This study was aimed to prepare canine parvovirus (CPV) VP2 protein polyclonal antibody.The recombinant expression vector pET28a-CPV-VP2 was constructed and transfromed into E.coli BL21 (DE3),the expression of recombinant proteins was induced by IPTG from which the fusion protein was identified by SDS-PAGE.The target protein was purified and emulsify with adjuvant,the prepared immunogen was inoculated into rabbit by subcutaneous injections to prepare of VP2 protein specific polyclonal antibody.The immuno-activity,titers,neutralization titers of the prepared polyclonal antibody were determined by immunoperoxidase monolayer assay (IPMA).The results showed that the expressed recombinant protein VP2 (rVP2) existed in the form of inclusion body with a molecular weight of 72 ku.The prepared polyclonal antibody titer was 1 600 dilution,the virus titer was 10⁷ TCID₅₀/mL,the neutralizing titer was 1∶2 884.The antibodies showed specific reaction with CPV.In conclusion,rVP2 specific polyclonal antibody showed wonderful immunocompetence,specificity and neutralizing activity,providing foundation for the development of genetic vaccine and clinical therapeutic method.

Myostain (MSTN) gene is a member of transforming growth factor-β (TGF-β) superfamily.In order to study the relationship between Luxi Huang cattle MSTN gene upstream sequence polymorphism and the transcriptional expression,DNA was extracted from leg muscle cells of Luxi Huang cattle and then sequenced.After analyzing MSTN upstream sequence, an evolutionary tree was constructed and a SNP site was found at 805 bp of MSTN upstream sequence of initiation codon.Two expression vectors contained the SNP site were constructed to transfect the C2C12 cells in vitro to study the influence of MSTN gene upstream sequence polymorphism on transcription activity.The results showed that single nucleotide change on MSTN gene upstream sequence affected the expression.It suggested that MSTN gene upstream sequence polymorphism of Luxi Huang cattle would affect transcription activity and play an important role in gene expression regulation.

We carried out the research of the effect of taurochenodeoxycholic acid (TCDCA) on the apoptosis of the mouse macrophage cell line RAW264.7 and expression of IAPs including CIAP-1,CIAP-2 and XIAP.The results showed that TCDCA with the dose of 0.05,0.10 and 10 μg/mL could extremely significantly prevent cells apoptosis that were induced by DEX (P < 0.01).TCDCA with the dose of 1 μg/mL could significantly promote mRNA expressions of CIAP-1 and XIAP (P < 0.05),while the dose of 10 μg/mL could significantly promote mRNA expressions of CIAP-1,CIAP-2 and XIAP (P < 0.05),TCDCA could also extremely significantly up-regulate the expression of CIAP-1,CIAP-2 and XIAP (P < 0.01) in RAW264.7 cell line by DEX,but this depended on different doses of TCDCA.The above research results showed that TCDCA could resist the cells apoptosis of the mouse macrophage cell line RAW264.7 that were induced by DEX,which was related with the influence on the mRNA expression of IAPs.

The study was aimed to clone and express LpxB gene,and perform the bioinformatics analysis of protein.The genomic DNA of Brucella melitensis M5-90 was used as template.According to the genome sequence of M5-90 on GenBank,a pair of primers was designed.LpxB gene,which was 1 188 bp,was amplified by PCR,and was ligated into pMD20-T vector.The constructed recombinant plasmid pMD20-T-LpxB was transformed into E.coli DH5α.The recombinant plasmid was confirmed by endonuclease digestion and sequencing.The coding region of LpxB from pMD20-T was digested by BamHⅠ and XhoⅠ.Then,the fragment was inserted into prokaryotic expression vector pET-28a,and the positive plasmid was named pET-28a-LpxB.The pET-28a-LpxB was transformed into E.coli BL21 (DE3).The expressed protein was identified by SDS-PAGE and Western blotting.DNAMAN and BioEdit softwares were used to analyze the sequence of amino acids encoded LpxB gene.The results showed that the CDS of LpxB was successfully cloned and expressed.The secondary structure of LpxB protein consisted structure α -helix,extended strand,β-turn and random coil which accounted for 52.41％,14.94％,8.10％ and 24.55％,respectively.

To optimize the loop-mediated isothermal amplification (LAMP) method to detect Mycobacterium tuberculosis in milk,the highly conservative 16S rRNA gene of Mycobacterium tuberculosis was selected as a target to design specific primers.Compared the modified method of CATB/NaCl,bacterial genome DNA extraction kit and thermal cracking method to extract the DNA of Mycobacterium tuberculosis,the best approach was chosen.Mycobacterium tuberculosis suspension liquid was diluted with sterilization milk to confirm the susceptibility of this assay.And then Brucella,Escherichia coli,Staphylococcus aureus,Listeria monocytogenes,Bacillus pasteurii and Salmonella were used for specificity detection.The results showed that the modified method of CATB/NaCl was better than the others.The sensitivity of LAMP was 3×10⁰ CFU/mL,and the specificity was 100%.The sensitivity of detecting Mycobacterium tuberculosis in milk by LAMP was 3×10¹ CFU/mL.

Hair follicle is capable of self-renew and its growth cycle is a dynamic process.In order to investigate the vary of differential expression genes in the Nei Mongol cashmere goat hair follicles during anagen,the mRNA expression levels of different genes in skin hair follicles were detected from June to October using Illumina technology.Evaluation of transcriptomes was undertaken to understand the time-course gene expression involved in both follicle types.The samples were collected from June to October.Most of differentially expressed genes appeared between July to August in primary hair follicle (PF) and between August to September in secondary hair follicle (SF).Seven differentially expressed genes,FAM83C,LOC102185501,FYCO1,CTTNBP2NL,C6orf132,KRTAP29-1 and CRYBG3 were identified.Their expression demonstrated similar trends at different months of cashmere growth cycle and the SF postpone for about one month.These findings indicated the hysteresis in SF gene regulation comparing with PF,and could provide the useful theoretical basis for the molecular mechanism of cashmere regeneration.

