In the study, we mainly did the research about efficient expression and characterization of bovine interferon-γ in suspension Sf21 cells. Bovine IFN-γ gene was amplified by RT-PCR, and inserted into the pFastBac^TM HTA to get the recombinant plasmid, named as pFastBac^TMHTA-IFN-γ. After transformation and transfection,the recombinant baculovirus was obtained. The target protein named rBovIFN-γ was successfully expressed in Sf21 insect cells during suspension culture. We identified the rBovIFN-γ with SDS-PAGE,Western blotting and mass spectrum(MS). Western blotting analysis showed that the recombinant protein had good activity. The protein MS identification results confirmed the recombinant bovine IFN-γ expression via the baculovirus expression system. MDBK/VSV antiviral test results showed that the rBovIFN-γ had high antiviral activity at 1.05×10⁵ IU/mg.

The Sip gene of the isolate M7 of Ⅰa serotype Streptococcus agalactiae from bovine was amplified using the PCR method,cloned into pGEM-T Easy vector and then sequenced. The molecular characteristics of Sip gene and its expression protein were analyzed using a variety of biological software. The results showed that Sip gene of M7 was 1305 bp and did not appear the gene deletion, the Sip gene of nucleotide sequence derived from M7 had a 98.0% to 100.0% of homology and a 97.2% to 99.8% amino acid homology among the corresponding sequences published in GenBank.Furthermore, the Sip gene of M7 shared of 100.0% nucleotide homology and 99.8% of amino acid identity with that of Ly2(FJ808732)strains in China and GB00549(FJ752159)in USA, respectively. The expression protein of Sip gene was a stable outer membrane protein,which was secretion protein and hydrophobic highly. The polypeptide contained a LysM functional domain which was related to immunoregulation between the N-terminal 52nd to 95th amino acid residues and a signal peptide consisting of 25 amino acids, which of cleavage site between 25th and 26th amino acids. Moreover there were a number of B cell and T cell epitopes on the protein. The results illustrated the Sip protein of M7 strain was conservative immune proteins without missing LysM superfamily domain.

To study the mutants of a highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV), HPPRRSV-JXA1 strain was continuously passed in Marc-145 cell cultures with or without anti-JXA1 pig sera for 30 passagesand named JXA1-Abf30 and JXA1-f30 respectively. Infected cells were collected for extraction of viral RNA every 10 passages,ORF3 and ORF5 genes were amplified,cloned and sequenced. Sequence analysis indicated that numbers of nucleotide mutations on ORF3 and ORF5 genes in JXA1-Abf30 and JXA1-f30 strain were 18 and 9, numbers of amino acids mutations were 15 and 3 respectively. The ratio of nucleotide mutative speed between the two strains was 186% , The ratio of amino acid mutative speed of two strains was 500%, non-synonymous VS synonymous mutation ratios (NS/S) of ORF3 and ORF5 genes in two strains were 5.0 and 0.5. Besides,there were 4 stable NS mutations in ORF5 gene and two of them (related to AA31 and AA190) were related to the known epitopes during passages with anti-PRRSV sera,which were not detected in passages without specific antibody. It indicated that mutations in PRRSV under antibody immune selection pressure were significantly faster than PRRSV without antibody immune selection pressure, anti-PRRSV sera had strong immune selective pressures on viral evolution in its ORF5 genes, but for ORF3 gene, mutation influence was less than ORF5 genes.

To enrich the basical genetic information and explore the sequence structure and the regulation mechanism of muscle differentiation in local excellent variety Longlin goat MyoG gene. A pair of primers was designed to amplify the sequence of MyoG gene from Longlin goat genome. The results showed that the sequence of MyoG gene with 2419 bp consisted of 3 exon, 2 intron and some of 5'and 3'untranslated regions. With 675 bp coding sequences, encoding a total of 224 amino acids, and the peptide chain had not signal peptide and transmembrane structure.Homology analysis showed, comparing the coding sequences and coding amino acids of Longlin goat MyoG with other species, the cluster result was similar to the phylogenetic tree. Both showed that Longlin goat and sheep, cattle, wild boar were closest relative, the homology of amino acid sequences between them was above 95%. So the mammalian MyoG gene was conservative in evolution, and cloning, sequence analysis and structure prediction of Longlin goat MyoG gene provided the biological information and theoretical basis for researching the gene expression and regulation, improving the muscle quality and developing goats breed.

In order to obtain protein expression map of nanoparticles copper(nano-copper) toxicity in rat liver, this study constructed the model of the nano-copper in rats, and obtained 2-DE maps of nano-copper toxicity in rats group and control group used by two-dimensional gel electrophoresis technique, PDQuest 8.0 analysis showed that the total 43 protein spots significantly differently expressed in rat liver of 200 mg/kg copper nanoparticles group which 27 spots were up and 16 spots were down. The toxicity of nano-copper in rats liver was studied by proteomics approach in the paper, the results would establish the foundation for clarifying the mechanism of the toxic effects of nano-copper at protein level.

