Haemophilus parasuis(HPS) is one of the main virulent bacteria that threating the pig industry worldwide.HPS infection typically causes serious inflammations，namely Glässer’s disease.But there are still many troubles in disease prevention and control.Though much progress has been made in the basic research fields of bacterial virulent factors and host immune responses(IR) recently，the mechanisms of host-bacillus interaction are largely unknown.As the different susceptibilities in pigs exist，genomic selection based on molecular breeding strategy will receiving increasing attention from scientists and commercial breeders.Here，some high through-out data from our and other labs are reviewed in brief and the model for the host IR is proposed.We hypothesize that the activation of macrophage during the early stage of IR may play pivotal roles in determining bacillus escape and/or host protective IRs.In addition，key molecules and pathways involved in the IR of macrophage are discussed，which we hope will be helpful for the marker identifications in objectives of molecular breeding.Still，there is much work to be done and some suggestions are proposed in the article.

Plant extracts are the perfect alternative product of antibiotics because of their lower toxic and side effects，unique natural character，nutrition and biological activity.Many of them could be used to manipulate gut function in both ruminant and non ruminant animals.This review considers the effect of 3 kinds of plant extracts，such as essential oils，saponins and garlic extracts，on rumen fermentation and discusses their mechanism of action and feasibility exploited as rumen manipulating agents.

 The formation and development of primordial follicle are the most important events in the ovary activation and exerting the optimum reproductive potency of an adult.Most researches about the follicular development were focused on the growing follicular，but not the primordial follicle.However，studies have shown that assembly timing of primordial follicle are differ between species.The signaling pathways，growth factor，transcription factors，hormone and other aspects collaborate to regulate the formation and development of primordial follicle.This paper will review the primordial follicle assembly，development and the influencing factors in the mammalian.

 This experiment was conducted to study the expression pattern of Krüppel-like factor 3(Gallus gallus KLF3，gKLF3) gene，and its effect on adipocyte differentiation.The expression pattern of gKLF3 in the adipose tissue and adipocytes of broilers were analyzed by using qRT-PCR.The effect of gKLF3 on adipocyte differentiation was analyzed by over-expression technology.The result showed that gKLF3 was consecutively expressed during the chicken adipose tissue development from 2- to 10-week-old.At 10-week-old，the gKLF3 expression in the abdominal fat tissues was significantly higher in fat line chicken than that in lean line chicken(P<0.05).Additionally，the significantly higher gKLF3 expression level was observed in preadipocytes in contrast to that in mature adipocytes(P<0.01).The expression levels of gKLF3 in preadipocytes induced by oleate were lower than that in control.Moreover，over-expression of gKLF3 inhibited adipocyte differentiation and down-regulated expression of FAS and C/EBPα (P<0.01).The result indicate that gKLF3 play an important role in the growth and development of chicken abdominal adipose tissue，and might restrain adipocyte differentiation by inhibiting the expression of C/EBPα and FAS.

This study was designed to investigate the role of L-BABP gene in lipid metabolism of hepatocytes and the effect of L-BABP gene on the genes related to lipid metabolism in chicken.L-BABP gene expression was downregulated by siRNA interference and upregulated via overexpression plasmids method.The genes expression level in chicken hepatocytes were detected at 24，36，48，60 and 72 h after transfection with the siRNAs and overexpression plasmids of L-BABP.The results showed that APOAΙ，APOB，PERILIPIN gene expression levels，the amount of cholesterol and triglyceride in the medium are significantly affected by L-BABP gene expression changes.Therefore，L-BABP may involve in fat deposition，metabolism of total cholesterol and the process of lipolysis lipid metabolism.