HOX genes express in nearly all eukaryotic cells such as yeast and human.To study tissue expression and bioinformatics of HOXC10 gene,the samples of heart,liver,spleen,lung,kidney,chest muscle,leg muscle,hypothalamus and ovary tissues were collected from 6 Jinghai Yellow chicken with 8-week-old to analyze the expression pattern of HOXC10 gene in different tissues by qRT-PCR method,and the different expression level were detected in ovary of 300 days old chickens in high yield group and low yield group.The bioinformatics of HOXC10 gene were analyzed by a variety of online softwares of Gallus gallus.The result showed that the expression level of HOXC10 gene in leg muscle was extremely significantly higher than that in other tissues of Jinghai Yellow chicken (P < 0.01);And it's expression level in low yield group was significantly higher than that in high yield group (P < 0.05);HOXC10 gene of Gallus gallus shared 77.4%,94.8%,98.6%,68.3%,67.9%,66.0%,59.0%,68.1% and 79.4% with Meleagris gallopavo,Columba,Anas platyrhynchos,Homo sapiens,Mus musculus,Bos taurus,Sus scrofa,Capra aegagrus hircus and Xenopus laevis,respectively;HOXC10 gene encoded 354 amino acids in which serine was the most abundant;There were 24 potential phosphorylation sites and a HD in 280 to 342 position of amino acid sequence;The subcellular of the encoding protein was mainly located in the cell nucleus,and it had no cross membrane structure.HOXC10 gene might play an important role in regulation network of growth and reproduction in Jinghai Yellow chicken.

To study the attenuation molecular basis of porcine reproductive and respiratory syndrome virus (PRRSV),whole genomes of PRRSV JXA1-5 (parental strain) and JXA1-86 (attenuated strain) were sequenced,compared and analyzed.And genomic sequences of JXA1/JXA1-86 were compared with the other two pairs of virulent parental/attenuated vaccine strains.The results showed that there was higher amino acid mutation rate in structural proteins among the three pairs of strains indicating that PRRSV JXA1 strain virulence changes were more related to structure proteins variations.Compared with JXA1,the amino acids encoding Nsp5,Nsp6,Nsp8,Nsp12,M and N proteins of JXA1-86 strain did not change,suggesting that virulence attenuating might not be relevant to those proteins.The genomic sequences of each generation PRRSV sequence were compared and analyzed,and the results showed that the variations among A₉₂₈V,I₁₁₅₅M,E₁₆₂₉D in ORF1a,I₁₁₈V in GP2,H₇₉N in GP3,I₁₂₄V in GP4 and K₅₉N in GP5 between 70th and 86th generations were found,which turned out that virulence attenuating might be associate with those variations.

In this study,a SYBR GreenⅠ dye based on Real-time quantitative PCR was established using the specific primers-pair according to gE gene characterization of porcine pseudorabies virus(PRV).The optimized results demonstrated that the detection assay with good linear determination range from 7.53×10¹ to 7.53×10⁶ copies per reaction.There was no cross-reaction occurred with nucleic acids extracted from the common porcine infectious diseases,such as porcine circovirus 2 (PCV2),porcine parvovirus (PPV),Haemophilus parasuis and Streptococcus susi,no amplification signals were detected.The results showed that the melting curve analysis with only one specific peaked with a melting temperature (Tm),which was (92.9±0.1)℃ at detecting PRV positive samples,also no specific peak could be detected for common porcine infectious diseases described above.Series experiments were carried out in order to assess the sensitivity,specificity and reproducibility for the method,following by the intra-assay and inter-assay CVs for Ct values obtained with the standard plasmids.The intra-assay and inter-assay statistics were 0.31% to 1.14% and 0.42% to 1.74%,respectively.All the results showed that the established method was sensitive,specific and reproducible,which meaned it could be used for the research of pathogenic mechanism of porcine pseudorabies virus.

In order to study the function of canine distemper virus (CDV) V protein,V gene fragment was inserted into the pGEX-6p-1 vector,and then pGEX-6p-1-CDV-V recombinant expression plasmid was obtained.Purified V protein immunized BALB/c mice to prepare the positive serum.Meanwhile,to confirm the distribution of V protein and subcellular localization with confocal laser technology,a eukaryotic expression recombinant plasmid pcDNA3.1-CDV-V was built,then transfected into Vero cells.The results showed that V protein was successfully expressed and the positive serum was prepared.The recombinant plasmid pcDNA3.1-CDV-V was expressed in Vero cells and mainly expressed in cytoplasm.The results laid a function for functional studies of CDV V protein.