In this study, two T cell epitopes genes of hamagglutinin conserved domains of avian influenza virus H9N2 strain and M2 gene of extracellular domain were synthetized and joined as HA-M2 multiple-epitopes fusion gene of avian influenza virus by RT-PCR. The recombinant eukaryotic expression plasmid of HA-M2 fusion gene containing enhanced green fluorescent protein (EGFP) report gene was constructed, and transfected by lipofectin to prepared duck embryo fibroblasts (DEF). The fluorescence expression was directly detected with fluorescence microscope, and the expression of HA-M2 was tested by RT-PCR and indirect immunofluorescence assay (IFA) respectively. The results indicated the successful construction of the eukaryotic expression plasmid HA-M2-pEGFP-N1, and showed that HA-M2 gene was expressed efficiently in transfected DEF. It provided a basis for the study of new universal influenza vaccines and further research on interaction of HA gene and its target cells and as for infection mechanism of AIV.

A set of primers used for reverse transcription loop-mediated isothermal amplification (RT-LAMP) detection was designed based on the conserved nucleoprotein gene of bovine parainfluenza virus 3(BPIV-3) complete genome sequence submitted in GenBank. The usefulness of RT-LAMP for rapid preclinical detection of BPIV-3 infection was evaluated. The reaction could be finished in 1 h under isothermal condition at 63 ℃. This RT-LAMP assay had a detection limit of 0.069 fg/μL per reaction, was higher sensitivities than that of RT-PCR. The specificity of this assay could be easily confirmed by agarose gel electrophoresis, color reaction or turbidity comparison. As a result, the RT-LAMP assay was an ideal method for detecting BPIV-3 in laboratory and primary diagnosis of clinical infection.

In order to construct of eukaryotic expression vector of hyaluronidase 2(Hyal-2) and its expression in bovine mammary epithelial cells, the full-length gene of Hyal-2 was amplified by overlapping PCR with two pairs of primers. Bringing HindⅢ restriction enzyme sites in upstream primer and BamHⅠ restriction enzyme sites in downstream primer. The expression recombiant plasmid was constructed by inserting the amplified gene into pEGFP-C1. The purpose gene was connected into eukaryotic expression vector pEGFP-C1 by connecting reaction. The vector was tranfected into bovine mammary gland epithelium cells by Lipofectamine 2000. The expression of Hyal-2 was verified by RT-PCR and Western blotting methods respectively. Sequencing analysis results showed that the nucleic acid homology was 99%, the amino acid homology was 99%. It was identified that the eukaryotic expression vector of Hyal-2 was successfully constructed and expressed in bovine mammary epithelial cells successfully.

To develop a detection method for glucoamylase in honey, BALB/c mice were immunized with the glucoamylase from Aspergillus niger. Splenocytes of the immunized mice were fused with SP2/0 cells with PEG. 12 monoclonal antibodies(McAbs) against the glucoamylase were obtained and identified. 10 hybridoma cell lines represented subtypes IgG1 and the other were IgG2b, and their light chains were κ chain. Western blotting analysis showed that the 12 McAbs specifically recognized glucoamylase. Hybridoma 2H4F9,6H9D8,8F2F11,8F2E9,1A8G6,1C4D5 excreting anti-glucoamylase McAbs were used to be injected into abdominal cavity of mice(ICR). The titers of the McAbs were all above 1∶1×10⁴. Six strains of monoclonal antibodies were confirmed to be specific for four different kinds of epitopes on glucoamylase competitive binding assay. Among them, McAb-6H9D8 and McAb-8F2F11 might direct to the same antigenic determinant on the GA, McAb-1A8G6 and McAb-1C4D5 to the second antigenic site, McAb-8F2E9 to the third antigenic site, McAb-2H4F9 to the fourth antigenic site.

Specific primers were designed according to ApoB100 gene sequence reported in GenBank in this study. Cloning was carried out by the pGM-T vector, recombined plasmid DNA was cut by BamHⅠand EcoRⅠenzymes and then sequencing. The recombinant expression vector pET-28a-ApoB100 was constructed with target gene and pET-28a vector, and transformed into Rosetta(DE3),then induced by IPTG. The expression product was identified by SDS-PAGE and Western blotting. The results showed that the homology of the cloned ApoB100 gene was 100% to that reported in GenBank. SDS-PAGE analysis showed that the molecular weight of recombinant protein pET-28a-ApoB100 was 35 ku. Western blotting positive results showed that the experiment successfully obtained the target protein.

The reovirus S1733 harvested from chicken embryos allantoic fluid was purified by sucrose gradient purification. The concentration was tested by protein testing kit. Then BALB/c mice were regularly immunized with purified reovirus S1733 contain 100 μg protein. After injecting four times, the spleen cells were collected and infused with myeloma cell SP2/0 follow ratio 5∶1. After selection in time and 3 times clone by limiting dilution. The two monoclonal antibodies were detected by indirect ELISA, Dot-ELISA, and immunofluorescence methods. The results showed that the purification protein was 43 mg/mL. Two hybridoma cells lines against reovirus named SF6-3K3 and SB2-1K3 were obtained. The titers were all above 10⁵ by indirect ELISA detection. It was not cross with others virus strains in this test by Dot-ELISA. The subtype was IgG1. It indicated that two hybridoma cell lines possessed the favorable specificity. Both of them could be used to detect reovirus in the future.

Infectious bovine rhinotracheitis (IBR) is a highly contagious, infectious disease that is caused by bovine herpes-virus-1(BHV-1). In addition to causing respiratory disease, this virus can cause conjunctivitis, abortions, encephalitis, andgeneralized systemic infections. IBR was originally recognized during the early 1950’s in Colorado feedlots. There is no direct treatment for the infection and the best way to control IBR is to vaccinate before a disease outbreak occurs. Nowadays, conventional IBR vaccines have been, and still are, routinely used in many countries, the glycoprotein E (gE)-deleted vaccines, either as a live or as a inactivated vaccine, have been commercialized in the European Union. In this review, we discussed the progress in development of gene-deleted vaccine, subunit vaccine, viral vector vaccine and DNA vaccine.