To deeply study the genetic diversity of chicken Interleukin 2 (IL-2) gene and provide a theoretic basis for the breeding for disease resistance，the genetic variation of IL-2 were studied in 448 individuals from 11 Chinese native，3 introduced and one cultivated chicken breeds by DNA sequencing method in this study.A total of 13 SNPs were identified by using DNA-pooling and sequencing.The results of analysis for a genome region including 5 single nucleotide polymorphisms(SNPs) among all the individuals showed that the frequencies of different alleles differed significantly among 15 populations，suggesting IL-2 had abundant genetic variations and provided promising gene resource for disease resistance.Analysis of several population genetic indexes indicated that the Jingxing Yellow chicken had the lowest level of genetic diversity,which was consistent with its breed formation and little differences of genetic diversity were observed among other breeds.Analysis of haplotypes revealed a total of 23 hapltoyepes.Among them，H1(CAACG)，H2(CAATT)，H4(TGTTG)，H9(TGTTG)，H11(TATCG) and H16(CATCG) were unique or dominant in Chinese native chicken breeds，suggesting these haplotypes may be associated with the disease-resistant ability.

The objective of this study was to identify the polymorphisms of Ihh gene and their correlation with production traits，then to detect the molecular markers related to production traits，and investigate distribution and expression in tissues detected in yak and cattle.At the same time，RT-PCR method was used to detect the expression of Ihh gene in different tissues in yak and cattle.BLAST and Chromas were used to screen the SNPs in Ihh gene by the wave height of sequencing map.The genotypes were determined by high-resolution melting curve(HRM) and the alleles frequency was estimated.The PHASE and SHEsis softwares were used to analyze matching chain disequilibrium and haplotype，respectively.The SPSS17.0 software was used for association analysis.Two SNPs(5855(C/T) and 6383(G/A)) of Ihh gene were indentified in the population，which were on exon 3.Through population genetics analysis，the results showed that 2 loci were at high polymorphic status(PIC>0.25).The χ² tests showed that the 2 loci were in the status of Hardy-Weinberg equilibrium(P＜0.05) in Gannan and Datong yaks; the 2 loci were not all in the status of Hardy-Weinberg equilibrium(P＞0.05) in Tianzhu yak.Analysis of matching chain disequilibrium and haplotype showed that there was a strengh linkage equilibrium between the 2 loci with 4 haplotype combinations and 3 genotype combinations.Analysis of association of polymorphism with body measurement traits at all mutation loci showed that 3 genotype combinations had significant effects on body height，body length，weight，tube girth and herrt girth(P＜0.05).Yak with CTGA genotype combination had higher body height，body length，weight，tube girth and herrt girth than individuals with CCGG and TTAA.The result of expression showed that Ihh gene was expressed in 8 different tissues.The results suggest that Ihh gene may have potential effects on production traits in the above mentioned yak populations and could be used for marker-assisted selection.

Based on the result of the previous genome-wide association study(GWAS) for milk production traits in Beijing Chinese Holstein population，trafficking protein particle complex 9(TRAPPC9) as a novel candidate gene for milk production traits was confirmed in new Chinese Holstein population.Promoter，exon and GWAS’s significant region was amplified and sequenced by screening the DNA pools to select single nucleotide polymorphisms(SNP) and analyze their association with milk production traits of Chinese Holstein population.The association analysis showed that the genotypes of SNP1 had significant effect on protein percentage(PP) and lactose percentge(LP)(P<0.01)；the genotypes of SNP2 had significant effect on PP and LP(P<0.05)；the genotypes of SNP3 had significant effect on fat percent(FP) and PP(P<0.01)；the genotypes of SNP4 had significant effect on FP(P<0.01)；the genotypes of SNP5 had significant effect on FP and LP(P<0.05).The expression of TRAPPC9 also had significant effect on milk production traits，but the DNA methylation of promoter region of TRAPPC9 had no direct effect.In conclusion，TRAPPC9 gene can be considered as a molecular marker applied to Chinese Holstein marker-assisted selection.