The experiment was carried out to investigate effects of three strains combinations of Lactobasillus buchneri (LAB),Aspergillus niger (AN),Candida tropicalis (CT),Bacillus subtilis (BS) and Lactobacillus plantarum (LAP) on fermentation quality of cassava residue.In this study,cassava residue were used as fermentative raw material,three strains were combined by 1∶1∶1 chosen from LAB,AN,CT,BS and LAP,getting five different combination.Combination 1:LAB+AN+LAP;Combination 2:LAB+CT+LAP;Combination 3:LAB+BS+LAP;Combination 4:AN+CT+LAP;Combination 5:CT+BS+LAP;1% urea or 1% urea+0.6% brown sugar was added to each combination;Blank group was without any additives;1% urea was added to control group Ⅰ and 1% urea+0.6% brown sugar were added to control groupⅡ.Experimental group Ⅰ was consisted of control group Ⅰ+different combination (1-5) while experimental group Ⅱ was consisted of control group Ⅱ+different combination (1-5).Water content in each group was about 65% by modulating with normal saline,each group was fermented in the vacuum of polyethylene film bag for 10 days.The results showed as follows:①pH of each experimental group was significantly lower than that of blank group and control group (P < 0.05),experimental group Ⅱ1 (LAB+AN+LAP+urea+brown sugar) could improve acetic acid content of cassava residue and reduce the NDF and ADF contents,and which were the lowest and were significantly lower than that of blank group and the control groups (P < 0.05);②Experimental group Ⅱ5 (CT+BS+LAP+urea+brown sugar) could improve the crude protein content in the cassava residue which was significantly higher than that of blank group and the control groups (P < 0.05);③It was favorable to propionic acid production by adding the corresponding strains+urea+brown sugar to the cassava residue fermentation;④Urea and brown sugar were added into cassava residue for improving the nutrition composition during the fermentation process was better than that only urea was added;⑤When the urea was added into the fermentation medium,the crude protein content in cassava residue was significantly increased,when cassava residue fermented quality was evaluate by butyric acid and acetic acid contents,all the groups got to the top of grade.In conclusion,solid-state fermentation using LAB+AN+LAP+urea+brown sugar or CT+BS+LAP+urea+brown sugar could offer an effective alternative to improve the fermentation quality of cassava residue.

In order to analyse the resistance to β-lactams antimicrobial agents and the prevalence of resistant genes of Salmonella in Guizhou province,130 Salmonella strains were isolated and identified from 9 different regions of scale pig farms.The drug sensitivity to 8 kinds of β-lactams antimicrobial agents were determined by using the broth microdilution method.All β-lactams resistant isolates were detected for the presences of TEM,OXA,CTX-M and SHV genes by PCR.The results showed that drug resistance of Salmonella to the commonly used β-lactams antimicrobial agents was very serious,and the resistance rate to ceftazidime was the highest (100%),followed by ampicillin and amoxicillin,were 76.15% and 80.77%,respectively.The resistance rates of ceftiofur and cephalexin were the lowest (46.15%).Salmonella strains were all of multiple drug resistance,of which double resistance was at lowest (2.31%),and eightfold resistance was highest (4.62%),multidrug resistance mainly concentrated in fourfold to sevenfold,accounted for 88.46%.PCR results showed that TEM,OXA,CTX-M genes detection rate were 85%,75% and 46%,respectively,while the SHV gene was not inspected.Resistant phenotype was basically consistent with resistant genes.The results indicated that the resistance of Salmonella stains from pig to β-lactams antimicrobial agents were widespread,and ceftazidime was particularly serious.The TEM,OXA and CTX-M genes were mainly carried β-lactams resistant genes in Salmonella isolates from Guizhou province.It had a great relationship between the prevalence of resistance genes and growth of antimicrobial resistance.

This study was conducted to compare the nutrient composition of longissimus dorsi between Diannan Small-ear pigs and DLY commercial pigs.53 Diannan Small-ear pigs and 58 DLY commercial pigs were chosen and longissimus dorsi samples were collected to determine the content of moisture,crude protein,crude fat,ash,amino acids and fatty acids.The results showed that the moisture content of Diannan Small-ear pigs was extremely significantly lower than DLY commercial pigs (P < 0.01).However,the content of crude protein and crude fat were extremely significantly higher than DLY commercial pigs (P < 0.01).And there was no significant difference in ash content (P > 0.05).The levels of threonine,valine,methionine,isoleucine,leucine,lysine and phenylalanine in longissimus dorsi of Diannan Small-ear pigs were extremely significantly higher than that of DLY commercial pigs (P < 0.01),and the content of myristic acid,palmitic acid,oleic acid,cis different oleic acid,11-cis-20 carbon olefine acid,saturated fatty acids,monounsaturated fatty acids and the total fatty acids in longissimus dorsi of Diannan Small-ear pigs were also extremely significantly higher than that of DLY commercial pigs (P < 0.01).In conclusion,Diannan Small-ear pigs had a better value in nutrient and development.

To study the effect of weaning stress on dairy buffalo calves,5 healthy calves were chosen and weaned,and the blood samples were collected at 1 day before weaning,and 1,14 and 21 days after weaning to analysis blood physiology and biochemistry indexes and immune function.The results showed that the count of white blood cell,lymphocyte,erythrocyte,monocytes and hematocrit,aspartate aminotransferase activity,blood glucose,cortisol and immunoglobulin G contents of dairy buffalo calves were no significant difference between before and after weaning treatments (P > 0.05).The count of neutrophils at 21 d after weaning were significantly lower than that of 1 d before weaning of dairy buffalo calves (P < 0.05).However,lactate dehydrogenase and alanine aminotransferase activity in weaned treatments were significantly higher than those of animals in the treatment before weaning (P < 0.05).The serum urea nitrogen of dairy buffalo calves at 21 d after weaning was significantly higher than that of 1 d before weaning (P < 0.05) and hemoglobin content at 21 d after weaning was significantly lower than that at 1 day before weaning (P < 0.05).It indicated that the physiological and immunity function of dairy buffalo calves were challenged under weaning stress.