In order to enrich the cytokine biological data of Guangxi Bama miniature pig, the lymphocytes of Bama miniature pig was separated from peripheral blood and the total RNA was extracted from the peripheral blood lymphocytes, respectively. The target genes of cytokine were amplified by RT-PCR and cloned into pMD18-T vector and transformed into E. coli DH5α, respectively. We chose positive clones and submitted them to the NCBI after sequencing the recombinant strain. Comparing homology of the cloned sequence with the known sequence, and using the software to analysis the cloned sequence. The results showed that the gene length of cloned cytokines IL-2, IL-12, IFN-γ, TNF-α of Bama miniature pig were 338, 377, 380, 402 bp,respectively. It was consistent with the expected length of target gene. The sequences analysis showed that the homology of the cloned genes was relatively high with known domestic pigs or wild boars. The sequence homology of IL-2, IL-12, IFN-γ,TNF-α were 98.9%, 99.5%, 99.2%, 100%, respectively. The research on cytokine gene sequence of Bama miniature pigs not only enriched the biological data of Bama miniature pigs, but also laid the foundation for research of the cytokine function. It also provided the basis for the quantitative detection of cytokine.

Glycosylation is the most common and most important protein post-translational modification of proteins in organism. It affects the characters and function of proteins significantly. In recent years, more and more drugs have been applied in clinic along with the development of glycobiology and glycosylation engineering. In the paper, we summarized the species of glycosylation modification and their effect on the protein properties. On the basis, the development and application of protein glycosylation engineering in the biological pharmacy were described. Protein glycosylation engineering will inevitably play an increasingly important role in the protein transformation as a new type of protein-modification means.

Ghrelin, a peptide lying widely in animal bodies, is the endogenous ligand for the growth hormone secretagogue receptor(GHSR). Ghrelin, after binding with GHSR in the hypothalamus and pituitary, not only promotes the GH releasing and increases appetite, but also adjusts the functions of digestive system and energy metabolism. This article mainly reviewed the recent developments on structure, distribution and biological effects of ghrelin as well as its potential applications in animal husbandry, which maybe promotes the further research of ghrelin.

Total RNA was extracted from bovine,the cDNA encoding PU.1 protein was cloned using reverse transcription polymerase chain reaction (RT-PCR). The PU.1 gene was subcloned into pIRES2 DsRed-Express2 vector and then the recombinant plasmids were transferred into H293T cells and bovine fetal fibroblasts cells by the method of lipofectaming. The results showed that we had successfully constructed the expression plasmid of PU.1 gene by fluorescence observation and identification of Western blotting,which layed the foundation for the differentiation of bovine blood monocytes and breeding for disease resistance in cattle.

A slaughter trial was conducted to evaluate the effects of dietary vitamin A supplementing dose on meat quality by using 16 Angus ? Qinchuan crosses steers at 24-month age in a complete randomized design after 4 months of finishing period with diets contained 0,1100,2200 and 4400 IU/kg DM. The results showed that VA dose did not affect subcutaneous fat thickness,mesenteric fat weight/living weight,longissimus area,dressing percentage,pure meat percentage and marbling score(P>0.05) which was better with the decrease of VA dose. But VA dose significantly affected final weight,daily gain,back fat thickness and shear force(P<0.05), back fat thickness and shear force were lowest at 1100 IU/kg DM, and average daily gain which had a tendency was numerically greater with VA dose increment when dietary VA dose was less than 2200 IU/kg DM.VA dose had no significant effect on the chemical composition of LD and GM except for matter and crude ash of GM,matter and crude protein of LD(P>0.05).VA dose had significant effect on drip loss, total pigment concentration and TBARS during aging(P<0.05), and different beef cuts responses to VA dose were not identical. Considering these results, the effect was the best when the Angus譗inchuan steers were fed with the diet contained VA 1100 IU/kg DM.

Eight female Small-tail Han×Polled Dorset lambs, with the average body weight of(8.90±0.38)kg, were fitted with the abomasum canula at the age of 40 days, and fed with diets 1, 2 and 3 during 60 to 80,81 to 120 and 121 to 240 days old respectively,and were supplemented with lysine at the dose of 1.2%,0.8% and 0.4% in diets 1, 2 and 3 respectively, to study the effect of dietary lysine supplementation on the nutrients arriving at the abomasum. The results showed that compared with that of the control group, supplementing lysine to the lambs from 60 to 240 old-days, increased the average amounts of DM arriving at abomasum by 7.01%(P＞0.05),11.59%(P＜0.01) and 10.65%(P＜0.05) respectively, increased the average amounts of OM by 8.48%(P＞0.05),13.01%(P＜0.05) and 12.14%(P＜0.05) respectively,increased the average amounts of CP by 11.12%(P＞0.05),21.73%(P＜0.01) and 12.44%(P＜0.05) respectively,increased the average amounts of microbial protein arriving at abomasum by 13.46%(P＜0.05),26.85%(P＜0.01) and 15.08%(P＜0.05) respectively,increased the average amounts of the total amino acids by 39.22%(P＜0.01),35.52%(P＜0.01) and 28.27%(P＜0.05) respectively,increased the average amounts of the total essential amino acids by 49.21%(P＜0.01),34.40%(P＜0.01) and 28.11%(P＜0.05) respectively,increased lysine by 43.77%(P＜0.01),37.25%(P＜0.01) and 34.38%(P＜0.05) respectively .The lysine arriving at abomasum mainly was combinative lysine both in the experimental and control groups, with less 1% of free lysine. Compared with the control group, by supplementing lysine to the lambs, the average amounts of voluntary intake of DM was increased by 12.57%(P＞0.05),19.38%(P＜0.01) and 16.29%(P＜0.01) respectively, the digested DM in forestomach was increased by 20.81%(P＞0.05),35.64%(P＜0.01) and 19.97%(P＜0.01) respectively,and the daily body weight gain was increased by 17.34%(P＜0.05), 21.30 %(P＜0.01) and 17.76%(P＜0.05) respectively. It was concluded that supplementing lysine to the 60 to 240 day’s lambs could increase the nutrients arriving at abomasum and the digested OM in forestomach, and as a result, increase the supplement of the nutrients to the lamb.