The present study was carried out to establish yak mammary epithelial cell (YMEC) line and identify its biological characteristic，which was separated using tissue explants method，purified by selective trypsinization，characterized by immunofluorescence and Western blot assay.Transfecting EGFP-N1 gene into cells using lipofection to examine whether foreign genes could be transfected into YMECs，fluorescence microscope was used to detect the expression of EGFP gene.Lastly，flow cytometry was used to evaluate the apoptosis level of cells with Annexin V/PI double staining and RT-PCR was used to examine gene expression of antibacterial peptide and apoptosis factors to determine the effect of Staphylococcus aureus infection on YMECs.Immunofluorescence assay results proved established cell line could express cytokeratin 18 rather than vimentin，indicating no fibroblast existing in epithelial cells.Positive expression of β-casein in YMECs demonstrated cultured cells could synthesize and secrete mammary- specific milk protein.Moreover，we detected EGFP protein in transfected YMECs by fluorescence microscope，showing EGFP gene was successfully transfected into the YMECs.After YMECs were challenged with for Staphylococcus aureus for 3 h，the apoptosis level of mammary epithelial cells was significantly up-regulated（P<0.05）.Furthermore，the expression of TAP（P<0.01），BNBD5（P<0.01）and BAX（P<0.01）genes increased; whereas that of BCL-2 gene decreased（P<0.05）.To conclude，we successfully established a stable yak mammary epithelial cell line，which can be used as a useful model in vitro for study of yak mammary gland development and differentiation.

This study aimed to investigate mRNA transcriptions of interferon stimulated genes ISG15 and OAS1 in the peripheral blood of Chinese Holstein dairy cows during early pregnancy，by which we hope to provide an insight into their applications in bovine early pregnancy diagnosis.A total of 27 heifers following spontaneous estrus(n=10) or oestrus synchronisation(n=17) were used for the study.Peripheral blood were collected at 0，14，18，21 and 28 d post artificial insemination(AI)；subsets of peripheral blood leukocytes(lymphocyte，monocyte，and natural killer cell) were isolated at 0，18 and 21 d；Real-time PCR was employed to detect the relative mRNA transcriptions of ISG15 and OAS1 in the whole white blood cells(WBC) and/or the leukocyte subpopulations.The results showed that bovine ISG15 and OAS1 in the peripheral blood of pregnant cows were significantly elevated versus non-pregnant cows at 18 d(P<0.05)；On 18 d post AI，ISG15 was up-regulated in pregnant cows with the average fold change(FC) of 3.92，whereas it was down-regulated in non-pregnant cows with the average FC of 1.46；OAS1 was similarly up-regulated(FC=3.49) in pregnant cows and down-regulated(FC=1.28) in non-pregnant cows.The relative transcriptions of ISG15 and OAS1 in pregnant cows following spontaneous estrus were significantly higher than those in non-pregnant cows at 18 d post AI；although the tendency in cows following oestrus synchronisation was the same，the difference was not statistically significant.Further analysis in the subpopulations of leukocytes showed that OAS1 in lymphocytes of pregnant cows was significantly elevated(+2.14)，whereas depressed in non-pregnant cows (-1.80) at 18 d post AI(P<0.05).The results indicated that regulations of ISG15 in peripheral blood of pregnant cows were more obvious than OAS1 at 18 d after AI；differential expressions of ISG15 and OAS1 between pregnant cows and non-pregnant cows were more obvious in cows following spontaneous estrus.In conclusion，ISG15 transcriptions in peripheral WBC could be a better indicator for bovine early pregnancy diagnosis(18 d post AI)，especially in cows following spontaneous estrus.Gene expression analysis of bovine ISG15 and OAS1 in the leukocyte subsets has provided additional insight into their roles during early maternal recognition of pregnancy.

The objective of this study was to discuss the effects of different dietary protein levels on insulin-like growth factor-1(IGF-1) and growth hormone(GH) secretion and mRNA expression in sheep，to provide a theoretical basis for scientific preparation of feed and study the growth in sheep.Eighteen 6-month-old prolific Suffolk rams were randomly allocated into 3 treatments with different dietary protein levels(low protein，medium protein and high protein).The concentrations in peripheral blood and mRNA expression levels in skin of IGF-1 and GH were detected at different growth stages(30，60，90 and 120 d) with different dietary protein levels by ELISA and SYBR Green Real-time PCR methods.The results showed that the average daily gain，the concentrations of IGF-1 and GH in peripheral blood，the mRNA expression of IGF-1 in skin were significantly affected by dietary protein levels.However，mRNA expression of GH gene did not change significantly.With the increased protein intake，the growth of sheep was faster，the concentration of IGF-1 was increased，but the concentration of GH was decreased in peripheral blood，mRNA expression of IGF-1 gene was up-regulated.