The experiment was aimed to investigate the effects of xylanases from different sources on growth performance and nutrition metabolizability of Yellow-feather broilers.540 of healthy 1-day-old Yellow-feather broilers were randomly divided into three groups with 6 replicates per group and 30 broilers per replicate,and the trial period was 42 d.The control group was fed a wheat-corn-soybean meal diet,and groups A and B fed the basal diet supplemented with 200 g/t Trichoderma xylanase (30 000 IU/g) and bacterial xylanase (30 000 IU/g), respectively.The results showed that,compared with the control group,the average daily gain,apparent metabolizability of crude fat,crude fiber,calcium and phosphorus,and apparent metabolizable energy were significantly higher (P < 0.05),the F/G were significantly decreased (P < 0.05) in both groups A and B,their apparent metabolizabilities of crude protein increased by 5.90% (P < 0.05) and 3.54% (P > 0.05), respectively.There were no significant differences for all the determined indexes between groups A and B (P > 0.05).The supplementation of Trichoderma xylanase or bacterial xylanase at 200 g/t could improve nutrient metabolizability,thus improve growth performance of broilers,while the effects of Trichoderma xylanase were better than that of bacterial xylanase.

The study was conducted to investigate the effects of medium-chain triglyceride replacing the soybean oil in the basal diet on growth performance and serum biochemical indices of broilers.One hundred and forty four healthy,similar body weights AA broilers at the age of 14 days were randomly assigned to four groups with six replicates ger group and six broilers per replicate.Groups A,B,C and D were respectively fed with 0,0.6%,1.2% and 2% medium-chain triglyceride substituted the equivalent amount of soybean oil (using corn flour to balance the energy levels in each group) .The test lasted for 28 days and the indexes were measured once every two weeks.The results showed that the average daily gain and average daily feed intake of groups B,C and D were higher than that of group A,and the F/G was lower than group A.The average daily gain of groups C and D at 28 to 42 days old and group D at 14 to 42 days old were significantly higher than that of group A (P < 0.05).The serum glucose,total content of amino acid,total protein and albumin of groups B,C and D at age of 28 and 42 days old were higher than that of group A,while the serum triglyceride,total cholesterol and urea nitrogen levels were lower.The total cholesterol content at 42 days old and triglyceride concentrations at 28 days old of group C were significantly lower than those of group A (P < 0.05).The results indicated that medium-chain triglyceride could raise average daily gain and reduce the total cholesterol content and triglyceride concentrations of broilers,which were useful for improving the growth performance.

This experiment was carried out to evaluate heterosis of the F1 hybrid of female Small-tail Han sheep and male Hu sheep in growth performance.The newborn lambs from Small-tail Han sheep and F1 hybrid of Small-tail Han sheep × Hu sheep were used in supplementary feeding (6 to 45 days old ) and fattening (46 to 150 days old) under the same conditions.The results showed that F1 hybrid were extremely significantly higher than Small-tail Han sheep in birth weight,weaning weight,three months weight and five months weight (P < 0.01),which were increased by 20.00%,25.17%,19.92% and 20.42%,respectively.The average daily gain of F1 were also extremely significantly higher than that of Small-tail Han sheep in both supplementary feeding period and fattening period (P < 0.01),the increase were 26.91% and 17.63%,respectively.At 5-month old,body height,chest circumference and cannon circumference of F1 hybrid were extremely significantly higher than that of Small-tail Han sheep (P < 0.01),but the body length had no significant difference (P > 0.05).The diarrhea rate of F1 hybrid was 8.33% lower than that of Small-tail Han sheep,and the survival rate was 5.55% higher.In addition,the economic benefit of F1 hybrid was 69.34% more than that of Small-tail Han sheep at the end of the experiment.The results suggested that the heterosis was obvious,meanwhile the survival rate and pre-slaughter live weight of F1 progenies could be significantly improved.

Milk proteins have great nutritional value,rich contain and balanced profile of amino acid,which provide nearly all the essential amino acids for the human body.However more than 90% of milk proteins are synthesed by the amino acids in the mammary gland.Furthermore,the amino acid is not only funcion as the substrates of milk protein synthesis,but also as the signaling molecules to regulate milk protein sysnthesis through mammalian target of rapamycin (mTOR) signaling pathways.It is essential to further study milk protein synthesis regulated by amino acids in the mammary gland and the molecular mechanism in dairy cows.The paper reviewed the supply of amino acids,metabolism of amino acids by mammary gland,factors affecting amino acid utilization and signal pathway for amino acid regulation in milk protein synthesis.

This study was aimed to investigate the effect of the Radix rehmanniae extract on immune organs indexes and H9 vaccine immune efficacy.1 200 1-day-old healthy chicks fed with basal diet were chosen and randomly devided into 6 Groups on 14-day-old:GroupⅠ,Ⅱ and Ⅲ adding 2‰,3‰ 0.4‰ Radix rehmanniae extract,respectively;Group Ⅳ adding 1‰ astragalus polysaccharide;Group Ⅴ immunned with H9 vaccine and fed with basal diet;Group Ⅵ was blank control group which didn't immunned with H9 vaccine and fed basal diet.After 7 days medicine-feeding,the samples were collected at 14,21,28,35,42,49 and 56 days and the immune organs indexes and the H9 antibody levels were determined.The results showed as follows:①Compared with group Ⅵ,the spleen and bursa indexes of groupⅠ,Ⅱ,Ⅲ and Ⅳ were significantly increased (P < 0.05),indicating that dietary supplementation with Radix rehmanniae extract and astragalus polysaccharide could improve spleen and bursa development (P < 0.05).Compared with group Ⅰand Ⅲ,the bursa and spleen indexes of group Ⅱ were significantly increased (P < 0.05),while there was no significant difference between group Ⅱ and Ⅳ (P > 0.05).②Dietary supplementation with Radix rehmanniae extract and astragalus polysaccharide could improve H9 antibody titers and hold it at a higher level.Compared with group Ⅰand Ⅲ,the H9 antibody titer of group Ⅱ was significantly increased from 42 days (P < 0.05),while there was no significant difference between group Ⅱ and Ⅳ (P > 0.05).In conclusion,the Radix rehmanniae extract could improve immune organs development and immune function of chicks,and keep antibody titers at high levels when matches with H9 vaccine.And the recommended supplemental level of Radix rehmanniae extract was 2‰ in diet for chicks.

