Since the 5'UTR gene (GenBank No.:AY278459.1) had 4 isolated regions,we designed a set of 4 LAMP primers to specifically recognize target gene sequences.This study developed a loop-mediated isothermal amplification (LAMP) method for detecting BVDV,using the pyrophosphate magnesium white precipitate for Real-time detection in LAMP reaction process of turbidity instrument,Real-time monitor liquid turbidity to determine result.The whole reaction lasted only 50 minutes at a constructed temperature of 63 ℃ to evaluate specificity,sensibility and repeatability of the method.The result demonstrated that the LAMP assay could only react with BVDV,its specificity was high;It could detect at least 10^-6-fold diluted samples,which was 100 more sensitive than PCR assay;And repeatability was good.The simple,rapid,high siensitivity and specificity LAMP assay was a potential tool for the detection of BVDV in field conditions.

To investigate genetic variation of avian infectious bronchitis virus (IBV) in Guangxi province,one strain of IBV was isolated from chicken.Two pairs of primers for amplifying the N and M genes of IBV were designed according to the sequences in GenBank.The N and M genes of the strain were amplified by RT-PCR,and they were proved to be the N and M genes of IBV by cloning,sequencing and compared with reference IBV strains published in GenBank.The results showed that the N gene from the IBV isolate consisted of 1 230 bp,coding 409 amino acids.The M gene from the IBV isolate consisted of 678 bp,coding 225 amino acids.The sequence analysis of N gene showed that it shared 87.2% to 93.3% nucleotide homologies and 90.0% to 94.4% deduced amino acid sequence homologies with IBV strains from GenBank.The M gene sequence analysis showed that it shared 83.6% to 91.0% nucleotide homologies and 82.7% to 92.9% deduced amino acid sequence homologies.The phylogenetic tree analysis showed that it was closely related to BJ and LX4 strains,and were clustered into one group;But with the distant relatives from other strains of IBV.These results suggested that the isolate was a new variant of IBV.

In order to study the causes of different cases,and analyze 16S rDNA homology of bacteria isolated from different cases,this study took the material of 10 different cases to isolate and culture bacteria,identify the isolates by microbiology.A pair of primers was designed to amplify the bacteria 16S rDNA gene.PCR products were sequenced and 16S rDNA homology analysis was conducted.The results showed that 10 strains bacterial were Escherichia coli through microbiology identification,6 strains of Escherichia coli from mammals cases,16S rDNA homologies among canine skin fester sores,canine pyometra,canine pneumonia,canine mastitis,dairy cow mastitis and calf diarrhea were 100.0% in 10 strains of Escherichia coli,4 strains of Escherichia coli from poultry animal cases,16S rDNA homologies among pigeon diarrhea,broiler diarrhea,pheasant diarrhea and white peacock diarrhea were also 100.0% in 10 strains of Escherichia coli;16S rDNA homologies in Escherichia coli of 10 different cases animal sources were 97.5% to 100.0%.This test proved that 10 different animals cases were caused by Escherichia coli infection,and had a high 16S rDNA homology among 10 strains of Escherichia coli.

The objective of this paper was to investigate the drug resistance of canine Escherichia coli (E.coli) strains and the carrying rates of four resistance genes,and explore the relationship between resistance phenotypes and resistance genes.This article chose the aminoglycoside antibiotics including gentamicin,amikacin,spectinomycin and tobramycin to carry out the antibiotics sensitivity test.According to the established PCR assays,we detected the molecular characteristics of the 156 strains of isolates.The positive fragments of four kinds of resistance genes were cloned and sequenced,and the relationships between antibiotics sensitivity test and the resistance genes were analyzed.The results showed that the resistance rates of canine E.coli strains to gentamycin,tobramycin,spectinomycin and amikacin were 55.8%,32.7%,25.0% and 20.5%,respectively.The detection rates of resistance genes aacC2,aphA3, aadA and aacC4 were 55.8%,26.3%,23.1% and 9.0%,respectively.Two strains carried all four kinds of resistance genes,eight strains carried two kinds of resistance genes,and the strains carried two or more resistance genes accounted for 40.4% (63/156).The sequence analysis showed that the amplified gene fragments had higher homology compared with the sequences from GenBank.The main resistance gene in canine E.coli was aacC2 and there was a positive correlation between resistance rates and resistance genes compliance rate.

The object of this study was to clone the Keratin-associated protein 8.2 (KAP8.2) gene from Small-tail Han sheep and Xinji Fine-wool sheep and test the tissues expression pattern.In this study,the total RNA were extracted from different tissues firstly,and KAP8.2 gene was cloned by RT-PCR.In the end,the different expression patterns of KAP8.2 gene in Small-tail Han sheep and Xinji Fine-wool sheep was detected by QRT-PCR.The cloning results showed that the KAP8.2 gene were cloned,and the length of KAP8.2 mRNA was 574 bp,containing a 192 bp ORF which encoded a protein of 63 amino acids.These KAP8.2 gene belonged the typical HGT-KAP family because the rate of glycine and tyrosine were 23.8% and 20.6%,respectively.The polymorphism analysis showed that there were some polymorphisms in KAP8.2 gene and almost SNPs located on both sides of CDS region.The region which located on the 3'UTR c192+19-39 of KAP8.2 from Small-tail Han sheep might be the target of fru-let-7j.The expression pattern analysis showed that KAP8.2 gene was a multiple organs expression gene,and there were significant differences in expression patterns of KAP8.2 gene in Small-tail Han and Xinji Fine-wool sheeps.In Small-tail Han sheep,KAP8.2 gene was over-expressed in spleen,liver and skin,but in Xinji Fine-wool sheep,KAP8.2 gene was over-expressed in skin and heart.In conclusion,there were significant differences in gene polymorphisms and expression pattern between Small-tail Han sheep and Xinji Fine-wool sheep,and the result suggested the KAP8.2 gene associated with some phenotypes of wool.

