One hundred and twenty piglets were used to investigate the effect of glutamine and glycyl-glutamine on the performance of piglets weaned at 28 days. The results suggested that supplementation of 1% glutamine and 0.2% glycyl-glutamine had no effects on the ADG and the feed efficiency. On the other hand, the rate of diarrhea was reduced, but the difference wasn't significantly (P>0.05).

This experiment was conducted to investigate the effects of prophase nutrition level on the performance of lambs in a two phase finishing scheme. Eighty Mongolian lambs(castrated lambs) were randomly divided into four equal groups:control group (CG) = fed ad libitum (ME:10.88 MJ/kg, CP:15%); moderate restricted groups (RG1 and RG2) = fed restricted (ME:10.88 MJ/kg, CP:10% and ME:9.41 MJ/kg, CP:10% respectively); severe restricted group (RG3) = fed restricted (ME:8.62 MJ/kg, CP:5.7%). The lambs were fed restricted diets for 60 days and followed by compensatory growth with the same diet (ME:9.75 MJ/kg, CP:12%) for each group. The changes of body weight,growth rate,dressing percentage, net percentage of carcass, meat-bone ratio, meat percentage of carcass,eye muscle area and GR value were measured during feed restricted and compensatory periods. The results showed that the BW of CG were significantly higher than those of RG1 and RG2(P＜0.05),and significantly higher than those of RG3(P＜0.01)by the end of feed restricted period.The dressing percentage,net percentage of carcass,meat-bone ratio,meat percentage of carcass,eye muscle area and GR value of CG were higher than those of RG1,RG2 and RG3 because of the feed restriction. The growth rate of RG3 were significantly higher than those of CG(P＜0.05)and the BW,dressing percentage,net percentage of carcass,meat-bone ratio,meat percentage of carcass of RG1,RG2 and RG3 gradually approuched to CG and the eye muscle area and GR value of four groups had no significant difference(P＞0.05)by the end of the feed compensation. The results indicated that the Mongolian lambs expressed different compensatory potency after nutrient restriction with defferent levels of energy and protein, suggesting that protein restriction had more negative effects than energy restriction in lambs. The lamb's growth were more affected by protein during feed restricted period, whereas the supplement of enegy was more important for lamb's growth during feed compensatory period.

This experiment was conducted to study the effect of spent coprinus comatus substrate in basal diet on growth performance and immune function in Tianfu ducks. Three hundred and twenty one-day-old Tianfu ducks were randomly divided into 4 groups,control group was fed basal diet,test group Ⅰ,Ⅱ and Ⅲ were fed basal diets added 2%,4% and 6% spent coprinus comatus substrate respectively.The results that compared with control group,spent coprinus comatus substrate in basal diet improved significantly average daily feed intake (P<0.05) and average daily weight gain of the first two weeks (P<0.05),had no significant effects on average daily weight gain of the last two weeks (P>0.05),decreased rate of feed to gain a little (P>0.05). There were no significant effects on immune organs indices and mitogenic response of lymphocyte (P>0.05).Therefore,basal diets supplemented with 2%,4% and 6% spent coprinus comatus substrate improved growth performance,but had no significant effects on immune function. 2% spent coprinus comatus substrate has the best effect by considered comprehensively.

200 one-day-old commercial generation AA broilers were randomly divided into four groups.In addition to the control group(Cu 11 mg/kg),three experimental groups were set up,they were groups of 110,330 and 550 mg/kg respectively.All of the experimental broilers were fed with corn-soybean meal for 50 days, and effect of high-copper diets on liver antioxidant function and liver cell apoptosis of broilers were assayed. The results showed that 3 high copper group of SOD and GSH-Px increased compensatorily,consequently, a performance of decline;and MDA compensatory increased. With time prolonging and the content of copper increasing, liver cell apoptosis rate rise and DNA fracture strip obviously appeared.It showed that daily ration of copper content 330 mg/kg or more could cause liver antioxidant function was impaired, liver cell apoptosis rate increased.

Milk is a natural source of kinds of fatty acids, which play important roles in the process of metabolism. FA in milk originated from de novo synthesis in mammary gland and blood directly. It varied with many factors, such as genetic, feed, lactation stage and age, season, farming, flock effect, etc. This article has reviewed fatty acids synthesis and correlation among them, of course, factors affected FA were also included.

This experiment was research on produced results of layers by single and composite probiotics preparations. Bacillus Subtilis preparations, Lactobacillus preparations and composite preparations were prepared ferment culture by Bacillus Subtilis, Lactobacillus and composite probiotics. Then these preparations were added 0.1% based on basis grain to 140 days of ageRoman brown commercial layers. The results showed that probiotics preparations improved arerage egg weight, day egg production, laying rate and decreased feed/egg ratio. composite probiotics preparations improved average egg weight by 1.87%, 1.91%, day egg production by 3.34%,2.77%, laying rate by 1.56%, 1.00%, A level egg by 9.16%, 5.91%, vitelline rate by 7.71%, 4.14%,each layer gross profit by 10.84%, 7.73%;decreased feed/egg ratio 1.99%, 1.49% respectively,while in the decreased unqualified egg ratio,death rate,average daily feed consumption,egg shape index and increased egg propotion,eggshell thickness,there was no significant difference. This experiment showed that composite probiotics preparations effect was better than single probiotics preparations.

