One pair of specific primers was designed and synthesized according to the sequence of Aleutian mink disease virus (AMDV) subbitted in the GenBank database, and then reaction requirements were optimized to develop a SYBR Green Ⅰ fluorescence quantitative PCR assay. The results showed that the fluorescence quantitative PCR assay could detect 10 copies/μL of plasmid DNA.Its specificity and reproducibility were very good when 10² copies/μL.Meanwhile, 8 field samples were detected with the methods and the positive rate was 75%. This study would lay the foundation for differentiation diagnosis and purification of the mink population.

The experiment was conducted to clone 5'-flanking region of bovine UCP3 gene to study regulatory mechanism of the specific transcriptional regulation region. The 5'-flanking region of bovine UCP3 gene was amplified with PCR, and then the 1080 bp 5'-flanking promoter sequence was obtained by product purification, ligation, transformation, and sequence comparison. The obtained cloning vector of 5'-flanking region of bovine UCP3 gene was analyzed by the promoter software in this study. The results indicated that there were 6 transcription start sites and 33 potential transcription factor binding promoter region of the gene, the highly conserved region was in -196～+100 bp of the upstream of transcription start sites, which might be conclude that the -200～+1 bp of the upstream of transcription start sites was the core. Comparing bovine with Panthera tigris, Felis catus, Bubalus bubalis, Pantholops hodgsonii, Capra hircus, Ovis aries,the homology of 5'-flanking promoter sequence were 82%, 82%, 98%, 95%, 96% and 98%, respectively. The 1080 bp 5'-flanking region of the bovine UCP3 gene was cloned successfully and the core promoter region was preliminary predicted in the gene.

The study was aimed at cloning, expressing the gene of outer membrance protein 2b (Omp2b) and analyzing the bioinformatics of Omp2b, according to the nucleotide sequence of Omp2b gene of Brucella melitensis M5-90, a pair of primers was designed, and Omp2b gene fragment was amplified from Brucella genome by PCR.The fragment was identified and cloned into prokaryotic expression vector pMD20-T. After sequencing, Omp2b fragment was ligated into pET28a to construct the recombinant expression plasmid pET28a-Omp2b, and then transformed into the E.coli BL21 (DE3) strains. IPTG was used to induce expression of Omp2b protein,and the protein was detected by SDS-PAGE and Western blotting. DNAMAN and BioEdit softwares were used to analyze the sequence of amino acids encoded by Omp2b gene. The results showed that Omp2b gene was cloned successfully; the ORF was 1041 bp, which encoded 347 amino acids; the expression vector pET28a-Omp2b was constructed; the molecular weight of the expression protein was about 38 ku; Omp2b protein secondary structure of alpha helix, extended strand, β turn and random coil were accounted for 20.17%, 26.22%, 5.76% and 47.84%, respectively.

This experiment was conducted to clone and express the nucleoprotein (N) gene of infectious bronchitis virus (IBV). A pair of specific primers was designed and synthesized according to the published sequence of IBV ZZ2004. RNA of IBV ZZ2004 strain was extracted and the fragment of about 1.23 kb was amplified by RT-PCR, which was correct fragment.The fragment was inserted into the cloning vector pGEM-T to construct pGEM-T-N, and nucleotide sequence was determined by dideoxy-mediated chain termination method. The pGEM-T-N was excised by EcoR Ⅰ and Xho Ⅰ, and inserted into the expression plasmid pGEX-6P-1 to obtain the recombinant expression vector pGEX-6P-1-N, which was used to transform into E.coli BL21 (DE3) competent cells. Then the recombinant pGEX-6P-1-N was induced by IPTG, SDS-PAGE and Western blotting results confirmed that the gene had been expressed in recombinant pGEX-6P-1-N, and the protein was soluble. The expression product could specially react to the anti-sera of IBV which showed that N protein had some biological activity. This study had laid a solid foundation for production of diagnostic reagents for testing IB and new type IB gene engineering vaccines.