In order to study of FBP1 gene expression under cold stimulating environment,75 AA broilers as research object were selected and randomly assigned to 5 groups,one of group was control group that raising in normal temperature,the remaining four groups were cold stimulation groups that raising temperature was 3 ℃ lower than the control group.The time of cold stimulation in four groups were 1,3,5 and 24 h,respectively.Cold stimulation started at 8 days of age,ended at 21 days of age.Broilers were slaughtered at 22 days of age and investigated FBP1 gene expression in each group.The results showed that the expression of FBP1 gene existed tissue specificity under cold stimulating environment.The relative expression level of FBP1 gene in liver was significantly higher than the other tissues in 1 h group (P< 0.05);The relative expression level of FBP1 gene in lung were significantly higher than the other tissues except of liver in 3 h group except for liver (P< 0.05);The relative expression level of FBP1 gene in liver were significantly higher than the other tissues in 5 h group (P< 0.05);The relative expression level of FBP1 gene in thymus were significantly higher than the other tissues in 24 h group (P< 0.05).FBP1 gene expression inside the heart was on the decline in the same organization different treatment groups as the rime of cold stimulating was increasing except for 3 h group.FBP1 gene expression of liver in 5 h group was significantly higher than other FBP1 gene expression in each treatment group.FBP1 gene expression of spleen was significant decreased in 1 h group,the other group extend a downward trend with the extension of cold stimulation time.FBP1 gene expression in lung was downward trend after rising first.FBP1 gene expression in kidney was significantly decreased in 24 h group,and the other groups had no obvious change.FBP1 gene expression of thymus in 24 h group was significantly higher than the other groups (P< 0.05),and there was no significant difference between control group and other four terament groups (P >0.05).

In order to establish a stable,simple and rapid detection method to estimate the activities of the recombinant chicken interferon-α/interleukin-2 fusion protein (rChIFN-α-Linker-ChIL-2,recombinant fusion protein) in vitro,the activities of rChIFN-α-Linker-ChIL-2 were estimated by detecting its specific immune response to monoclonal antibody (MAb) against ChIFN-α and ChIL-2 by ELISA assay.The antiviral activities of rChIFN-α-Linker-ChIL-2 protein were tested by inhibiting the 50% appearance of cytopathic effect (CPE) of vesicular stomatitis virus (VSV) and infectious bursal disease virus (IBDV) on the passage cell lines DF1.The promoting proliferation activities of lymphocytes in the chicken peripheral blood and spleen of recombinant fusion protein were tested by MTS method.The results showed that rChIFN-α-Linker-ChIL-2 protein had the ability of specific immune response to anti-ChIFN-α MAb and anti-ChIL-2 MAb,respectively.The antiviral activity of rChIFN-α-Linker-ChIL-2 protein inhibiting the reproduction of VSV on DF1 cell line was higher than IBDV,and both of the antiviral activities of recombinant fusion protein against VSV and IBDV were much higher than the recombinant ChIFN-α protein (rChIFN-α) control.The recombinant fusion protein had apparent promoting proliferation activity of lymphocytes in the chicken peripheral blood and spleen,which were much higher than that of the rChIFN-α control.The study suggested that the activities detection and estimation methods of the recombinant fusion protein in vitro were successfully established,which laid the foundation for the further study of the synergy activity of recombinant fusion protein in vivo.

In the study,an indirect ELISA was developed using the purified thymidine kinase (TK) protein as antigens to detect the antibody of duck plague virus (DPV).The TK gene of DPV was amplified by PCR using specific primers designed according to the sequence of TK gene in GenBank.Using gene recombination technology,the TK gene was cloned and inserted into prokaryotic expression vector pET-32a and the recombinant plasmid was identified by PCR,double enzyme digestion and sequence analysis.The positive recombinant plasmid was then transformed into E.coli BL21 (DE3) and induced by IPTG.The expressed protein was purified by gel extraction and analyzed by Western blotting.Then an indirect ELISA was established to detect DPV antibody by using the purified recombinant TK as the coating antigen.The other assay conditions were also optimized.The recombinant plasmid was constructed successfully and Western blotting detected the target protein,TK recombinant protein could be recognized by positive serum,and the result indicated that the recombinant protein had good antigenicity.The optimum reaction conditions were as follow:The optimal dilution of enzyme labelled antibody was 1:200,the optimal dilutions of antigen and antibody were 1:400 and 1:200,reaction time was 60 min,most prefer blocking solution was 5% BSA,closed for 60 min,chromogenic time was 10 min.The method had good stability and high sensitivity,and it could provide a reliable method for the clinical detection,immunization surveillance,and prevention and cure of DP.

The internal transcribed spacer (ITS) and 5.8S ribosomal DNA (5.8S rDNA) of roudworm (Ascaridia galli, A.galli) samples collected from Liangshan,Sichuan were amplified by PCR using a pair of conserved primers (BD1 and BD2),then sequenced and analyzed the sequences,phylogenetic trees based on ITS-1 and ITS-2 were reconstructed using Mega 5.0 software.The results showed that ITS and 5.8S rDNA sequences of Ascaridia galli were 974 to 989 bp,with homology among these sequences was 98.9% to 100.0%.The sequences of ITS-1,5.8S rDNA and ITS-2 were 473 to 481,157 and 337 to 359 bp,respectively.With homologies of 98.5% to 100.0%,100.0% and 98.5% to 100.0%,respectively.The phylogenetic trees showed that all the Ascaridia galli isolates were formed the same cluster,and could identify with other Ascaridida.The results indicated that there was no significant variation in ITS-1 and ITS-2 sequences within Ascaridia galli,while inter-species difference was obvious,so they could be used as genetic marker to identify the species of Ascaridia galli,and provided the foundation for further research on molecular classification,epidemiological survey and population genetics of Ascaridia galli.