For searching the differences of amino acids content between the different breeds of sheep or the different strains of the same breed,the content of 17 amino acids of wool in 4 breeds,including Hetian sheep (Pure plain Hetian,Pure mountain Hetian and New Hetian strains),Karakul sheep (Silkworm,girdle-type and rib-type three flower-types),Duolang sheep and Chinese Merino was determined by high performance liquid chromatography (HPLC). The results showed that the total amino acid content of wool from Hetian sheep was higher than that of Duolang sheep,and the difference was statistically significant(P＜0.05). The differences in amino acids content were not statistically significant between the other breeds(P＞0.05). Those amino acids in Duolang sheep such as Asp,Ser,Glu,Arg,Thr,Ala,Pro,Val,Lys,Ile,Leu,Phe were lower than that of the other breeds. The content of Cys in Karakul sheep was higher than that of the other breeds,but Ser,Gly,Thr,Pro,Tyr were lower than that of the other breeds. The ratio of α-helix promoting/α-helix inhibiting amino acids in the wool,Chinese Merino was 0.90,while Karakul sheep was 0.79,Hetian sheep was 0.77 and Duolang sheep was 0.75.The results showed that, total amino acids of Hetian sheep wool was higher than that of other species, the helix ratio of Chinese Merino wool was higher than that of other species,the cystines of Karakul and Hetian sheep wool were higher than that of other species.

This article was conducted to study the factors of isolation buffalo small antral follicles,so that to establish the effective isolation method of buffalo small antral follicles. The results showed that the method of mechanical combine with enzyme harvested significantly more small antral follicles (5.64) than mechanical method (3.12,P＜0.05). The average number of small antral follicles harvested from ovary was increased as enhancing the digesting time of enzyme,and the groups of 12 min and 15 min were significantly higher than that of digesting for 5 and 10 min(P＜0.05),but when digesting time of enzyme was prolonged to 20 min,the theca of follicles was broken out that could not isolate the intact small antral follicles. Most small antral follicles harvested from ovary which surface had less than 2 mm follicles but no corpora luteum significantly higher than that of ovary which surface had corpora luteum but no follicles(7.50 and 2.25,P＜0.05). These results indicated that selecting the ovary which surface had less than 2 mm follicles but no corpora luteum and adopting the method of mechanical combine with enzyme to digest 12 to 15 min was fit for isolating the buffalo small antral follicles.

This study was designed to establish amniotic fluid-derived stem cells line and explore the potential to differentiate into osteoblasts. The amniotic fluid-derived stem cells were cultured under the condition of 15% FBS and detected the embryoid bodies forming ability and investigated the expressions of the stem cell markers (Oct-4,SH2,SSEA-1,Nanog,HLA-ABC,CD117) in different passage cell by RT-PCR. Furthermore,amniotic fluid-derived stem cells were induced by dexamethasone,β-glycerophosphate,ascorbic acid. Amniotic fluid-derived stem cells in vitro were able to expand extensively,form the embryoid body which expressed marker genes of three embryonic germ layers,and differentiate along osteoblasts. The results showed that we successfully isolated the Mongolian sheep amniotic fluid-derived stem cells and established cells line which can be induced to differentiate into osteoblasts.

To investigate regulating effects of taurochenodeoxycholic acid (TCDCA) on immune function in mice under heat stress,50 Kunming mice were divided randomly into five groups: normal control group,heat stress group,TCDCA1 group (50 mg/kg),TCDCA2 group (100 mg/kg),and TCDCA3 group (200 mg/kg). The mice of normal control group were fed in the temperature of 22 ℃,and the mice of other groups were treated with temperature of 35.5 ℃±0.5 ℃ and 60% humidity for 7 days. The spleen index,the change of T cell subgroup and B lymphocyte of peripheral blood,the contents of intestinal sIgA and serum IgG in mice were measured. The results showed that spleen index in mice of heat stress group was significantly lowered compared to the normal control group(P＜0.05);the contents of blood serum IgG and Intestinal sIgA were significantly (P＜0.05) higher;the percentage of CD8⁺ and CD19⁺ lymphocytes decreased significantly (P＜0.05),and the CD4⁺/CD8⁺ value increased significantly (P＜0.05),which demonstrated that the immune functions of mice under heat stress falls into disorders. The results also indicated that TCDCA have no effects on spleen index in heat stress,TCDCA enhanced the percentage of CD8⁺ and CD19⁺ lymphocytes in peripheral blood remarkably (P＜0.05),TCDCA recovered the CD4⁺/CD8⁺ value,remarkably and TCDCA decreased concentration of IgG in blood serum and sIgA in intestinal remarkably (P＜0.05). These findings suggested that the immunity of mice in heat-stress could be destroyed and leaded to disorder,and can be accumulated by TCDCA.

Vitamin E is a kind of fat-soluble vitamins which is of great importance to animals. Vitamin E is utilized by intestinal absorption,lipoprotein transport,storage and regulation in the liver and the uptake and metabolism in varies of tissues. The mechanism is involved in those processes,including the transport of various transporters,catalysis of enzymes and the interaction between different nutrients. Understanding of the mechanism would better to discover the key point in vitamin E utilization and therefore to facilitate vitamin E utilization by regulating the key point. This review briefly summarizes the research advances of the mechanisms involved in vitamin E absorption,transport and metabolism.

