the detection of QTL about reproductive traits.

The purpose was to investigate the association between the polymorphisms of PRLR gene exon 10 and reproductive traits in Songliao Black pig and Landrace pig. Polymorphisms of PRLR gene exon 10 were detected by PCR-RFLP in Songliao Black pig and Landrace pig, then their genetic effects on reproductive traits were analyzed by the least square method. The results showed that there were two alleles (A and B) and three genotypes (AA, AB and BB) in the two pig species, and the frequencies of the dominant allele A were 0.551 and 0.578 in Songliao Black pig and Landrace pig respectively.The weaning piglets number, the number of nipples of individuals with AA genotype were significantly higher than the other two genotypes (P＜0.05). The TNB、NBA of individuals with AA genotype in primiparous Songliao Black pig were significantly higher than other genotypes (P＜0.05), other reproductive traits were not significantly affected by genotype, but showed the genotype AA＞AB＞BB trend.

As one of lately developed small RNA molecule, MicroRNAs(miRNAs) play crucial roles during a variety of physiological and pathological processes of Eukaryotic cells, by means of post-transcriptional regulation. Animal meat productive trait is a quantitative trait. Muscle’s unit is muscle fiber, which is mainly composed of myosin, but myosin’s composite is regulated by internal and external environment. In this article, the recent studies of miRNAs in main livestock and poultry’s muscle growth and development are reviewed, so as to offer useful suggestion for miRNAs research.

This experiment aims to investigate Chinese medicinal compound polysaccharide’s (CMCP) influence on mice blood index with different components and different degrees of purity. In our experiment firstly we select cyclophosphamide to build mice immunity-inhibition model and then we respectively gavage CMCP with different components and different degrees of purity, compound total polysaccharide and compound water decoction into mice to measure their effects on mice’s peripheral hemogram. The result shows that polysaccharide A2, purity polysaccharide B2, compound total polysaccharide and compound water decoction can significantly increase the number of erythrocytes, the number of leukocytes, neutrophilic granulocyte’s percentage in leukocyte classification number and the percentage of lymphocytes in mice. The result also shows a very significant difference from the model group (P<0.01). Therefore it can be seen that polysaccharide A2, purity polysaccharide B2, compound total polysaccharide and compound water decoction can improve reduced peripheral hemogram in mice blood index, which is caused by cyclophosphamide, to regulate immune imbalance.

Porcine epidemic diarrhea was a kind of serious disease in the pig industry, a variety of reasons might cause diarrhea, which brought to the difficulty of clinical diagnosis. In this study, nested PCR technology successfully used in the rapid diagnosis of clinical cases of porcine epidemic diarrhea, which provided a new way of thinking for the rapid diagnosis of clinical cases of porcine epidemic diarrhea.

Porcine reproductive and respiratory syndrome(PRRS) is one of serious swine diseases that has been devastating the swine industry. Since 2006, HP-PRRS has been outbroken from south to north in China,fast and accurate diagnostic methods and techniques to detect the pathogen play decisive roles in the entire PRRS prevention and control. This paper describe the PRRSV diagnostic methods and the latest technology in order to provide technical support and reference for PRRS diagnosis and prevention.

One local strain of porcine parvovirus was isolated from the dead fetus in Fujian province. The isolated virus could cause typical cytopathogenic effect (CPE) in PK-15 cell at hour 36 after infection such symptoms as shrunk, aggregated, and detached. The isolated virus could be neutralized by the positive sera. The DNA extracted from the virus isolate was subjected to polymerase chain reaction (PCR) analysis, a set of primer which can be used to amplify 531 bp fragment in the gene of PPV was designed. The homology of VP2 gene sequence between the isolated viruses and NADL-2 from GenBank was 99.2%, it was indicated that the isolated virus was porcine parvovirus (PPV). In coclusion, this study laid the foundction for furthen development of the pathogenic mechanism of virus, epidemiology, diagnostic studies and vaccine.

A virus strain was isolated from the liver of a raccoon dog, which was suspected as the canine distemper. It was identified to be a canine distemper virus by morphology test, hemagglutination test, indirect immunofluorescence assay and molecular virology test. The successful isolation of the virus founded the base of studying the molecular biological characteristics and pathogenic mechanism of the virus and construction of gene engineering vaccine.

To measure organ weight and organ coefficient of outbred Wuzhishan mini-pigs (WZSPs), and analyze the factors that affect organ weight and organ coefficient.74 common WZSPs(43 males and 31 females) were chosen for this study, and the body weights and 6 organs weights were determined, then the organ coefficients were counted. Based on ages and body weights, difference, correlation, and regression analysis in organ weights and organ coefficients were performed respectively. Only few organ weights of different aged animals showed significant differences between males and females (P<0.05), but there was no significant difference for the organ coefficients(P＞0.05). Results of ANOVA showed that organ weights increased significantly in accordance with the growths and increases of body weighs(P<0.0001); and the organ coefficients of heart, liver, lung, and stomach decreased significantly with the growth of animals(P<0.0001), but the decreases of spleen and kidney coefficients were not significant(P＞0.05); while all the organ coefficients decreased significantly with increases of body weights(P<0.01). Multiple regression analysis demonstrated that organ weight and organ coefficient of the same organ were affected by body weight on different levels. Among the effect factors on organ weights and organ coefficients, the effects of sex differences were mild, while age and body weight showed great effects, and body weights were the key factors. Organ weights increased significantly and organ coefficients turned opposition in accordance with the increases of body weighs.

