This study aimed at developing a novel technique of isolating the Oct-4 expressing cells from the sheep amniotic fluid and fetal renal tissues, respectively, and comparatively analyzing the difference in the molecular characteristic of these two differently derived cells with Real-time PCR. The results showed that the cells from the sheep amniotic fluid and fetal renal tissue were both positive for Oct-4, embryonic germ cells gene SSEA-1, the main organization compatibility antigen gene MHC-Ⅱ, apoptosis gene Bax, and inhibition gene of apoptosis Bcl-2, negative for the symbol marker of MHC-Ⅰ, Nanog and TERT, but there were marked differences in the relative expression levels of Oct-4， Bax and Bcl-2 genes. Furthermore, AFSC and RTSC were both capable of forming embryoid bodies through the suspension culture in vitro. In conclusion, our study successfully established a new procedure of isolating and culturing Oct-4 expressing cells derived from the sheep amniotic fluid and fetal renal tissues, which might have the same origin.

4 of E.coli strains were isolated from the soil of chicken farm on the EMB medium, and further identified as Escherichia coli by 16S rDNA sequence analysis. One virulent phage ΦLY4 was isolated when propagated on the E.coli LY4 as host cells. The phage had the hexagonal head and tail after observing with negatively stained method by electron microscope. The drastic diameter of this phage was 2.5 to 3.8 mm in size, and its phage titer was about 2.0×10¹⁰ PFU/mL, the optimal multiplicity of infection was 0.0001. It kept high stability at the temperature of －20 to 37 ℃ and pH at 5.0 to 10.0. Its viability was lost completely at the simulated gastric conditions （pH 1.2）. The growth curve showed that its incubation and burst phase was 20 min, and the lysis ability was strong. All of these properties showed that the phage ΦLY4 had a potential application in the control of environmental pollution.

In order to establish an indirect ELISA method for detection of equine herpesviruses type 4 （EHV4） antibody, the glycoprotein G （gG） protein with specific epitope worked as a detection antigen. After optimizing conditions, the indirect ELISA method was developed successfully，specificity and repeatability tests were determined. The result was that the EHV4 gG only reacted with antibodies against EHV4;but not with antibodies against EHV1; both the intro-batch and inter-batch variation coefficiencies were lower than 10%.Concordance of the indirect ELISA relative to commercial EHV1/4 antibody kit was above 90%.The results indicated that the indirect ELISA method could be used for the detection and epidemiological surveys of EHV4 infection.

To study immediate early 180 （IE180） gene of pseudorabies virus (PRV) and its function，IE180 gene was amplified by PCR method with designed specific primers, and then IE180 cloning vector was successfully constructed. After sequence determination, IE 180 gene sequence and amino acid sequence were compared and analyzed by using DNAStar software among seven published PRV reference strains. The results showed that the CDS region of IE 180 gene consisted of 4425 bp, coded 1474 amino acids，the gene and amino acid sequence homologies between HB-98 strain and reference strains were 98.4% to 99.1% and 97.8% to 98.4%. Phylogenetic tree analysis demonstrated that HB-98 strain was genetically closely related to TNL strain（AF352564.2）.

To lay the foundation for investigating the role of smurf2 in aristolochic acid nephropathy,we optimized the best transfection condition of Smurf2 siRNA,and screenned the best siRNA fragment,then realized the Smurf2 transient gene silencing. We cultured renal tubular epithelial cells (RTECs),Lipofectamine^TM 2000 was taken for transfection medium,transfecting Smurf 2 siRNA into RTECs,the transfection condition was optimized by observing expressions of green fluorescences and undertaking Real-time PCR reactions.After transferring the Smurf2 siRNA, mRNA inhibition ratios were detected by Real-time PCR.We detected the effects of compound Smurf2 siRNA on RTECs activity by CCK-8;at the same time,we detected the expression of Smurf2 under different sites of Smurf2 siRNA by Real-time PCR and Western blotting. The results showed that the best transfection condition was 1.5 μL Lipofectamine^TM 2000 and 30 pmol siRNA,the transfection efficiency was 70% to 80%.Smurf2-619 siRNA was proved with the most obvious interference effect.Smurf2 protein levels decreased significantly in the group of Smurf2 siRNA compared with the normal group （P<0.05）.We determined the best transfection condition of Smurf2 siRNA, Smurf2-619 siRNA had the highest inhibition rate for the expression of Smurf2.

Based on the MC1R nucleotide and amino acid sequences of Canis familiaris, Vulpes vulpes, Alopex lagopus and Nyctereutes procyonoides published in GenBank, sequence variation and genetic evolution of MC1R gene were analyzed by means of bioinformatics softwares. The results showed that， the length of Canidae family MC1R gene coding sequence was 954 bp, the base composition was C＞G＞T＞A and the content of G+C was higher than that of A+T; A total of 20 polymorphic sites, including 16 single mutation sites and 4 parsimony informative sites were detected from four Canidae familes. In addition, the base conversion was more than transversion types. There were higher similarity of nucleotide and amino acid between four Canidae families; The phylogenetic distance between the Vulpes vulpes and Alopex lagopus was shorter than that between Vulpes vulpes and Canis familiaris. The result was basically consistent in molecular phylogenetic tree constructed by three methods which were NJ, ME and UPGMA, the Vulpes vulpes was clustered together with Alopex lagopus, Nyctereutes procyonoides was earlier divergent than the other three Canidae families.

The infectious bursal disease virus (IBDV) strains，Gx (very virulent strain), F9 (intermediate strain) and Gt (attenuated strain) had highly similar genetic background, but their biological characteristics were significantly different. To investigate molecular details of the interaction between various IBDV and their host cells, the genes of VP2 capsid protein of Gx, F9 and Gt were cloned into vector pGBKT7, and the recombinant bait vectors pGBGxVP2, pGBF9VP2 and pGBGtVP2 were constructed. Then the self-activation and toxicity of the bait vectors were tested using the Matchmaker Gold Yeast Two-hybrid System. The results showed that three bait vectors all had no self-activations and toxicities to yeast cells. This research benefited to further investigate the interaction between IBDV and its host cell using the yeast two-hybrid system.