The purpose of this study was to know the effects of different species bovine sera on the proliferation and differentiation of 3T3-L1 preadipocytes. In order to provide the possible method of early selection for marbling,we use cell model in vitro culture to simulate the environment of bovine adipose tissue. Collected the bovine sera of different species from seventeen Qinchuan cattle and twenty-eight Qinchuan- Angus hybrid cattle. One part of sera were heat inactivated. The other were activated. The relatively cell number,which stands for proliferation of 3T3-L1 cells,was assayed by MTT colorimetry. Fat content and enzyme activities,which contain glycerol phosphate dehydrogenase (GPDH) and fatty acid synthetase (FAS),represented the differentiation. Oil red O staining was used to determine the fat content,and enzyme activities were determined by colorimetry after inducting adipogenesis. The results showed as follow. First,heat inactivated sera were quite significantly higher than activated ones on relatively cell number of 3T3-L1 cell at day 2 and 4,and it's the same results with fat content at day 2,4,6 and 8 (P＜0.01). Second,Qinchuan-Angus hybrid bovine sera were significantly higher than Qinchuan bovine sera on relatively cell number at day 2 and 4 in cell culture (P＜0.05). Qinchuan sera were significantly higher than hybrid sera on lipid content at day 8 (P＜0.05). However,there were no significant different at other days on cell number and lipid content (P＞0.05). Moreover,there were no significantly different between two species of cattle sera on GPDH and FAS activities in cell culture at day 8 (P＞0.05). Self-made bovine sera within heat inactivation promoted proliferation and differentiation of 3T3-L1 cell. Bovine sera is one of the factors that affect proliferation and differentiation of 3T3-L1 cell. Qinchuan- Angus hybrid bovine sera may be good for proliferation,wahile Qinchuan bovine sera may be good for differentiation. More studys are needed.

To investigate the effect of AAⅠ in inducing apoptosis of the renal tubular epithelial cells(RTECs) in mouse, mouse primary RTECs were cultured and identificated by morphology observation. After RTECs treated with or without AAⅠ, DNA ladder, flow cytometry analysis (FCAS) the apoptosis of the RTECs. Mouse RTECs could be successfully cultured, with the increase of AAⅠ, cells lost epithelial characteristics. After 24 h, 20 to 80 μg/mL of AAⅠ stimulated RTECs, cells apparence late apoptosis. The apoptosis of mouse origin RTECs could be induced by AAⅠ, the degree of apoptosis is along with the increasing of AAⅠ action time and density.

To investigate the inclusion of Trimethoprim (TMP) with Hydroxypropyl-β-cyclodextrin (HP-β-CD) in aqueous soliutions,change of thermodynamic parameters during the inclusion process and inclusion molar ratio of the host and guest molecules. The measurements of apparent inclusion constant of TMP with HP-β-CD, the change of entropy,enthalpy and free energy of the reaction, inclusion molar ratio were carried out by the following methods separately were phase solubility method,equimolar series method and molar ratio method,respectively. That all the phase solubility diagrams showed a typical A_L-type in various temperature aqueous solutions, suggested the formation of a inclusion complex of 1∶1 molar ratio.The change in the thermodynamic parameters suggested that the inclusion could proceed spontaneously along with the release of heat and the increase of entropy. An 1∶1 molar ratio inclusion complex of TMP with HP-β-CD could be formed spontaneously and the solubility of TMP in aqueous solution increased. Appropriate temperature probably favor the progress of inclusion procedure.

The experiment was conducted to study the antimicrobial activity of essential oil from Wedelia trilobata(L.) Hitchc. to the common pathogenic bacterias on livestock and aquaculture.The essential oil was extracted from Wedelia trilobata(L.) Hitchc. by steam distillation, and then its antimicrobial activity and chemical composition were assayed. The results showed the essential oil of Wedelia trilobata(L.) Hitchc. had significant inhibitory effect on 7 kinds of bacteria tested,and when the concentration of the essential oil was higher, the antibacterial activity was enhanced.Forty-eight ingredients were identified from Wedelia trilobata(L.) Hitchc.by GC-MS, the maor components were Tricycloheptane,1,3,3-trimethyl-3-Carene,alpha.-Phllandrene,Caryophyllene oxide,etc.

For developing the typical testing standard of transgenic mammals and its derived food and the technical support for entry﹠exit animal quarantine, detection of the transgenic component in transgenic cow by multiplex PCR was developed in this study. The endogenous mtDNA gene specific for cow identification, exotic hLF gene,marker NPTⅡ gene were amplified according to different primers. The process of the multiplex PCR reaction was optimized. The system described herein represent simple, accurate, and sensitive detection methods in which only one reaction was necessary to detect multiple target sequences that can be reliably used for the identification of specific lines of transgenic cows.

The canine distemper virus (CDV) isoladed from lesser panda and domesticated by the laboratory of genome sequence determination,genetic characteristics and genetic stability of the H gene. According to full-length genome sequence of canine distemper virus published in GenBank, 17 pairs of specific primers were designed and synthesized to amplify full-length cDNA as template with total RNA of lesser panda canine distemper virus, using the method of RT-PCR, and cloned into pEASY-Blunt Simple vector for determination of full-length cDNA sequences.The results showed that the homology of whole genome sequence of lesser panda source canine distemper virus and 12 different strains AF014953, AY445077, AY542312, AY466011, AY386316, AF164967, EU716337, EU726268, AY443350, AB474397, GU138403 and AY649446 is 86.6%, 92.4%, 92.5%, 92.5%, 94.3%, 96.3%, 95.9%, 87.1%, 94.5%, 92.3%, 87.4% and 94.6%, respectively. It had the closest genetic relationship with the standard virulent strain A75/17, gene homology was 96.3%, but compared the vaccine strain Onderstepoort and it had distant genetic relationship, only 86.6% homology.Phylogenetic analysis of H gene of lesser panda source canine distemper virus and other thirty CDV published in GenBank in different districts showed that, canine distemper virus of lesser panda source belonged to the AsiaⅠ, the potential glycosylation sites of H protein gene were formed by amino acid from 309 to 311 of H protein, the wild strains had it common but the vaccine strain didn't, and may be associated with the virus immunogenic. Therefore, attenuated panda source CDV in the prevention of immune targeted may be better than the existing vaccines strain.

