This experiment was conducted to study effects of rumen protected Lysine and Methionine supplements on ruminant fermentation and roughages degradability of small-tail han sheep.A total of 10 small-tail han sheep((45±2.6) kg of body weight) with permanent rumen cannula were selected for the study.The study was conducted in three replicates,each of which lasted for 15 days(12 days of pre-testing period and 3 days of formal test period).Different supplements of protected l ysine and methionine were added into basal diet.Protected amino acids supplements had no effects on rumen pH,NH₃-N concentration and VFA concentration(P>0.05).Compared to control group,mycoprotein content in group I was higher(P<0.01).Appropriate protected amino acids supplements could significantly improve the DM,ADF rumen degradation rate(P<0.05) but not the CF,NDF ruminal degradation rates(P>0.05).It could be concluded from results that the appropriate supplement of protected Lysine and Methionine was 8.4,2.4 g/d per sheep under the experimental conditions,respectively.

To search for the molecular markers associated with meat traits in yaks, CAPN4 mutations in the promoter region were investigated in different populations and their effects on carcass and meat quality were also analyzed.A total of eight hundred and thirty yaks which sourced from Gannan yak, Tianzhu white yak and Datong yak were used in this study.PCR-SSCP method was used to detect the CAPN4 mutations in the promoter region.Associations between mutations and the carcass and meat quality traits of Gannan yak were analyzed using the general linear model (GLM) in SPSS 19.0 program.The mutation (a.-1222G>A) in the promoter region was identified, which led to three PCR-SSCP patterns (AA, AB and BB).The results of the statistical analysis indicated that the genotypes and allele effected carcass and meat quality in Gannan yak with different ages.At the ages of four and six, yaks possessing the genotype BB had higher cooking rate than those with possessing the genotype AA and AB (P<0.05).At the ages of three and six, individuals with possessing the genotype AB had higher water loss rate than those with possessing the genotype BB and AA(P<0.05).However, yaks at the age of five, individuals with possessing the genotype BB had higher water loss rate than those with the genotype AA (P<0.05).At age of three and five, individuals with the genotype BB had bigger eye muscle area than those with possessing the genotype AA and BB (P<0.05).Yaks with possessing allele A had negative effect on cooking rate at the ages of four and six, as well as water-holding capacity and eye muscle area in all yaks.Excepting these yaks at the age of five, individuals with the presence of allele A had lower carcass weight (P<0.05), the mutation had no effects on carcass weight and tenderness.The mutation (a.-1222G>A) in the promoter of CAPN4 had an effect on carcass and meat quality partly, it might be used as a genetic marker to improve those traits in Gannan yak.

Based on the gE and gH genes of pseudorabies virus (PRV), two pairs of special primers were de- signed, respectively, single PCR assays for gE and gH genes were established. Under the optimized amplification conditions, the duplex PCR method was established to differentiate virulent and vaccine strains of pseudorabies vi- rus. In a duplex PCR assay, two fragments of 429 bp for gE gene and 355 bp for gH gene were simultaneously am- plified from pseudorabies virus virulent strains DNA, only a 355 bp fragment for gH gene was amplified from pseud- orabies virus vaccine strains DNA, whereas no PCR products were amplified from porcine circovirus type 2 and por- cine parvovirus. Then the detection limit of duplex PCR was estimated to be 1 × 10²TCID₅₀ /0. 1 mL. This method is suitable for differentiation of wild-type pseudorabies virus and vaccine strains in clinical specimen.

Two hundred and forty 1-d-old Arbor Acres broiler chicks were randomly divided into four groups each including 60 broilers. The four groups were respectively fed with adding 0.0% inulin, 0.3% inulin, 0.1% bacillus and 0.3% inulin + 0.1% bacillus in broiler diets. On 21 and 42 day, the number of cecal bacteria was detection by Counting bacteria method and ammonia of broiler??s fecal excreted was measured by Nessler??s reagent method. The resultsshowed that on 21 day, the addit ion of inulin and bacillus to broiler diets decreased the number of E. coli and Salmonella, especialy in the adding 0.3% inulin + 0.1% bacillus group, but there was no significant variat ion in the amounts of aerobic and Lactobacillus; On 42 day, the action of making the number of E. coli and Salmonella decreased was moresignificant than 21 day, though the quantity of aerobic didn??t vary, the amount of Lactobacillus was increased, whicould make microflora system better. Furthermore the adding of inulin and bacillus to broiler diets could reduce dissem-ination of ammonia in excretion of broilers, especialy in the adding 0.3% inulin + 0.1% bacillus group, which wasbenefit for improving hen house environment.