To detect porcine epidemic diarrhea virus (PEDV) accurately and sensitively, two pairs of specific primers amplifying M gene were designed and the nested RT-PCR method was established. The lowest detective value of the nested RT-PCR as proved by the sensitivity test was 50 fg of PEDV RNA template, however,that of the conventional RT-PCR was 50 pg. The nested RT-PCR primers designed in the study could not be used to amplify the PRRSV, CSFV, TGEV and GARV RNA templates in the specificity test. The results showed that the nested RT-PCR was a rapid, sensitive and cost-effective method for detecting trace amounts of PEDV nucleic acid.

Bacterial ghost technology is a novel method for preparing the inactivated vaccine. This nondenaturing inactivated method keeps many antigenic determinants which serves as a candidate vaccine to prevent pathogenic bacteria. This experiment took lysis E gene from phage PhiX174 DNA by PCR and inserted E gene into pBV220 vector, then cloned protein lysis component (PLC) from expression vector pBV220+E which we had constructed before, this PLC contained repressor protein cI857, lysis E gene and terminator sequence rrnbT1T2, as well inserted PLC into the broad-host shuttle vector pBBR1MCS-2. Finally, the broad-host lysis plasmid pBBR+PLC was electrical transformed into Brucella canis RM6/66. The results showed that the broad-host lysis plasmid had highly killing effect for Brucella canis RM6/66 after 42 ℃ induction and its lysis rate was 100%, meanwhile Brucella canis RM6/66 ghosts were successfully generated. This study used bacterial ghost technology to prepare the Brucella canis ghost, which played an important role in the prevention of canine brucellosis, simultaneously provided a new strategy for human and animals’ vaccine against brucellosis.

This test was intended to construct eukaryotic expression vector of porcine growth hormone secretagogue receptor (pGHS-R), and observe its expression in transiently transfected HEK293T cells. The coding DNA sequence of GSH-R gene was amplified by SOE-PCR technology from porcine genomic DNA and cloned into pcDNA3.1(+) vector, then inserted myc-tag. The positive clone was confirmed by enzyme digestion and sequencing. The expression vector was transiently transfected into HEK293T cells. The expression of pGHS-R protein was identified by Western blotting. The results showed that the CDS of pGHS-R was successfully cloned; the construction of pcDNA3.1(+)-myc/pGHS-R was correct and its expression of pGHS-R protein was confirmed by Western blotting. The results showed that the eukaryotic expression vector pcDNA3.1(+)-myc/pGHS-R was successfully constructed and pGHS-R protein was successfully expressed. The study laid a foundation for further study on GHS-R gene function in vitro.

To establish a PCR method of Theirelia annulata （T. annulata）, a pair of primers was designed to specifically amplify a 393 bp high immunization fragment based on T. annulata surface protein （TaSP） gene conserved region，T. annulata, T. sinensis, T. sergenti, T. uilenbergi, T. luwenshuni, T. ovis, T. equi, B. caballi and B. bovis were tested by PCR with the primers. While T. annulata genome templates were amplified that different concentrations diluted in order to determine the sensitivity of the experiment. 150 bovine blood samples were detected using PCR and microscopic. The PCR results of specificity assay showed that only T. annulata genome template amplified specific fragment. The sensitivity result showed that the minimum dose of T. annulata that could be detected by PCR assay was 10^-10. 150 blood samples were detected by PCR assay and microscopic examination of Giemsa-stained blood semars,the results showed that PCR assay was sensitive and specific. It was suitable for the detection of T. annulata.

A loop-mediated isothermal amplification (LAMP) assay was developed for detection of Salmonlla in fecal samples of experimental monkeys. According to the specific sequences (fimY) of Salmonlla in GenBank, one set of primers was selected and the reaction condition was optimized. The results showed that the detection limit of LAMP method was 1.35×10¹ and 1.35×10³ CFU/mL in Salmonella pure culture and clinical samples,respectively,which was the same as routine PCR. The amplification could complete in one hour, and the result could be distinguished by naked eyes. There was non-specific amplification of other pathogens. These results suggested that this LAMP assay was a simple and specific method for rapid detection of Salmonlla in fecal samples.

To investigate the relationship between the expression level of soluble stem cell factor （SCF） and goat coat color. The expression level of soluble SCF in goat skins with black and white coat color was analyzed with QRT-PCR technique and statistics software. The relative expression level of soluble SCF in skin with black coat color was 0.27 times higher than that with white coat color normalization with GAPDH gene. It suggested that the expression level of SCF gene might not be related to coat color in goat.

The purpose of this experiment was to construct the oligonucleotide library used in the selection of aptamer and optimize the PCR conditions in the selection process. We constructed the oligonucleotide library and designed primers according to the immobilized sequence of amphi-oligonucleotide using Primer Premier 5.0. The proportion of upstream and downstream primers, quantity of template, anneal temperature and cycle numbers of the symmetric and asymmetric PCR were optimized and their repeatabilities were verified. After optimizing, we set up the best symmetric and asymmetric PCR conditions achieved the desirable dsDNA and ssDNA. The optimized PCR showed high reproducibility. We constructed the oligonucleotide library and optimized PCR condition successfully. This study provided a reference for selection of aptamer and laid a foundation for further study.

The primers were designed and synthesized according to the sequences of S gene of transmissible gastroenteritis virus （TGEV）, and then reaction requirements were optimized to develop a SYBR Green Ⅰ fluorescence quantitative RT-PCR assay. Meanwhile, 12 field samples were detected and the results were compared with that of routine RT-PCR. It was showed that the fluorescence quantitative RT-PCR assay could detect 43.07 copies/μL of plasmid DNA and its specificity and reproducibility were very good, while the sensitivity of the routine RT-PCR was 4.307×10³ copies/μL.The results of field test also showed that its sensitivity was higher than that of routine RT-PCR. The SYBR Green Ⅰ fluorescent quantitative RT-PCR assay for detecting S gene of TGEV was developed for the basis of early， rapid detection and quantitative analysis of the infection intensity of TGEV in this experiment.