Use the genome DNA of pseudorabies (PRV) virus SD strain as template,the major epitope of PRV envelop glycoprotein gD was amplified by polymerase chain reaction (PCR)and cloned into pGEM-T vector,then inserted into the downstream of T7 promoter to construct an expression plasmid pET-32a. After induction by IPTG,the fusion protein was highly expressed in Escherichia coli BL21(DE3) in the form of inclusion bodies. The recombinant protein was purified with His-bind affinity chromatography. SDS-PAGE and Western blotting analyses revealed that the recombinant protein with the expected 45.2 ku could react with pig serum containing antibody against PRV. All results indicated that the recombinant fusion protein can be used as an antigen of diagnostic assay to detect the PRV antibody in pig serum.

Two hundred,24 week-old health cage layers were chosen,and divided randomly into four groups : the control group (containing calcium 3.55% in based diet),the low calcium group(containing calcium 1.27% in low-calcium diet),the test group 1 (adding 8 mg/kg ipriflavone to low-calcium diet) and test group 2 (adding 20 mg/kg ipriflavone to low-calcium diet). The effects of ipriflavone on relevant hormone of cage layer osteoporosis (CLO) were observed. The results showed that serum PTH levels showed an upward trend in each group. On 60 days,compared with the low-calcium group,PTH of test group 1,2 significantly reduced 6.12% and 7.05%(P＜0.01) . On 60 days,compared with the low-calcium group,the activity of serum AKP of test group 1,2 significantly reduced 23.72% and 25.08% (P＜0.01). In each group,the serum CT levels showed the upward trend in the 15 to 30 d,and the declining trend in the 30 to 60 d. The entire experimental period,compare with the control group,the serum calcium levels of experimental group and low calcium group decreased,but the difference was not significant. The serum E₂ levels were not significant difference in each group. The results suggested that the low calcium diets supplemented with ipriflavone can prominently adjust relevant hormone levels of calcium metabolism,and promote deposition of calcium ion to the bone,reduce bone demineralization and mitigate the degree of osteoporosis layers.

The PCR assay and PCR-RFLP were applied to study the function of aroA gene on the detection and genotyping of Haemophilus parasuis. 18 strains of Haemophilus parasuis isolated from swine in Guangxi were detected successfully by using the PCR assay with primers targeted to aroA gene, and the sensitive test showed that the lowest amount of Hps detectable by this PCR-based test was 10². The aroA gene of the 18 strains was sequenced, and enzyme restriction analysis of the aroA sequences available was performed to define which restriction enzymes more practical. HindⅢ and FokⅠ were chosen to do RFLP analysis in all 18 Haemophilus parasuis strains and one reference strain. The result of the analysis showed that there were three patterns among the groups. The study suggests that the PCR assay and PCR-RFLP applied to detection and genotyping are useful to further research the biological characteristics and gene function of Haemophilus parasuis.

Two porcine reproductive and respiratory syndrome virus (PRRSV) strains which named YN-1 strain and YN-2 strain, were isolated by inoculating into Marc-145 cell. The two isolated strains can induce Marc-145 cell stack together, pull net, formation of plaque, and other typical lesions for 4 blind passage, with TCID₅₀ 10^-3.6/0.1 mL. RT-PCR based on NSP2 gene showed that there were 90 nucleotides deletion than Ch-1a strain, the typical gene characteristic of highly pathogenic PRRSV. Genetic evolution of ORF5 showed the two isolated strains were in a small branch with high identity 99.5%. They were in a branch with Shandong strain JN-HS,Hennan-1 and Vietnam 347-T-KSA strain with identity 99.2% to 99.8%. The two isolated strains were in a different branch with Ch-1a and VR-2332 strains,the identity were 94.4% to 94.5%, and the identity 98% to 99% with other Chinese highly pathogenic strains, the two isolated strains belonged to the high-pathogenic strain.

This study from different areas of Guangdong province inspection disease material success in the identification of four strains Haemophilus parasuis and according to Haemophilus parasuis 16S rRNA specificity gene detection of Hps by PCR and genetic gene sequencing allogenic analysis, through which GenBank online than analysis, with domestic and international strains isolated strains 16S rRNA sequence homology in 98.2% above, separation between homology strains in between 99.6% and 100%, which demonstrates that the outbreak strain is pair of Haemophilus parasuis.

The porcine interleukin-6 (PoIL-6) mature peptide gene was amplified from the total cellular RNA of porcine spleen cells induced by bacterial lipopolysaccharides (LPS) by RT-PCR, and the PCR production was subsequently cloned, sequenced and sub-cloned into the prokaryotic expression vector pQE30. The PoIL-6 mature peptide gene was expressed in E.coli JM109 and the expressed fusion recombinant protein (rPoIL-6) was purified under the innovated recombinant fusion protein purification method of denaturing by 8 mol/L urea, refolding by a self-innovative renaturation buffer and dialyzing by PBS buffer etc. The porcine IL-6 ELISA assay was used to detect the specifically immunological activity of the rPoIL-6 protein, and the cell proliferation activity induced by rPoIL-6 protein was assessed by MTS assay. The result showed that the PoIL-6 mature peptide gene with a length of 555 bp was successfully cloned, expressed, and purified with the molecular mass of about 20 ku and more than 95% pure on SDS-PAGE, which indicated the correct PoIL-6 fusion protein had been obtained. The rPoIL-6 protein could specifically react with McAb aginast rPoIL-6 and significantly promote the proliferation of porcine spleen lymphoblast cells.