This experiment was conducted to study the effect of increasing the ratio of non-fiber carbohydrate to neutral detergent fiber on bacteria flora，endotoxin and histamine content in rumen and plasma of dairy goats.Six rumen-cannulated lactating Guanzhong goats were used to investigate the effects of increasing dietary NFC/NDF ratio (i.e.non-fiber carbohydrate to neutral detergent fiber) on bacteria flora，and endotoxin and histamine contents in the rumen and the plasma of dairy goats.Induction of subacute ruminal acidosis (SARA) was performed by increasing dietary NFC/NDF ratio from 1.02(stage Ⅰ) to 1.24(stage Ⅱ)，1.63(stage Ⅲ) and 2.58(stage Ⅳ).Each stage lasted 10 days.A dynamic pH monitoring system and a rolling-tube method were used to assess changes in ruminal pH and the numbers of total bacteria，amylolytic bacteria and Fusobacterium necrophorum in rumen.Limulus test and fluorescence spectrophotometry were used to assess the contents of endotoxin and histamine.Our results showed that as the dietary NFC/NDF ratio increased from 1.02 to 2.58，the rumen pH decreased significantly (P<0.05) at accelerated rates，while the endotoxin and histamine contents in the rumen and the numbers of amylolytic bacteria and necrotic fusiform bacilli increased significantly (P<0.01 and P<0.05 respectively).We conclude that abnormal increases of endotoxin，histamine and F.necrophorum played an important role in the occurrence and development of SARA.

The objective of this study was to evaluate the effects of lactic acid bacteria and propionic acid on the fermentation quality and aerobic stability of total mixed ration(TMR) silage prepared with whole-crop corn in Tibet.The treatments were as follow：control(no additives)，inoculated lactic acid bacteria with 1 g•kg^-1 or propionic acid adding with 0.4% on fresh matter basis of TMR.All silos per treatment were opened after 45 days of ensiling，5 silos per treatment were used for fermentation quality determination，and then sampled on 6，9 and 12 days after exposure to air which were subjected to aerobic stability evaluation，5 replicates for each treatment at every time point.The results showed that the fermentation quality of control was good as indicated by the appropriate dry matter content，abundance of lactic acid bacteria count and water soluble carbohydrate content.The addition of lactic acid bacteria further improved fermentation quality.Although the addition of propionic acid inhibited the lactic acid fermentation，the fermentation quality was also good.During aerobic exposure stage，control and lactic acid bacteria addition silages showed decrease in lactic acid content，increase in pH，ammonia nitrogen/total nitrogen and the number of yeast.Lactic acid content in propionic acid silages increased during the first 9 days of aerobic exposure，and then significantly(P<0.05) decreased.The pH and the number of yeast in propionic acid silages were significantly(P<0.05) lower than those of control and lactic acid bacteria addition silages.Adding lactic acid bacteria improved fermentation quality but did not affect the aerobic stability；adding propionic acid inhibited lactic acid production during fermentation，while delayed aerobic deterioration of TMR silages，thus adding propionic acid improved the aerobic stability of whole-crop corn total mixed ration silage.

