Bovine mastitis，especially subclinical mastitis that induced by Staphylococcus aureus (S.aureus)，is the most common disease，which is very difficult to prevent and control in dairy industries.In recent five years，our research group has collected 1 112 milk samples from nine dairy farms in Northern China，which included lactating cows with high and low somatic cell counts(SCC) as well as clinical mastitis cows within eleven sampling batches.A total of 191 strains of S.aureus were isolated and identified in the detected Northern dairy cattle and the average infectious ratio of mastitis infected by S.aureus was 11.7%(the lactating cattle was 12.4% and the clinical mastitis cattle was 10.8%).Antimicrobial susceptibility test and toxic genes detection revealed that Chloramphenicol，Amikacin and Cefoperazone had strong inhibitory effects on S.aureus mastitis， the highest detection rate of the toxic genes was panton-valentine leukocidin gene(70.2%).Pulsed-field gel electrophoresis (PFGE) analysis revealed the epidemical patterns of S.aureus in mastitis cattle in Northern and Eastern China.Combined with the related studies from domestic and foreign countries，the review systematically compared and analyzed the infectious ratio，the antibiotic resistance，the main toxic genes，the epidemical patterns of S.aureus mastitis.The strategies to prevent and control the incidence of S.aureus mastitis in dairy cattle were also discussed.

 Based on Web of Science，this paper used CiteSpace software to perform knowledge mapping analysis on 2 382 international scientific literature on thermal stress of dairy cattle，to show the current research situation and focus the future trend and the existing problems，important papers，authors and institutions in the field.This paper can be considered as a supplement for the experimental papers and review papers in the related area，helping readers to understand the research situation and knowledge structure，then quickly position primary focus and key literature.

In order to further determine the C/EBPα binding site of L-FABP and analyze its role in the regulation of L-FABP transcription，the C/EBPα binding site in L-FABP promoter was effectively mutated using site-directed mutagenesis in the current study.The results showed that the expression of L-FABP was significantly repressed by C/EBPα，but the L-FABP promoter activity was significantly promoted after the mutation of C/EBPα binding site.In a conclusion，the expression of L-FABP was significantly repressed by C/EBPα，and C/EBPα probably involved in the regulation of L-FABP expression through this binding site.These results will provide a foundation for further research on the regulatory mechanism and function of chicken C/EBPα on L-FABP gene.

To scan SNPs and candidate genes affecting body composition traits and provide new methods for further genetic improvement of Jinghai Yellow chicken，a genome-wide association study on 13 body composition traits was carried out using the Illumina chicken 60K SNP Beadchip in Jinghai Yellow chicken in the present study.The result showed that a total of 13 SNPs reached 5％ Bonferroni genome-wide significant level(P<1.8E-6) and 130 SNPs reached “suggestive” genome-wide significant level(P<3.59E-5) with body composition traits.The 13 significant SNPs were located nearby or in 12 candidate genes including GRIK1，NCAPG， KCNIP4，CACNA2D2，and so on，among which 6 SNPs were located in a region approximately 1.6 Mb in length on chicken chromosome 4(74.3-75.9 Mb).Among the 130 “suggestive” significant SNPs，25 SNPs were located in a region 7.4 Mb(71.5-78.9 Mb) in length on chicken chromosome 4.5 650 haplotpyes were established and 14 of them were found to be associated with 6 body composition traits.Nine out of 14 haplotypes were located in the region of 74.3-75.9 Mb on chicken chromosome 4.Five candidate genes of LCORL，QDPR，KCNIP4，LDB2 and FAM184B were located in this region.The present study suggested that genes located in the region of 71.5-78.9 Mb on chicken chromosome 4 and GRIK1，NCAPG，KCNIP4，CACNA2D2，LCORL，QDPR，KCNIP4，LDB2，FAM184B genes might play important role in regulation of body composition of Jinghai Yellow chicken.