Porcine reproductive and respiratory syndrome virus (PRRSV) were one of the worldwide causative agent for the disease of pigs and caused heavy losses every year.In this study, a TaqMan-LNA based Real-time RT-PCR assay and a reverse transcription loop-mediated isothermal amplification （RT-LAMP） assay were established.The detection limits were 10² and 10¹ copies/μL for Real-time RT-PCR and RT-LAMP assay， respectively. Two methods both showed good specificity.Futhermore both methods were analysed with clinical samples, and they exhibited high sensitivity and specificity.The Real-time RT-PCR and RT-LAMP assays would provide useful tools for the detection of PRRSV.

miRNA is a short RNA researched widely in recent years，the main function is to regulate the expression of individual growth, development, disease related gene, involved in a series of important processes of life，including early development, cell proliferation, cell apoptosis, lipid metabolism, cell differentiation and so on. In this paper, we summarized the main regulation of poultry miRNA on embryonic development,gonad development, muscle growth, fat metabolism and briefly discussed future research on poultry miRNA,in order to provide reference for further related research.

In order to compare the sensitivity to classical swine fever virus E2 protein targeting dendritic cell vaccine between rabbits and BALB/c mice, we firstly inoculated rabbits and BALB/c mice with glycosylated E2 protein and E2 protein (2 mg in rabbits and 20 μg in mice) according to the body weight percentage of rabbits and BALB/c mice. The IgG levels in the serum were determined by ELISA method in different time. The results showed that the BALB/c mice generated mild immune response only, and the rabbits had a strong and lasting immune protection. Secondly, the research on inoculation ammount in BALB/c mice were explored further. The results indicated that the immune effects were obviously better after inoculation with 100 or 50 μg than that of 20 μg inoculation amount. The IgG level in serum was high and stable after the BALB/c mice inoculated with 50 μg glycosylated E2 protein. However, the mice emerged immune suppression in initial stage after inoculation with 100 μg glycosylated E2 protein, and the IgG level increased to high level on 21 d. Surprisingly, the IgG level was low and only slightly fluctuated after the mice inoculated with 100 μg E2 protein. The above results proved that the sensitivity of rabbits was significantly higher than that of BALB/c mice to the glycosyated E2 protein and E2 protein. Furthermore, the immune effect of glycosylated E2 protein was better than that of E2 protein, and the best inoculation dose was 50 μg in BALB/c mice.

The Muscovy duck were used to clone the sequence of fatty acid synthase (FAS) gene by RT-PCR，the sequence was analyzed by related softwares and on-line tools, the tissue-specific expression and the effect of overfeeding on FAS gene mRNA level were performed by Real-time PCR. The results showed that the cloned sequence was 1122 bp and encoded 374 amino acids. The nucleotide homology of Muscovy duck FAS was high similar with other avain species from 92% to 99%. The tissue-specific expression result showed that the mRNA expression level of Muscovy duck FAS gene was the higher in liver and adipose tissues and was not detected or little in other 10 tissues. It revealed that Muscovy duck FAS gene was tissue specific.The mRNA expression of Muscovy duck FAS gene in liver was significantly increased after overfeeding （P<0.05）.The results indicated that FAS gene could play an important role in duck liver steatosis.

This trial was conducted to evaluate the effects of Ligustrum lucidum fruits extracts on productive performance and immune function of layer after laying peak period. A total of 180 47-week Hyline brown layers were randomly divided into 4 groups with 3 replicates per group and 15 birds per replicate in a single-factor completely randomized experimental design. The control group were fed a basal diet, the experimental groups Ⅰ, Ⅱ and Ⅲ were fed the basal diet supplemented with 0.05%, 0.10% and 0.15% Ligustrum lucidum fruits extracts, respectively. The results showed as follows， compared with the control group, the daily egg production and laying rate of layer in the three experimental groups were extremely significantly increased (P＜0.01) respectively, the feed/egg in experimental group Ⅲ was extremely significantly decreased by 10.9% (P＜0.01), but among 4 groups the average egg weight and rate of broken or soft egg did not significantly change (P＞0.05); and the content of testosterone in experimental groups Ⅱ and Ⅲ were extremely significantly decreased (P＜0.01), the contents of progesterone, estradiol, immunoglobulins (IgM, IgA) and interleukin-2 (IL-2) in three experimental groups were all extremely significantly increased respectively compared with the control group (P＜0.01). In conclusion, supplement with the Ligustrum lucidum fruits extracts in the basal diet could significantly increase the productive performance and immune function of layer after laying peak period.

A total of 224 5-week-old healthy Yellow-feathered broilers with similar boby weight（367.9±4.53）g were randomly allocated into 4 groups (Ⅰ，Ⅱ，Ⅲ and Ⅳ) with 4 replicates per group and 14 broilers in each replicate. Group Ⅰ was the control group and fed the basal diet；Groups Ⅱ，Ⅲ，Ⅳ were experimental groups which fed the basal diets supplemented with 3.75‰，5.00‰ and 6.25‰ metabolic organic acids （based short-chain keto acid and organic)，respectively.The results showed that compared with the control group，the average daily gain of group Ⅲ was significantly increased (P＜0.05)，the feed-gain ratio of group Ⅳ was significantly decreased (P＜0.05). There were no significant effects on breast meat rate，leg meat rate and abdominal fat ratio in each group (P＞0.05)，however，it could improve serum total protein and albumin levels，reduce alanine aminotransferase content of 8-week-old broilers.Therefore，it could be seen that the product of metabolic organic acids could effectively improve the average daily gain，lower feed-gain ratio of broilers，and promote the growth of Yellow-feathered broilers by regulating the level of serum proteins and enzymes，its appropriate level of adding in diet was 5.00‰.