Swine Japanese encephalitis virus (JEV) is one of pathogens that causing sow abortion.In this experiment，the DNA replicon vector was constructed and exogenous gene was expressed.Based on the JEV SA14-14-2 sequences，viral structural protein encoding gene prM and parts of E gene sequences were removed by mutant PCR methods，which retained nonstructural protein genes and non-coding region sequences，subgenomic replicon pAC-JEV that inserted FMDV-2A and HDV ribozyme sequences was constructed.In order to identify its abilities of self-replication，expression of nonstructural protein (NS3) were detected by western blotting and indirect immunofluorescence assay.The results showed that NS3 protein level increased gradually with extension of transfection times.To explore JEV replicon ability about expressing foreign proteins，we inserted an enhanced green fluorescence protein (EGFP) gene，HA gene of swine influenza virus and GP5 gene of porcine reproductive and respiratory syndrome virus into the multiple cloning site of pAC-JEV(pAC-JEV-EGFP，pAC-JEV-HA，pAC-JEV-GP5) and mutation of NS5 gene was deleted (pAC-JEV-ΔNS5).The plasmids were transfected into BHK-21 cell，EGFP mRNA level was significantly increased at 48，72，96 h in pAC-JEV-EGFP by real-time PCR，the EGFP fluorescence was directly detected by the fluorescence microscope at 12 h，and EGFP fluorescence signal lasted for 8 days.HA of swine influenza virus and GP5 of porcine reproductive and respiratory syndrome virus were detected using mouse anti-HA and GP5 antibodies by Western blotting.The results show that HA and GP5 can be expressed in BHK-21 cells.This JEV subgenomic replicon pAC-JEV can be used to study JEV gene function and viral protein interaction with cellular proteins；it also is a valuable tool for DNA vaccine research.

The aim of this study was to understand the pathological characteristics of duck reovirus naturally infection in Hubei province.Pathologic autopsy，HE staining，and the immunohistochemical analysis were used to study the ducks naturally infected with duck reovirus.Results were as follows：At necropsy，large yellowish-white necrotic foci were observed in the spleens and livers.By histopathology，hemorrhagic，necrotic lesions and granulomas were seen in both organs.Immunohistochemical detection showed that all spleen samples had positive signals，but in liver and bursa only part of the samples were positive.Taken together，our results reveal that the liver and spleen necrosis is the characteristics of duck reovirus infection in Hubei province.

In this study，we cloned BoIFN-α0 mature peptide gene and expressed it in the eukaryocyte， and also studied the activity of this protein.To achieve secretary expression of bovine interferon-α0 (BoIFN-α0 ) in Pichia pastoris，the BoIFN-α0 mature peptide gene was synthesized by PCR and inserted into the secreted expression vector pPICZαA.The Pme Ⅰ linearized resultant recombinant plasmid was transformed into P.pastoris GS115 by electroporation and the positive recombinant strains P.pastoris were selected by 100 μg•mL^-1 of zeocin^TM and identified by PCR with the AOX1 primers and the specific primers.The positive recombinant strains were induced with methanol and achieved secretory expression in P.pastoris Yeast expression system.The products in culture medium were detected by SDS-PAGE and Western blot.The SDS-PAGE results showed that the recombinant protein was expressed in the supernatant with molecular of 18 and 21 kDa (with the difference of glycosylation).Furthermore，the BoIFN-α0 was able to efficiently inhibiting virus replication in vesicular stomatitis virus (VSV)/MDBK cells detection system and its antiviral activity was about 5.72×10⁶ U•mg^-1.These results provided a basis for further application of BoIFN-α.

 A study on ampicillin (AMP) residue depletion was conducted in chicken tissues.AMP was extracted from chicken tissues with sodium dihydrogen phosphate solution.After being deproteinizated with acetonitrile and degreased in n-hexane saturated，the analyte was extracted by saturated dichloromethane.The supernatant was reacted with formaldehyde under acidic and boiling conditions.Finally，AMP was determinated by high performance liquid chromatography (HPLC) with fluorescence detector.The excitation wavelength was set at 327 nm and emission wavelength was set at 409 nm.The average recovery in chicken tissue was 75.31%-85.87% with coefficients of variation (CV) lower than 10.20%.The limit of detection (LOD) was 3.5 μg•kg^－1 (S/N=3) and the limit of quantitation (LOQ） was 10.0 μg•kg^－1(S/N=10).After the chickens were orally administered successively AMP of 120 mg•kg^－1 and 240 mg•kg^-1 of body weight one time every day for 7 days，the maximum residues of AMP in chicken tissues were detected at 4h after final administration.Concentrations of AMP fell sharply at the 3th withdrawal day，all lower than MRL (50 μg•kg^－1) at the 5th withdrawal day，and all lower than LOD (5.0 μg•kg^－1) at the 9th withdrawal day.At the same time definition，the concentration of AMP in muscle was lowest，but highest in liver.However，AMP in kidney needed more time for depletion.The residues of AMP in chicken tissues were all positively correlated with AMP orally administered doses.The concentration-time dates were analyzed by WT1.4 software to get the withdrawal time (WT) with 95% upper tolerance limit，the WT advised in normal dose group was 4 d，and in double dose group was 5 d.