According to the gene sequences in GenBank of alpha-toxin,one pair of specific primer was designed for amplifying the specific fragments of alpha-toxin gene. After optimization of annealing temperature and primers concentrations,PCR assay was established for simultaneous detection of the Clostridium perfringens. The specific bands of 255 bp was amplified.The sensitivity and specificity tests showed that the PCR were sensitive in 0.39,0.62,0.52,0.87,0.92 ng/L A,B,C,D,EC.perfringens. No band was amplified from Escherichia coli,Streptococcus,Staphyloccocus aureus and Pasteurella multocida. 54 clinical samples were detected by PCR.The results revealed that the established PCR assay was sensitive,specific and reproducible and it could be used to detect rapidly Clostridium perfringens isolated from sheep.

Heavy metals cannot be rendered harmless by chemical or biological remediation processes and can persist in body of human and animals for long periods. Through the food chain heavy metals may deposit into human and animals body and even a little quantity can cause serious health problems which could transfer to the next generation on the genetic level. So in environmental and agricultural food monitoring it is very important to quantitatively analyze heavy metals. Traditionary analysis methods for detecting heavy metal residues include UV-visible spectrophotometry,atomic absorption spectrometry and mass spectrometry,et al. And immunoassay is a new alternative method for detecting heavy metal residues.

A loop-mediated isothermal amplification(LAMP) assay for rapid detection of Babesia bigemina was developed using primers specific to six distinct regions of Cytochrome b gene of Babesia bigemina. LAMP was performed using Bst DNA polymerase at 65 ℃ in water bath for 60 min and amplification results was visualized with SYBR Green Ⅰ. Specific bands could be detected for Babesia bigemina,but not for others. The detection limit of the assay was 0.085 fg/μL for Babesia bigemina,which was up to 1000 times higher than that of PCR methods. The LAMP methods was rapid and could be easily performed at a low cost.

Single nucleotide polymorphisms (SNPs) have been always the focus of most molecular biologists. However,the core of genome research in organism is to understand the molecular mechanisms of phenotypic differences so that a complete understanding of all the kinds of variations is necessary besides SNPs. Copy number variations (CNVs) are DNA segments ranging from kb to Mb. Many of CNVs are big enough to encompass whole genes so that they are more likely to affect organismal fitness compared to SNPs. There are now multiple studies demonstrating a strong link between copy-number differences at specific genes and phenotypic differences. In this paper,we review the outline of CNVs,focusing on the methods used to detect them and the molecular mechanisms responsible for generating this type of variation. In addition,research of CNVs in livestock and poultry was introduced.

Information on transgenic livestock which had been successfully produced in the world since 1991 was collected from literature database of NCBI and CNKI websites. Based on these information and technology of ASP.NET,information database of transgenic livestock was constructed (http://www.gmanimal. com.cn/sjk/ index.asp). The database contained information on species,the names of the exogenous genes,transgenic methods,phenotypic characteristics,research institutions,countries or regions,literature link and so on. To date,there were 79 records in the database,which included 37 records of transgenic pig,16 records of transgenic cattle,12 records of transgenic sheep and 14 records of transgenic goat. This database was open and free for public,users could query and download relevant information,which achieved share on information of transgenic livestock.

In order to make sure whether the DNA methylation reprogramming is efficient in SCNT animals,we analyzed the DNA methylation status of IGF2/H19 imprinting control region (DMR1,DMR2,DMR3) in heart,liver,spleen,lung and kidney of cloned pig using bisulfite sequencing analysis. The results demonstrated that the methylation level of DMR1 and DMR3 in dead cloned pig tissues were different from that of normal pig,but there was no significant differences among them (P＞0.05);The DMR2 showed hypermethylation in the lung of the dead cloned pig,and was significantly higher than that in the control(P＜0.01).Also,the tested CpGs sites from 4 to 9 and 12 to 17 exhibited full methylation. There was no significant differences among other tissues in the DMR2 (P＞0.05). Results showed that the abnormal DNA methylation proflies of DMR2 may occurred in the lung of cloned pig,which may be one of the factors for the death of cloned animals.

The possibility of intraspecific and interspecific fertilization between buffalo and cattle was explored in this study. Buffalo and bovine oocytes matured in vitro for 22 to 24 h were fertilized by injection of buffalo or bovine frozen-thawed spermatozoa. At 16 to 18 h after injection,a portion of ICSI oocytes were fixed in acetic ethanol to examine the pronuclear formation,others were cultured in vitro to evaluate their cleavage rate and embryonic developmental ability. There was no difference in the two pronuclear formation rate (47.73% and 36.67%),cleavage rate (68.00% and 72.64%) and blastocyst development(16.44% and 22.39%) of buffalo oocytes fertilized with buffalo (intraspecies) and bovine sperm (interspecies) (P＞0.05). The two pronuclear formation rate (60.00%),cleavage rate (73.33%) and blastocyst rate (28.57%) of bovine oocytes fertilized with bovine sperm (intraspecies) were also not different with the embryos from those (45.28%,71.31% and 33.70%,P＞0.05) of bovine oocytes fertilized with buffalo sperm (interspecies). This results indicated that: ① Buffalo oocytes fertilized with bovine sperm and bovine oocytes fertilized with buffalo sperm could develop to blastocysts, ②The developmental capacity of bovine oocytes was higher than buffalo oocytes as evidenced by their proportion cleaved and developed to blastocysts after intraspecific and interspecific fertilization by ICSI.