This experiment was designed to study the polymorphisms of Myostatin(MSTN) gene and its effects on body weight，body measurement and carcass traits in goats.The polymorphisms of 5′-regulatory region and exon 3 of MSTN gene in some goat populations were detected by PCR-RFLP and PCR-SSCP in the present study，respectively.The results showed that：The transition(C→T) at －662 bp in the 5′- regulatory region leaded to polymorphisms of TaqⅠ and there were CC，CT and TT genotypes in the 7 populations.The frequency of genotype CC in Boer goat was higher than that in the 3 indigenous goat populations，but the frequency of genotype TT in Boer goat was lower than that in the 3 indigenous goat populations.The frequency of allele C was higher than that of allele T in the Boer goats，while it was contrary in the 3 indigenous goat populations.Except for Boer goat populations，5 other populations were at Hardy-Weinberg nonequilibrium(P<0.01).Three genotypes NN，NM and MM were found in exon 3 of MSTN gene in 7 populations.The individuals with genotype CT-NN and CT-NM were not detected in the 7 populations.The individuals with genotype TT-NN and TT-NM were not detected in Boer goats，and the individuals with genotype TT-NM were also not found in F1.Furthermore，individuals with genotype TT-MM were also not observed in 3 indigenous goat populations.The correlations between the polymorphisms of MSTN gene 5′-regulatory region and weaning weight，meat weight and hindquarters meat weight were very significant(P<0.01).The average values of weaning weight and meat weight of individuals with genotype CC were significantly higher than that of individuals with genotype TT(P<0.01 in weaning weight，P<0.05 in meat weight) and the average values of hindquarters meat weight were not significantly different among different genotypes(P>0.05).The average values of weaning weight，meat weight and hindquarters meat weight in genotypes CC，CT and TT decreased in turn.At the same time，the significant correlations between the polymorphisms of MSTN gene 5′-regulatory region and liver weight was found(P<0.05).The average values of liver weight of individuals with genotype CT was significantly higher than that of individuals with genotype TT and the significant difference in liver weight among other genotypes were not observed(P>0.05).The significant correlations between the polymorphisms of exon 3 and meat weight，hindquarters meat weight and liver weight were observed(P<0.01).The genetic effects for genotypes were MM>NM>NN and the average values of meat weight，hindquarters meat weight and liver weight of individuals with genotype MM were significantly higher than that of individuals with genotype NN(P<0.01 in meat weight，P<0.05 in hindquarters meat weight and liver weight).The genetic interaction between exon 3 and 5′-regulatory region affected significantly weaning weight，hindquarters meat weight and liver weight(P<0.05).The average values of the 3 indexes of individuals with genotype CC-MM were the highest among the compound genotypes.Therefore，the different MSTN genotypes might be used as molecular genetic marker to select body weight，body measurement or carcass traits in goats.

The objective of this study was to elucidate the characteristics in histomorphology and the potential molecular mechanisms underlying the dermal hyperpigmentation in Youzhou Dark-skin goat.First，we analyzed the characteristics of histomorphology in 100 d-fetal skin in Youzhou Dark-skin goat(dark skin) and Yudong White goat(normal skin).Then we examined the mRNA expression of Pax3，Mitf and Tyr in skin and B16 cells in the 2 breeds.The results showed that：1) There were significant differences in the skin pigmentation between breeds at fetus skin rapid development stage；2) The skin mRNA levels of Pax3 and Tyr were significantly higher in Youzhou Dark-skin goat than that in Yudong White goat(P<0.001)，whereas no significant difference was observed in Mitf expression between the 2 breeds(P>0.05)；3) The expression of Pax3 was decreased(P<0.001)，Tyr was increased(P<0.01) during the B16 cell differentiation，as compared with the proliferating B16 cells.The results indicate that：1) There might be diversity in migration pathway of the dermal melanocytes between the dark-skin and normal skin goats during the early stage of skin development differentiation；2) Downregulation of Pax3 and upregulation of Tyr are indispensible for melanogenesis in melanocytes；3) The high expression of Pax3 and Tyr are associated with the dermal hyperpigmentation in Youzhou Dark-skin goat.

The transcription level and methylation in promoter regions of Hoxa6 and Hoxa10 genes in multi-costa properties and common yak were investigated to explore the mechanism of transcriptional regulation of Hox gene in the process of multi-costa properties.The mRNA expression leves of Hoxa6 and Hoxa10 genes in multi-costa properties and common yak were detected by real-time fluorescence quantitative PCR(qRT-PCR)，and the methylation of Hoxa6 and Hoxa10 genes promoter regions were analyzed by bisulfite-sequencing PCR(BSP)，which included modifying，cloning and sequencing the promoter regions.The results showed that the expression level of Hoxa6 gene was significantly higher in multi-costa properties yak than that in common yak(P<0.05)，while the expression level of Hoxa10 gene was significantly lower in multi-costa properties yak(P<0.05).The methylation status of CpG2 island in the Hoxa6 promoter region was significantly different between multi-costa properties and common yak，especially on No.3，4，8，20 and 21 CpG sites.However，the methylation status of CpG1 island in the Hoxa10 promoter region was significantly higher in multi-costa properties yak than that in common yak(P<0.05)，and there was almost no methylation on No.9 and 12 CpG sites in common yak.All above results suggested that the high DNA hypermethylation of Hoxa10 gene in multi-costa properties yak inhibited the expression of Hoxa10 gene in a certain degree，and the low DNA hypomethylation of Hoxa6 gene enhanced the expression of Hoxa6 gene.It was concluded that methylation in Hox promoter regions may have some influence on transcription regulation of Hox genes，and there may be some correlation with the formation of multi-costa properties.