To investigate the transcriptional regulation mechanism and expression pattern of porcine Single-minded 1 (SIM1) gene.5' promoter fragment was amplified with PCR method and the potential transcription factor binding sites were predicted with bioinformatics method;Eight promoter segments with different length were obtained by promoter deletion analysis and cloned into pGL3-Enhancer vector, then the promoter activities were determined in transiently transfected 293T, MGC803 and Bel7402 cells by the dual-luciferase assay system, respectively.Meanwhile, SIM1 protein expression pattern was detected in seven tissues using Western Blot (WB) method.The results showed that all these SIM1 promoters presented activities in 293T, MGC803 and Bel7402 cells, and the lowest in 293T cells.The region of -699 — -489 bp contained the key cis-regulatory elements and further bioinformatics analysis found some key transcription factor binding sits, such as Smad, CEBPα and PAX6.WB experiments revealed that, besides its known expression in central nervous system, SIM1 protein could be detected in all examined tissues.Grayscale analysis showed a strong SIM1 protein expression in brain, hypothalamus and testes, a weak to moderate expression in muscle, thyroid gland, subcutaneous fat and liver.These results indicated that SIM1 gene may play important role in nerve cell development, fatness metabolism and gonads development.

The objectives of the present study were to elucidate the polymorphism of GnRHR gene and its relationship with litter size of Laoshan dairy goat.Seven pairs of primers were designed to detect single nucleotide polymorphism (SNP) of GnRHR gene in Laoshan dairy goat with different kids by PCR-RFLP and PCR-SSCP.The results showed two mutations (A261G and G55A) were detected.For A261G locus in exon1,the does with genotype GA had 0.06 and 0.04 kids more than those with genotype AA and GG in primipara ewes,but the differences were not significant (P>0.05);the does with genotype GA had 0.35 and 0.45 kids more than those with genotype AA and GG in multiparity ewes (P>0.05).For G55A locus,two genotypes (GG and GA) were detected,the does with genotype GG had 0.13 and 0.19 more than those with genotype GA in primipara ewes and multiparity ewes,respectively.However the differences were not significant (P>0.05).These results preliminarily indicated that A261G locus of GnRHR gene is a potential molecular marker for improving litter size in dairy goats.

The objective is to obtain expression profile of Chinese Simmental cattle ucp1,ucp2,ucp3 genes,to reveals the relationship between the ucp1,ucp2,ucp3 gene and energy metabolism,to explore the expression rule of key gene of fat deposition,and to screen the candidate gene affecting fat deposition.Total RNA were extracted from fifteen tissues(liver,lymph,intercostals muscle,shoulder muscle,spleen,kidney,rib eye muscle,lung,heart,gut fat,greater omental fat,testicle,small intestine,pancreas and backfat)of twenty animals from Chinese Simmental cattle.The primers of ucp1,ucp2,ucp3 genes were designed and synthesized,and the data were normalized using the β-actin gene as internal control.The specific expression of ucp1,ucp2,ucp3 genes in different tissues were investigated by real-time quantitative PCR.There were different degree expression of ucp1,ucp2,ucp3 genes mRNA in the tissues and organs of testicle,small intestine,heart,pancreas and so on.The expression of ucp1 was highest in the backfat,middle in the pancreas,small intestine,testicle,greater omental fat,gut fat,and heart,and lowest in the other tissues.The highest level expression of ucp 2 was detected in Testicle,and there were also higher expression of ucp2 in the lung,liver,and backfat than in other tissues.The highest level expression of ucp3 was in rib eye muscle,Intercostal muscle,and shoulder muscle,middle in the lung,and the lowest in other tissues.Correlation analyses showed that the expression level of ucp3 gene in rib eye had no significant relation with intramuscular fat and backfat thickness,but had positive relation to eye lean area(r= 0.788).It is concluded that the expression level of ucp1,ucp2,ucp3 genes had tissue specific might be related to the various functions on different tissues of cattle.The ucp3 gene can be used as a candidate gene related to carcass traits.

A coding region sequence of yak VEGF-A was cloned by molecular cloning technique in this study. Some characters of the VEGF-A gene and encoded protein sequences were predicted and analyzed by the methods of bioinformatics in the following aspects as the general physical and chemical properties, hydrophobicity, signal pep- tide and secondary structure. Results showed that the full-length of yak VEGF-A contains a complete ORF（573 bp） encoding 190 amino acid. The protein encoded by yak VEGF-A has obvious signal peptide,it is a hydrophilic pro- tein. The secondary structures are mainly composed of Helix and Coli. The VEGF-A and related proteins of Bos grunnien,Bos taurus,Ovis aries,Capra hircus are close in the phylogenetic tree,which showed highly homology.

The aim of this present study are the investigation of the MITF-M gene coding region sequences,the prediction of its translated protein structures and the dictation of its transcription mRNA expression level in skin tissues of yaks with black and white skin color.This study was aim to reveal of the molecular mechanism of skin color formation in yak.In this study,a coding region sequence of yak MITF-M was cloned by RT-PCR technique.Characters of the MITF-M gene encoded protein sequences were predicted and analyzed by the methods of bioinformatics in basically chemical properties and secondary structure.The expression levels of MITF-M mRNA in skin tissues of yak were detected by semi-quantitative PCR.Quantitative PCR was employed to examine the expression levels of MITF-M mRNA in different color yak skin tissues.As results,MITF-M gene was cloned,which contains a complete ORF of 1 242 bp.This ORF encoding a peptides chain which contain 413 amino acids.The second structure of this peptides chain was contain helix,coli and extended strand.And the expression level of MITF-M mRNA in skin tissues between dark yak and white yak were extremely remarkable (P<0.01).