According to the method of the national feed hygienic standards, the experiment detected and safetly evaluated the different growth stages of Pleurotus ostreatus medium matrix component.In order to evaluate the feed value of the Pleurotus ostreatus and realize reasonable utilization of spent mushroom substrate(SMS) resources by detecting on the nutrient components of spent mushroom substrate after different harvest times,we found that the fourth harvest of SMS used as feed had a higher nutrition value and lower production cost. The crude protein content was 10.10% at this moment. Compared with original substrate,the crude protein content was increased by 59.31%,crude fiber content was reduced by 29.57%,and the degradation rate of ligin, hemicellulose and cellulose were increased by 63.62%, 46.60% and 77.06%. Based on the national feed hygiene standards,the content of free gossypol, aflatoxin B₁ and heavy metal in SMS (the fourth harvest) were measured. The results showed that the free gossypol content was only 67.90 mg/kg,aflatoxin B₁ was not detected (＜10 μg/kg).And the content of lead,fluorine,chromium,cadmium and mercury all met the national feed hygiene standards. So it was feasible to use SMS(the fourth harvest) as a new non-conventional roughage resource for ruminants.

Along with the development of swine industry to scale and intensive in recent years, how to improve the reproductive performance of sows has become the hot issues in pig industry. But reproductive performance of sows is influenced by inheritance, nutrition, environment, health and other factors, among the nutritional factors which are one of the most important factors. Taking some simple and feasible nutriology method during different phases in sow raising management can markedly improve sow reproductive performance. In this article many nutritional regulation measures will be put forward based on analyzing and discussing the nutritional factors affecting reproductive performance of sows.

The purpose of this paper was to study the effects of the different levels of germination corn on rumen fermentation parameters, blood components, production and quality of milk in Holstein dairy cows. 15 lactating Holstein cows were selected as the experimental animals and were randomly divided into 3 groups. Each group had five dairy cows. The control group was fed the basal diet(TMR+alfalfa hay). T1, T2 groups were fed with 20% germination corn and 20% fermented germination corn in the basal diet. The results showed that the supplement of germination corn and fermented germination corn could increase the concentration of VFA and NH₃-N in rumen,and decrease pH in the rumen(P＞0.05). The 4% fat corrected milk of T1 and T2 were significant higher compared with the control (P＜0.01),the feed efficiency of the treatment 2 was significantly improved and reduced somatic cell counts compared with the control (P＜0.05). T1, T2 groups could decrease content of cholesterol of milk,and had no obvious affect in blood components. In conclusions,the supplementation of germination corn and fermented germination corn could improve rumen fermentation parameters,milk performance,feed efficiency and signicantly decrease somatic cell counts, and content of cholesterol of milk had a dectrawing trend.

The assay was aimed to investigate the law of trace elements (Cu, Fe, Mn, Zn) and interleukin (IL-1, IL-6) contents in serum using Cyst-regulating powder to treat infertility caused by ovarian disease of dairy cattle.Flame atomic absorption spectrometry and enzyme-linked immunization absorb assay were used to detect levels of the trace elements and interleukin contents in serum from the experimental and normal group before and after treatment.The results were showed as following.①After treatment, the effective rate of follicular cyst was 83.33%, the effective rate of delayed ovulation was 90.91%.②In follicular cyst body, Mn, Zn contents were significantly lower than the normal group in pre-treatment (P<0.05), Fe content was significantly higher than the normal group(P<0.05), after treatment, their content levels were closed to the normal group, and the difference was not significant(P>0.05), the Cu, IL-1, IL-6 contents were closed to the normal group before and after treatment. ③In delayed ovulation, Fe content was signifcantly higher than the normal group(P<0.05),Mn content was extremly significantly lower than the normal group in pre-treatment(P<0.01), Zn content was signifcantly higher than the normal group (P<0.05), after treatment, the contents was not gradually significant(P>0.05),but Cu, IL-1, IL-6 content levels were not significantly compared with the normal group before and after treatment(P>0.05). In conclusion, the effect of trace elements and interleukin in serum can’t be ignored by using Cyst-regulating powder to treat infertility caused by ovarian disease of dairy cattle.