With the development of animal husbandry and the widespread use of feed additives,the indiscriminate use and abuse of feed additives had become a problem to be settled.In the study,the detection techniques of feed additives and the shortages of feed additive detection technology at present were summarized.At the same time,the development trend of feed additives for high-throughput screening technology was described,aiming to apply the technical supports for the green and safe production.

In order to select the best tracer dye for gastric emptying and intestinal propulsion experiment,three kinds of dyes including of dextran blue 2000,direct blue 53 and ink were selected.The absorbance values of the series of concentration of dyes were tested in a range of wavelength by micro double dilution method.Moreover,the maximum absorption wavelength and measurable concentration range of three dyes were determined.According to measurable range of three kinds of dyes,the optimal gavage doses were obtained in mice.The gastric residual rates,small intestinal propulsion rates and residual rates in four parts of equal small intestine were tested.The results showed that the maximum absorption wavelengths of dextran blue 2000,direct blue 53 and ink were 630,630 and 405 nm,respectively.Their measurable concentration ranges were 0.16 to 5.00 mg/mL (R²=0.9863),0.78 to 6.25 μg/mL (R²=0.9984) and 0.02% to 0.16% (R²=0.9979).The difference of gastric emptying rates among the three dyes was not significant (P > 0.05),and intestinal propulsion rate of direct blue was significantly higher than those of the other two dyes (P < 0.05).Total residual rates of dextran blue 2000,ink and direct blue were 93.6%±4.5%,71.5%±8.5% and 18.7%±2.8%,respectively.The results suggested that ink was the best tracer dye if only the intestinal propulsion rate in vivo needed to be determined,while dextran blue 2000 is appropriate if a precise determination of the gastric and intestinal emptying needed to be done.

In order to detect the clinical efficacy of oligosaccharide sulfate(DEAE) and dextran sulfate (DS-4000) against bovine mastitis,56 cows with bovine mastitis were divided into 5 groups (DEAE group,DS-4000 group,DEAE＋penicillin group,DS-4000＋penicillin group and penicillin group),and their breasts were perfusion with different drugs twice one day for 3 days.The effective rates of drugs were observed by clinic symptoms.Milk samples were collected before and after treatment of drugs,and the bacterial negative conversion rate of each group were detected by microbial examination.The results showed that total effective rates of 5 groups were 10.00%,20.00%,58.33%,75.00% and 50.00%,respectively,and the bacterial negative conversion rates were 0,12.50%,50.00%,54.55% and 44.44%,respectively.In conclusion,the curative efficiencies of the polysaccharides against bovine mastitis were weak,but it could be improved by combination with penicillin.

Milk fat globule membrane protein is packaged outside the fat granule during mammary gland lactating. Milk fat globule membrane has three-layered structure composed of phospholipids,sphingolipids and proteins. Milk fat globule membrane protein contains more than one hundred kinds of proteins,of which there are the most abundant of eight kinds of proteins including mucin 1,xanthine oxidoreductase,mucin 15,CD36,butyrophilin,lactadherin,adipophilin,fatty acid binding protein .Milk fat globule membrane protein has been shown to have special value which reflects the physiological state of cow,affects the growth of calf and maternal immune as a source of nutrition and immunity for calves,and affects the human and animal's immunity regulation and physical fitness,it may be applied to treat diseases in the future. This article summaries the study of milk fat globule membrane protein in recent years aimed at describing the source,composition of the milk fat globule membrane proteins,introducing a novel established structure of milk fat globule membrane protein,explaining the physiological characteristics of some main proteins and progress in milk fat globule membrane proteins research and making recommendations for future research directions.

The animal breeding industry is faced with a variety of adverse factors such as disease,stress and the negative effects of using antibiotics,like intestinal flora imbalance,drug residues and resistance.These influences have become a threat to food safety,and cause many other issues and potential risks.Therefore,the development of new green additive is inevitable for the sustainable development of animal husbandry.Essential oil is a kind of volatile liquid which is extracted from different organizations of plants such as fruit,root,leaf or flower,with strong fragrance and sweet smell.Essential oil is regarded as an "efficient,safe,stable and controllable" plant-derived drug,it has the functions of antibacterial,antiviral,anti-parasite,antifungal,enhancing the animal immune function,anti-stress ability and improving animals production performance,etc.Essential oil is one of the focus and hot spot of current research and development,it can be used as a substitute for antibiotics in the production of livestock and poultry.This review summarized the research status of essential oils,focused on their functions about antibacterial,growth promoting,anti oxidation and immunity enhancement.It is wished to prove reference for the research and application of essential oil.

The purpose of the study was to analyze the association between MUC13 gene polymorphisms and piglet diarrhea in Huanjiang miniature pigs.93 diarrhea piglets and 95 healthy controls were included in the study.The MUC13 gene polymorphisms were determined using high resolution melting (HRM),and the results of HRM were verified by direct sequencing.Mutations were compared between diarrhea piglets and healthy controls.SHEsis online platform was used to calculate the haplotypes.There were statistically significant association on G224A site showed significant difference between diarrhea piglets and healthy controls by allele frequencies (P < 0.05),and there were also statistically significant differences between the two groups by genotypes among GG,GA,GG+GA and AA (P < 0.05).But no statistically significant association was found between polymorphisms of A112G,T170C and G164C sites and piglet diarrhea risk (P > 0.05).However,haplotype Ⅹ (GTAC) in the distribution of diarrhea group was extremely significantly higher (P < 0.01).The present data revealed that MUC13 gene G224A site and the haplotypes Ⅹ (GTAC) were associated with reduce risk of piglet diarrhea.