In order to develop genetic engineering subunit vaccine,milk samples of cows with subclinic mastitis were collected.THB(Todd-Hewitt Broth)solid selective medium and pigment test were adopted,combined with species specific gene cfb to isolate and identify Streptococcus agalactiae.ORF and B cell epitope analysis were carried out according to the cfb gene sequences published in GenBank.Primers designed with Primer 5 in cfb's B cell epitope enriched sequence and eukaryotic expression elements and restriction enzyme sites were added.Genome DNA of the isolates were extracted and served as PCR templates for cfb amplification,prior to link with the T-A cloning vector for sequencing.Correctly cloned cfb sequence were then linked to the pcDNA^TM 3.1 V5-His A(pcDNA-cfb)and transformed to DH5α competent cell for plasmid proliferation and sequencing.Results showed 8 strains of Streptococcus agalactiaee were characterized from 89 milk samples,and the pcDNA-cfb was successfully constructed.Primary selection by THB selective medium and pigment test could be used for quick isolation of Streptococcus agalactiae.Analysis of the cfb sequence by Bcepred and Lasergene-Protean software showed the cloned cfb sequence contained multiple dominant B cell epitope,thus had the potential to serve as target gene sequence for genetic engineering subunit vaccine of Streptococcus agalactiae.This work could served as the basis for further developing genetic engineering subunit vaccine for Streptococcus agalactiae.

Heat shock protein 70( HSP70) gene was regarded as a candidate gene for thermo resistance of cows.In this study,a 549 bp fragment of HSP70 gene of Chinese Holstein was amplified by polymerase chain reaction( PCR), the polymorphisms of three loci of HSP70 gene were detected by PCR-SSCP technique.By sequencing, three point mutations, G/C at nucleotide 6 400,C/T at nucleotide 6 600 and G/A at nucleotide 6 601,were identified in HSP70 gene sequence.χ2 test indicated that three polymorphisms in the population unfitted with Hardy-Weinberg equilibrium( P <0.01).Meanwhile, the relationship between milk performance and heat tolerance was analyzed.The results showed that in the three loci, the different genotypes of HSP70 gene had no significant influence on milk protein percent and fat percent,but had significant influence on 305-day milk yield.Individuals with genotype BB showed higher 305-day milk yield than those with genotypes AA and CC( P <0.05).And the cows with genotype BB showed lower potassium content in erythrocytes ( PCE) and decrease rate of milk yield than those with genotypes AA and CC.It indicated that BB genotype was the advantageous genotype.These mutations could be used in MAS for anti-heat stress cows in our breeding program．

In order to investigate the effect of overexpression of cdc20 during cashmere goat oocyte maturation in vitro ,gMyc- cdc20 vector was constructed and overexpressed in the oocyte by microinjection after transcripted into cRNA in vitro.The results showed that the normal oocytes needed 8 h from GV stage to GVBD stage,and 18 h to MⅠ.Twenty-four hours later,75% of these oocytes extruded first polar body.Then microinjected gMyc- cdc20 cRNA into GV stage oocytes and cultured them in vitro.MYC-CDC20 fusion protein could be detected by Western Blotting after 10 h.However,both of the development time and the ratio of oocyte maturation were not significantly different compared with those of control.It could conclude that overexpression of cdc20 alone had no effect on the oocyte maturation of cashmere goat in vitro.

To observe the effect of orexin A on the expression of OX 1 R in ovine luteinized granulosa cells in vitro,the cells were treated with different concentrations (0.05,0.2,0.5,1 μg/mL) of orexins A.Then the total RNA was extracted from the cells after treated in 0,2,6,12 h,and the mRNA expression levels of OX1R were eval-uated using reverse transcription quantitative PCR.In result,orexin A induced release of OX1R in ovine luteinized granulosa cells.In high concentration (1 μg/mL) group,the OX1R mRNA expression level was increased to the highest in 2 h(P<0.01) and gradually weakened since then;in low concentration(0.05,0.2,0.5μg/mL) group, the OX1R mRNA expression level was increased in 12 h and the expression level of 0.2 μg/mL group was in-creased to the highest in 12 h(P<0.01).Orexin A induced expressions of OX1R mRNA,the group of 1 μg/mL was more rapid up regulation compared with the low concentration (0.05,0.2,0.5 μg/mL) group.

Adipocyte fatty acid-binding protein（ A-FABP）, as a member of the fatty acid binding protein（ FAB- Ps） family,play a pivotal role in regulating intracellular fat concentration and then affect intramuscular fat（IMF） in mammalian. So A-FABP was suggested as a candidate gene of IMF in some specie,s of livestock and poultry. In this study, single nucleotide polymorphisms （SNPs） was investigated at A-FABP intron 3, exon4 and 3＇-UTR in Gannan yak, Qinghai yak and Tianzhu white yak（Bos grunniens） by polymerase chain reaction-single strand conformational polymorphism（PCR-SSCP） so as to analyse their molecular genetic characteristics. Five novel SSCP patterns,repre- senting five different alleles A-E were identified in three yak populations. With alignment of the bovine （ Bos tauras） and yak A-FABP allelic sequences, six SNPs including a synonymous mutation （ c. 4222A 〉 G） at exon 4 were checked in yak. SNP c. ＊ 94T 〉 A in 3＇-UTR of A-FABP which was only detected in yak population represented one of genetic characteristics of yak that differ from the bovine. Alleles distributed disproportionably among three yak populations, such as only alleles A-C were observed in Tianzhu white yak and alleles A-D in Qinghai yak. This may be related to geographical distribution and breeding methods of different yak populations. The mutations in amplified region of yak A-FABP would become a potential locus for genetic markers of meat quality traits of yak as it showed moderate polymorphism with PIC in the range of 0.29 -0.36 and higher effective number of alleles（Ne）.