 The research was conducted to clone the sheep CYP19 gene ovarian promoter fragments and detect the tissue-specific expression in cells level.According to the known sequence the specific prmiers were designed，1.1 and 0.5 kb of sheep CYP19 gene ovarian promoter fragment were amplified by PCR，then the sequences were analyzed by software with published sequences.After eukaryotic expression vector pCYP19-1.1-EGFP-N₂ and pCYP19-0.5-EGFP-N₂ were constructed by cloning promoter fragments into the pEGFP-N₂ vector without CMV，then the recombinant plasmids were transfected into sheep granular cells and fetal fibroblast cells by liposome Lipofectamine ^TM LTX+PLUS.The EGFP fluorescence expression was observed under the microscope after transfection for 24，48 and 72 h.The sequenced results showed that sheep CYP19 gene promoter fragments which were cloned were 1.1 and 0.5 kb length and had highly homologous with the published sequences.The sequence analyzing with transcription factor binding sites prediction software indicated that the promoter fragment contained a core promoter cis-element smilar with TATA box and transcription factor binding sites.24 h after transferring with pCYP19-1.1-EGFP-N₂，the EGFP-expressing positive granulosa cells could be observed，48 h after transferring，the EGFP-expressing positive granulosa was increased，72 h after transfection the EGFP-expressing positive granulosa increased to the most.But after transfection the EGFP which was expressed in sheep fetal fibroblast cells was little.The results also showed that no EGFP-expressing positive granulosa cells and fetal fibroblast cells were observed at 24，48 and 72 h after transfection with pCYP19-0.5-EGFP-N₂.The sheep CYP19 promoter 1.1 kb can drive foreign gene expressing in sheep granulosa cells and it can be used for the functional studies of fecundity-related genes and transgenic animal research，but it is not ovarian specific expression promoter.

In order to reprogram the Japanese Black cattle fetal fibroblasts(JBCFF)，the Xenopus laevis oocyte extracts was used to incubation with the cells in present study.After 24 hours of culture in medium，there was no significant difference in acetylation of histone H3K9 between the JBCFF treated with extracts and the cells untreated by immunofluorescence assay.The well-defined colony structures of JBCFF treated with extracts were formed with the continuous culture for 5-6 d and the alkaline phosphatase staining of the cells colony was positive.Similarly，the OCT4 protein was detected in the cells colony.Additionally，Oct4 and Nanog mRNA expression，two pluripotency marker genes，were detected in those cells by RT-PCR，but Sox2 gene expression was not detected.Furthermore，qPCR results showed that the expression levels of Oct4 and Nanog genes in colony cells increased with days of culture(4，5，6 d ) after extracts treatment.In conclusion，the Xenopus laevis oocyte extracts could induce the partial reprogramming of the JBCFF into a low differentiated state.

This study aimed to evaluate the effects of guanidinoacetic acid(GAA) and combination of GAA and betaine on muscle energy metabolism and meat quality in finishing pigs.A total of 180 cross castrated male pigs(Duroc×Landrace×White) were randomly assigned to 3 experimental diet groups：control(CON，basal diet)，GAA group(GAA；basal diet supplemented with 1 g of GAA per kg of feed)，combination group(CGB；basal diet supplemented with 1 g of GAA and 0.5 g of betaine per kg of feed).Each treatment was replicated in 3 pens with 20 pigs each.The experimental period lasted 15 d.Compared with the control group，GAA and CGB supplementation significantly reduced drip loss(P<0.01)，hardness，cooking loss and shear force(P<0.05)，increased pH_{45 min} (P<0.05)，pH_{24 h} (P<0.01).In addition，dietary GAA and CGB increased the contents of creatine(P<0.05)，phosphocreatine(P<0.01) and adenosine triphosphate(ATP)(P<0.001) in muscle.The results indicated that the dietary supplementation with GAA and CGB could improve meat quality via regulating muscle energy metabolism of finishing pigs.