With the wide application of the artificial insemination (AI) in animal husbandry,the utilization rate of excellent sire got greatly improved.The key point of AI is predicting the time of ovulation and insemination accurately.It is also the point which the animal husbandry technologists highly concerned to improve the application effect of AI.Estrus and ovulation are regulated minutely by reproductive hormones in vivo in farm animals.Timed artificial insemination (TAI) is based on the synchronization of estrus and ovulation,both of which are triggered by reproductive hormones.So the optimal time for AI could be determined from the reproductive hormone treatment.The purpose of TAI is to improve the reproduction performance of farm animals,reduce the workload of identification of estrus,and promote batch management.This review mainly summerized the principles of TAI and its applications status,and the prospects was also discussed.It would provide a reference for the research and application of new reproductive technology in farm animals.

The objective of this study was to investigate the correlation of ecological traits on body size traits and economic traits (body size traits and slaughter traits) in Chinese native chicken breeds and reveal the essential connection of three group traits.In this study,simple correlation analysis and canonical correlation analysis (CCA) were applied to estimate the relationship between local 6 ecological traits and 8 body size traits,6 ecological traits and 8 slaughter traits,8 body size traits and 8 slaughter traits of Chinese native chicken breeds.The result of ecological traits and slaughter traits suggested that the first canonical correlation coefficient was 0.536 (P < 0.01),the value of pct was 55.046% for the total correlation information.It showed that the correlation between two sets of traits mainly by mean annual temperature and dressed weight closely related to the cause,the lower mean annual temperature,the heavier dressed weight.The result of ecological traits and body size traits suggested that the first canonical correlation coefficient was 0.679 (P < 0.01),the value of pct was 54.118%,for the total correlation information.It showed that the correlation between two sets of traits mainly by latitude and shank girth closely related to the cause,the higher latitude,the smaller shank girth.The second canonical correlation coefficient was 0.556 (P < 0.05),the value of pct was 28.251% for the total correlation information.It showed that the correlation between two sets of traits mainly by forst free period and chest bone length closely related to the cause,the longer forst free period,the longer chest bone length.The result of body size traits and slaughter traits suggested that the first canonical correlation coefficient was 0.901 (P < 0.01),the value of pct was 81.063% for the total correlation information.It showed that the correlation between two sets of traits mainly by chest bone length and dressed weight closely related to the cause,the longer chest bone length,the greater dressed weight.

To find genotype with fast growth rates in Ningxia hibrid Simmental cattle,the genetic variation of UCP2 gene were detected by PCR-SSCP method and DNA sequencing,then analyzed UCP2 gene polymorphism related with some growth traits.The results showed that there were three SNPs (T632C,A655G and A689C) in UCP2 gene exon 1,which divided into AA,BB and AB genotypes by PCR-SSCP method.Individuals with different genotypes in different months of age,body weight,body height, chest circumference and body length were AA > AB > BB.AA genotype in different age of body height and chest circumference were significantly different than BB genotype (P < 0.05;P < 0.01),and were no significant difference with AB genotype (P > 0.05).The results demonstrated that T632C locus had significant meaning for improving weight trait breeding of Ningxia hibrid Simmental cattle,AA genotype could be the breeding genotype.

In this study,we aimed to determine whether bear had different blood group systems.Whole blood samples were collected from 40 Asiatic Black bears in Fujian province,China.Tube method was used to detect antibodies in plasma,and antibody isotype was determined with 2-mercaptoethanol.Plasma was further analyzed by mass spectrometry.The plasma from four bears had antibodies,possibly IgM isoform,which could agglutinate RBCs from 30 bears.Blood samples from 10 bears were tested by human blood typing reagents.The results showed that 4 bears had blood type like human type O,while 6 bears had like human type B.Plasma protein had extensive homology to serum albumin-like isoform 1 found in giant panda (Ailuropodamelanoleuca).We suggested that Asiatic Black bear might have at least one blood group system with two blood types.If the sick bear needs blood transfusion,a cross-matching test was necessary.Moreover,giant panda might receive blood from Asiatic Black bear in case of emergency.

A pigeon paramyxovirus serotypeⅠ(PPMV-1) isolate was recovered from samples collected from dead pigeons and initial isolation of the virus was performed in 9-day-old specific-pathogen-free (SPF) chicken embryos.The isolate was identified by the hemagglutination (HA) test,hemagglutination-inhibition (HI) assays,RT-PCR,sequencing and analysis of partial of F gene of the virus.The virulence and pathogenicity of the isolate were determined by mean death time (MDT),intracerebral pathogenicity index (ICPI) in 1-day-old chicks and animal regression experiment.The results indicated that one PPMV-1 was isolated from dead pigeon and named as GXP120012.The F gene sequence analysis of PPMV-1 GXP120012 strain showed that the cleavage sites of the virus was ¹¹²R-R-Q-K-R-F¹¹⁷ motif in accordance with characteristic of velogenic strains;The nucleotide sequence homologies of F gene between strain GXP120012 and PPMV-1 reference strain pi/CH/LLN/110713 were high and reached 98.9%.They located the same branch on phylogenetic tree and fell into genotype Ⅵ.The MDT and ICPI values of isolate were 102 h and 0,respectively.The results of animal regression experiment showed that GXP120012 isolate was virulent for pigeons and avirulent for chickens.The present study provided useful material and theoretical basis for the study of molecular epidemiology and prevention and control of Newcastle disease in pigeon in future.