This research was conducted to determine the effect of different hormones or treatments methods on lamb's follicular oocytes development, and the difference of oocytes' maturation process in vitro between superovulated lamb and normal adult ewe was studied.Besides, the maturing rate of lamb's oocytes in vitro based on different prescription of culture solution was determined. The results suggested as following: the optimal age for early using of Liaoning Cashmere goats' lamb was 5 to 6 weeks, with the body weight between 5.6 to 7.5 kg, the optimal method of lamb's superovulation was injecting FSH 40 mg three times and PMSG 400 IU once in sequence with the interval of 12 hours, adding 10% FBS (fetal bovine serum) into culture solution can be replaced by 12 mg/mL BSA, which influenced nothing to the maturation of lamb's oocytes. However, the matured rate of oocytes in vitro of superovulated lamb's was significantly lower than normal adult ewe's.

Pregnancy-associated glycoprotein (PAG) is a pregnancy specific protein which found in the peripheral blood of pregnant ruminant animal. PAG plays an important role during pregnancy. Currently,detecting the PAG concentration in the serum can used for pregnancy diagnose. This article reviews the discovery,purification,metabolism of PAG,and the application of PAG in early pregnancy diagnosis.

In the production of poultry,more female chick are needed to the egg laying poultry when incubating while more male chick are needed to the meat poultry. What's more,the treatment of male chick spends much in the egg laying poultry. Not only the cost of incubation,but also the costs of the treatment to waste chick can be saved if the sex of the early embryo can be identified earlier when incubating. Several methods of sex identification of poultry's embryo are introduced in this paper,so as to supply reference for the research in this aspect.

Fleckvieh beef cattle and Japanese Black beef were used as donor,local cattle as receptors in this study,then estrus synchronization and superovulation were conducted, and 234 embryos were obtained,of which were divided into four groups,including 113 fresh whole embryos,55 fresh double half embryos ,56 frozen whole embryos and 10 frozen double half embryos. The transplantation effect of whole embryo and double half embryo were detected. The results indicated that the pregnancy rate of fresh double half embryo was 65.45%,which was higher than fresh whole embryo group(63.72%),but no significant difference(P＞0.05).Frozen double half embryo group was 20.00%,which was not significantly lower than the corresponding whole embryo group(39.29%)(P＞0.05).However,both frozon groups were significantly lower than fresh groups. The calving rate of fresh double half embryo was 95.56%,of which 35.55% were twins and 72.09% were survived. The calving rate of fresh whole group was 63.16%,which was significantly lower than the fresh double half group(P＜0.05),but the surviving rate was significantly higher than the fresh double half group(90.00% and 72.09%)(P＜0.05). As for the birth weight,there was no significant difference among all of the four groups.

To study the effects of foot-and-mouth disease vaccine on sperm quality including the average collected volume, the average density, the average activity,the rate of semen abandon and the number of frozen semen tubules in bulls of different months. 30 bulls were divided into 3 groups according to their age and they were injected foot-and-mouth disease vaccine simultaneously. The results showed that after injecting foot-and-mouth disease vaccine, the sperm collected volume was no significant differences (P＞0.05) among the three groups, and the average sperm density was also no differences (P＞0.05) between bulls aged 24 and 32 months besides the 42 months bulls whose density of sperm was significant differences(P＜0.01) after injection. At the same time, the sperm activity in all experimental bulls was significantly lower (P＜0.01) than that before injection, and the time of the influence on 32-month-old bulls was the shortest. Otherwise, the rate of semen abandon of bulls aged 32 months was no significant differences (P＞0.05), and which was increased (P＜0.01) significantly in 24 and 42 months bulls. The number of frozen semen tubules was also no significant differences (P＞0.05) in 32 months bulls and which was obvious differences(P＜0.05) in another two groups in the later period of injection. It was concluded that injection of foot-and-mouth disease vaccine affected differently sperm quality of Fleckvieh bulls of different months, especially youth bulls.

At present,coccidiosis is still mainly controlled by the use of chemotherapeutic agents;thus,new drugs are urgently needed due to the rising problem of drug-resistant strains of Eimeria. To discover and select suitable drug target was the precondition of drug development as well as the key point of drug screening and oriented synthesis. This article reviews the progress in research of the drug target screening.

This paper outlined yak major parasitic diseases,its clinical symptoms and drug therapy,meanwhile,this paper also pointed out that the current lack of parasitic diseases control of yak and drug development direction for yak disease parasite control.

Acariasis is one of the diseases caused to reduce production performance of domestic animals, which affects the digestion and absorption, even results in death of animal and threatens human health. Nowadays, chemical drug is widely used for prevention and control acariasis in domestic animals. Those drugs have a bad influence on ecological environment. Therefore, looking for low-toxic, high efficiency natural plant acaricidal drug are concerned. The paper reviewed situation of the research of the acaricidal plants and existing problem.