To assess the effect of cassava meal adding level and conditioning temperature on growth performance，nutrients digestibility and utilization of broilers，a 4×3 factorial design was used with 4 levels of cassava meal at 0，15%，30%，45%，and conditioning temperature at 60，75，90°C.A total of 1 920 Cobb male chicks of 1-day-old were randomly allocated to 12 treatments，and 8 replicates for each of 20 chicks per replicate.The diets were fed to birds for 21 days.The results showed that the effect of interaction between cassava meal adding level and conditioning temperatures on hardness and durability of diets(P<0.05)，starch digestibility at gizzard and starch digestion rate were significant(P<0.01).Hardness and durability of diets were significant decreased with the increasing of cassava meal level(P<0.05 or P<0.01)，and the reverse case for the starch digestibility at proximal jejumum and starch digestion rate(P<0.01).Group containing 45% cassava meal was higher at G/F than that of control(P<0.01) and 15% group(P<0.05)，but the dry matter(DM) apparent digestibility and apparent utilization and AME were lower than those of control(P<0.05)，and DM apparent digestibility was also lower than that of 15% group(P<0.05)，and the AME was lower than that of both 15% and 30% groups(P<0.05).Group containing 45% cassava meal had higher starch digestibility at gizzard，distal jejumum and proximal ileum than that of 15% group and control(P<0.01).Starch digestibility at proximal jejumum and starch digestion rate were significant improved with the increasing of cassava levels(P<0.01).Hardness and durability of diets were significantly increased with the increasing of conditioning temperature (P<0.01 or P<0.05).Average body weight daily gain of broilers fed diet conditioned at 75 ℃ was lower than that of others(P<0.05)，and the daily feed intake was lower than that of birds fed diets conditioned at 90 ℃(P<0.05)，and the same case for the starch digestibility at gizzard and distal jejunum and starch digestion rate(P<0.05).Diet conditioned at 60 ℃ had the lower G/F，starch digestibility at gizzard，proximal and distal jejunum，and starch digestion rate than those of diet conditioned at 90 ℃(P<0.05)，but the reverse case for the DM apparent digestibility and apparent utilization(P<0.05) and AME(P<0.01).The results suggest that，under this trial conditions，the optimum cassava inclusion level of diet and conditioning temperature is 30% and 75 ℃，respectively.

This experiment was conducted to evaluate and compare the accuracy of 8 published models to predict the enteric methane emissions in dairy cows，and analyze the factors that could affect the accuracy of prediction.28 Chinese Holstein dairy cows were selected in the Wangcheng Bairuopu dairy farm，Hunan province，to determine the enteric methane emissions，body weight，milk production，feed intake and other nutrients intake(acid detergent fiber，neutral detergent fiber and gross energy) and etc.The difference between the predicted and observed values of enteric methane emissions from dairy cows was estimated and compared based on the mean squared prediction error（MSPE）and concordance correlation coefficient（CCC）methods，and the factors influencing the accuracy of predictions of 8 published models were analyzed.The results showed that the highest accuracy of prediction for model 8，medium accuracy of prediction for model 1，2，3 and 6，and the lowest accuracy of prediction for model 4，5 and 7.The errors influencing the accuracy of model 1 and 2 were mainly caused by deviation of regression slope from unity；the errors influencing the accuracy of model 3，6 and 7 were mainly caused by the overall bias；the errors influencing the accuracy of model 4 and 5 were caused by both deviation of regression slope from unity and overall bias.The predicting errors of model 1 and 2 were attributed to the difference of calculated Ym and IPCC default Ym；the predicting errors of model 3 was due to that enteric methane emissions for the model 3 was higher than that in this study under the same DMI；the predicting errors of the model 4，5 and 6 were mainly caused for not considering the rumen feed digestion of carbohydrate，ruminal passage rate of dietary nutrient and etc；the predicting errors of model 7 was due to that rice straw intake couldn’t represent real dietary forage intake.Model 8 with 2 variables of milk yield（kg•d^-1）and BW（kg）had the highest accurancy of prediction in this trial.Experiments are still needed to collect more data to develop accurate and reliable methods of models to predict enteric methane emissions in dairy cows.

This study was conducted to investigate the effect of T-2 toxin exposed to BALB/c mice on pH and moisture content in feces，apparent digestibility of nutrients，mineral elements and amino acids，as well as morphological structure in small intestine.A total of 80 BALB/c mice weighted about(20±2) g were randomly allotted to 4 groups(20 mice per group).The mice were fed one of four kinds of diets(0，0.4，1.0 and 2.5 mg•kg^-1•BW T-2 toxin exposed daily via intragastric administration for 28 d).The results showed as follows：the pH in feces in 1.0 mg•kg^-1•BW group was higher than that in control and 0.4 mg•kg^-1•BW groups(P<0.05)，pH in feces in 2.5 mg•kg^-1•BW group was the highest among all 4 groups(P<0.05 or P<0.01).The apparent digestibility of crude protein，ash，crude fiber and ether extract were greatly decreased with the increasing of T-2 toxin addition，and there were significant difference among different treatment groups(P<0.05 or P<0.01).The apparent digestibility of mineral elements(including Ca，Fe，Mg，Na and P) of 0.4 mg•kg^-1•BW group was largely reduced compared with control group(P<0.01).The mineral elements(including Ca，Zn，Mg，K，Mn and P) in 1.0 and 2.5 mg•kg^-1•BW groups were lower than those of control and 0.4 mg•kg^-1•BW groups(P>0.05 and P<0.01)，and 2.5 mg•kg^-1•BW group was significantly lower than 1.0 mg•kg^-1•BW group(P<0.01).The apparent digestibility of amino acid(including Asp，Thr，Ser，Glu，Gly，Ala，Val，Ile，Leu，Phe，Lys，His，Arg and Pro) in 1.0 mg•kg^-1•BW group were also observably declined compared with control or 0.4 mg•kg^-1•BW groups(P<0.05 and P<0.01)，and the digestibility of all kinds of amino acid in 2.5 mg•kg^-1•BW group were the lowest among all the 4 groups(P<0.01).Moreover，the duodenum，jejunum and ileum presented mucosal damage and microvilli exfoliated when the dose of T-2 toxin was more than 1.0 mg•kg^-1•BW.In addition，reduction of villous number and villous length，increase of crypt depth，decrease of the ratio of villus length to crypt depth were also observed in 1.0 and 2.5 mg•kg^-1•BW groups，and there were significant differences between 1.0,2.5 mg•kg^-1•BW groups,and control,0.4 mg•kg^-1•BW groups(P<0.05 and P<0.01)，respectively.It was concluded that the supplementation of T-2 toxin could damage intestinal mucosal，induce the increase of pH，decrease the apparent digestibility of nutrient，mineral elements and amino acid，and all these damage and decreasing were associated with dose-effect relationship of T-2 toxin.