In order to prepare the monoclonal antibodies against duck retinoic acid inducible-gene Ⅰ(RIG-Ⅰ) full-length protein, the N-terminal a (1 to 900 bp)and b (751 to 1650 bp) segments of RIG-Ⅰ gene were amplified from cDNA of duck spleen and then cloned into the prokaryotic expression vector pET-30a. The recombinant plasmid was transformed into BL21 cells and expressed by IPTG inducing,then immuned BALB/c mice by the protein, and fused the spleen lymphocytes with SP2/0 myeloma cell, detected by indirect ELISA. 17 strains hybridoma cell lines which had good reactivity with duck RIG-Ⅰ protein were screened; the titer of monoclonal antibody supernatant was 1:512; 2 strains monoclonal antibody 10H7C3 and 2A10A2 had good reactivity with duck RIG-Ⅰfull-length protein detected by indirect immunofluorescence (IFA) and Western blotting. The specific murine anti-duck RIG-Ⅰ monoclonal antibodies that prepared in the study laid a foundation for further research of RIG-Ⅰ gene function.

According to the multiple alignments identified major histocompatibility complex Ⅰ (MHC Ⅰ) gene conserved sequence registered in GenBank from the family ducks (Anatidae) anser waterfowl, a pairs of specific primers for the fragments of MHCⅠgene of goose F1 from fast-growth lines were designed and synthesized by Primer Premier 5.0. Using the genome DNA of goose F1 from fast-growth lines, the target gene fragment was obtained by PCR. To conduct sequencing of the fragments of MHCⅠgene of goose F1 from fast-growth lines and make sequence alignment and analysis of protein structure and function by bioinformatics, and research the characteristics of MHCⅠgene of goose F1 and the physicochemical properties of the protein. Bioinformatics was analyzed the nucleic acid data, deduced amino acid sequence and phylogenetic trees. The result of sequence analysis showed that the fragments of MHCⅠgene of goose F1 from fast-growth lines was 1036 bp in length, which coded 96 amino acids polyprotein. The homology were 93% and 83% with MHC Ⅰ gene and coding sequence of Wulong goose in NCBI respectively. There were 72 different bases sequence and 16 amino acids change. There also was higher homology with other poultry, and existed genetic relationship of Siji goose > chickens > ducks.The homology segment sequences corresponding to the fragments of MHCⅠ gene of goose F1 coded 96 amino acids protein, which molecular weight, PI, positively or negatively charged amino acid, estimated half-life, instability index, aliphatic index and average hydrophobicity were 11.342 ku, 5.32, 14, 17, 2.8 h, 34.92, 42.81, -1.066, respectively, and appeared 9 B cell epitopes, but contained no signal peptide. These results indicated that the protein for hydrophilic non-secreted proteins, had the high immunogenicity. In addition, The protein structure study indicated that alpha-helix, beta-sheet, beta-turn and random coil were 31.25%,16.67%, 14.58% and 37.50%, respectively. There existed amino terminal domain and carboxyl terminal domain in the tertiary structure. Therefore, MHC gene had significant difference between species and populations of individuals by the pathogen pressure in environment, and there were the interaction between polymorphism of MHC molecules and the diversity of antigenic peptide. MHC determined the differences of individual susceptibility to disease, and could be treated as a candidate gene for disease resistance.

In order to research on the production process of competitive ELISA test kit for peste des petits ruminants virus (PPRV), in this test, nucleoprotein (N) protein expressed in insect cell and monoclonal antibody of PPRV were used to establish a high specificity, sensitivity and strong competitive ELISA method for detection of PPRV, and the competitive ELISA procedures were optimized. 977 sheep and cattle serum samples collected from Xizang and Inner Mongolia were assayed with this established competitive ELISA and rC-ELISA antibody kit for PPRV of the OIE at the same time. The detecting results displayed that specificity, sensitivity and accuracy were 99.89%,93.82% and 99.38%. The competitive ELISA method of PPRV established in this assay would provide the basis for research on the production process of ELISA test kit as well as a reference for prevention scientifically.

The objective of this study was to establish a rapid detection method of loop-mediated isothermal amplification assay (LAMP) for equine herpesvirus-1 (EHV-1) and evaluate the reliability of the method. Designing several pairs of LAMP primers targeting to the conserved sequence of gBgene of EHV-1, using the LAMP Real Time Turbidimeter LA-320 meter to monitor the reaction so as to screen the primers and optimize the conditions, we established a LAMP method which could amplify specific DNA of EHV-1, then added SYBR Green Ⅰ to judge the result by eye. Specific DNA of EHV-1 was amplified efficient under 65 ℃ in 50 min. There was no cross reaction to other similar viruses such as EHV-4, and the reaction system also had very high sensitivity, 10^-4 dilution multiple target could be detected, the established LAMP was 10 times higher than ordinary PCR. After the reaction, add SYBR Green Ⅰ to observe results, the results were consistent with LAMP Real Time Turbidimeter LA-320 meter. Compared LAMP method result with PCR method result of four clinical samples, coincidence rate was 100%. The LAMP method in this test was rapid, specific, sensitive, easy operation and low equipment requirement, and had application prospects.