According to the conservative region of H5 AIV HA gene,the primers were devised and synthesized. A serial 10 fold dilutions positive plasmid was prepared and used for standard. The standard curve revealed the linear relationship between Ct (cycle threshold) and template concentration with a good correlation (R²＝0.995). The RRT-PCR method for the detection of AIV was highly specific and sensitive,and it could be used for rapid quantitative detection of AIV H5 subtype. No cross-reaction was detected against other avian disease viruses. Sensitivity was 8 copies of AIV genome. The total meet rate with traditional virus isolation method was more than 90.0% in detecting 178 clinical cloacal swab samples. It showed that a real-time RT-PCR method for detection of H5 subtype avian influenza virus had been established in this study,which had properties such as high speed,specific,sensitive,cheap,high-throughput,and good biological safety as well as displaying good prospects in the rapid screening of clinical samples and epidemiological monitoring of avian influenza virus H5 subtype.

To understand the mechanisms of DPV infection and immunity,the IFN-γ mRNA expression was investigated after infection with virulent and attenuated duck plague virus. The mRNA levels of IFN-γ in liver and PBL were evaluated dynamically after inoculation with virulent and attenuated DPV respectively by real-time PCR. The virus loads were also assessed. Results:①After infection by virulent DPV, IFN-γ mRNA levels in liver didn't show regulatory changes while high level of IFN-γ mRNA levels in PBL and few virus DNA could be detected in the early stage of infection (1-12 h). IFN-γ mRNA expression decreased dramatically after 6 h post infection and persisted in a very low level until death (144 h).Virus load increased gradually and reached peak at 144 h. ②After vaccination with attenuated DPV strain, there were persist and high levels of IFN-γ mRNA expression in liver,reached peak at 12 h (8 times higher than control), while the level in PBL was lower and unstable. The attenuated DPV loads increased dramatically with a relative low copy number in liver. However, the virus DNA could not be detected in PBL. IFN-γ possibly plays an important role in protecting ducks against virulent virus infection. It is also suggested that high level expression of IFN-γ in liver induced by attenuated DPV could enhance immunity against DPV infection.

Mastitis is one of the common diseases in dairy cows, especially subclinical mastitis has caused huge economic losses. This paper reviewed mastitis treatment of Chinese herbal medicine in dairy cows, such as mastitis pathogenesis, the shortcomings of traditional treatment, the progress on Chinese herbal medicine control mastitis in dairy cows.

Mycoplasma ovipneumoniae, the cause of a typical pneumonia of sheep and goats, is responsible for important economic losses in ovine industry worldwide. The aim of this study was to develop an inactivated Mycoplasma ovipneumoniae vaccine. Mycoplasma ovipneumoniae isolate SC02 was screened and cultured in modified Thiaucourt's broth media and harvested when the titers reached 10⁹ CCU/mL. The cultures were concentrated 20 folds as antigens for oil adjuvant vaccine preparation. Six-month-old goats which without Mycoplasma ovipneumoniae antibody were injected subcutaneously with 5.0ml(2×10¹⁰ CCU/mL) the vaccine. 21 days after immunity, the goats, including tested and controlled goats, were tracheally inoculated with 5.0 mL(≥2×10¹⁰ CCU/mL) highly virulent Mycoplasma ovipneumoniae strain Y98,and the temperature and clinical symptoms were observed daily.All goats were dissected to observe pathological changes post 30 days of injection of Y98 strain. The results showed that all five goats inoculated vaccine did not appear clinical symptoms and pathological changes; conversely, the control goats had developed clinical symptoms with cough, fever, runny nose. Moreover, typically lung pathological changes of Mycoplasma ovipneumoniae were observed in controll ainmals in anatomical examinations. The results showed that the vaccine developed in this study can protect goats effectively against Mycoplasma ovipneumoniae.

The pharmacokinetics was investigated and compared in 50 healthy rabbits following single subcutaneous (5 mg/kg body weight) administration of 5 different ivermectin injection. The ivermectin concentrations in plasma samples were determined by HPLC-UV. Pharmacokinetic analysis of plasma drug concentration-time data for ivermectin was carried out with a computer program 3P97. The ivermectin concentration-time data were fitted to one-compartment open model after single subcutaneous administration of ivermectin injection. The main pharmacokinetics parameters(E) were as T_1/2(Ka)(8.10±3.68)h,T_1/2(Ke)(36.34±14.63) h,T_max(22.38±8.63) h,C_max(354.01±57.75) ng/mL,AUC(25843±3029) ng/(mL·h). The statistics analyzed by SPSS results showed there were no significant differences (P>0.05) in T_1/2(Ka),T_1/2(Ke).However, pharmacokinetic of C_max was significant differences (P<0.05). Pharmacokinetics of AUC was higher than others, the results of present studies showed that the ivermectin injection demonstrated rapid absorption, rapid elimination, highly bioavailability. The safety of ivermectin in rabbits indicated that ivermectin injection satisfied clinical requirements cluded that supplementation of bacillus preparation in diets increased the antioxidant capacity and improve intestine microbial flora of layers.

The vaccine immunization is an important tool to prevention and control of porcine reproductive and respiratory syndrome currently. The traditional PRRS vaccines played a major role in preventing and controlling porcine reproductive and respiratory syndrome,variation with the acceleration of PRRSV,the traditional PRRSV vaccines can not afford full protection in recent years. With the development of genetic engineering technology,a number of new vaccines developed very rapidly,some vaccines have been commercialized on the market and widely applied to production,it gives human and animal disease prevention and treatment has brought new hope. Although the new vaccine is also flawed,but as molecular biology,immunology and other disciplines,the development of new vaccines through constant transformation,will better serve the sustainable development of humans and livestock.