Porcine circovirus type 2 (PCV2) caused significant economic losses of swine industry,it's very important to establish a economy and rapid detection method.To strengthen the prevention and control of porcine circovirus disease,rapid and high sesitive detection method of PCV2 was established using nested PCR.According to PCV2 ORF2 gene conservative region,we designed two pairs of nested PCR primers,constructed ORF2 gene recombinant plasmid for standard template and optimizated the PCR reaction conditions to establish nested PCR detection method of PCV2 and detect the clinical samples of pig farms.Our results showed that the sensitivity and repeatability of this method were very high,the minimum detection level was 10 copies/μL and detection rate of clinical samples was 97.3%.In brief,the established nested PCR detection method of PCV2 in this study provided powerful means for quick and efficient detection of PCV2,and prevention and control of porcine circovirus disease in the swine industry.

Proteomics is a discipline which study proteome using the technology of protein isolation,identification and quantitative.Proteomics technology also has greatly developed,and proteomics technologies based on mass spectrometry have been widely used in the related field.In this review,we mainly introduced the advantages and disadvantages of some proteomic technologies based on mass spectrometry on the studies of protein-protein interactions,differential proteomics and quantitative proteomics,reviewed the advance of hair follicles and focused on the proteomic technologies application in wool-sheep and cashmere-goat.Finally,some prospects of proteomics technologies application in the hair follicles study of wool-sheep and cashmere-goat were presented.

miRNA is a kind of 22 nt non-coding small RNA,which recognizes the target mRNA by complementary base pairing,and regulates the gene posttranscriptional expression using RNAi silencing mechanism.miRNA exists in animals,plants,viruses and various microorganism,and its expression is under the strict spatial-temporal regulation.Previous studies have showed the miRNA plays a key role in the regulation of tissue development,cell differentiation,apoptosis and cancer.The canonical biogenesis pathways include the process of the microprocessor in nucleus,nuclear export,Dicer dicing in cytoplasm and the generation of the miRISC complex.Besides,there are other synthetic pathways such as mirtron pathway,tRNA pathway and endogenous siRNA pathway.For providing a reference for the research and the application of the miRNA,this review summarizes the biosynthetic pathways of the miRNA.

56 Small-tail Han sheep lambs with birth body weight (3.38±0.33)kg were randomly divided into two groups (n=28 each group),fed with milk powder substitute or fish meal substitute since 4 days of age respectively and after 21 days of age,eight lambs in each group were turned to starting feed.At the age of 7,14,21,28 and 35 days,four lambs were slaughtered from each group respectively.The contents of small intestine were collected,determining the α-amylase activity,lactase activity,trypsinase activity,chymotrypsin activity of chyme supernatant from the small intestine,to investigate the enzyme activity in small intestine at different ages with different milk substitutes.The results showed that the activity of α-amylase,lactase,trypsinase,chymotrypsin of chyme supernatant from small intestine of the lamb fed with milk powder substitute was linearly correlated with age,and the average daily gain was 157.2 g/d,without significant change with age.At different ages,the activity of α-amylase,lactase,trypsinase,chymotrypsin of chyme supernatant from small intestine of the lambs fed with fish meal substitute was inferior to those fed with milk powder substitute,43.7%,46.0%,35.2% and 10.2% on average,respectively,and the average daily gain was reduced,especially after 21 days of age.It was revealed that the growth of the lamb fed with fish meal substitute might be influenced by the immune response of fish meal protein.After the lambs were given starting feed,the activity of α-amylase,lactase,trypsinase,chymotrypsin of chyme supernatant from small intestine decreased,but recovered later.The activity of α-amylase,lactase,trypsinase of the lamb fed with milk powder substitute before 21 days of age reduced more than the lambs fed with fish meal substitute,but the activity was still high.It was concluded that the activity of α-amylase,trypsinase,chymotrypsin of chyme supernatant from small intestine of the lamb increased with age,the activity of lactase decreased with age,while the activity of digestive enzyme in the small intestine of the lamb fed with fish meal substitute was significantly reduced.After the lambs were fed with starter,digestive obstacle occurred,but disappeared later.

To study the effect of nutrient restriction and compensation on weight,serum indexes and gene expression of chest fat adipokine in Mongolian lamb,thirty male lambs were randomly divided into control group (20.26 kg± 3.63 kg) and feed restricted group (21.29 kg±3.00 kg).The experiment lasted for 120 days,including 60 days for feed restriction and 60 days for compensatory feed.At the end of each stage,five lambs from each group were slaughtered and chest fat was collected.Serum concentration of triglycerides,non-esterified fatty acids (NEFA),total cholesterol were measured.Quantitative Real-time PCR were used to determine Adiponectin,Visfatin,PPAR-γ and ACC genes expression.The results showed that:①The lamb body weight of restricted group kept maintain level and that of control group showed a increase tendency.During the compensatory period,the average daily gain (ADG) of restricted group was significantly higher than control group (P< 0.05).②During the restriction period,serum triglycerides concentration of lambs in restricted group was significantly decreased at day 30 and 60 (P< 0.05),and NEFA concentration was significantly increased at day 30 (P< 0.05).After 60 days of nutritional compensation,all serum parameters we tested were not significantly different between restrict group and control group (P >0.05).③During the restriction period,PPAR-γ and ACC gene expression in restricted group were significantly lower than that of control group (P< 0.05),while no significant differences were found during the compensation period (P>0.05).Adiponectin and Visfatin genes expression in chest fat was not significantly changed throughout the experiment (P >0.05).We could know that nutritional restriction reduced fatty acid synthesis and adipocyte differentiation ability,adipose tissue was mobilized energy metabolism,and total cholesterol was recoveried growth on the compensation.However,the ACC and PPAR-γ genes in restricted group were significantly decreased with the body weight increased rapidly during the compensatory stage.Adiponectin and Visfatin were not significantly different between restricted and compensatory stages.