To study the effect of sulfated yeast β-glucan on mRNA transcription and hemagglutination of H1N1 swine influenza virus in vitro, influenza virus infected MDCK was taken as model, SYBR Green Real-time PCR and HI test was replication adopted to measure the effect of sulfated yeast β-glucan on mRNA transcription and hemagglutination.The results showed that the sulfated β-glucan could inhabit the hemagglutination at the concentration of over 1.25 mg/mL, while it could inhibit influenza virus's mRNA transcription at the concentration of 5 mg/mL.Sulfated yeast β-glucan had the ability to inhabit reproduction of H1N1 SIV in vitro, and the ability to inhabit hemagglutination and mRNA transcription may be targets of sulfated yeast β-glucan to anti-SIV.

The research is to study the autophagy induced by porcine circovirus-like virus P1.PK15 cells(Both P1 infection and control group) were collected at various time periods(14,24,48 h) and used to detect autophagy by transmission electron microscopy.Compared with the control groups,the typical morphological features of autophagy was detected in PK-15 cells infected with P1.The result suggests that porcine circovirus-like virus P1 can induce the autophagy of PK-15 cells.

The non-structural protein p70 gene of PoSaV CH430 strain was cloned by RT-PCR and then sequenced in our study.The results showed that the full length of p70 gene was 1 995 nt encoding a protein of 665 aa.Homology alignment and evolutionary tree showed that PoSaV CH430 strain belong to GⅢ.The bioinformatics analysis demonstrated that the isoelectric point and molecular weight of non-structural protein p70 were 5.96 and 72 876.6 u.The protein had no signal peptide and transmenbrane domain.There were 25 phosphorylation sites including 6 Sers, 13 Thrs and 6 Tyrs.Protein phosphorylation was concerned with signal transduction, so this protein may be a signaling molecule.There were Trypsin-like serine proteases and RdRp functional domain.Main structure of trypsin-like serine proteases functional domain was a column structure and an incomplete β-barrel.Main structure of RdRp functional domain was a fold whose organisation has been likened to the shape of a right hand with three subdomains termed fingers, palm and thumb.

Improving the grouth level and meat production of Luxi Yellow Cattle,this paper use PCR-RFLP technology to analysis the correlation of Pit-1 gene polymorphism and growth traits of 111 hybrid cattle of Luxi Yellow Cattle and Limousin.The results show that the Pit-1-HinfⅠ loci of 451 bp′s PCR products is polymorphism after enzymes Hinf Ⅰ enzyme,the allele frequency of A and B were 0.32 and 0.68,it is not fit for Hardy-Weinberg balance.The rump length of AA genotype is higher than AB and BB genotype within population(P0.01),it can be used one of candidate genes;The hip width of AB genotype is higher than AA and BB genotype(P0.05) within the group;But there are no differences among 3 genotypes in the head length,forehead width,chest circumference,chest deep and wide,body length,rump length,waist angular wide,shank circumference,body height,the cross department high,the hip higher(P0.05)

To improve the in vitro maturation rate of bovine oocytes,the present study was carried out to determine the effects of cumulus cell on in vitro maturation rate of cumulus-oocytes-complexs(COCS)and artificial denuded oocytes(DOs),and its pathway and strength was also investigated.The results showed that co-culture with discrete cumulus cells significantly improved the in vitro maturation rate of denudes oocytes(57.98±14.27 vs 35.53± 14.00,P<0.05),but had no effects on COCS(76.66±5.77 vs 66.76±9.46,P>0.05);Co-culture with surrounding cumulus cell did not significantly improve the in vitro maturation rate of denudes oocytes(35.83±18.32 vs 35.53±14.00,P>0.05).The analysis results showed that cumulus cell promoted the in vitro maturation of oocytes by the pathway of gap-junction,however discrete cumulus cells worked by the pathway of paracrine;Three or more layers cumulus improved the mean maturation ability of their surrounded oocytes by 85.34%,added discrete cumulus cells separately improved the mean maturation ability of COCs and DOs by 16.18% and 60.61%.