Porcine reproductive disturbance diseases have affected the development of pig industry seriously. In order to investigate the epidemics of those diseases in Shanxi province, pigs from 42 breeding farms in 8 cities were selected. It was found that piglet death rate accounted for 13.77% because of sow reproductive disturbance diseases. Then 257 tissues and serum from 1776 diseased pigs at different ages were got and the pathogens and antibodies for swine fever, swine blue ear disease, swine pseudorabies and swine circovirus type 2 were analyzed by PCR and ELISA. The results showed that the swine fever virus infection rate was 28.4% and antibodies positive rate was 57.55%; porcine reproductive and respiratory syndrome virus and its variants infection rate was 3.89% and 39.69%, respectively, and antibody positive rate was 44.99%; swine pseudorabies virus infection rate was 10.12%, gE antibody positive rate was 2.98% and gB antibody positive rate was 76.35%; swine circovirus type 2 infection rate was 15.18% and antibody-positive rate was 56.98%. It was suggested that single and mixed infections of those four kinds of diseases were occurred seriously in pig breeding farms in Shanxi province, which supplied the evidence for those diseases prevention and control in Shanxi province.

The duck infectious serositis is one of the most acute and septic infectious with a high mortality,which severely affect the development of ducks industry. All kinds of conditions were screened in study on infecting method of serositis in duck. Ducks of six-week old were injected with Pasteurella anatipestifer by abdominal cavit. On the basis of the experiment of bacteria dose of infection,the most suitable experimental infectious dose of bacteria was 0.5 mL (4.5×10⁹CFU/mL). Injected ducks began to sicken after five hours,and the mortality came to 80% in 3 to 5 days. The clinical symptoms,the pathology examination and etiology identification are in line with the requirements of disease model replication. The method can provide theoretical basis for pathology study and screening for new drugs.

The idiopathy cobra in vivo separation's bacterium after the morphology observation, the raise characteristic and the biochemistry experiment, appraises for the backwoods E.coli. This backwoods E.coli performs the animal experiment, may cause the white mouse death, explained that it has certain toxicity. The commonly used 14 kinds of antibiotic slip of paper to separates the strain with clinical on to enter takes a walk the sensitive experiment, screens the sensitive medicine to use in the clinical care obtaining the good results, active control cobra large intestine bacillosis's unpopularity.

Global warming has become the focal topic nowadays,greenhouse gases (GHG) emissions caused by animal husbandry takes 18% of the wlobal's total emission,which has become the main cause of global warming. Therefore,the study of GHG emissions mechanism from various livestock and find out the influencing factors which impact on GHG emission are of great significance on reducing GHG emission. Reviewed the mechanism of greenhouse gas emissions though livestock and put forward the measures on reducing greenhouse gas emissions,provided reference for exploring energy saving,low greenhouse gas emissions of environmental breeding.

The aim is to reconstruct the BHK-21 cells contaminoted by Escherichia coli using Enrofloxacin.This study tested the MIC of Escherichia coli and BHK-21 cells,found the safe concentration of Enrofloxacin was 10 mg/L, it didn't effect the growth of BHK-21 cells, and could inhibit the Escherichia coli. By physical and chemical method, the contaminated BHK-21 cells were treated with the MEM growing solution of the same concentration of Enrofloxacin circularly. The Escherichia coli were cleaned successfully after 3 cycles and 3 passages, the BHK-21 cells were recovered. The method offered a reference for saving the contaminated cells.

A strain of Eimeria coccidia was isolated from suspensions of oocysts by the use of the single oocyst isolation technique. The oocysts of the strain were elliptical in shape and had average dimensions of (22.51±0.4790) μm×(15.94±0.5599)μm. Its prepatent period was 120 h and 50 min, and the peak of oocyst production occurred on the 6^th post infection. Comparing the data of our observation with literature of Eimeria coccidia the isolated strain of Eimeria coccidia was identified as Eimeria. brunetti and was named as Eimeria. brunetti-GXNN. By further researching the characteristics of the postcocious line we would use it as an immunity candidate strain of Eimeria coccidia.

Blood serum epidemic research which to find the immune effect of partial hogpen from 2009 to 2010 in Yunnan province after major porcine infectious disease vaccine injection had been taken by ELISA test and latex agglutination test. The result showed the positive rate of porcine antibody respectively against CSFV, PRRSV, PCV2, PRV, PPV and Japanese encephalitis virus made up 84.33%(1211/1436), 74.84%(803/1073), 0.00%(0/80), 62.26%(287/461), 54.25%(83/153) and 63.33%(76/120).After considering all the results presented above, we can safely predict that it's high time we prevent and lessen sow aborted by improving PRRSV, PRV, PPV and Japanese encephalitis virus vaccine using effect.

The high level harmful gas in poultry house is the key factor that induces the respiratory systemic disease of poultry. We reviewed the kind of gas and the harm of which to poultry. Furthermore, we reviewed the measures that have been taken to control the gas.

All field isolates of Bordetella bronchiseptica (Bb) in this study were derived from samples of lung or nose swab of pigs with respiratory disease from pig farms in Henan province. These strains were identified base on culture characteristics,biochemical assay and polymerase chain reaction (PCR) targeting the specific flaB gene of Bb. The result showed that 8 strains were isolated. The drug sensitivity test results showed that these Bb strains were highly sensitive to doxycycline,chloramphenicol,Flubendazole,Ofloxacin,tetracycline,kanamycin,gentamicin,Nalidixic acid and amikacin.