To determine the pathogenic bacterium infecting the small tail Han Sheep，three strains，named LK-1，LK-2 and LK-3，were isolated from the heart，liver，spleen，lungs，kidney and intestines of three dead cases on a sheep farm in Henan province.Identification by the Biolog Microbial Identification System，and further 16S rRNA sequence and phylogenetic analysis demonstrated that the three isolates were Citrobacter freundii.Infection with the bacterial suspension to the healthy small tail Han Sheep could reproduce the diseased symptoms as occurred naturally and the same bacterium could be recovered from these infected sheep.Three isolates also have certain pathogenicity to the BALB/c mice，with the lethal doses of 5.0×10⁷，6.0×10⁷，5.8×10⁷ CFU，respectively.The susceptibility test to antibiotics demonstrated that three isolates LK-1，LK-2 and LK-3 were susceptible to 14 drugs，including cefotaxime，gentamicin，tetracycline，chloramphenicol，enrofloxacin，etc.Three isolates have similar antimicrobial resistance profiles to 17 of 21 drugs in this study.But they have different profiles to 4 drugs，including amoxicillin，cefaclor，streptomycin and azithromycin.In this study，we reported the pathogenicity on Small Tail Han sheep of Citrobacter freundii at home and abroadfor the first time.Citrobacter freundii is the pathogen of the dead small tail Han Sheep cases.

In order to investigate the relationship between apoptosis and transcription and expression of several major cytokines during Mycobacterium tuberculosis (MTB)infection，THP-1 derived macrophage was selected to establish in vitro model.Infected cells with different virulence MTB (H37Rv，BCG)，flow cytometry was used to detect macrophage apoptosis at six time points，quantitative PCR was used to detect transcription level of cytokines TNF-α，IL-10，IL-6，ELISA was used to detect the secretion of cell supernatants of three cytokines.The results showed：the early apoptotic rate of BCG and H37Rv group were both higher than control group and the difference was statistically significant （P<0.05）.Transcription levels of cytokines，TNF-α，IL-6 were consistent with the level of secretion.There was an increase with time on apoptosis of low virulent strain，and there is a same trend on secretion of TNF-α，an opposite tendency on secretion of IL-6.Apoptosis of virulent strains was less than attenuated vaccine strain.Secretion of virulent strains was complex，like increase first and then suppression.The relationship between cytokines and apoptosis of cells at different time points may provide new ideas for study of macrophage apoptosis mechanisms.

A lentogenic Newcastle disease virus (NDV)，SD22/13，was isolated from heron.The biology and pathogenicity were characterized in order to understand the evolution of NDV better.F gene of SD22/13 was sequenced and compared with that of reference strains.The antigenicity of the virus was compared with that of LaSota and other genotype Ⅶ virulent NDV strains via cross hemagglutination inhibition (HI).The pathogenicity of SD22/13 to SPF chicken was also conducted.The results indicated that SD22/13 was classified as a class Ⅰ genotype 3 NDV based on a partial sequence of the F gene.The antigenicity correlation between SD22/13 and LaSota was 56.1%，and the correlation of which with genotype Ⅶ NDV strains was less than 50%.Experiments on animals demonstrate that the isolate SD22/13 can replicate well in SPF chicken and induce a high immune antibody and had slight pathogenic to immune organ and kidney.Chickens inoculated with SD22/13 were challenged with genotype Ⅶ virulence NDV strain on 30dpi，and there had no death in chickens infected with SD22/13.The results demonstrated that SD22/13 stain had significant differences with genotypeⅦ NDV in biological characterization and pathogenic to chickens.Although the antigenic correlation was low between SD22/13 and genotype Ⅶ NDV，class I viruses can provide 100% protection rate，but it cannot block virus shedding upon challenge with genotypeⅦ virulent NDV.