This experiment was conducted to research the effects of diet added cellulose and active dry yeast on apparent digestibility，nitrogen and energy metabolism of Grassland Red cattle.Four healthy Grassland Red cattles with an average age of eight months and an average body weight of (170±10) kg was selected，and a 4×4 Latin square design of experiment was adopted. Guinea grass and mix beneficiated burden material were used as basic diet. The experiment was divided into 4 periods and each period included 37 days. Group Ⅰ was added basic diet didn’t add cellulose and active dry yeast， which was regarded as the control group,group Ⅱ was added cellulose for 190 mg/kg in basic diet,group Ⅲ was added active dry yeast for 200 mg/kg in basic diet,group Ⅳ was added active dry yeast for 200 mg/kg and cellulose for 190 mg/kg in basic diet. The results showed as follows：① Groups Ⅲ and Ⅳ had significant effect on dry matter digestibility （P <0.05）,and compared with group Ⅰ, dry matter digestibility from group Ⅲ was increased by 9.01%.② Group Ⅲ had significant effect on apparent digestibility of organic matter，nitrogen free extract，neutral detergent fiber，crude protein, acid detergent fiber，cellulose and hemicellulose in diet (P＜0.05)，and compared with group Ⅰ，the digestibility of cellulose and hemicellulose from group Ⅲ was separately increased by 14.65% and 18.81%.③ Group Ⅲ had significant effect on digestible energy and metabolic energy(P＜0.05)， and compared with group Ⅰ，metabolic energy from group Ⅳ was increased by 16.63%.④ Every indicators of cellulose and active dry yeast mixed added has no seen superposition effect.These results indicated that the active dry yeast could improve the utilization ratio of Grassland Red cattle diet and reduce energy losses，mixed addition had not appeared superposition effect.

Betaine and its hydrochlorides was played an important role in the methyl metabolism as a high efficient methyl donors，which could adjusted the osmotic stress，promoted both fat and amino acid metabolism，enhanced protein synthesis and improved meat quality，so it was widely used in feed industry and livestock production as a novel and safe nutritional additive.The objective of this paper was to review the recent reports about physiological function of betaine at the moment，and its application in ruminants and single-stomached animal production.

Mycotoxin was widely existed which done a serious threat to animal and human health，and it could be degraded by the mycotoxin degradation agents and reduced its toxicity which done damage to animals and humans，the degradation agents of mycotoxin were deeply studied at the moment.This article summarized from the dangers of mycotoxin，the classification degradation agents of mycotoxin，the degradation of mycotoxin applied in feed and provide relevant informations for the research of the mycotoxin.

The experiment was conducted to study the effects of compound probiotics on growth performance, apparent digestibility and serum biochemical traits of finishing sheep. One hundred and eighty finishing sheep with an average body weight of (20.45±0.77)kg were randomly divided into 3 groups with 3 replicates per group of 20 sheep per replicate.The control group was fed the basal diet, the groups Ⅰ and Ⅱ were fed the basal diet in which 0.1% and 0.2% compound probiotics were supplemented. The experiment lasted for 30 d. The results showed that the average daily gain of the experiment groups Ⅰ and Ⅱ were both higher than that of the control group, higher above 12.88％ （P<0.05） and 7.36％ （P>0.05） respectively, and the average daily feed intake in experiment groups was higher than that of the control group（P＞0.05).The number of F/G in experiment groups Ⅰ and Ⅱ were lower 6.50% and 6.00% than that of the control group（P<0.05）. The apparent digestibility of organic matter, crude protein and acid detergent fiber were significantly increased in the experiment groups (P＜0.05). The BUN in the serum of the experiment groups Ⅰ and Ⅱ were significantly lower than that of the control group(P＜0.05), the Glu were significantly higher than that of the control group(P＜0.05). But the effect on other blood indices was similar to that of the control group (P＞0.05). It concluded that feeding compound probiotics could improve the growth performance and apparent digestibility of nutrients, reduce the contents of BUN in serum, increase the contents of Glu in serum of finishing sheep, and better effect was obtained in the group of 0.1% compound probiotics.

To investigate and establish the methods of in vitro culturing and identification of endothelial progenitor cells (EPCs) of Bama Minipigs. The mononuclear cells were gained from Bama Minipigs bone marrow by density gradient centrifugation，inoculated （2×10⁷） in the culture plates，which were coated with HFN and induced differentiation to EPCs.The EPCs were observed by inverted microscope, compare the proliferation status of both derived EPCs by observing the growth curve，MTT method，and the express of EPCs-related antigen ac-LDL, UEA-1, AC133 and vWF by immunofluorescence staining.MTT results showed that the proliferation of the cells were obvious at 3 to 5 d.FACs test results showed that vWF adherent cells twice percentage of positive cells was 74.91%, AC133-positive cell percentage was 79.54%, while the secondary without adherent cells induced vWF and AC133-positive rate were 33.65% and 39.87%. The cells were identified by immunofluorescence expression of ac-LDL, UEA-1, AC133 and vWF were positive. Thus, the cells isolated and cultured from Bama Minipigs bone marrow were shown that strong ability of self-renewal proliferation and multipotency of differentiation which identified as the endothelial progenitor cells.

Cell apoptosis is an autonomic ordered programmed cell death in order to maintain homeostasis, which is controlled by multiple genes. Cell apoptosis plays a key role in the tissue differentiation, organ development and individual homeostasis in multicellular organisms. Cell apoptosis can promote the cells death which are infected or injuried,and remove them out of the body; clinically cell apoptosis and many diseases are directly related. The proteins of Bcl-2 family that involved in cell apoptosis are important regulatory molecules, can both suppress and promote cell apoptosis. So the members of Bcl-2 protein family and its structure feature, the relationship between Bcl-2 protein family and cell apoptosis, and its biological significance are mainly discussed.