 By measuring the content of Nitric Oxide and the Activity of Nitric Oxide synthase， this experiment was conducted to study the influence of lamotrigine and L-NNA on the change and investigate the mechanism of the action of NO on the cerebral hemorrhageand the protective effect of those two pharmaceuticals in the temporal lobe epilepsy rabbit induced by pilocarpine.Sixty healthy Haerbin rabbits，2.0-2.5 kg，were divided into the group of physiological saline，the epilepsy model group，and the treatment group of lamotrigine and L-NNA.Nitric Oxide （NO） content and the activity of NOS in the hippocampus and temporal lobe after epilepsy were determined by using nitrate enzyme reduction method.We found that the activity of NOS increased at 6 hours after epilpsy， and increased to the highest at 1 day， then decreased gradually and basically returned to normal level after 7 days，but it was still higher significantly than that of other groups （P<0.05）.The level of NOS in two therapy groups between L-NNA and lamotrigine were lower than that in model group at every time extremely notable difference or notable difference （P<0.01 or P<0.05），and the decreasing level of NOS in L-NNA group was lower than that of group of lamotrigine between 6 h to 1 day （P<0.05）.The change of the NO content and the activity of NOS were accordant primitively and directly correlation.SABC immunohistochemical method was used to detect the neuronal NOS positive neurons in hippocampus，the density of neuronal NOS positive neurons in hippocampus’ CA3 region increased，and the cytoplasm stained darker， soma-sectional area and the length of tuber became smaller; After the treatment of lamotrigine and L-NNA，the density of neuron decreased，and the soma-sectional area and the length of tuber became significant longer （P<0.05）.The results indicate that NO is involved in the initiating process of the temporal lobe seizures，and the high concentration of NO can damage the neurons；L-NNA and the novel therapeutic drug lamotrigine can inhibit the concentration of NO after epilepsy seizures，which have a better therapeutic effect on the epilepsy seizures.

 To study the effects of ginseng stem and leaf polysaccharide (GSLP) on small intestinal mucous tissue development and immune cells of chicken，600 of 7 days healthy Hy-line Variety Brown chicken (52.79±4.58 g) were freely divided into 10 treatments，60 chicken per treatment (3 replications，20 chicken per replication)，5 treatments were included in normal fed group and others were in immune depressed group.The additive levels of GSLP were 0，0.2，0.4，0.8 and 1.6 g•kg^－1 in each group based on basal diet.After feeding stage，9 chicken of each treatment (3 per replication) were randomly selected for samples collection of jejunum and ileum.The results showed that：(1) GSLP improved villus width and surface area of jejunum (P<0.01)，0.8 and 1.6 g•kg^－1 GSLP decreased villus height and recess depth of ileum (P<0.01)，but GSLP had no significant effects on villus to gland ratio；(2) The interactions of GSLP with immune state were observed in jejunum villus height (P<0.05) and mucous thickness (P<0.01)，and ileum villus height (P<0.05)；(3) 0.4，0.8 and 1.6 g•kg^－1 GSLP increased the amounts of CD3⁺ T lymphocyte(P<0.05)，and 1.6 g•kg^－1 GSLP increased the amounts of mast cell (P<0.05) in jejunum，and dosage effects were existed in the influence of GSLP on immune cells.In conclusion，GSLP had somewhat improvement on small intestinal mucous tissue development，but had no significant influence on nutrients absorptive function；0.4 to 1.6 g•kg^－1 GSLP increased the level of jejunum mucous immunization；and GSLP could improve the development of jejunum villus height and mucous thickness depressed by cyclophosphamide in early trial stage.