This study was conducted to find out the avian reovirus (ARV) proteins which can activate the phosphatidylinositol 3-Kinase-dependent Akt(PI3K/Akt) pathway.According to the analysis of amino acid sequence of ARV proteins，the σA，σNS，μA，μB and μNS were selected as the putative proteins which mediated the activation of PI3K/Akt pathway.Reconbinant plasmids，σA-pcAGEN，σNS-pcAGEN，μA-pcAGEN，μB-pcAGEN and μNS-pcAGEN were constructed and transfected into Vero cells，and the expression of the target genes were identified by immunofluorescence test and Western blot.The phosphorylated Akt (P-Akt) profile of transfected cells were examined by flow cytometry and Western blot.The results showed that σA，σNS，μA，μB and μNS genes were expressed in the Vero cells，and the expression of P-Akt of σA-pcAGEN and σNS-pcAGEN groups increased markedly.The PI3K inhibitor LY294002 could inhibit the expression of the P-Akt of σA-pcAGEN and σNS-pcAGEN groups.The results indicate that σA and σNS protein could activate the PI3K/Akt pathway.

The research aimed to understand the main pathogens of pigs diarrhea virus found in Inner Mongolia in order to provide theoretical basis for pig diarrhea control and prevention.Through the Multiple RT-PCR method，the porcine epidemic diarrhea virus (PEDV)，transmissible gastroenteritis virus (TGEV)，and porcine rotavirus (PoRV) from 394 tissue samples collected from the pig farms in 8 cities of Inner Mongolia were tested.M gene of the PEDV positive materials were cloned and analyzed along with the sequence alignment and phylogenetic trees of 48 M gene sequences from other domestic and international.The tested results indicated that the positive rates of PEDV and PoRV were 63.96% and 2.03%，respectively，yet the TGEV tested negative，The Mixed infection rate of PEDV and PoRV was 2.03%.Phylogenetic analysis indicated that CH/NMG/ XLGL strain stayed close relative with classic strains such as CV777，located at G2.The rest of the strains stayed close relatives with the domestic many provinces before 2012，and Thailand，South Korea，Russia，Vietnam and other neighboring countries were same branch，G1-1.United States，in particular，the nucleotide sequence homology was 99.5%-100%.From 2013 until now，the main pathogeny for pig diarrhea in Inner Mongolia was PEDV，followed by PoRV.The infection degree was different in varying cities，and pigs of all ages can be infected，especially the suckle pigs and the sow.Most of the prevalent strains exhibited higher affinity with the strains isolated from the most provinces and cities domestic since 2012，but the stains were far apart from Inner Mongolia strains isolated before 2007，vaccine strains and classic strains，and some sites appeared base mutations.

As a key enzyme in the glycolytic pathway，glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has wide applications (vaccine candidate and drug target) against some parasites，but until now，few studies have been carried out in tapeworms.In order to determine its molecular character and practical value，Echinococcus granulosus GAPDH (EgGAPDH) was cloned，expressed and analyzed.The EgGAPDH gene isolated from E.granulosus cDNA was 1 011 bp in length，encoding 336 amino acids.The quaternary structure of this protein consisted of four chemically identical subunits，O，P，Q，and R.Each subunit consisted of an NAD+-binding domain (residues 4-151 and 315-336)，a catalytic domain (residues 156-314) and one enzyme active site (residues 149-156).Recombinant EgGAPDH was expressed and purified from Escherichia coli.Specific immunogenicity of EgGAPDH was shown by Western blot，ELISA analysis，and epitope prediction (16 B cell epitopes and 9 cytotoxic T lymphocyte epitopes).EgGAPDH gene was successfully amplified，cloned and expressed，EgGAPDH has three functional areas and a number of epitope prediction，combing with the results of Western blot and ELISA，it may be serve as a vaccine candidate and drug target against larval of E.granulosus.

In order to clarify distribution of Nosema germplasm in China，68 samples of adult honey bee from 16 provinces of China were collected.All of the samples which with clear signs of population depletion and positive to microsporidian spores using light microscopy were selected for molecular germplasm identification by method of multiplex PCR.The sequenced products of all of the 68 samples were identical to the corresponding Nosema ceranae sequence.Only one of samples collected from Shandong was found that had infected by Nosema apis.The results indicated that vast majority species of nosema infect Apis mellifera in China is Nosema ceranae，Nosema apis is very rare within Apis mellifera.