Proteins are the chief performers for physiological functions in a cell, and often function as protein complexes or interact with nucleic acids. A proteome represents all the proteins expressed by a cell, a tissue, or an organism at a defined time point under certain circumstances. Therefore, proteomes will undergo dynamic changes with alterations in time and space. The aim of proteomics is to globally study structures, functions and expression modes of proteome as well as interactions among proteins in a proteome. Based on the contents of study, proteomics can be classified as expression proteomics, structural proteomics and functional proteomics,respectively. The study of proteomes will help understand structures of proteins, functions of cells, nature and active rules of life, and will provide scientific basis for diagnosis and treatment of diseases as well as development of novel drugs. The techniques commonly used in proteomic studies include two-dimentional polyacrylamide gel electrophoresis, mass spectrometry, yeast two-hybrid system and protein chips. This article will provide a brief overview of proteomics and techniques frequently utilized for proteomic studies.

To study the antigenic relationship among the original canine parvovirus type (CPV) 2 (CPV-2) and the variants CPV-2a,the strain of 10⁵ TCID₅₀/100 μL from infected Tibetan mastiff which turns out the gene was CPV-2a and CPV-2 strain from vaccine was picked out to be generally amplified in the F81 cells. After analyzing the strains with HA and protein concentration with PEG6000, the viral suspension was emulsified with the adjuvant to immune two groups of each three New Zealand rabbits in every two weeks and four times. Serum antibody titer was detected in each preimmune. After four times immunity, serum of each group was collected to make separated gene polyclonal antibody. Then the antibody titer was tested by SN and HI with CPV-2 and CPV-2a virus and protection data was analyzed by SPSS 17.0. And two polyclonal antibodies were separated treated 2 dogs which was inoculation CPV-2a and infected. Blood samples were collected every day during treatment,then WBC was analyzed by SPSS 17.0. The results showed that the CPV-2a polyclonal antibody was more significant to CPV-2a virus than the other one by HI and SN (P<0.01).But counteract CPV-2 strain was not significant (P>0.05). CPV-2a polyclonal antibody was given more protection against CPV-2a and WBC was faster recovery to normal level than the other in experimental treatment (P<0.05).It showed that CPV-2a polyclonal antibody have higher homologous protection than the antibody induced by CPV-2 vaccine in Chinese canine parvovirus control.

The fusion VP3 gene sequence of goose parvovirus (GPV) was amplified by PCR using a pair of specific primers from GPV FY89 strain.The PCR product was cloned into pFastBac/HBM-TOPO vector and transformed into DH10Bac cells.Recombinant Bacmid-VP3 was transfected into Sf9 insect cells，and a fusion protein about 61 ku was expressed and identified by SDS-PAGE analysis.The antigenicity of the recombinant VP3 protein was confirmed by Western blotting.

A pair of specific primers was designed to amplify the goose parvovirus (GPV) and Muscovy duck parvovirus (MDPV) and establish a polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method for rapid detection of the infection status of waterfowl.An EcoRⅠ restriction enzyme was used which could digest MDPV PCR fragments into two different parts, with gene-lengthes of 530 and 280 bp, while GPV PCR fragments couldn't be digested by EcoRⅠ restriction enzyme. We established a fast differential diagnosis for GPV and MDPV using only one pair of primers and a restriction enzyme.

The whole proteins of 6 Mycoplasma bovis (M.bovis) isolates (W70 isolate,1738 isolate,Q3 isolate,Q1 isolate,JX02 isolate and 677 isolate),which isolated from different parts of the country was extracted by lysate,were analysed by SDS-PAGE and Western blotting with homemade antiserum.The results showed that there were differences on polypeptide quantity and definition of the whole proteins of 6 isolates:W70 isolate,1738 isolate,Q3 isolate and Q1 isolate were better than JX02 isolate and 677 isolate,which molecular weight were 23.2 to 130.8 ku.6 isolates were appeared 55 and 43 ku band when analysed by Western blotting.In conclusion,there were differences on polypeptide composition of whole proteins of the different M.bovis isolates,and 55 and 43 ku were the main immunogenicity protein of M.bovis.These results provided the theoretical basis for serology diagnosis,molecular diagnosis and the vaccine development of the M.bovis disease.