The aim of this study was to investigate the effects of Ginsenoside Rb1(GSRb1) induce Hegu acupoint microvascular vasomotion and the effect on sensitive threshold of Rat myocardial microvascular endothelial cells(RMMECs).Microvascular vasomotion was detected by laser doppler blood flow monitor combined with iontophoresis device.The changes of calcium in the RMMECs were measured by laser scanning confocal microscopy combined with calcium fluorescent probe (Fluo-3AM).The results were as follows：The minimum amount of ATP could induce calcium transient increase in RMMECs was 0.5 μg•mL^-1，the dose of significant difference was 0.7 μg•mL^-1 and the minimum amounts of GSRb1 inducing calcium changes in RMMECs was 8 μg•mL^-1.After incubation 8 μg•mL^-1 of GSRb1，the minimum amount of ATP could induce calcium transient increase in RMMECs was 1.5 μg•mL^-1.Eight μg•mL^-1 of GSRb1 can inhibit the ATP-induced intracellular calcium transient rise，that meant，GSRb1 increased the sensitive threshold of RMMECs to ATP.

The research was conducted to screen the influence factors on swainsonine production in fungal endophyte，Undifilum oxytropis，from locoweed.U.oxytropis were inoculated in liquid medium of different pH or containing different concentration of PEG，L-pipecolic acid，L-lysine and ɑ-ketoglutaric acid，respectively.To estimate the influence of different factors on swainsonine production in U.oxytropis，the swainsonine of cultured fungal mycelia and zymotic fluid were extracted and detected after shaking culture.The results showed that low pH (pH4.5) inhibit significantly (P<0.05) the synthesis of swainsonine in fungi.With the increase of PEG in culture medium，the production of swainsonine in fungi was also inhibited significantly (P<0.01).When the fungi were inoculated respectively in liquid medium that contained equivalent concentration (10^-3 mol•L^-1) of L-pipecolic acid，L-lysine or ɑ-ketoglutaric acid，the production of swainsonine was inhibited significantly (P<0.05) in the group added L-pipecolic acid.However，the yield of swainsonine were increased significantly(P<0.05) in the group added L-lysine.When the initial concentration of L-pipecolic acid was 10^-3 and 10^-2 mol•L^-1，the production of swainsonine in U.oxytropis was inhibited significantly (P<0.05).However，when the initial concentration of L-pipecolic acid was 10^-4 mol•L^-1，the yield of swainsonine in U.oxytropis was increased significantly (P<0.01).When the initial concentration of L-lysine was 10^-1，10^-2 and 10^-4 mol•L^-1，the production of swainsonine in U.oxytropis was inhibited significantly (P<0.05).When the initial concentration of ɑ-ketoglutaric acid was 10^-1，10^-2 and 10^-3 mol•L^-1，the production of swainsonine in U.oxytropis was inhibited significantly(P<0.05).Low pH or PEG added to medium could significantly inhibit the production of swainsonine in U.oxytropis.It was significant that the influence of L-pipecolic acid，L-lysine and ɑ-ketoglutaric acid on the swainsonine producing of fungal endophyte.However，the extent of influence on fungal swainsonine producing was highly correlated with the concentration of L-pipecolic acid，L-lysine and ɑ-ketoglutaric in liquid media.

The aim of the present study was to establish a pyrosequencing method targeting S gene for detection and identification of schmallenberg virus (SBV).In alignment with published more than 10 viruses S gene sequences which belong to Bunyaviridae and are highly homogenous with SBV.Adaptive sequence for pyrosequencing was synthesized and in vitro transcripted to cRNA as the RT-PCR templates.After RT-PCR reaction，the products were detected with forward and reverse pyrosequencing and identified by genomic alignment.Results showed that the RT-PCR products were 166 bp.The sensitivity of the method that the minimum copies could be detected was 104 copies.The specificity that sequencing length by pyrosequencing was around 100 bp which could make a definite diagnosis for SBV.The results presented here demonstrate that the detection and identification method for SBV by pyrosequencing was established.The method can be used in fast diagnosis for SBV.