To investigate the effect of adding mannan oligosaccharide to diets with different concentrate to forage ratios on the degradability of neutral detergent fiber (NDF) and acid detergent fiber(ADF) in rumen of sheep by nylon bags technique,6 wethers (White Suffolk♂×Small-tailed Han sheep♀) installed permanent rumen fistula were used for offering rumen fluid.The 4×6 two-factor experimental design was chosen and the two factors were the ratio of concentrate to forage (A1(20:80),A2(30:70),A3(40:60),A4(50:50)) and the dose of mannan oligosaccharide (B1 (0),B2 (0.4%),B3 (0.8%),B4 (1.2%),B5 (1.6%),B6 (2.0%)).The results showed that the rumen degradability of NDF and ADF in different times,degradation parameters and effective degradability were influenced by the factor A significantly (P< 0.05).The rumen degradability of NDF and ADF in each time of group A1 were higher than group A3 and A4 (P< 0.05).Expect for 24 h,the rumen degradability of ADF were influenced significantly by the factor B,and the rumen degradability of ADF in group B5 was higher than group B1 and B2 (P< 0.05).The rumen effective degradability of NDF in group A3 was significantly higher than groups A1 and A2 (P< 0.05), and a little higher than group A4 (P >0.05),which showed that the rumen effective degradability of NDF and ADF were higher when the ratio of concentrate to forage was 40:60.

The assay was aimed to investigate the effect of different microbial inoculant combinations of Candida tropicalis (CT),Bacillus subtilis (BS),Lactobacillus plantarum (LAP) and Lactobacillus buchneri (LAB) on the quality and aerobic stability of sugarcane tops silage.There were 10 groups:Sugarcane tops silages with no additives served as control group (C);GroupsⅠ1,Ⅰ2 and Ⅰ3 were treated with CT+BS at 10,20 and 30 mL/kg,respectively;Groups Ⅱ1,Ⅱ2 and Ⅱ3 were treated with CT+BS+LAP at 10,20 and 30 mL/kg,respectively;Groups Ⅲ1,Ⅲ2 and Ⅲ3 were treated with CT+BS+LAB at 10,20 and 30 mL/kg,respectively.Each treatment was treated with 0.5% fresh forge urea.The results showed that:①Compared to control group,CT+BS treatment groups had significantly higher levels of pH and acetic acid (P< 0.05),dry matter recovery (DMR) and lactic acid content were significantly decreased (P< 0.05),and the aerobic stability was lower.②Compared to control group,CT+BS+LAP treatment groups had significantly higher levels of pH and lactic acid (P< 0.05),and the aerobic stability was lower.③In CT+BS+LAB treatment groups,the levels of acetic acid were significantly higher and DMR was significantly lower than control group (P< 0.05),and the CP content and aerobic stability were highest in all groups.So the quality of sugarcane tops silage was lower in CT+BS treatment groups and CT+BS+LAB treatment could improve the quality and aerobic stability of sugarcane tops silage.

The experiment was conducted to study the effects of phytooestrogens supplementation on blood biochemical indexes and antioxidant capacity of Yili maria during lactation.A total of 24 Yili maria during lactation were divided into four groups with similar age,parity,lactation months,i.e.groups Ⅰ,Ⅱ,Ⅲ and control group.The control group and trial groups fed with the same based diet,groups Ⅰ,Ⅱ and Ⅲ were supplemented with 0.7 g soybean isoflavone,0.7 g genistein,0.35 g soybean isoflavone and 0.35 g genistein respectively.The total duration of the experiment was 90 d,the first 10 d of the experiment was the preliminary period and the formal experiment lasted for 80 d.The blood samples were collected by jugular vein in the experiment of 0,20,40,60,80 days,respectively,3 500 r/min centrifuged 15 min separate plasma and -20 ℃ saved for later use.The results showed that compared with control group,total protein,albumin content and GSH-Px,SOD,CAT activity of blood in trail groups were no significant difference (P >0.05);But CAT,T-AOC activity of blood in trail groups had a upward trend.GLB,BUN content of blood in group Ⅱ were significantly higher than control group and group Ⅰ (P< 0.05);no significant difference was observed among other groups (P >0.05).Compared with control group,the MDA content of blood in groups Ⅰ and Ⅱ were trends to decrease,and group Ⅰ reached significant level (P< 0.05).Therefore,phytooestrogens supplementation had enhanced immune function,improved the body's antioxidant capacity of Yili maria during lactation,when supplementary feeding 0.7 g soybean isoflavone on antioxidant capacity of Yili maria during lactation was more obvious effect.

Good food intake behavior is the basis of maintaining the growth and development of animal and improving production performance.Olfaction and taste are two important senses in the progress of animal food intake.They are determined by olfactory receptor genes and taste receptor genes respectively,which are relatively large gene family in mammalian genomes.They express not only in the sense organs,but also widely in the respiratory,digestive,reproductive and other systems.This paper analyzed the consumption of mutton and sheep breeding in China.It gave an overview of the latest research progress on the olfaction,taste and their receptor gene.Finally,the function and mechanism of olfaction and taste in animal food intake,and the utilization in livestock production were expounded.This paper provided a theoretical reference for the study of the sense of olfaction and taste in animal food feeding especially in sheep.

To determine the apparent digestibility of cassava residue (CR) nutrient and the effects of CR on growth performance when replaced the different levels corn in diet,two experiments were designed.In the first experiment,8 Duroc×Landrace×Yorkshire crossbred barrow pigs with the similar weight (22.46 kg±1.08 kg) were chosen for determining the apparent digestibility of CR.The pre-test period lasted for 10 days,and the trial period lasted for 5 days.In the second experiment,160 Duroc×Landrace×Yorkshire crossbred pigs with the similar weight(21.09 kg±0.72 kg) were randomly divided into five groups with four replicates per group and eight pigs per replicate.The control group was fed with basal diet while the experimental groups were fed diets containing 5%,10%,15% and 20% levels of substitution of corn by CR,respectively.The pre-test period lasted for 5 days,and the trial period lasted for 30 days.The results showed that the content of CP and EE in CR were 2.73% and 0.68%,but CF and total energy were 15.34% and 13.55 MJ/kg.The content of Lys and Thr were 0.11% and 0.08%,respectively.Calcium and phosphorus apparent digestibility were low,just only 40.24% and 34.76%,respectively,while apparent total energy digestibility was 65.62%.The apparent digestibility of Arg,Glu,Tyr,Met,Leu and Val were higher than 80%.The average daily gain was reduced with increasing dose of CR,which the group of 15% and 20% addition were significantly lower than that of control group (P< 0.05).However,the average daily gain of 5% and 10% groups were no significant difference with the control group(P >0.05).The feed cost of 5% and 10% addition groups were decreased by 0.01 and 0.05 yuan.In conclusion,under this experimental condition,the appropriate addition of CR for growing-fishing pig (20 to 50 kg ) was 5% to 10% in the diet.