The objective of this study was designed to investigate the effectiveness of mast cell pattern recognition of recombinant VP1-VP4 of foot-and-mouth disease virus(FMDV).To this end，murine mast cell line P815 were pulsed with the recombinant VP1-VP4 of FMDV after inhibitors of Toll-like receptor 2/4(OxPAPC)，mannose receptors(Mannan)，and scavenger receptors(Sennoside B) were administered to murine mast cell line，respectively.Degranulation of P815 mast cells stained with toluidine blue was observed at the given time.And the levels of TNF-α in P815 mast cell culture supernatants were determined by ELISA at different timepoints.It was showed that the number of degranulated P815 mast cells with administration of scavenger receptor inhibitor was significantly lower than that of cells pulsed with recombinant VP1-VP4 protein at 15 min and 30 min(P<0.001).Intriguingly，the TNF-α release from the mast cell line activated by recombinant VP1-VP4 at 24 h was significantly blocked with three inhibitors(P<0.01)，of which，mannan，a mannose receptor inhibitor，showed the strongest inhibition on TNF-α release.Our data indicate that murine mast cell line P815 degranulation triggered by recombinant VP1-VP4 was predominantly mediated via the recognition of scavenger receptors,while the TNF-α release from the activated mast cells was constitutively initiated with the recognition of mannose receptors，TLR2，and TLR4.Of note，recognition of recombinant VP1-VP4 through mannose receptors plays a predominant role in triggering TNF-α release from P815 mast cells activated by recombinant VP1-VP4.

The aim of the present study was to explore the signaling relationship between the chemokine regulated on activation，normal T cell expressed and secreted (RANTES) and its receptors［chemokine receptor 1(CCR1) and chemokine receptor 5(CCR5)］.The molecular cloning techniques and Western blotting were used to construct eukaryotic expression vectors，and then used dual luciferase report system tests to detect the expression of firefly luciferase after receptors activated or not with RANTES，to indirectly reflect the amount of cAMP response element(CRE) and serum response element(SRE) expression level.The results showed that CCR1 activated with RANTES could reduce the level of CRE expression and enhance the level of SRE expression in HEK293T cell，and CCR5 activated with RANTES could enhance the level of CRE expression and SRE expression.The results proved RANTES interacted with CCR1 could stimulate the level of SRE expression and reduce the level of CRE expression increased by Forskolin，with CCR5 could enhance the expression level of CRE and SRE.

This study was conducted to characterize the structure and distribution of extracellular matrix(ECM) proteins of testis in aging yak testis.The testis from 9 aging and 10 young adult yak were prepared for distribution investigation of laminin(LN)，type IV collagen(ColⅣ) and heparan sulfate proteoglycans(HSPG) by immunohistochemistry and histochemistry methods.The observations made with the light microscope showed obvious morphological changes of aging，there were partly progressive regressed in the seminiferous epithelium and the volume of interstitial tissues has further increased and abundant with collagen and reticular fiber，and also there were AB-PAS positive stain in aging yak seminiferous basement and blood capillary.Statistical data showed that the number of Sertoli cells and Leydig cells in aging yak testis were reduced obviously，but the cross-sectional area of the seminiferous tubules and average interstitial tissue areas were more larger than which in the young yak testis.Immunostaining analysis appeared that the LN was present similarity in Sertoli cells and peritubular myoid cells in the aging and young adult yak.The relative expression of LN in spermatogenic cells was decreased and almost no detected in aging yak Leydig cells(P<0.01).By contrast，the distribution of Col Ⅳ without statistical differences in yak testis at different ages.The relative expression of HSPG in the aging yak testis tissue was more lower than which in young adult yak，and the strong immunoreactivity for HSPG was seen in the Leydig cells(P<0.01).In addition，the average absorbance of LN，Col Ⅳand HSPG expression was no obvious difference between different ages.All the elements of the intralobular interstitial space may undergo degenerative changes in aging yak which live in plateau environment，the secretion of Col Ⅳ increased company with the synthesis of collagen fiber reinforced，and the distribution of ECM proteins LN and HSPG were significantly decreased may closely related with the alterations of Leydig cells′ synthesis and secretion ability.

This study was conducted to determine variation of left and right quarter skin temperature and their temperature difference in order to evaluate the possibility of temperature difference of left and right quarter as indicator of subclinical mastitis detection by determination of left and right quarter skin temperature difference range.Thermal images of rear left and rear right quarters of 587 Chinese Holstein dairy cows were collected by infrared thermograph technology(IRT).The cows were chosen randomly.The udder skin temperature was calculated using image analysis software.The result demonstrated that rear left and rear right quarter skin temperature were characteristic of normal distribution.The variation of rear left and rear right quarter skin temperature were （35.57±1.31）℃ and （35.51±1.34）℃ respectively.Their difference was not significant(P>0.05) and was distributed symmetrically.Mean somatic cell count(SCC) in milk had the rising trend as left and right quarter temperature difference increasing.Mean milk SCC was(298 ± 110)×10³ mL^-1 when rear left and rear right quarter skin surface temperature difference was above 1.5 ℃ and left and right quarter skin temperature was significantly different(P<0.05).This study showed that subclinical mastitis could be detected initially when skin temperature difference between left and right quarters was over 1.5 ℃ and SCC was above 3×10⁵ mL^-1 for the detection of subclinical mastitis.