Lactose is a special component of milk and is closely related to milk yield. The factors of influencing lactose synthesis are extensive. The paper reviews the related genes and cell signaling pathways, including glucose transporters, intracellular catalytic lactose synthesis enzymes, hormones (insulin, prolactin, growth hormone, leptin, glucocorticoid ), cell factors(IGF-Ⅰ,IGF-Ⅱ,EGF) and its main cell signal transduction pathway.

The liver is the largest digestive gland in poultry, and its growth and development are vital for playing functions. In the current study we observed the development of broiler liver tissue with the histological method, and compared the differences of the development of liver tissue between fat and lean lines during embryonic period. We found that the lobes of liver become bigger with developing, and the hepatic lobule structure was integrity when chicken embryo was 12-day-old. The hyperplastic and hypertrophic growth lipid droplets in liver cells significantly increased in the later stage of the embryogenesis, and the proliferation activity of liver cells located around was stronger than those which located in the center of the lobes of liver. During the whole embryonic period, there was no difference on the deposition of lipid droplets between the two lines.

The differrnces of goat corneal limbal epithelial cells cultured respectively on intact amniotic membrane, denuded amniotic membrane and cryopreserved denuded amniotic membrane treated in different freezing time were assessed to find a optimum treatment method of amniotic membrane in this study. Limbal explant were cultured on the intact amniotic membrane, denuded amniotic membrane, uncryopreserved denuded amniotic membrane, 30-day cryopreserved denuded amniotic membrane, 90-day cryopreserved denuded amniotic membrane and 180-day cryopreserved denuded amniotic membrane respectively. The contrasts were performed among the growth characteristics and morphologies of corneal limbal epithelial cells seeded on different treated amniotic membrane. The results showed that the adherence and proliferation of cells cultured on denuded amniotic membrane were better than that of cells cultured on intact amniotic membrane. The cells cultured on both of the amniotic membranes could grow and form multilayer cellular structure. However, the keratinization of superficial cells cultured on the intact amniotic membrane was more than that of superficial cells cultured on denuded amniotic membrane. The adherence and proliferation of cells cultured on 30-day cryopreserved denuded amniotic membrane were better than that of cells cultured on uncryopreserved denuded amniotic membrane, 90-day cryopreserved denuded amniotic membrane and 180-day cryopreserved denuded amniotic membrane. The cells on all different treated amniotic membrane could grow and form multilayer cellular structure. However, no significant differences were found among the cellular structure of the different treated amniotic membrane. The 30-day cryopreserved denuded amniotic membrane was the optimum choice which could be used for reconstructing tissue engineering artificial cornea.

Two hundred and eighty 50-weeks Green-eggshell chickens were randomly allotted to 4 treatments, each group was assigned 5 replications and 14 layers per replication. The control group was provided with corn-soybean meal basal diet, the trial groups were respectively fed the basal diet supplemented with 0.5%, 1.0% and 1.5% Leonurus heterophyllus Sweet, in order to observe the influence to biochemical parameters, hormone and ovary in Green-eggshell chickens. The results were showed as follows: compared with the control group, 1.0% and 1.5% Leonurus heterophyllus Sweet added to Green-eggshell chickens could significantly decrease TC,TG and LDL,apparently increase HDL,E₂,FSH and ovarian relative weight. Leonurus heterophyllus Sweet could affect fat metabolism, reduce the content of blood lipid. For saving cost,the most appropriate volume was 1.0%.

Apoptosis is a process that cells autonomicly go to death, so that to maintain homeostasis and adapt to living environment. Apoptosis involves a series of gene activation, expression and regulation. It has been observed that there are the occurrence of apoptosis during preimplantation embryos development, but if the degree of apoptosis is beyond the normal range, and it is not conducive to the development of embryos. This paper provides a review of the current state of knowledge on apoptosis during preimplantation embryo development, in order to provide a theoretical basis for improving the quality of embryo in vitro culture.

Cell suspension culture is a key process technology in biological production by large-scale animal cell culture in the bioreactor, this technology becomes the mainstream mode in global biological production process today. The development of microcarrier, the basic principles and application status of bioreactors, existing problems of suspension culture, the challenges and prospects of suspension culture in China are reviewed in the article.

To provide evidence for screening the poisonous components and physiologically active alkaloids, this study was performed to analyze primarily the alkaloids of Swainsona salsula. Preliminary tests, thin layer chromatographic analysis and gas chromatography-mass spectrum analysis were used to determine the alkaloid components extracted from Swainsona salsula in Ningxia. The results of the thin layer chromatographic analysis showed that the alkaloids of Swainsona salsula and matrine shared the same TLC spot, TLC retention Rf value. Gas chromatography-mass spectrum analysis also indicated that Swainsona salsula alkaloids and matrine shared the same GC retention time and mass spectra. However,Swainsonine in these samples could not be detected by thin layer chromatographic and gas chromatography-mass spectrum. The results showed that Swainsona salsula collected from Ningxia was ascertained to contain substantial anounts of matrine, but no Swainsonine was presence in Swainsona salsula or the Swinasonine level of the plant was low the detectable limit in this experiment.

Cationic antimicrobial peptides were a class of small peptides with anti-extrogenous pathogen invasion. As an important component of congenital immune defense system against infections, they were widely distributed in vivo. It exhibited potent and broad-spectrum activities against both Gram-positive and Gram-negative bacteria, fungi, viruses, protozoa, and cancer cells,and normal eukaryotic cells with low toxicity. It was an ideal alternative to a new generation of antibiotics. However, the same as antibiotics, some bacteria were resistant to certain antimicrobial peptides. The antibacterial and drug-resistant mechanism of the cationic antimicrobial peptides were summarized in the article to provide certain reference.