The study was conducted to determine the relative bioavailability (RBV) of 1α-hydroxycholecalciferol (1α-OH-D₃) to 25-hydroxycholecalciferol (25-OH-D₃) in Ross 308 broiler chickens from 1 to 42 days of age.300 one-day-old Ross 308 broilers were randomly assigned into 6 groups with 5 replicates per group and 10 chicks per replicate.The six groups were fed with basal diets in which 2.5,5.0,10.0 μg/kg 25-OH-D₃ and 1.25,2.5,5.0 μg/kg 1α-OH-D₃ were added,respectively.The RBV of 1α-OH-D₃ to 25-OH-D₃ was determined by the slope ratio method.The results showed that average body weight gain (ABWG),feed to gain ratio (F/G),plasma phosphorus (P) were improved with the 1α-OH-D₃ and 25-OH-D₃ addition and the tibia and femur weight,length,and ash weight were linearly increased with the levels of 1α-OH-D₃ or 25-OH-D₃ increasing (P< 0.05).Using ABWG,F/G,and plasma P as the criteria,RBV of 1α-OH-D₃ to 25-OH-D₃ were 2.16,2.56,and 2.77.Using tibia weight,length,width,and ash weight as the criteria,RBV of 1α-OH-D₃ to 25-OH-D₃ were 1.80,1.88,2.42,and 2.04.Using femur weight,length,and ash weight as the criteria,RBV of 1α-OH-D₃ to 25-OH-D₃ were 1.89,1.98,and 2.18.These results indicated that the activity of 1α-OH-D₃ was approximately 2.17 times higher than that of 25-OH-D₃ in promoting growth performance and bone mineralization in broiler chickens from 1 to 42 days of age.

Fatty acids are important compositions of nutrient,including short-chain fatty acids,medium-chain fatty acids and polyunsaturated fatty acids.They play a role in optimizing intestinal microflora,modulating nutrient metabolism,improving immunology and reproductive performance of pigs except for providing energy.This paper reviewed fatty acid profile of feed ingredients,basic biological functions of different variety of fatty acids and its application in swine industry to provide evidences for better using lipids and fatty acid in pig diet.

This experiment was aimed to study the protease properties produced by Bacillus subtilis JNB001 via the index of protease activity to investigate the effects of pH,temperature and other factors on the enzymatic reaction,and the ability of the crude enzyme solution to hydrolyze various animal protein was measured by the degree of hydrolysis.The results showed that the most suitable pH of reaction was 8.0 and the temperature was 45 ℃.The crude enzyme solution in the range of pH 5.5 to 9.0 was relatively stable,while it was not suitable for the preservation of the enzyme solution when pH was over 9.0.At 45 ℃ it had good thermal stability when exceed 50 ℃ for 10 min, the residual activity had been less than 60%.Metal ions such as Ca²⁺ and Mg²⁺,reducing agent like DTT,surfactant as Tween-80,etc.could greatly improve the enzyme activity.On the opposite,Fe²⁺ and Cu²⁺ as well as other ions,metal chelator EDTA,denaturant SDS and other materials could inhibit the protease activity.The K_m was 6.43 mg/mL,and V_max was 47.39 μg/min.It had a range of reaction pH from 7.0 to 9.0 and temperature from 40 to 50 ℃ in the situation for different protein substrates.The crude enzyme solution could hydrolyze pig hair,pig nail,hair removal of cowhide,gelatin and other hard protein substances.The results provided the strong date support for the Bacillus subtilis JNB001 which could be used in dead livestock harmless biological treatment system and other aspects of industrial applications.

The standard curve of two antibiotics (colistin sulfate and bacitracin zinc) was carried out to assess antimicrobial activity of bacteriocin,which obtained from our earlier work,using cup-dish method.In addition,we also observed the effects of the reaction temperature on the antibacterial activity of the bacteriocin.Results showed that 125 mg/mL bacteriocin displayed similar antibacterial activity to 26 mg/mL colistin sulfate or 11.9 mg/mL bacitracin zinc.Furthermore,the bacteriocin remained good thermal stability under high temperature (nearly 60 to 80 ℃) in 15 minutes.Overall,this research built a direct relationship between this bacteriocin and antibiotics,and testified good stablility of this bacteriocin,providing theoretical support and technology reference to promote its large scale production.

This study was aimed to detect the genetic similarity and groups homozygosity of Xinjiang Brown cattle inbred group.Use 13 pairs of microsatellite primers to structure of genetics in the comparative analysis by the two Xinjiang Brown cattle over 40 years completely closed breeding out 55 Xinjiang Brown cattles group and 25 Xinjiang Brown cattles in control group,in order to carry out the identification of inbred groups.For Xinjiang Brown inbred group,the homozygosity was 0.9064,the polymorphism information content was 0.0873,it was found that the groups had a low polymorphism;The homozygosity was 0.4647,and polymorphic information content was 0.4882 in Xinjiang Brown cow of control group.According to the coat color of the inbreeding groups was divided into four subgroups A,B,C and D.The genetic purity of each subgroup were 0.9029,0.9632,0.9208 and 0.9551.The results showed that the detection of Xinjiang Brown cattle inbred group had higher homozygosity and lower heterozygosity,had became a high genetic purity of inbreeding group.The closure of Xinjiang Brown cattle inbred group was rare breeds of cattle,and was valuable genetic resources.