The assay was aimed to investigate the immune effects of Radix astragalus fermented by Lactobacillus and Bacillus. Eighty-four mice were divided into six groups that were physiological saline group (negative control group), non-fermented Radix astragalus group (positive control group, 150 mg/mL), group Ⅰ(60 mg/mL), group Ⅱ (90 mg/mL), group Ⅲ (120 mg/mL), group Ⅳ (150 mg/mL). Mice were given medicine by gavage for 30 d and then they were killed to detect the weight, the immune organ (thymus and spleen) index and immune factor (IgG, IL-6, sIgA). The results indicated that the weight of experimental groups apparently rose; the immune organ index of group Ⅰ was extremely significantly higher than that of the control group (P＜0.01), but the immune factor （IgG/sIgA） of groups Ⅲ and Ⅳ extremely significantly exceeded the control group (P＜0.01). Finally, there were obviously promotional effects of Radix astragalus fermented by Lactobacillus and Bacillus on immune function of mice.

To explore the effects of chicken transfer factor liposome on mouse macrophage phagocytosis, digestion and absorption, 30 SPF female mice with almost the same size and weight were selected and divided into 3 groups randomly and each group with 10 mice. Each group was intraperitoneal injected with CTF, CTFL and normal saline respectively every other day and 5 times with 2 mL for each one, with the normal saline group as the control. After injecting 11 and 21 d, mouse macrophage phagocytosis rate was detected. At the same time, the duodenum, jejunum and ileum of each mouse were taken out and fixed in formalin liquid, for paraffin section, HE stain, then examined under a microscope and the length of intestinal villi were carefully measured. The results showed that macrophage phagocytosis rates of groups CTF and CTFL were extremely significantly different comparing with the control group (P＜0.01), while groups CTF and CTFL had no significant difference (P＞0.05). At the 11th and 21th day of the experiment, compared with the control group, the average length of duodenum of groups CTF and CTFL were significantly different (P＜0.05), the average length of duodenum of group CTFL was longer than that of group CTF, while the average length of jejunum and ileum of all groups had no significant difference (P＞0.05).The results indicated that both CTF and CTFL could increase the immune function of the mouse and CTFL was better for promoting digestion and absorption.

This assay was aimed to study the pharmacological effects of carboxymethyl glucan (CMG) injection，so as to provide experimental bases for clinical diseases use. The mice were used to study acute toxicity assay, the ability against Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli. In addition, lymphocyte transformation rates of dairy cows were determined by MTT method, and the clinical application of CMG injection was also carried out. The results indicated that the maximum tolerance dose of mice to CMG was above 240 mg per mouse, and in the 4 mg per mouse, the rates of protection against Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli were 60%, 62.5%, 87.5%. CMG could apparently increase the lymphocyte transformation rate of dairy cows. In clinical application, the mastitis incidence of experimental group decreased by 26.8% compared with the blank control. All the results above showed that CMG injection could be used as a potential new immunopotentiator for clinical application in animals.

Rumen epithelium is the major site for the rumen microorganism adhering and growing，nutrient absorption，transport and metabolism.In addition，it acts as a physical barrier between rumen environment and blood circulation system.Therefore，maintaining integrity of structure and function of rumen epithelium is very important for healthy and production performance of ruminant.This paper reviewed the basic structures and cautions of circulating Ussing chamber system，and evaluating methods on rumen epithelial mechanical barrier function in ruminant，which aims to provide references for maintaining rumen epithelial barrier function.

To establish a quality standard for Zhike Pingchuan granules, Atarian aster, Cortex mori radices, Radix stemonae, Loquat leaf and Houttuynia in the components were identified by TLC, and the content of Shionone was detected by HPLC.The results showed that the TLC identification was highly specific and the spot was clear; the HPLC separation was carried out on C₁₈ column, the mixture of acetonitrile-water was used as mobile phase. The detection wavelength was 200 nm, the calibration curve of Shionone showed a good linearity over the range of 10.2 to 510.0 μg/mL, the regression equation was A＝11481C＋36617(R²＝1,n=6), the average recovery was 99.89%, RSD=1.57%. The method established by this experiment was simple, accurate and focused with good reproducibility, so it could be used to control the quality of the preparation effectively.

The assay was aimed to study the pathology and toxicology changes of compound Agaricus blazei Murill polysaccharide oral liquid impacting on rats through continuous gavage.Divide rats into high dose group,middle dose group,low dose group and solvent control group.Give rats a continuous gavage for 9 weeks with 5.6,2.8,1.4 g/kg compound Agaricus blazei Murill polysaccharide oral liquid and same capacity of distilled water (NS).Observe and detect the clinical symptoms,hematology,blood biochemical and pathologic changes of viscera of rats.The results were that in the clinical symptoms and pathology,there were no conspicuous differences.In body weight,there were no significant differences among the groups (P＞0.05).Compared with control group,heart index,liver index,kidney index of high dose group, and heart index and liver index of middle dose group all had significant differences (P＜0.05).Compared with control group,WBC,LYM,MID,GRA and PLT of high dose group， and WBC,GRA and PLT of middle dose group all had significant differences (P＜0.05).Compared with control group, Cre and TG of high dose group, and TG of middle and low dose groups all had significant differences (P＜0.05).The results showed that the middle and low dose groups of compound Agaricus blazei Murill polysaccharide oral liquid had no obvious toxicity.

Polyunsaturated fatty acid is a materials with a specific biological function, which have played an important role in immune regulation, cell growth, gene regulation, lipid metabolism and cancer on organism.This article outlined the biological functions of polyunsaturated fatty acids and the research progress of its application on chicken production, including the regulation of production performance on chickens, eggs, meat quality and other aspects of the immune.