This study was conducted to optimize the culture and cryopreservation conditions for buffalo hand-made clone (HMC) embryos. Microdrops (MD), well of the wells (WOW), flat surface (FS) were adopted to culture HMC reconstructed embryos;and 40% EG, 25% EG (ethylene glycol, EG) + 25% DMSO (dimethylsulphoxide, DMSO) and 20% EG + 20% DMSO + 0.5 mol/L sucrose were adopted to vitrify HMC blastocysts. Furthermore, the efficiency of HMC and traditional somatic cell nuclear transfer (SCNT) were also compared. The results showed that the cleavage rate of embryos cultured in WOW was higher than those cultured on FS (P<0.05) and in MD(P<0.01);the blastocyst rate of embryos derived from WOW system (40.0%) was also higher than those derived from FS (19.8%) and MD (8.3%) systems (P<0.01);the cryosurvival rate of blastocysts vitrified with 20% EG + 20% DMSO + 0.5 mol/L sucrose was higher than those vitrified with 40% EG (P<0.01);both the fusion rate and blasotycst rate of HMC reconstructed embryos were higher than those of SCNT reconstructed embryos (P<0.05;P<0.01), while the cryotolerances of HMC and SCNT blastocysts were not different from each other. These results indicated that HMC could be an alternative to traditional SCNT in buffalo, WOW was most suitable for culture of HMC embryos, and vitrification of HMC blastocysts with 20% EG＋20% DMSO＋0.5 mol/L sucrose could get a reasonable cryosurvival rate.

Stallion semen cryopreservation is a technology which adds cryoprotectants in semen, then a series of processing and stores in ultra-low temperature(liquid nitrogen or dry ice).In recent years, as the frozen semen gradually applied in many countries on horse breeding, dramatic progress has been made on the cryopreservation technology. This article reviews stallion semen cryopreservation of semen dilution, centrifugation, dilution again, cooling balance, packing, frozen and stored in liquid nitrogen. It designed to help stallion semen cryopreservation for further research and provide a reference in order to promote healthy and sustainable development of our modern equine industry.

Concentration changes in each reproductive hormones during sheep oestrus cycle are closely related with follicular development, maturation and ovulation. In order to study the dynamic changes of the serum reproductive hormones of the Bamei Mutton sheep and their relationship with ovulation rate, the electrochemical method was used to measure serum concentrations of two steroid hormones (E₂ and P₄) for 12 adult ewes during estrus, and the variance analysis procedures of the SAS 9.0 were used to analyze the relationship between hormone concentration and ovulation rate. The results showed that variation of the two hormones were different between the laying-single-egg group and the laying-double-eggs group sheep. Concentration of E₂ firstly decreased and then increased in laying-single-egg group, and continuously declined in laying-double-eggs group. Concentration of P₄ continuously increased in laying-single-egg group, and firstly increased and then decreased in laying-double-eggs group. Concentration differences of E₂ and P₄ at each time point between the two groups were not significant(P＞0.05)。

For improving the freezing and de-freezing methods for boar semen further, it was aimed to investigate the effect of different freezing frequency(-100 ℃ 10 min,-120 ℃ 10 min,-140 ℃ 10 min) and thawing temperature(37 ℃ 30 s,45 ℃ 30 s,52 ℃ 30 s,60 ℃ 30 s) on cryopreservation of boar semen. The results showed that the sperm motility, plasma integrity, normal acrosome rate by freezing treatment with -120 ℃ 10 min, were all significantly higher than those of the other three groups respectively(P＜0.05). The sperm motility, plasma integrity, normal acrosome rate by thawing treatment with 37 ℃ 30 s, were not significantly higher than those of the other three groups respectively(P＞0.05), but the difference of the sperm abnormality in the treatment with 60 ℃ 30 s was significant(P＜0.05), but with 45 ℃ 30 s and 52 ℃ 30 s were not significant(P＞0.05). Therefore, it could be concluded that the combination of freezing by -120 ℃ 10 min and thawing by 37 ℃ 30 s were appropriate to frozen-thawed 0.25 mL pipettes of boar semen.

In order to improve the frozen porcine sperms motility and fertilizing ability after thawed, sperms were extended in basic cryoprotection extender with 0, 10%, 20%, 30% seminal plasma respectively before freezing. Sperm motility, membrane integrity, fertilization rate and blastocyst rate of IVF embryos and artificial insemination results were chosen as criteria for assessing the sperm quality. The results showed that the motility, membrane integrity of sperms extended in cryoprotection extender containing 10% seminal plasma were improved significantly (P<0.05) after thawed compared with the control group. The blastocyst rate of IVF embryos derived from these sperms was also increased significantly. Artificial insemination with the thawed sperms from the group extended in cryoprotection extender containing 10% seminal plasma resulted in a significantly lower pregnancy rates, litter size and number of live littermate compared with fresh sperm insemination(P<0.05). These results indicated that cryoprotectant with 10% seminal plasma could increase the motility of thawed porcine sperm after cryopreservation and the blastocyst rate after IVF, but for artificial insemination it was still need to be improved.