In this study, the high sulfur keratin B2A promoter of Tianzhu White Yak gene was cloned and got sequence, and analyzed the bioinformatics.Comparison of forecasting results by subcellular localization, phosphorylation analysis, secondary structure of proteins, tertiary structure, MEGA 5.10 software by NJ method and phylogenetic tree construction and MegAlign software to analyzed the difference between B2A gene and KAP1.1 gene. B2A gene CDS region was 471 bp, encoding a protein with 156 amino acids, the secondary structures were mainly composed of extended strand, beta turn and random coil.B2A and KAP1.1 protein of subcellular localization, phosphorylation analysis, conserved domains, secondary structure, tertiary structure and homology were different.But from the function prediction results showed that the two protein's function were exactly same, and Clustal X software was used to compare the amino acid sequence.B2A protein compared with KAP1.1 protein has ten amino acids deletion at position 33, the remaining sequence has very high conserved between two proteins.So it was proved that the B2A gene was KAP1.1 gene sub gene.

12 Hetian sheep were divided into 4 groups(the control group and experimental groupⅠ,Ⅱ,Ⅲ) to investigate the effects of flavonoid of Oxytropis glabra DC feeding on microbial number in rumen,reveal the utilization of O.glabra DC in future.The dried plant of the O.glabra DC was comminuted,and three experimental groups were fed with flavonoid of O.glabra DC at 10,30,50 mg/kg everyday respectively during the test period.The sheep consumed the forages freely until typical symptoms were observed.The test period was 35 d.Before challenge test and at intervals of 7 days in this test,rumen liquid was collected and the microbial number were test.The results demonstrated that the number of total bacteria in experimental group Ⅰ and experimental group Ⅱ were significantly higher than normal control group from 21 d(P<0.05),and the number of total bacteria in experimental group Ⅲ were not obvious in brainstem(P>0.05).The correlations between flavonoid of O.glabra DC and number of Anaerobic fungi and protozoa in rumen were poor. The number of Fiber degrading bacteria in rumen of experimental group Ⅰ and experimental group Ⅱ had no significant changes,The number of Ruminncnccus albus and Ruminococcus flavefaciens in experimental group Ⅲ were lower than normal control group,but had not obvious in brainstem(P>0.05).The results showed that the amount dose of flavonoid of O.glabra DC had no bad effects on microbial number in sheep.

The present study aimed to establish ear skin fibroblasts(cGSF)from a cloned Huanghuai white goat and explore the biological characteristics of the established cGSF cell line.The results were as follows:Both explants adherence culture(A method)and trypsin digestion(B method)could result in the successful growth of primary fibroblasts.However,after 24 h in culture,adherence rate of cells harvested from A method was higher than that of B method.The amount of harvested cells using A method was also greater than using B method.As for the growing characteristics,the growth curve of cGSF cells was typical "S" type.For the time spent on reaching to the exponential growth phase(Day 3 Vs.Day 4)and the platform phase(Day 6 Vs.Day 8),cGSF was earlier than control(non-cloned goat fibroblast cells). We also found that rate of chromosomal abnormalities in cells from cloned goats was higher than those from non-cloned.In order to test feasibility of transfection using cGSF,pEGFP-N1 plasmid was subjected to be transfected with cultured cGSF,and we found that cGSF cells were more easier to form clonies after G418 screen.Therefore,if cells from cloned animals are employed in transgenic research,appropriate readjustment in cell isolation, in vitro culture,transfection and positive cell screen and enrichment should be done accordingly.In all,cells in vitro derived form cloned goat show differences in biological characteristics from cell lines of non-cloned goat.

In yak,Agouti gene is a potential candidate gene in yak skin's color formation and regulation,investigate in Agouti gene sequence polymophism lay the foundation for revealing yak skin and coat color genetic mechanisms.The presented study using DNA cloning techniques to obtain yak Agouti gene encoding sequences.Using bioinformatics tools to forecast the protein's basic physical,chemical properties,hydrophobicity,signal peptide and secondary structure.The polymorphisms of Agouti gene CDS were identificated by DNA pools and direct sequencing in Yak.The results showed that the obtained Agouti was a 593 bp DNA sequence.Gannan black yak Agouti gene was identical with that of Tianzhu white yak.Also,yak Agouti gene was similar with Bos.The yak Agouti gene was named YAK ASIP and registered number in NCBI was KJ630463.This gene encode a hydrophilic protein which containing 133 amino acids.It contains a clear signal peptide and several phosphorylation sites.The secondary structure of the protein was α helix secondary structure and random coil.The Agouti and related proteins of Bos grunniens,Bos taurus,Ovis aries are close in the phylogenetic tree,which showed highly homology.The gene sequence detection showed that the yakAgouti gene was without any polymorphisms.