PCR-SSCP and DNA direct sequencing technology were used in this study to identify the polymorphisms of CDS 130—587 bp region of DRD1 gene in individuals of Cherry Valley duck and Xingyi duck, the association between SNPs and growth traits were also performed subsequently. The results showed that 2 alleles named A, B that resulted in 3 genotypes included AA, BB and AB respectively were confirmed in this study. AA genetype was predominant in two duck breeds with frequencies of 0.7416 and 0.5643, respectively, moreover, allele A was predominant compared to allele B for two duck breeds with frequencies of 0.8539 and 0.7204. The polymorphism information content (PIC) were 0.2184 and 0.3217 which belonged to low polymorphism and moderate polymorphism. Chi-square test demonstrated that the genotypes distribution of SNPs locus in the population of Cherry Valley duck was agreement with Hardy-Weinberg equilibrium (P>0.05) and deviated significantly from Hardy-Weinberg equilibrium (P<0.05) in the population of Xingyi duck. The results showed that two SNPs of c.189 T＞C and c.507 T＞A which belonged to synonymous mutation and missense mutation that resulted in the change from serine to arginine. Association analysis showed that the weight of 8 weeks and body length of 12 weeks of Cherry Valley duck for BB genotype individuals were significantly higher than AB genotype (P<0.05). Weight of 10 and 12 weeks and chest depth of 12 weeks of BB genotype individuals were significantly higher than AB and AA genotypes (P<0.05). Results disclosed that the SNPs locus might be an important marker of marker assisted breeding for growth trait of duck.

In order to research genetic diversity and phylogenetic for Jlo and Hongyuan and Ola the three groups of Tibetan sheep, Xinjiang fine-wool sheep and Aletai sheep, to sequence and analyze the complete mtDNA D-loop sequences of 89 inviduals from 5 groups sheep.The results displayed that the 89 inviduals from 5 groups sheep had 168 polymorphic loci, 82 were singleton variable sites,86 were parsimony informative sites, the average contents of A+T were 62.5%, significantly higher than the 37.5% of C+G. Identified 70 haplotype, nucleotide diversity(Pi) and haplotype diversity (Hd) are between 0.01040 to 0.02892 and 0.640 to 1.000, the average number of nucleotide differences (K) were 18.340, Tajima’s test found that the five groups were not significant. By building NJ phylogenetic tree to indicate that Tibetan sheep of Hongyuan subbreed was closely with Ola, then Jlo groups got together with Hongyuan and Ola a class,then, cluster with the Xinjiang fine-wool sheep and European mouflon and Urial. Argali and Argali Ovis ammon alone for a class.The 5 groups of sheep had three origins.

The study is to determine the DNA methylation status of porcine Lbx1 and Lbx2 genes. Bisulfite sequencing（BS）technique was used to analyze the specific methylation on promoter and exon 1 regions of Lbx1 and Lbx2 genes in longissimus dorsi muscle.The results showed that the differentially methylated region was found in a CpG island of Lbx1 gene exon 1, the differentially methylated region was outside of the CpG island of Lbx2 gene promoter region. These results suggested that the high DNA methylation in CpG island might be involved in the down regulation of Lbx1 gene in longissimus dorsi muscle.

Kiss-1 gene played important roles in regulation of sex gland axis in mammals, and also regulated other reproductive function. This paper summarized the research progress of Kiss-1 gene in male reproductive system, and the research trend in this field was also analyzed.

Tibet chicken from Linzhi were used to study their mitosis observation and karyotype by preparing chromosomes with direct bone marrow.Through a microscope the mitotic progression including the interphase, early protophase, protophase, metaphase, anaphase and telophase of Tibet chicken and the change of mitotic progression could be observed. By comparing various periods， the chromosome in early protophase stretched the longest and the bases exposed to the most.The early protophase mitotic progression of Tibet chicken was better suit for gene mapping. Meanwhile,the karyotype of Tibet chicken was studied. Experimental results showed that the chromosome number （2n） of Tibet chicken was 78 and there were three pairs of metacentric chromosomes,a pair of submetacentric chromosomes and two pairs of telocentric chromosomes in five pairs of chromosomes and a pair of sex chromosomes which had been analyzed. The results were different from previous, and causes of the result needed more researches.

The study was aimed to identify the virulent strain CCGGD201101 isolated from dead chicks of a chick farm in Jilin province and determine its pathogenicity. The suspected pathogens were identified by means of physiological and biochemical tests and 16S rDNA sequencing. The median lethal dose to Kunming mice was determined to verify bacterial virulence.The results showed that this strain was Acinetobacter baumannii. The half lethal dose was determined with Acinetobacter baumannii CCGGD201101 as target while Acinetobacter baumannii standard strain (ATCC 19606) as control. The pathogenicity of Acinetobacter baumannii strain CCGGD201101 was further confirmed.

To determine the antimicrobial resistance and the phylogenetic background of E.coli strains in Guangdong province, a total of 264 E.coli isolates were collected from diseased swine in 2011, which contained 131 strains from feces and 133 strains from liver. Antimicrobial susceptibility test was performed with agar dilution method and the phylogenetic background was determined by multiplex PCR. Statistical methods were used to analyze the relationship between antimicrobial resistance and the distribution of phylogenetic group among the E. coli strains isolated from feces and liver. The results showed that the multi-drug resistance (MDR) of the isolates was serious and there were 128 kinds of drug-resistant spectrum. The resistance rates of the isolates to ampicillin, sulfamethoxazole/trimethoprim, nalidixic acid and tetracycline were very high, and the isolates were sensitive to colistin, ceftazidime and amikacin. The majority of isolates belonged to phylogenetic groups A and B1 (91.66%). There were no significant differences in distribution of phylogenetic group between the E.coli strains isolated from feces and liver (P＞0.05), but the resistance rates to certain antimicrobials were significantly different in different phylogenetic groups (P＜0.05). The study was to investigate antimicrobial resistance and the distribution of phylogenetic groups of the E.coli isolates so as to provide a foundation for the use of antimicrobials in veterinary clinical medication.

Shiga toxin-producing Escherichia coli (STEC) are serious foodborne pathogens,which can cause severe human diseases such as hemorrhagic colitis,hemolytic uremic syndrome,etc,cattle,sheep and other ruminants are major hosts that cause lead to disease in humans. This paper summarizes the epidemiological situation of Shiga toxin-producing E. coli,and make a review on the research progress of virulence factors that locate on Shiga toxin-producing E. coli chromosome and virulence plasmid.