To analyze the effect of different genotypes in Mx1 gene exon 14 on the reproductive performance in Meishan pigs,PCR-RFLP method was used to amplify DNA and detect the polymorphisms of Mx1 gene exon 14, and the relationship between different genotypes and the reproductive performance was further analyzed by generalized linear model (GLM) in this study.The results of PCR-RFLP showed that there was a G36T mutation on Hin6 Ⅰ restriction site in Mx1 gene exon 14 and three genotypes were detected named as AA,AB and BB,in which BB was a dominant genotype.The results of correlation analysis showed that the reproductive performance of three genotypes in Mx1 gene presented the increasing tendency of BB >AB >AA in total number born (TNB),number born alive (NBA),little size at weaning (LBW),birth weight (BW) and weaning weight (WW) in gilts sows on the whole.Moreover,there was significant difference in TNB,NBA,LBW and WW in gilts sows (P< 0.05).However,there was no significant difference among all reproductive performance in multiparous sows (P >0.05).The results of this study indicated that the polymorphisms of Mx1 gene exon 14 had significantly genetic effects on production performance in Meishan gilts sows instead of multiparous sows.

The growth hormone (GH) and growth hormone releasing hormone (GHRH) contents in serum of 3 weights (90,110 and 130 kg) of Chuanzang Black swine hybrid and DLY pigs were detected by enzyme-linked immune sorbent assay (ELISA),their relationship with cross sectional area of muscle fiber (CSA) and intramuscular fat (IMF) content were also analyzed.The results showed that the GH contents of Chuanzang Black swine hybrid and DLY pigs of 3 weights were 14.25,14.61,14.11 and 17.86,16.98,16.77 μg/L,respectively,while the GHRH contents of which were 22.88,23.98,24.33 and 27.72,27.47,28.39 μg/L,respectively.There was no significant difference in different weight of the same breeds (P >0.05) except the significant difference between 90 and 130 kg of DLY pigs (P<0.05).The GH and GHRH contents of Chuanzang Black swine hybrid were both significant lower than DLY pigs of the same bodyweight (P< 0.05) except GHRH content of 110 kg weight (P >0.05).Relevance analysis showed that GH and GHRH contents had a significant positive correlation with CSA (P< 0.05),and an extremely significant negative correlation with IMF content (P< 0.01),which prompted that GH and GHRH might regulate the growth of muscle fiber and IMF deposition through blood.

The association of Chinese Merino sheep eNOS gene exon8 polymorphisms with brucellosis susceptibility was studied in this research.The sequences of human and Chinese Merino sheep eNOS gene exon8 were aligned by the bioinformatics methods,and the polymorphisms of human eNOS gene in the NCBI SNP database were statistically analyzed.The polymorphisms of 101 Chinese Merino sheep negative samples and 61 Chinese Merino sheep positive samples were detected by PCR-SSCP method,and then the PCR products of different alleles were sequenced,aiming to determine exon8's polymorphism loci,and the allele frequency and genotype frequency of SNP loci were statistically analyzed.A novel SNP locus (ss974768653:A142G) was detected at the 142 bp of eNOS gene exon8,and the locus had no difference in allele frequency and each genotype between negative and positive groups (P >0.05).There might be no correlation between the A142G polymorphism locus of Chinese Merino sheep eNOS gene exon8 and brucellosis susceptibility.

It aimed to investigate the sex differences and correlation of body size and slaughter traits in Wei-ning chicken,the research measured the body size and slaughter traits of 65 Wei-ning chicken(♀:33;♂:32 ) at the age of 180 days according to the nation livestock body size and slaughter measurement standards,and analysis on the correlative between body size and slaughter traits.The results showed that the lean body length was significant difference in the different sex chicken groups (P< 0.05 ),and the cocks were higher than hens,the comb hight and keel bone length of cocks were extremely significant higher than hens (P< 0.01 ).All of the body size and slaughter trait showed difference between cocks and hens,moreover,the Wei-ning chicken also showed excellent appearance,hold the strong body constitution character,good meat performance and adaptability for other native chickens,the body constitution and meat performance of cocks were better than hens.Besides,the different body size,most of slaughter traits and between body size and slaughter traits were positively correlaton,and most of indexes showed the significant or extremely significant correlation levels (P< 0.05;P< 0.01),these data would be crucial insight for the breeding,especial early breeding of Wei-ning chicken.

In order to explore the effect of different L-lysine levels added in semen dilution on the fresh Dorper sheep quality stored at liquid state,3 health Dorper ram were used to collect semen which were equal-packaged and added to solution containing different levels (0,0.1,0.2,0.3,0.4 and 0.5 g) L-lysine in 120 mL diluent and at 0,6,24,30,48,54,72 and 78 h,the sperm motility,membrane integrity and acrosome integrity were detected.The results showed that the sperm motility in groups added 0,0.1 and 0.2 g L-lysine were higher than other groups,and that in 0.4 and 0.5 g L-lysine groups decreased faster than other groups;The sperm motility in 0.5 g L-lysine group reduced to 0 at 30 h,and it reduced to 0 at 48 h in the group added 0.4 g L-lysine;The effective survival time and survival index in the group added 0.1 g L-lysine was higher than other groups (P< 0.05);The membrane integrity of 0.1 g group was the highest,that had no significant difference with control group from 0 to 54 h (P >0.05),while after 72 h the two groups had significant differences (P< 0.05).The membrane integrity in 0.4 and 0.5 g groups were the worst,and that was significant different with other groups (P< 0.05);The acrosome integrity of 0.1 g group was the highest which had no significant difference with control group (P >0.05),0.4 and 0.5 g groups were the worst,the difference between them was not significant (P >0.05),while was extremely significant difference with other groups except for 0 and 6 h (P< 0.01).The results suggested that dilution added a high concentration of L-lysine could inhibit sperm quality,when the count of L-lysine was more than 0.2 g,sperm quality was significantly decreased,adding 0.1 g L-lysine could improve Dorper sheep fresh sperm quality stored at liquid state.