"This trial was conducted to study the effects of garlic stem powder and oregano leaf powder on antioxidant capacity,non-specific immune performance and meat quality of the mirror carp(Cyprinus specularis).A total of seven trial treatment groups:group 1 was control group,group 2 added 10 mg/kg Flavomycin,group 3 and 4 added 0.5% and 2.5% garlic stem powder respectively,group 5 and 6 added 0.1% and 0.5% oregano leaf powder respectively,group 7 added 0.5% garlic stem powder and 0.5% garlic stem powder.Each treatment had 3 replicates of 10 fish with initial body weight(201.45±16.25)g.The feeding trail was conducted for 8 weeks.The results showed:Compared with the control group,SOD content of G3,G4 and G6 was increased in hepatopancreas significantly(P0.05);MDA Content of G3 and G4 was reduced in serum significantly(P0.05),MDA content of G5 and G6 was reduced in hepatopancreas significantly(P0.05),but SOD activity of the garlic and oregano test groups was inhibited in serum significantly(P0.05),there was no significant difference with SOD and MDA content of each test group in muscle(P 0.05).Compared with the control group,the lysozyme activity of G5,G6 and G7 was increased in spleen significantly(P0.05),the lysozyme activity of G3,G4 and G5,G6 was increased in serum significantly(P0.05);the serum C3,C4 activity of G3 and G5 were increased significantly(P0.05),the renal lysozyme of the test group was no difference significantly(P0.05).Compared with the control group,the crude protein of the fish muscle in G3 and G4 was increased significantly(P0.05),the crude protein was increased and the crude ash was reduced of muscle in G5 and G6 significantly(P0.05).The water-holding capacity of muscle in G4 and G7 was increased significantly(P0.05).Conclusion:feed was added garlic stem powder or oregano leaf powder which can increase anti-oxidation capacity,non-specific immune activity and improve meat quality of the mirror carp."

In the research,polymorphsm analysis on genes of reproduction(FSH ?,ESR and PRLR),meat quality(Haln) and dseaseresistance(FUT I ) traits of 606samples of 8 pg breeds (Lawu black,Dapulan black,Licha black,Yimeng black,Yantablack,Wuli an Black,Changwewte and Luyan white) from Shandong province were detected by the measures of PCR or PCRRFLP.The result ndicated that there were gh polymorphisms of the 5 genesmeasured in the eght pg breeds.Especially,the favorable gene?frequences of reproducton trat were rather gh,whch were the basis of hgh reproducton trat.There are fferent gene dstrbutng characterstics of Halnand FUT I n Chnese ndi genouspg breeds and exotc pig breeds.Through comparison of gene frequenci esof Halnand FUT I,mpacts of exotc pig breeds on pig breeds of Shandong province were analyzed pmarly.It wll provde useful scentfic data to preservaton and utilzaton of pig resource of Shandong province.

The objective of the current study was to characterize the concentrations of immunoglobulins(IgA,IgG,IgM) and lactoferrin in different bovine milk. Sandwich enzyme}inked immunosorbent assay (ELISA) was used to quantify the immunoglobulins and lactoferrin(Lf) contents in Yak milk,Buffalo milk and Holstein cow milk samAles,and correlation analysis was completed. The results showed that immunoglobulins and lactoferrin in Yak milk,Buffalo milk and Holstein cow milk samples had a good linear relationsship between concentration and absorbance values,and the standard curve equation R was greater than 0.99. There were significant differences of IgA,IgG IgM,and Lf in different milk ( P<0.05).The contents of IgA,IgG,IgM and Lf of species showed remarkable individual differences. IgA and IgM in Holstein cow milk showed a negative correlation,while the others species had a positive correlation. The results indicated that the content of immunoglobulins and lactoferrin in the milk were influenced by genetic factors and had each characteristic. According to the correlation of immunoglobulins and lactoferrin, forecasting the content of immunoglobulins and lactoferrin within species may be possible.

"Cell culture is the basis technique of cell biology;it is widely used in cell biology,molecular biology,and endocrinology.Currently in vitro the bovine mammary epithelial cells culture methods include enzyme digestion method,mammary tissue culture method,mechanical disruption and centrifugal method from milk based on their morphology.The application and the future trend of the bovine mammary epithelial cells culture in vitro methods were reviewed in this article. "

This study deals mainly with porcine circovirus}ike agnet Pl antigen purified by immuno affinity chromatography. The purified Pl virus, which came from elution buffers from adsorption column coupled antill epitope-ontaining peptide antibody to either Epoxy}etivated agarose or NHS activated agarose,was confirmed by transmission electron microscopic observations. The study suggests that purified Pl antigen by immuno-ffinity chromatography is an effective way.

This current study was conducted to investigate the biological characteristics of chicken IECs(Intestinal epithelial stem cells)by isolating and culturing in vitro.IECs were isolated from small intestine of 18 day-old chicken embryos.The suitable culture medium for IECs was DMEM/F12.Then its proliferation was analyzed by subculture and the growth curve.The purified chicken intestinal epithelial stem cells were identified by morphology,immunohistochemical methods and RT-PCR.The results showed that chicken IECs can be cultured in vitro and have been passed to 11 generation.The purified IECs were positive by morphology,immunohistochemical methods and RT-PCR.To sum up,IECs can be isolated from small intestine,which can proliferate rapidly in vitro.