The aim of this study was to reveal the effect of Hericium erinaceus polysaccharide(HEP) on the intestinal mucosal immune and antioxidant capacity of ducklings which were infected with muscovy duck reovirus(MDRV).Through the establishment of simulation model which was similar with MDRV natural infection，200 of 1-day-old healthy ducklings were randomly divided into blank control group，HEP control group，cohabitation infected control group and HEP prevention group.The samples from ducklings in each experimental group were collected and determined in the first day，the third day，the sixth day，the tenth day，the fifteenth day and the twenty-first day after cohabitation infection.The results showed that，HEP could promote the secretion of SIgA，IFN-γ and IL-4 on intestinal of infected MDRV ducklings in different degree，also with the maintenance of intestinal mucosal immune balanced effect，and increased SOD activity and GSH content，decreased the content of MDA significantly，it turned out to have the function of improving antioxidase activity，removing free radicals and inhibiting lipid peroxidation.These results indicated that HEP had certain effect on regulating intestinal mucosal immunity of the ducklings and also improving the ability of anti-oxidation，reducing the intestinal barrier function which was damaged by MDRV effectively.

The aim of this study was to clone porcine STARD3NAL(STARD3 N-terminal like protein)，and analyze the structure of STARD3NAL，and investigate its expression in porcine different tissues.The porcine STARD3NAL gene was cloned by 5′，3′-RACE and RT-PCR，the protein structure of porcine STARD3NL was analyzed by bioinformatics methods.The expression patterns in different tissues were detected by real-time RT-PCR.The results showed that the porcine STARD3NL cDNA was 1 319 bp(GenBank accession No.：HQ634946) in length，which coded 235 amino acids and shared highly identity with those of other species.The phylogenetic tree analysis indicated the porcine STARD3NL was closely related to cattle and sheep，but distantly related to the zebrafish.The porcine STARD3NL protein contained conserved MENTAL domain and was hydrophobicity protein and there were 4 transmembrane helices in the STARD3NL protein.The secondary structure of STARD3NL protein showed that the STARD3NL protein fold for 60% into α-helix.This polypeptide contained no signal peptide in its amino acids，and was a nonsecretory protein.The residing probability of STARD3NL in the endoplasmic reticulum was 39.1%.By real-time PCR，mRNA expression of porcine STARD3NL were detected in all 14 tissues investigated，and STARD3NL mRNA expression levels in liver were the highest and in longissimus dorsi were the lowest.In addition，no significant difference of STARD3NL mRNA expression levels in lung tissues was detected between Laiwu pigs(LW) and YL pigs(Yorshire♂×Landrace♀) in infected circovirus group(P>0.05).These results will provide the data for further research of STARD3NL gene structures and function，breeding for disease resistance.

In order to explore the mechanisms of ulcerative colitis in animals and its impact on body，10-day-old SPF chicks were used to build 2，4，6-trinitrobenzene sulfonic acid(TNBS) -induced model by clystering with TNBS in dose of 50，100，and 150 mg•kg^-1，diluting with 50% ethanol，that can help us to analyzes the dose effect of TNBS-induced ulcerative colitis.Further observation and testing about the changes of the chicks’ clinical symptoms，pathology and body weight showed that the ideal medicine dose was 100 mg•kg^-1 of TNBS，which can induce ideal changes of clinical symptoms and pathology in experimental animals without uncontrolled injury and massive death.This study successfully established chicks’ pathological model of ulcerative colitis which laid a foundation for deep research of the development of ulcerative colitis.

 In present study，to investigate the relationship between polymorphism of Agouti gene and coat color in fox，the whole exon 2，whole intron 2 and partial exon 3 sequence of the Agouti gene were sequenced from 58 fox samples，belonging to 5 species of 2 genera.A total of 6 SNPs(g.269G>T，g.295C>T，g.325T>G，g.518G>A，g.608C>A and g.846G>A) in intron 2 were detected firstly，revealing high genetic diversity.All individuals(n=25) of Vulpes vulpe were homozygous at the 6 sites，and the genotype were in order of GG，CC，TT，GG，CC and GG.All individuals(n=33) Alopex lagopus were another homozygous at the 6 sites，and the genotype were in order of TT，TT，GG，AA，AA and AA.Total 2 haplotypes(H1：GCTGCG and H2：TTGAAA) were found at the 6 mutation sites and their frequencies were 42.1% and 57.9%，respectively.H1 was widely distributed in all tested individuals of Vulpes vulpes，but H2 was widely distributed in all tested individuals of Alopex lagopus.Further analysis showed that the detected SNPs and haplotypes were not significant relation to coat color，but speculated that they may be the important functional sites and haplotypes to distinguish Vulpes vulpe and Alopex lagopus.