In this study,we collected 502 california mastitis test (CMT) positive milk samples from several cattle farms and then the bacteria were isolated and identified from them.After that,we used multiplex-PCR for screening out methicillin-resistant Staphylococcus aureus (MRSA).Then we investigated the MRSA's antibiotics susceptibility to 19 kinds of antibiotics by discdiffusion method.The results showed that 338 strains of Staphylococci including 89 strains of MRSA were detected out from the 452 milk samples with bacteria.Additionally,most MRSA showed high sensitivity to vancomycin,fosfomycin,chloramphenicol and clarithromycin,while resistant to penicillin,polymyxin,spectinomycin and ceftazidime.The results provided certain factual basis and method for guiding the use of antibiotics in bovine mastitis,rapid identification of MRSA strains and prevention or control of MRSA infection.

To research on polysaccharides from Lagenaria siceraria (Molina) Standl.against Newcastle disease virus (NDV).Total polysaccharide and four different solvent extractions of Lagenaria siceraria (Molina) Standl.were extracted by the methods of hot water extraction and ethanol precipition in this study.The content of polysaccharide was measured by phenol-sulfuric acid method and infrared spectroscopy method.The safe concentration and growth of chick embryo fibroblast (CEF) were assayed by MTT method,in order to facilitated the comparison under the same level,the safe concentration was united as 78.125 μg/mL.Under the safety range of concentration,detected the block-virus activity,anti-virus activity and virus-killing activity of polysaccharides through the ways of pre-adding polysaccharides,post-adding polysaccharides and adding polysaccharides with NDV.The results showed that the direct inactivation and propagation inhibition activity of total polysaccharide and four different solvent extractions were stronger than anti-absorption function.Anti-virus inhibition rate of 70%,80% gradient alcohol precipitation of polysaccharides from Lagenaria siceraria (Molina) Standl.(LSP₇₀,LSP₈₀) and total polysaccharide (LSP_t)were 40.41%,44.54% and 61.85%,virus-killing inhibition rate were 44.74%,58.76% and 59.38%.LSP₈₀ had the highest virus inhibition rate as 37.14% in the block-virus activity of those five polysaccharides.In summary,70%,80% gradient alcohol precipitation and total polysaccharide in polysaccharides possessed better activity and would be as the materials for further research.

The study was aimed to express CAP protein of porcine circovirus type 2 (PCV2) by eukaryotic expression system.The PCV2 TZ0601 strain was the template,the CAP protein with or without the signal peptide of PCV2 coding sequence were cloned into pOET3 vector.The constructed plasmid were confirmed by restriction enzyme digestion and DNA sequencing,then sf9 insect cells were transfected with recombinant plasmid pOET3-CAP and pOET3-CAP-X.The test was designed to express the CAP protein with or without the signal peptide of PCV2 by flashBAC baculovirus expression system.Expression of PCV2 gene were confirmed by indirect immunofluorescent assay (IFA),SDS-PAGE and Western blotting.The results showed that the eukaryotic expression plasmids of pOET3-CAP and pOET3-CAP-X were constructed successfully and the gene was highly expressed in sf9 insect cells.After expression,we could see our target band about 25 to 35 ku with SDS-PAGE and Western blotting.The immuno-reactivity of the protein was confirmed by antisera against PCV2.This would lay a foundation for a further study of PCV2 subunit vaccine and diagnostic antigen kit.

As an excellent pharmaceutical carrier,liposomes exhibit characteristics of wide drug loading range,high efficiency and low toxicity.Besides increasing stability and solubility of loading drugs,liposomes can ascribe targeting and sustained release features to loading drugs.Meanwhile,it can improve bioavailability of loading drugs.Based on the above characteristics,liposomes is becoming hot spot of research and application.This article reviewed construction methods of liposomes based on structure,particle design and chosen of preparation method.Combined with application and demands of veterinary medicine,research progress on application of liposome in the field of veterinary medicine was submitted.It is expected that this article will provide reference for the development of new drug delivery systems used in the field of veterinary medicine.

The experiment was done in order to understand the effect of four kinds of Tibetan veterinary drug extracts on the antibacterial diarrhea,and to guide the clinical treatment of the disease.Using K-B disk diffusion method and broth dilution method,the antibacterial effect of drug extracts of Veronica ciliate Fisch,Usneadiffracta Vain.,Sophoraflavescens var.flavescens and Lamiophlomisrotata (Benth.) Kudo on fourteen kinds of common pathogenic bacteria were detected.Four kinds of Tibetan veterinary drug prescription were optimized by orthogonal design software and the mice model was established by clinical isolates of Salmonella blegdam.According to the LD₅₀of four kinds of Tibetan veterinary drug optimization prescription,it was divided into three groups of 0.060,0.030 and 0.020 g/mL,respectively.The results showed that the four kinds of Tibetan veterinary drug on most trials of pathogenic bacteria had good inhibitory effect.The Usneadiffracta Vain had good inhibitory effect on all the test bacteria.Four Tibetan veterinary drug optimization prescription of high dose on mice with bacterial diarrhea showed the best therapeutic effects,there was conducive to the treatment of the clinical disease.

In order to explore the incidence,pathogenic factors and the formation mechanism of cystic ovarian disease (COD) in Beijing scale dairy farm,we used the rectal examination and ultrasound diagnosis,combined with the dairy parity,calving season and serum biochemical indices and hormone levels to investigate the distribution,influencing factors and formation cause of cow ovarian cysts.The results showed that the incidence of cystic ovarian disease (COD) was 10.73% in the testing dairy herds and the incidence of COD was associated with the parities.The incidence rate of ovarian cyst in the cows calving in the summer (51.06%) was higher than those calving in the spring (8.51%) and winter (17.02%).Insulin,IGF-1 and E₂ were closely related with the formation of the cow ovarian cyst.The average concentrations of serum insulin and IGF-1 in cows with ovarian cysts were significantly lower than those of normal cows(P < 0.05),but the average concentration of serum E₂ was significantly higher than that of normal cows(P < 0.05).This study provided reliable basis for the prevention and treatment of cow ovarian cyst.