Single nucleotide polymorphism of the α-inhibin (INHA) and βA-inhibin(INHBA) genes were detected in Bamei mutton sheep by PCR-SSCP. The results indicated that there were polymorphism in the amplified region for three primer pairs (1，4 and 6)，no polymorphism in the amplified region for other three primer pairs. For?primer 1，two genotypes (AA and AB) were detected in Bamei mutton sheep. The ewes with mutation heterozygous?genotype AB had 0. 17 (P ＜0. 01) lambs more than those with Mutation homozygous genotype AA in Bamei mutton?sheep. For primer 4，three genotypes (AA，AB and BB) were detected in Bamei mutton sheep. Sequencing revealed?a mutation(G→A )at 857 bp of Exon 2 of the INHBA gene in genotype DD compared with genotype CC. This mutation did not result amino acid change. The ewes with mutation heterozygous genotype DD had 0. 44 (P ＜ 0. 01)?lambs more than those with Mutation homozygous genotype CC. For primer 6，three genotypes (EE，FF and EF)?were detected. Sequencing revealed a mutation(A→T)at 288 bp of Exon 1 of the INHA gene in genotype FF compared with genotype EE. This mutation resulted amino acid change of Met→Lys. The ewes with mutation heterozygous genotype EE had 0. 21 (P ＜0. 01) lambs more than those with Mutation homozygous genotype FF. These results preliminarily show that the INHA and INHBA genes were important genes that influence the prolificacy in Bamei mutton sheep.

Intron 3 of the ovine GH gene mutation of Gansu alpine fine wool sheep growth traits may have effects and the gene markers to provide reference of Gansu alpine fine wool sheep growth candidate.Gansu alpine merino and Texel merino,Bond merino,Australia merino crossbred sheep GH gene intron 3 polymorphism was detected by PCR-SSCP technique,analysis of GH gene mutation in intron 3 of lamb birth weight,three-month weight,weaning weight and daily gain influence.The results showed that the first mutation between the Gansu alpine merino and crossbred lamb was GH gene intron 3 of 98 bp a C>T mutation,and this mutation gives AA,AB and BB three genotypes,Gansu alpine merino and crossbred sheep in the control group that the PIC values were between 0.25-0.50,was moderate polymorphism.The dominant genotype of which was BB.Alleles associated with the presence or absence of body weight and daily gain analysis showed the presence of allele B of Gansu alpine merino 3-month weight,weaning weight and daily gain were significantly higher than the individual's lack of allele B(P<0.05),deletion allele a special Texel Gansu alpine merino in birth weight,weaning weight and average daily gain was significantly higher than that of the existence of areas a allele individuals(P<0.05).Bond Gansu alpine merino of allele B 3-month weight,weaning weight,average daily gain were significantly higher than the B allele missing individuals.Correlation analysis showed that different genotypes of Gansu alpine merino correlation between birth weight was not significant,BB type Gansu alpine merino of 3-month weight and weaning weight were significantly higher than AA genotype(P<0.05),average daily gain of AA and BB genotypes Gansu alpine merino were also significantly correlated(P<0.05).In the control group,Bond Gansu alpine merino BB type of birth weight,weaning weight and average daily gain were significantly higher than those of AA genotype(P<0.01).Bond Gansu alpine merino,Australian merino,Gansu alpine merino different genotypes with body weight and average daily gain were no correlation(P>0.05).The results showed that the intron 3 of GH gene could be used as a marker of growth traits of Gansu Alpine Merino candidate gene.

A total of 360 one- day- old female broiler chickens of the Arber Acres strain were randomly assigned to 3 groups，fed with iso- energy and iso- nitrogen diets containing corn，wheat or rice as sole starch source， respec- tively， for 28 days．The technique of terminal- restriction fragment length polymorphism(T- RFLP) was used to evalu- ate the effects of dietary types on intestinal microbial community diversity of broiler chickens．The results showed that the microflora represented by the restricted fragments of T- RF160，T- RF253 and T- RF292 were the dominant bacterial communities in duodenum，jejunum and ileum of broiler chickens．However，the dominance degree of flora represented by the restricted fragments of T- RF253 and T- RF292 in broiler chickens fed with rice diet was slightly lower than that of birds fed with corn diet and wheat diet．The profile and content of dominant colonies presented larger difference between cecum and small intestine．The different diets had no obvious significance on Shannon- weiner index，Simpson index and Evenness index of intestinal microflora in broiler chickens(P ＞ 0． 05)．There was a trend of higher degree of microbial community diversity and less dominant population in rice diet group than that in corn diet group and wheat diet group．With the intestinal segment passing towards the back，microbe species in il- eum and cecum were richer than that in duodenum and jejunum，and gradually tended to stabilization．This revealed that the microbial community structure among different intestinal segments had great difference，and the dietary grain types had a certain degree of influence on intestinal microbial community profile and diversity in broiler chickens．

The objectives of this study were to clone and analysis the coding region of KAP1.4 gene and to predict KAP1.4 protein structure in order to provide basic molecular data for the study of wool and cashmere traits. The result showed that the coding region of KAP1.4 gene in cashmere goat and sheep was 579, 549 bp long and espectively encoded 192 amino acids, 182 amino acids. Bioinformatic analysis showed that there was no significant difference in the basic physical and chemical characteristics of KAP1.4 protein of cashmere goat and sheep. They was a non-transmembrane protein and possessed the same length and the signal peptide cleavage point, having a protein kinase C phosphorylation site, three casein kinase Ⅱ phosphorylation sites and six N-terminal acylation sites. The protein secondary structure was constructed by β-sheet, β turn and random coil. Tertiary structure prediction showed that there were significant differences of three-dimensional structure in space between cashmere goat and sheep KAP1.4 protein. It would further affect the function and become an impact factor on the regulation of wool or cashmere traits.