The Bacillus cereus spore had a strong resistance against the environment and physical and chemical factors. Appropriate conditions of temperature, moisture and nutrition could promote the spore germination and affect resistance of spore to conventional sterilization method. This research selected Bacillus cereus as the research object, made it spore form under the appropriate condition, and study the resistance of germinated spore under the conditions of high temperature、H₂O₂ and chlorine disinfectants. The results confirmed that appropriate conditions (37 ℃,humidity 40%, pH 7.4, 30 min) could effectively promote the Bacillus cereus spore germination. And the resistance of germinated spore decreased apparently under the conditions such as high temperature (80 ℃,2 h), H₂O₂ (1.5 mol/L,2 h) or chlorine disinfectants (available chlorine content was 800 mg/L,2 h). The relatively effective killing rates were 98.4%, 98.9% and 99.1%, respectively. And the effective kill rates were 88.3%, 88.7% and 88.9%, respectively. The results had important reference meanings to further research on sterilization conditions of Bacillus genus bacillus.

The assay was aimed to study the safety of recombinant porcine interferon alpha 1. Twenty healthy piglets were randomly divided into four groups (three experimental groups and one control group), three experimental groups were intramuscular injection of low, middle and high dose recombinant porcine interferon alpha 1. Each group continuously infused recombinant porcine interferon alpha 1 for 7 days. Animal vital signs, blood routine examination and tissue pathology indicators were observed. The appearance and behavior characteristics of each experimental group were normal; changes in body temperature and weight were also within the normal range; by HE staining assay,there were no organ lesions in important organs such as brain, lung, liver, heart and spleen; blood routine examination and biochemical indexes were also normal. The results showed that the recombinant porcine interferon alpha 1 had no significant side effects on piglets and was safe to use.

The purpose of the research was to make preparations for establishing the disease resistance evaluation system as well as laying the basis for disease-resistance breeding of the Jinghai Yellow chicken. Chicken were divided into infection and control groups, the infection group was orally inoculated with Eimeria tenella oocysts (2.5×10⁴ oocysts per bird). 10 resistance associated blood indexes were measured through the method of ELISA. The results showed，the difference of indexes 3rd day after inoculation was not significant （P＞0.05） which suggested that it was hard to detect any significant change within these indexes in this period of infection;3rd to 5th day after inoculation, the body weight gain was extremly significantly higher than that of control group （P＜0.01）;the concentration of IFN-γ 8th day after inoculation was significantly lower than that of the control group, the concentration of SOD 8th day after inoculation was significantly higher than that of the control group （P＜0.05）,and the concentrations of MDA,GSH-Px,CAT were extremely significantly higher than those of the control group （P＜0.01）；the concentrations of IL-2,IL-16,IL-17,NO and β-carotene had no significant difference with control group （P＞0.05）. 3rd to 5th day after inoculation，the body weight gain, the concentrations of the 4 antioxidant enzymes (CAT, GSH-Px, MDA, SOD) and IFN-γ could be taken as the candidate indexes for the disease resistance evaluation system of the Jinghai Yellow chicken.

The aim of the study was to investigate the biological characteristics and antibiotic resistance of E.coli isolated from bovine mastitis in Shihezi area in Xinjiang so as to raise therapeutic effect and decrease drug residue.The milk samples of bovine mastitis were collected from Shihezi and a total of 21 E.coli isolates were further identified by microbiological tested，and antibiotic resistance of 21 strains were also studied.The results showed that among the 21 strains，the resistant rates of 6 out of 21 antibacterial drugs exceeded 50%，the E.coli could resist 15 kinds of antibacterial drugs at most and 5 kinds of antibacterial drugs at least，and the E.coli strains which were resistant 6 or more antibacterial drugs occupied 76.19%.So we concluded that E.coli isolates had produced different level resistance to many kinds of antibacterial drugs and presented serious multiple antibiotic resistance.

The effects of 2 Bacillus subtilis strains，Q4 and Q10 inhibiting the growth of E. coli and Salmonella.Regarded E.coli and Salmonella as indicators，investigated by the co-incubation， pre-incubation and post-incubation method.The results revealed that 2 Bacillus subtilis strains showed different ability to inhibit the growth of E. coli and Salmonella after incubating with Q4 and Q10， respectively.Having inoculated Bacillus subtilis and then co-incubation pathogens inhibition were most obvious in means of the strongest inhibitory effect at 24 h，at the same time，pre-incubation Bacillus subtilis and then co-incubation pathogen the inhibition was the weakest of all，the pathogens and Bacillus subtilis co-incubation were the second. From the former consequences that 2 Bacillus subtilis strains of Q4 and Q10 had inhibitory effect on the pathogen，but there were differences in inhibition，and related to the duration of Bacillus subtilis and pathogen inoculation.

The infection model of avian reovirus in chicken was established, which layed the ground for vaccine efficacy test.The 49-day-old chickens were inoculated with 10^2.0, 10^3.0, 10^3.5, 10^4.5and 10^5.5 ELD₅₀/0.2 mL of two strains of ARV, T98 and AV2311, respectively, in foot pad inoculation method. Clinical signs were observed ten days and recorded daily. 6 days after inoculation, serum samples were taken. 10 days after inoculation, chickens were sacrificed for necropsy. Serum antibody was detected by ELISA. The results indicated that the morbidity of 10^5.5and 10^4.5 ELD₅₀/0.2 mL were 100%, and the foot pads of chicken were swelled severely, which were dark red or purple. Symptoms of AV2311 set was a bit lighter than that of T98 strain. The morbidity of 10^3.5 ELD₅₀/0.2 mL was 90% and the morbidity of 10^3.0and 10^2.0 ELD₅₀/0.2 mL were 80%. The ELISA result indicated that only the serum efficacy of 10^4.5and 10^5.5 ELD₅₀/0.2 mL set of two strains of ARV were positive.The experiment proved that the virulence of ARV T98 strain was strong, and had good immunogenicity. The best inoculated dose of ARV T98 strain was 10^4.0 ELD₅₀/0.2 mL, which provided the important basis for researching the quality standard of chicken viral arthrilis vaccine.