This experiment was conducted to investigate the liver-protective effect of Chinese herbal compound probiotics (CHCP) on acute liver injury layers.One hundred and eight 1 day old hens were divided into 4 groups with 3 replicates in each group and 9 layers per replicate in trial 1.The layers in model groups Ⅰ to Ⅲ were gavaged with 10% (V/V) soybean oil solution of carbon tetrachloride (SCCl₄) according to 1,2 and 4 mL/(kg·BW) at 14,28 and 35 d,respectively.The layers in control group were gavaged with 2 mL/(kg·BW) soybean oil.In trial 2,sixty 1 d layers were divided into 5 groups:Control group (soybean oil),model control group(SCCl₄)and low-dose,middle-dose and high-dose CHCP group (SCCl₄+1‰ CHCP,SCCl₄+2‰ CHCP and SCCl₄+4‰ CHCP respectively).CHCP were used by drinking water since 7 days.SCCl₄ were gavaged according to 2 mL/(kg·BW) at 14 and 28 d.The results showed as follows:The model of layers liver damage could be built by intragastric administration of 2 mL/(kg·BW) 10%(V/V) SCCl₄ at 14,28 d respectively,with the signs of hepatic steatosis,severe vacuolar degeneration,nuclear condensation and necrosis.Compared to the model control group,the serum AST levels in low,medium and high dose CHCP groups were decreased by 4.35% (P > 0.05),7.57% (P > 0.05) and 9.79% (P < 0.05),the serum ALT levels in medium and high dose CHCP groups were decreased by 34.92% (P < 0.01),36.51% (P < 0.01),the serum total bilirubin content in medium and high dose CHCP groups were decreased by 25.49% (P < 0.01),27.45% (P < 0.01).The liver cell congestion was reduced to varying degrees in different dose CHCP groups,and the liver cell had no vacuoles,arranged in neat rows,abundant cytoplasm and uniform in different dose CHCP groups.In conclusion,2‰,4‰ CHCP could reduce hepatocyte necrosis,decrease the serum activities of ALT,AST and total bilirubin levels,and had protective effect on hepatic injury induced by SCCl₄.

The present study was designed to analyse the factors influencing fatty liver weight.Taking the overfeeding results in 2014 and the local temperature together,effects of gender,temperature and body weight on fatty liver weight were studied and the results would provide reference information for improving fatty liver production and breeding work.Totally 415 geese at 10 weeks old were overfed for 28 days.Body weight before and after overfeeding,fatty liver weights,abdominal adipose weights were measured.Blood samples of geese were collected for DNA extraction to determine the gender.The results showed that body weight before and after overfeeding and gain weight during overfeeding period were all associated with liver weight extremely significantly and the correlation coefficient were 0.40,0.62 and 0.46,respectively (P < 0.01).The body weight after overfeeding showed extremely significant effect on fatty liver weight (P < 0.01).Gender showed no significant effect on liver weight when the body weight were similar between female and male geese (P > 0.05).Effect of temperature on liver weight was extremely significant (P < 0.01).Overfeeding work should be conducted between 13.6 to 25.2 ℃ and the optimal temperature was 20.3 ℃.Higher temperature would result in reduction of fatty liver weight.In summary,temperature was a fundamental guarantee for fatty liver production and body weight was an indirect selecting index when cultivate a breed specific for fatty liver.Further,gender showed no significant effects on fatty liver weight.

L-malic acid exists widely in apples,pears and other fruits.It is one of the important members of tricarboxylic acid cycle.Its taste is close to the natural apple's sour,and it has some characteristics such as high acidity,soft,long residence time.L-malic acid has been widely used in some quality beverages,food industry at present.In animal production,L-malic acid is used as acidifier and an inhibitor of pathogenic microorganism.L-malic acid by promoting the utilization of S.ruminantium on lactic acid,the production of propionic acid and the concentration of CO₂ to prevent ruminal acidosis,and then to improve the utilization ratio of dietary energy and animal production performance.This article is a summary of the physiological functions of L-malic acid and its application in the production of ruminant in order to provide scientific theoretical basis for the rational and efficient use of L-malic acid in ruminant production.

The experiment was conducted to study the best extract process of mulberry leaf polysaccharide,three factors and three levels orthogonal test was designed to optimize the extraction process.Three factors were considered,including ratio of solid to liquid,decoction time,the time of extracting and the yield of mulberry leaf polysaccharide was determined with phenol-sulfurie acid colorimetry as the evaluate index.Meanwhile,the influence of ethanol concentrations on mulberry leaf polysaccharide extract was studied.The results showed that decoction times could greatest affect the extraction percent of the polysaccharides.The extraction percent of polysaccharide was the highest (4.94%) when the rate of solid to liquid was 25,the time of extracting was 2 h and decoction times was 2.While the most appropriate process was 25 times for the rate of sdid to liquid,2 h for extracting and decoction times was 3,considering with practical situations.The concentration of ethanol would significantly affect polysaccharides yield and purity.When the concentration of ethanol was 70%,the yield of mulberry leaf polysaccharid was highest.When the concentration of ethanol was 40%,the content of mulberry leaf polysaccharide was highest (41.5%),significantly higher than other ethanol concentration (P < 0.05).It suggested that the higher the ethanol concentration,the lower the polysaccharide content,but the higher the yield.The optimum extraction of mulberry leaf polysaccharides were that the rate of solid to liquid was 25,extractign for 2 h,decoction 3 times and 70% ethanol precipitation.