The occurrence and life history in Aromia bungii(Faldermann) were observed.The results showed that A.bungii occurrenced once on every 4 years in Shijiazhuang,Hebei Province.The imago occurrenced in the end of June or the bengining of August.The ratio of the female and the male was 1∶1.31.Eggs hatching began in the bengining of July,reached the peak at the end of July or the beginning of August,and finished in the middle of August.The hatching larave began to damaged in the middle or the end of Sepetember,and overwintered in the tunnels of trunks in the end of December.From the middle of March to the end of June or the begining of July in the seconed years,the overwintering period began to be over.Then the larave in the trunk xylme began to damage again,and reached overwintering in the beginning or the middle of sepetember or the beginning of October.The larave damaged once more in the begining of April of the third year, and stopped damaging in the begining or the middle of June.Until the begining or the middle of May in the forth year the larave began to pupate.The resting stage was 290.7 d averagely.The larave pupated in the pupal chamb for 17.3 d averagely,then came out.

This work would help for wheat breeding that we could get some low phosphorus tolerance germplasm resources via the methods of the genetic transformation of AeBiPAP1 into wheat cultivals.In this study, PAP1 gene was cloned from Aegilop bicornis with designed primers according the sequence of PAP1 genes from wheat and slen der falsebrome based on homologous cloning method and named AeBiPAP1.The cloned AeBiPAP1 gene was 1.124 kb in size and encoded 335 amino acids, with molecular weight of 38.131 kDa and isoelectric point 5.77.Sequence alignment of AeBiPAP1 with PAP1 of wheat showed that there were 35 single nucleotide polymorphisms, 22 locus transitions and 13 locus transversion and AeBiPAP1 had high homology to PAP1 of wheat with the identities of 95.52%.Amino acid sequence alignment showed that 15 deduced amino acids of AeBiPAP1 were different from PAP1 of wheat.Bioinformatics analysis of AeBiPAP1 gene encoding protein found that the protein has a signal peptide but have not the across-membrane structure and the AeBiPAP1 coding protein with tertiary structure prediction found that the encoded protein contains the metal ions center predicted it belongs to the metal protein.Besides, its possibility of localization in the plasma membrane is higher than being secreted to outside of the cell, the phylogenic tree results showed that PAP1 gene from Aegilop bicornis shared high similarities to the previously reported PAP1 genes from other plants, such as Triticum aestivum, Brachypodium distachyon, and Setaria italica, among them, AeBiPAP1 gene shared the highest similarities to PAP1 gene from Triticum aestivum.

In order to learn more about the complete genome and molecular evolution regular of Japanese encephalitis virus(JEV) strains isolated from swine,the full-length genome sequences of three JEV isolates,namely as BSF.ZZ-1,BSF.ZZ-3 and CSF.XZ-2D isolated from swine were analyzed in this paper.The results show that the length of the complete genomes of BSF.ZZ-1,BSF.ZZ-3 and CSF.XZ-2D was 10 977,10 977,10 976 nt,respectively.There is an insert base of ＂G＂ at the 10 701 nt site in the genomes of both BSF.ZZ-1 and BSF.ZZ-3.Compared to the virulent strain SA14,the homologies of nucleotide and amino acid sequences of BSF.ZZ-1,CSF.XZ-2D and BSF.ZZ-3 are 99.4% and 99.0%,respectively.Compared to the isolates HW(HW1),HWe(HW2) and SH0601 from swine,the homologies of the present three isolates are all more than 98.0%.Phylogenetic analysises with other 30 reference isolates covering all five JEV genotypes indicated that BSF.ZZ-1,CSF.XZ-2D and BSF.ZZ-3 located in a same evolutionary branch,belonging to the genotype III.

In order to investigate the effect of orexins A on the energy metabolism,growth and development of sheep ovary,the luteinising granulosa cells were isolated and cultured in vitro.The passage luteinising granulosa cells were treated with 0.05,0.10 μg/mL of orexins A,and the total proteins of the cells were extracted at 0,2,6,12,24 h treatment,respectively.The relative expression of P-AMPK and P-ERK were determined by Western Blot.The results showed that the expression of P-AMPK of the cells treated with 0.05,0.10 μg/mL of orexins A 2 h after treatment were significantly upregulated compared with that of control (P<0.01),respectively.The expression of P-AMPK of the cells treated with 0.05,0.10 μg/mL of orexins A 6 h treatment reached the extreme value (P<0.01),respectively,and expression of P-AMPK of the cells treated with 0.10 μg/mL of orexins A were significantly higher than that of the 0.05 μg/mL treated group.The expression of P-ERK of the cells treated with 0.05,0.10 μg/mL of orexins A 2 h treatment were significantly upregulated compared with that of control (P<0.01),respectively.The expression of P-ERK of the cells treated with 0.10 μg/mL of orexins A 24 h treatment were significantly higher than that of the plateau phase (6-12 h) and reached the extreme value(P<0.01).It was indicated that the orexins A was involved in the regulation of reproductive function by regulating the energy metabolism,growth and development of sheep throughout the signaling pathway of P-AMPK and P-ERK.