In order to study molecular genetic characters of Xiangxi cattle and search molecular markers related growth traits of Xiangxi cattle, the polymorphism of bovine SST gene was detected by PCR-SSCP, and association between polymorphism of g.934G>A locus in exon 1 of somatostatin (SST) gene and growth traits of Xiangxi cattle were analyzed. The results showed that there were two alleles (G and A) and three genotypes (GG, AG and AA) on g.934G>A locus of SST gene in Xiangxi cattle, which G allele frequency was more than that of A allele. The locus of g.934G>A was in moderate polymorphism, and fitted in Hardy-Weinberg equilibrium (P>0.05). The individuals with GG genotype had larger withers heigh and body length than AA genotype (P<0.05), and the heart girth and body weight of individuals with GG and AG genotypes were significantly larger than AA genotype(P<0.01). G allele had positively effect on withers heigh, body length, heart girth and body weight of Xiangxi cattle. The results of this study suggested that g.934G>A of exon 1 in SST gene might be effect on growth traits of Xiangxi cattle, and it might be used as the molecular genetic marker for breeding.

According to Haemophilus parasuis which was isolated from the department,the strain of poison strong,stable growth and virulence stability was selected as the original strain. The original strain was induced to cause a mutation by chemical agents,then after toxicity test,significantly less virulent strains were selected. The safety trials found that these attenuated strains had a high security, and layed the foundation for the development of attenuated vaccine.

To detect the healthy sheep’s nasal swab, anal swab, silage, soil,in part areas of Yili, 5 strains LM were identified from 280 samples by cultural character, biochemical character, and PCR amplification,and their surrounding environment is 1.78%. Furthermore, 5 strains LM didn’t have the same fingerprint diagram different from standard LM with ERIC-PCR,and these isolates were identified as LM belonged to the different gene type.

In this experiment, cell culture and MTT detecting techniques were used to examine the influence of probiotics on the T and B lymphocyte proliferative function to ConA and PMA in peripheral blood and immune organs of 1 day old ND-vaccinated chicks. The results were as follows: After vaccinated, the lymphopoiesis function in peripheral blood and immune organs increased in different extent compared with those of control chicks, probiotics and ND-vaccinated group was better than only ND-vaccinated group; after challenged with vNDV, the lymphopoiesis function in peripheral blood and immune organs and the immune protection rate increased in different extent compared with the control group, probiotics and ND-vaccinated group was better than only vaccine group. The results showed that probiotics could enhance the effect and the immune protection rate of ND vaccine to vNDV.

This case is by surgical repair of traumatic diaphragmatic hernia and diaphragmatic hernia causes and surgical success factors are summarized for the implementation of the type of surgery and prognosis, with further accumulation of experience.

This experiment was to analysis on the infection of atrophic rhinitis in some large-scale pig farms of Jilin by aetiological investigation. 148 positive blenna narium were collected from 6 large-scale pig farm in some areas, carried out isolation and identification of pathogen of swine atrophic rhiniti. The results showed that 84 pigs were infected by Bb (56.76%), 53 pigs were infected by Pm (35.81%), 11 bigs were polyinfected (7.43%). This results indicated that the atrophic rhinitis was mainly infected by Bb, polyinfection was light in large-scale pig farms of Jilin.

Torque teno sus virus(TTSuV) was first reported by American researcher in 1999, subsequent studies confirmed the virus prevalence in the pig population of some countres. The paper reviewed on epidemic area, genetic variation, susceptible animals, route of transmission and pathogenicity of TTSuV.

Cytokines as immunoadjuvant could enhance cellular immunologic response and humoral immunoresponse of animals, thus enhance specific immunity response of vaccine. Cytokines had crucial role in bacterium, virus, parasite and tumor vaccine’s researched. This cytokines include interleukin, interferon and granulocyte-macrophage colony-stimulating factor and so on. Recent advances were reviewed in the effective mechanisms and application of cytokines, which could provide important references for veterinary clinic application.

The highly pathogenic porcine reproductive and respiratory syndrome was an acute highly pathogenic infectious diseases, which caused by PRRSV mutant in recent years, the pig industry has caused huge losses by this disease, the latest fashion features, diagnosis, prevention and control of the highly pathogenic porcine reproductive and respiratory syndrome were introduced in this paper.

The brain tissue of piglet was obtained and the 217 bp long fragment in gp50 gene was amplified by the polymerase chain reaction (PCR). Therefore, the piglet was confirmed as porcine pseudorabies virus infection. Subsequently, the porcine pseudorabies virus was isolated on BHK-21 cells after 5 generations. The cells displayed typical cell lesions and were identified as porcine pseudorabies virus by PCR. The virus infectivety of the pseudorabies virus was 10^8.68 TCID₅₀/0.1 mL by Reed-Muench. Finally, the rabbits were inoculated with 10^7.0 TCID₅₀/mL virus. On 48 h the injection sites showed typical itching, skin damage and other symptoms. The rabbits were dead after 72 h by the inoculation of pseudorabies virus. Consequently, the results indicated that the isolated pseudorabies virus was highly pathogenicity to susceptible animals. It would be the research foundation of the virus epidemiology, pathogenesis,vaccine and diagnositic.