To preliminary investigate the mechanism of drug resistance and drug-resistant elimination of swine E.coli O157:H7,sub-inhibitory concentration in vitro drug-induced method was used in this study,for inducing two strains of E.coli O157:H7 becoming florfenicol highly resistant strains,and eliminated florfenicol resistance of florfenicol-resistant strains by continuous cultured without florfenicol pressure.The drug sensitivity and resistant genes of florfenicol-induced strains and florfenicol-resistance eliminated strains were detected.The results showed that compared with original strains,after drug-resistant induction by florfenicol,swine E.coli O157:H7 were resistant to florfenicol,amoxicillin,cefazolin,cephradine and cephalothin from high sensitivity,and resistant to cefotaxime,ofloxacin,ciprofloxacin and azithromycin from moderate sensitivity.However,after florfenicol-resistance eliminated,swine E.coli O157:H7 restored sensitivity to these drugs.In addition,florfenicol resistance gene,quinolone resistance genes and β-lactamase genes were found in the plasmids of swine E.coli O157:H7,the resistance genes were consistent with the resistance phenotype.According to the results,with the long-term presence of florfenicol pressure,swine E.coli O157:H7 resistant to florfenicol,and acquired penicillins,cephalosporins and quinolones cross-resistance;While swine E.coli O157:H7 was eliminated partial drug-resistance by continuous culture without florfenicol pressure.

In order to study mutations of bovine breast cancer 1 (BRCA1) gene and its association with somatic cell count and somatic cell score in Holstein dairy cows in Beijing.SNPs of BRCA1 gene exon 13 and 14 were obtained by cloning,sequence alignment and searching for known mutations.In our research,25025 T>A of exon 9 and 46126 G>T of exon 13 in BRCA1 were detected by SNaPshot in Holstein dairy cows raised on the outskirts of Beijing city,as well as the association between SNPs and somatic cell count (SCC) or somatic cell score (SCS) were also analysed.The results showed that both 25025 T>A and 46126 G>T were detected in Holstein dairy cows.TT genotype at locus 25025 T>A and GT genotype at locus 46126 G>T were dominant genotypes,respectively.At locus 25025 T>A,least squares mean of SCC and SCS in individuals with AA genotype was significantly lower than that with TT and TA genotypes (P<0.05;P<0.01).Least squares mean of SCC in individuals with TT genotype was significantly lower than that with GG and GT genotypes (P<0.05),while SCS of different genotypes showed no difference at locus 46126 G>T (P>0.05).These results preliminarily indicated that the 25025 T>A and 46126 G>T mutations of BRCA1 gene could be used in molecular marker-assisted selection programs of mastitis resistance in Holstein dairy cows.

From 2012 to 2014,300 chicken cloacal swab samples were collected from a broiler farm in Liaoning province,152 strains of chicken-derived Escherichia coli were isolated and identified.The 152 strains of chicken-derived Escherichia coli were detected on 14 kinds of veterinary antimicrobial drugs and 16 kinds of antimicrobial drugs for human by using trace dilution method,and reference to CLSI enterobacter decision criteria to determine.The results showed that chicken-derived Escherichia coli had obvious resistance to veterinary antimicrobial drugs (amoxicillin,ampicillin,enrofloxacin,trimethoprim-sulfamethoxazole,sulfafurazole,tetracycline) and antimicrobial drugs for human (piperacillin,ticarcillin,ciprofloxacin),the resistance rates were high.The resistance test results of isolated chicken-derived Escherichia coli comparing with the MIC value distribution of EUCAST wild type strains were same,but through comparing MIC value distribution,the results showed the isolates of spectinomycin,ampicillin,enrofloxacin,trimethoprim-sulfamethoxazole,cefalotin,cefuroxime,ciprofloxacin and tobramycin drugs MIC value moved to the right obviously,indicated that the antibiotic resistant strength of Escherichia coli increased gradually.The process of breeding and strict biosecurity measures should be taken scientifically to control diseases,and the antibacterial agents should be applied reasonably to control broiler farm production and propagation of bacterial drug resistance.

One-day-old moderate maternal antibody Jinghong chickens and SPF chickens were vaccinated with infectious bursal disease (IBD) immune complex (IC) vaccine and BX strain live vaccine,respectively,pathological changes of bursa of fabricius in chickens were observed at the 9th day after immunization.On the 28th day after immunization,blood samples were taken and the IBDV neutralizing antibody were tested,meanwhile experimental chickens were challenged with high virulent IBDV,the protective rates of experimental groups were calculated.The results showed that at the 9th day after immunization,the bursa of fabricius in SPF and Jinghong chickens were normal in IC vaccine group,however atrophic changes of bursa of fabricius in BX strain live vaccine group were shown.At the 28th day after immunization,the IBDV neutralizing antibody of IC vaccine group and BX strain live vaccine group in SPF and Jinghong chickens were 9.2log2,8.5log2,7.2log2 and 4.4log2,respectively,while 100% protection rates were provided.The results provided that IC vaccine had better immunity effect on one-day-old moderate maternal antibody chicken,high levels of neutralizing antibodies against IBDV were produced and protection rates were 100%.

Chicken immunosuppressive diseases can not only make chicken immunity decline,seriously affect the growth,and also cause some other vaccine failure and have great effects on the chicken breeding.The reasons causing immunosuppression are varies,including viral factor,chemical factor,irritability factor,nutrition factor and so on,of which the viral factor occupies the important position.This paper summarized the progress of etiology characteristics,virus detection and prevention of several common viral immunosuppressive diseases on chicken,so as to provide certain reference for research,prevention and treatment of these diseases.