In recent years, with the rapid development of large-scale pig farms and the continuous improvement of intensification, the prevalence of swine disease is increasingly severe. With its safety, no toxicity and no side effects, as a new immunostimulants and immune adjuvant, astragalus polysaccharides are gradually known to most farmers and studied by researchers. This review summarizes the progress in the research of immune fuction of astragalus polysaccharides in piglets, providing a reference for the prevention of swine in veterinary clinical practice.

21st,with the development of human life and the level of health, the demand of green food (meat, eggs, milk) is growing, consumers are more concerned about food safety. Countries in the world, carry out the study of drug substitute for feed additives especially in Europe, Japan and other developed countries. Research and extension of green safe feed additive are encouraged and promoted actively, probiotics attract the world’s attention. But scientists still need explore the relationship between stability and continuity of such product and physiological, biochemical, nutritional aspects. Domestic and foreign scholars are working on the study of bacteriocin (antibacterid peptides or peptide antibiotics), with the progress of study and in-depth at home and abroad, people have found that these peptides not only have anti-bacterial, anti-fungal effect, but also have anti-parasite, virus, cancer and other functions, in medicine and food preservation technology, it becomes a hot research. In today’s food safety, it is possible that bacteriocin will do as a effective and safe feed additive,they need intensity and be developed, bacteriocin genetic engineering, protein engineering and molecular screening methods will continue to research and breakthroughs, it will alternant antibiotics and product green food for human finally.

One virus strain was isolated from the samples of brain and internal organs of a death piglet from a hogpen in Beijing. ST cell line shrunk, aggregated and detached from the culture system in 24 h approximately. The isolate could be neutralized by PRV standard positive serum. Rabbits were injected with the isolate and the typical clinical syndrome could be observed, such as extra tickle. According to the gD gene sequences of PRV in GenBank, a pair of primers was designed, a specific fragment was amplified by PCR, the results of sequencing analysis showed that the amplified fragment was the gene sequence of the PRV. All of the evidences indicated that the isolated virus was pseudorabies virus, and was named Beijing strain in accordance with separate location.

6 strains of 120 90-day-old Daheng quality broilers were collected, every strain including 10 females and 10 males, their contents of IMP and amino acids in the chest muscle were determined. The results indicated that: ①there were different IMP content among strains, and the highest content was strain S08; IMP content between strains were significantly different (P＜0.05), except between S03 and S08, S02 and S06, S02 and S08×S01, S06 and S08×S01. ②The content of glutamic acid, glycine, aspartic acid, alanine and arginine showed differences among these strains. The hybrid strain S08×S01 showed the highest content of five amino acids. ③There were no significant differences between cocks and hens for most strains of IMP(P＞0.05). There were significant differences between cocks and hens for most strains five amino acids content(P＜0.05).

The pigs fed with high grade corn or moldy corn were checked for swine fever antibodies and disease respectively. The pigs fed moldy corn had low levels of antibodies against swine fever, and there was zearalenone poisoning secondary for swine fever.The pigs fed with high grade corn had qualified levels of antibodies against swine fever, and no swine disease occured.

Through selecting the kinds of adjuvant and its dosage, investigating pharmaceutical technology and stability in granules preparation procedure, the assay was aimend to establish a optimum granules preparation technology of traditional Chinese medicine compound ‘Modified Pulsatilla’. Taking quality of soft material as appraisal indice, investigating the quantity ratio between medicine solution and adjuvant, taking appearance, clear degrees, water content, loss rate of granules quality as appraisal indices to study of the dosage of soluble starch, the dosage of α-lactose, the dosage of dextrin and the concentration of HPMC, and at last to establish a optimum granules preparation technology. When the ratio was 5∶1 between medicine solution and adjuvant, the quality of soft material was good. When the adhesive phase was 0.3% HPMC and the ratio was 3∶1∶1 among soluble starch, α-lactose and dextrin, every appraisal indice of granules was best. Through the test, it was proved that the traditional Chinese medicine compound ‘Modified Pulsatilla’ granules could be preparated in the condition of laboratory. The preparation technology was simple and clear, the granules were forming effective, high stability and up to specification which was in the pharmacopeia of China.

Sampling methods are an essential part of research in veterinary science. Based on the agricultural professional standard of the People’s Republic of China which is sampling methods for laboratory investigation of animal diseases and clinical practice and research experience, this article describes the types, quantities and operations of the sample collection for the content and purpose of different studies. Also, collection, preservation and processing methods of sample are introduced. It is paid more attention to the latest domestic and international progress on the sampling methods. Thus, these would provide a reference for veterinary scientific research in the future.