In order to investigate the infective situation of leptospirosis on dogs and cats, 20 pet disease monitoring sites were established in the Beijing area in 2011. 396 serum and urine samples of dogs and cats were collected at random for the investigation. Survey results showed that 106 antibody-positive samples in 200 serums by the microscope agglutination test, with positive rate of 53%. Among these positive samples, there were 20 derived from cats with positive rate of 27.8%, and others were dog serums with positive rate of 67.2%. A total of 196 urine samples were tested by RT-PCR, there were 54 positive ones founded with the positive rate of 27.6%, of which 21 copies of the cat urine with 4 positive samples, the positive rate was 19%. Dog positive urine sample were 50 copies in 175, the positive rate of 28.6%.

One strain of bacteria was isolated from Liaoning samples of pigs with suspected multiple subserosal inflammation and arthritis in 2011. We used the lungs, pericardial fluid, blood from diseased samples via bacterial isolation and culture techniques, biochemisty tests in addition to PCR identification finally indicated that the bacteria belonged to Hemophilus parasuis. Then drug sensitive test and analysis were performed on the strain named HPSLN111013. The results showed that the strain on some aspects such as morpha, cultural character and biochemisty tests were in accord with separate strains from other areas in China. Then drug sensitive test revealed that the isolate was the most sensitive to florfenicol, ceftiofur and lomefloxacin. These results supplied a foundation for the future research of Hemophilus parasuis’s serological and gene typing. Ascertaining the infection source and genetic variation of Hemophilus parasuis provided a basis of prevention and control measures of this disease in Liaoning province.

Body condition scoring, fecal scoring and walking scoring, the indispensable indicators to predict the health of cows and monitor the level of feeding management of cows in the actual production, are becoming increasingly important, which are summarized in this paper based on previous studies.

Optimization of Bacillus licheniformis M109 culture medium in this research based on the orthogonal test, carbon source, nitrogen source and carbon nitrogen ratio were determine as necessary for fermentation medium of Bacillus licheniformis M109. In order to enhance Bacillus licheniformis M109 fermentation level, growth curve of Bacillus licheniformis M109 and trend of pH, dissolved oxygen(DO) were studied. Though studying the fermentation process of growth, the optimal pH and inoculation amount were determined. The results found that DO was a key factor that limited the growth of Bacillus licheniformis M109. It was studied regulation rotate speed of stirring to improve dissolved oxygen and the fermentation density in culture medium. Another study found that Bacillus licheniformis M109 fermentation level could be improved by adding fresh medium. Through optimization of culture medium and conditions, the fermentation level of Bacillus licheniformis M109 incneased to 1.2?10¹⁰ CFU/mL from the inital 1.0?10⁹ CFU/mL and the rate of bacillus arrived to 88%.

To research the air quality of the scaled pig farm and evaluate the influence of the air quality on animal and human beings’health, the experiment adopts the environmental detection method to get the statistics about the air quality of sucking piglets pens,nursery pens and fattening pens. The experiment shows that in the pigs houses the average total bacteria amount of the pigs’houses in four seasons are 118.77×10³,28.51×10³,102.04×10³,62.71×10³ CFU/m³.The concentration of NH₃ in four seasons are 9.67,10.87,9.95,12.74 mg/m³.The concentration of H₂S in four seasons are 2.81,1.95,2.68,2.78 mg/m³. The total bacteria amount of outside the pigs’houses in four seasons are 13.57×10³,13.20×10³,16.70×10³,11.85×10³ CFU/m³. The concentration of NH₃ are 1.01,1.44,1.07,0.42 mg/m³. The concentration of H₂S are 0.06,0.08,0.03,0.01 mg/m³. The total amount of bacteria in four seasons are the spring＞autumn＞winter＞summer(P＜0.05). The total bacteria count inside the house are bigger than that outside the house(P＜0.05). The total amount of bacteria in four seasons are all beyond the national standard except the summer. The concentration of NH₃ and H₂S are within the national standard.The temperature, relative humidity and wind speed are within the national standard.

Elaphurus davidianus is one of the special species in Chinese deer family, belonging to 1st class of protected animals. Because of irregular hunting and environmental changes, it has been become extinct in China since 1900. The reintroduction of Elaphurus davidianus is the first large-size species reintroduction project in China. At present, three large Elaphurus davidianus populations have been established, including Beijing Milu Park, Dafeng Milu National Nature Reserve in Jiangsu and Shishou Milu Nature National Reserve in Hubei, which represent three types of captive breeding, semi free ranging and free ranging. This paper systematically reviewed the biological characteristics, ecological habitat, population, genetic reproduction, feeding and medical value of Elaphurus davidianus. It proposed that the scientific monitoring systems and network management system should be established to promote the protection and healthy reproduction of the Elaphurus davidianus population.

To find the nutritive incentives of kidney stones on beef cattle in Ningxia, the nutrient content of diet, the element of the kidney stone and the blood biochemical index were measured. The results showed that there were 23 beef cattle detected with kidney stones within 40 samples. The incidence of a disease was 57.5%. The forage to concentrate ratio, calcium content and phosphorus content in the diet were too high, but calcium-phosphorus ratio in the diet was too low. Stone energy spectrum analysis showed that the main chemical components were phosphorus and magnesium, preliminary judgment for ammonium magnesium phosphate or potassium phosphate magnesium. The serum phosphorus and the serum magnesium were higher than reference standard, while the serum calcium was lower. High forage to concentrate ratio, low calcium-phosphorus ratio and diet with high calcium and high phosphorus were the nutritive incentives of kidney stones on beef cattle in Ningxia. The magnesium content in diet might also have a positive effect on formulation of kidney stone.