To determine the drug resistance of E.coli strains from swine and chicken in organic farm,collection of stool specimens was conducted in one organic farm in Conghua city,Guangdong province in 2014,in which 118 E.coli strains were separated,including 55 from chicken and 63 from swine.The sensitivity of the E.coli strains from swine and chicken to 18 kinds of drugs was detected through agar dilution method.Similar properties in drug resistance were observed both in E.coli strains from swine and chicken in the same organic farm.In consideration of the resistance rate,samples were relatively sensitive to ceftazidime,cefquinome,cefoxitin,amikacin,apramycin,gentamicin,neomycin,florfenicol,chloramphenicol,imipenem,ciprofloxacin and olaquindox (chicken 0 to 21.8%;swine 0 to 14.3%).These isolates showed moderate resistance to ampicillin and streptomycin (chicken 29.1% to 38.2%;swine 23.8% to 27.0%).About 56.4% chicken isolates and 47.6% swine isolates showed resistant to more than 3 kinds of drugs.However,severe resistance was observed in trimethoprim-sulphamethoxazole,tetracyclineand and doxycycline (chicken 45.5%,76.4% and 72.7%;swine 50.8%,81.0% and 68.3%).The results also indicated that under organic farming mode,the drug resistance rates of E.coli strains from swine and chicken were relatively low in general.Multi-drug resistance existed but was relatively slight.Although the drug resistance rate to several specific antimicrobials was high,it was still lower than that in conventional farms.Since most studies focused on the drug resistance of the bacteria under conventional farming model,this study filled the blank of the research of drug resistance under organic farming model.

Rifaximin is a kind of new drug for the treatment of dairy cow disease which antimicrobial spectrum is wide.It has a strong antibacterial activity and unique antibacterial mechanism.Rifaximin is suit for local administration because it is no mutagenic,abnormal toxicity,carcinogenicity and reproduction toxicity and so on.In this paper,the physical and chemical properties,action mechanism,pharmacokinetic,pharmacology and toxicology properties,clinical application were investigated and reviewed,so as to provide a theoretical basis for the application in prevention and treatment of animal diseases,and make veterinary workers have a comprehensive understanding of rifaximin and guide the clinical application.

In December 2014,a group of 4 weeks old broilers died in a large-scale chicken farm in Hebei province.The clinical symptoms of the ill and diseased chicken were depression,dyspnea,joint swelling and claudication.In order to diagnose the disease,a systematic analysis was conducted by laboratory diagnostic methods that included histopathological observation,bacterial isolation,PCR detection,drug sensitive test and animal pathogenicity experiment.The histopathological observation results showed that hemorrhage in the lung,the edema of heart and liver,a large number of heterophil granulocyte infiltration.Bipolar stained brevibacterium was observed under the microscopy.The PCR amplification results of Pasteurella multocida KMT-1 gene and avian reovirus σB gene were positive by using specific primers.The isolated Pasteurella could be lethal to mice and the isolate was highly sensitive to ceftriaxone and cefoperazone.Combination with the above laboratory test results and the clinical symptoms,we confirmed that the cases were mixed infection of avian reovirus and Pasteurella.

Antibiotic resistance of bacteria is a major problem in the clinical treatment of infectious diseases,and it has received the extensive attention.Bacteria producing inactivated enzyme or passivated enzyme is a mainly way to acquire resistance.Besides,the mechanisms of cell wall permeability barrier,efflux pump and target changes are existed,these mechanisms interact to the level of the bacterial antibiotic resistance.With the clinical application of new antibiotic,the new resistant mechanisms emerge and the antibiotic-resistant bacteria are widespread.The study of bacterial resistance mechanisms has instructional meaning for antibiotic-resistant bacteria control and new drug development.We elaborated the bacterial resistance from the origin of the resistance,mechanisms,characteristics and test method in this article.

African swine fever (ASF) is mainly occurred in Africa.It invades into Europe and America by chance,then invades in to Eastern Europe and the Caucasus area.And it has not been controlled until now.African swine fever virus (ASFV) can spread by soft tick vectors and affect wild and domestic pigs.The ability of the virus to survive within a particular ecosystem is defined by the ecology of its wild host populations and the characteristics of livestock production systems.African swine fever is viral disease of domestic and wild pigs which leads high mortality and causes great economic losses due to absence of available vaccine and treatment.Apart from prevention and culling,there are no other control measures.Prevention and control of the infection require good understanding of its epidemiology,so that targeted measures can be carried out.

To investigate the epidemiology of multi-pathogens related with post-weaning multi-systemic wasting syndrome (PMWS) in Jiangsu province and surrounding areas,125 healthy samples and 261 diseased samples which obtained in Jiangsu,Anhui and Zhejiang provinces from the January 2014 to May 2015 were screened by PCR for the presence of PCV2,PRRSV,PPV,TTV and P1.These results revealed the following infection rates of PCV2,PRRSV,PPV,TTV1,TTV2 and P1 were 39.38%,21.76%,3.11%,15.80%,16.32% and 10.10%,respectively.In comparison,the co-infection of several viruses occurred in the diseased pigs,and the main co-infections were PCV2+TTV2 (15.32%) and PCV2+PRRSV (11.87%).These observations indicated that the phenomenon of co-infection with multi-pathogens had been widely existing in swine,which increased the difficulty of prevention and controlling of the PMWS.

Japanese encephalitis B (JE) is a zoonosis caused by Japanese encephalitis virus which invades the central nervous system,infected by mosquito.Various animals could be the sower and reservoir of JE including swine,horse,cattle and avian.JE could cause a major public health trouble and a horrible crisis of the farm all in Asia and the Western Pacific region.Therefore,it is necessary to diagnose it rapidly and accurately.This review article made a sketch of approaches for the rapid serological assays of JE,including traditional method,emerging technologies and specific improvements of the assay,so as to provide references for the study of rapid,simple and accurate JE diagnose methods.