The assay was to study the sensitivity of the pathogenic Escherichia coli from ostrich to ciprofloxacin combined with baicalin. First, ciprofloxacin and baicalin against Escherichia coli from ostrich were detected by double dilution method, the resistant Escherichia coli from ostrich were T1 to T4, T7,T8,T10,T12,T21,T25. Then Escherichia coli from ostrich were induced broth dilution checker-board method, when ciprofloxacin and baicalin were used alone, the MIC were 64, 512 or 1024 μg/mL,respectively, when the use of ciprofloxacin and baicalin were combined, the fractional inhibitory concentrations (FIC) of Escherichia coli from ostrich (T1，T2，T3，T7，T8，T10，T12 and T21) were all 0.375, the FICs of Escherichia coli from ostrich (T4，T25) were both 0.25, all were below 0.5. The results suggested that the combination of ciprofloxacin and baicalin demonstrated synergistic effect on the Escherichia coli from ostrich.

In order to fulfill more complicated and precise technological requirements in designing immunity and disinfection program for a scale-size pig farm，based on the Shanhei pig farm breeding data network platform，combining with its immunization and disinfection plan for breeding and used-to-meat pigs，this study pointed out data management standards of pigs immunization and disinfection plan，and developed a intelligent awaking and amount statistics analysis module system on pigs immunization and disinfection action. The system was based on .Net 2005 program language and SQL Server 2005 network database. This system showed that the system realized dynamic immune action reminding in different type pig herds for different vaccination types，as well as a awaking of disinfection action in different pig sheds in a future duration，and improved efficiency and comprehensive statistical analysis ability in planning immunization and disinfection events and was helpful to human arrangement and drugs rationing.In addition， the system can be transferred for application in other pig’s farm，by means of the changes of immunization program and disinfection plan.Furthermore，it is a major precondition to guarantee system to analyze and compute immunization and disinfection action plan accurately and timely that both timely and targeted adjustment in immunization and disinfection plan and dynamically complete pig’s individual information.

This study was planned to investigate the distribution and antibiotic resistance of pathogens involve in cow endometritis，which will be helpful to guide clinical rational drug use.All 47 cow endometritis samples collected from three large-scale farms located in Lanzhou were used to isolate and identify pathogens. Besides，K-B paper disk method was performed to measure the antibiotic resistance of the four isolates. The results demonstrated that Arcanobacterium pyogenes (37.1%)，Escherichia coli (24.2%)，Enterococcus faecium (22.6%) and Streptococcus agalactiae (16.1%) were the predominent agent causing cow endometritis in Lanzhou.In addition，the four pathogens were resistant to penicillin，sulfamethoxazole compound，ofoxacin and bacitracin，and the drug resistance rates reached 60% to 100%. However，These pathogens were sensitive to ceftazidime，cephalothin V and florfenicol，and the antibiotic sensitive rates were 100%.

β-lactam antibiotics has been applicated for prevention and control of animal infection in animal husbandry.The excessive level of such antibiotics caused by its abuse and nonobservance with the withdrawal period，human health has been widely threatened by the antibiotic residual in animal-derived food，so it needed rigidly control the antibiotics residues in animal-derived food.In this paper the current detection methods of β-lactam antibiotic residues have been introduced，including microbial detection，immunological detection，physical and chemical testing，the advantages and disadvantages of each method has been analyzed. The purpose of this paper will provide a valuable reference for the improvement of the antibiotic residue detection technology in animal-derived food.

The pregnancy rates at 20 and 50 days were tested by sampling 91 hybridization sows more than 16 days using the intuitive, fast, accurate and non-invasive portable veterinary B ultrasound instrument in the pig breeding farm had 300 sows. The results showed that the nonpregnant diagnostic accuracy rate was 100% at 18 to 20 days, and the pregnant diagnostic accuracy rate was 100% at 40 to 50 days. According the diagnostic results, the pregnancy, disease, estrus, nonpregnancy and non-estrused sows were took the appropriate measures such as nursing, treatment, supplementary breeding and sensuality, the non-productive days of the individuals and groups of sows were improved by above measures. The study built a intuitive, fast, accurate sows early pregnancy diagnosis technology, which improved the fertility of the individuals and groups of sows increased the economic benefit of pig farm.

Excessive amount of antibiotic residues in milk，would seriously endanger the health of consumers.Currently，the maturity and development of high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) technology which has became a powerful tool to solve above problem. This technology combined separation，qualitative analysis and quantitative analysis，which has also been fast analysis, strong selectivity，high sensitivity and good repeatability，and could detect multiple medicines in one sample simultaneously.This paper summarized the recent research progress on determination of antibiotic residues in milk by HPLC-MS/MS technology，and elaborated the pretreatment method，chromatography and mass spectrometry conditions and determination results of HPLC-MS/MS technology.

To investigate the effects of NaHS, an exogenous hydrogen sulfide donor, on skeletal muscular fatigue. Toad gastrocnemius was stimulated by pulse electric curent. The results showed that NaHS at concentrations of 5×10^-4 mol/L (in vitro, P < 0.01), 1×10^-3 mol/L (in vitro, P < 0.01), 2×10^-3 mol/L (in vitro, P < 0.01) and 5.6 mg/(kg·BW) (in vivo, P < 0.01) could prolong the time of contraction amplitude decreasing to 90%, 50% and 10% of the maximum contraction amplitude. The study showed that exogenous hydrogen sulfide had effect on antifatigue.

A HPLC method was developed with immunoaffinity chromatography columns（IAC）as purify process to detect the residual of avermectin，ivermectin，doramectin and eprinomectin in bovine muscle tissue.Samples were extracted with methanol,purified by IAC and followed by derivatization with acetic anhydride and 1-methylimidazole in acetonitrile.The optimum time required for the completion of derivation reaction of the 4 drugs was 100 min at 96 ℃.Recoveries ranged from 77.3 % to 119.5% with CV from 1.5% to 18.9% when these drugs were spiked in bovine muscle at levels of 5，10，50 and 100 ng/g.The limit of detection of the method was 0.8 ng/g in bovine muscle tissue.This method could be used to detect the residual of avermectins in bovine muscle tissue.