To investigate the prevalence and molecular characterization of class Ⅰ integron in Escherichia coli isolated from beef cattle, and analyze the relationship between integron and antimicrobial resistance, susceptibilities to 11 antimicrobials were conducted on 92 isolates, the presence and characterization of class Ⅰ integrons and inserted gene cassettes were performed using PCR combined with sequencing analysis. The results showed that 29 isolates had been detected positive for class Ⅰ integron integrase gene (intⅠ1) among 92 isolates, and aadA1 and dfrA17＋aadA5 were the most prevalent gene cassette arrays detected. The resistance rates of 92 isolates to ampicillin, streptomycin, sulfisoxazole and tetracycline were all more than 45.0%. As revealed by analyzing the association between resistance phenotypes and class Ⅰ integron, isolates that contained the class Ⅰ integron were significantly highly resistant to ampicillin, chloramphenicol, streptomycin, sulfisoxazole and sulfamethoxazole-trimethoprim (P<0.05), but not to quinolones and amoxicillin/clavulanic acid. The conclusion was that E.coli isolated from beef cattle were seriously resistant to antimicrobials,and integron/cassettes widely existed. The presence of integrons and the association of antimicrobial resistance determinants with transferable elements might play a crucial role in the dissemination of antibiotic resistance among E.coli. Data reported here clearly emphasized the need for a stricter application of antimicrobials restriction policies in feedlot setting.

In view of the harm of bluetongue (BT) and the importance of RNA quality control,pTrcMS vector was constructed by which p-MS2 plasmid was introduced 6His-tags between codons 15 and 16 of MS2 bacteriophage coat protein (CP), using a site-directed mutagenesis method.pMD19-T-BTV plasmid was digested with KpnⅠ/HindⅢ and ligated into pTrcMS to create the recombinant plasmid pTrcMS-BTV.pTrcMS-BTV in E.coli was induced with 0.5 mmol/L IPTG,then the Armord-RNA(AR) particles were captured with MagneHis^TMNi-Particles from bacterial lysates,and verified by Real-time RT-PCR method.The PCR results indicated that the His-tagged AR particles were highly pure without genome DNA.A TEM photograph showed that the AR particles had the shape of a round particle that was 26 nm in diameter.The rarefied AR particles were able to withstand RNase A and extremely stable.The limit of detection was determined to be 2.5×10² copies/mL by Real-time PCR.The His-tagged AR particles would provide quality-control for BT molecular detection.

The nutritional ingredients,including crude protein (CP),ether extract (EE),crude fiber (CF),crude ash,nitrogen free extract (NFE),total starch,amylopectine,amylase,amino acid (AA),non-starch polosaccharides (NSP) and minerals content of three kinds of cassava products (CPS) and corn control,and in vitro dry matter (DM) and starch digestibility,and starch profiles were studied.The results showed that CP and AA of the CPS were lower than that of corn,and the quality of its CP was poor.Cassava pellets (CE) had higher EE,crude ash and CF (P<0.05).Cassava residue (CR) was lower at NFE than that of cassava chips (CC) (P<0.05),but the CF and crude ash of it were higher than those of CC and corn (P<0.05).The total starch content was similar among CC,CE and corn,and the amylose/amylopectin of CPS was lower than that of corn (P<0.05).The resistant starch of CR was higher than that of others (P<0.05).The starch granule shape of corn was irregular polyhedron and cassava starch granule shape was a ball.The Ca, K, Fe and Mn of CPS was higher than those of corn,and reverse case for the P and Zn (P<0.05).Corn was lower at SNSP than that of CPS,and SNSP of CR was higher than that of CC and CE (P<0.05).Insoluble NSP of CR was higher than that of CC and CE(P<0.05),and insoluble galactose was higher than that of others (P<0.05).DM and starch digestibility of CE was higher than that of others (P<0.05),and DM and starch digestibility of CR was lower than that of CC and corn (P<0.05).The evaluation results have certain guiding significance for the future application of cassava products in feed industry.

Methods of in vitro culture technique and orthogonal experimental design were using in this experiment to identify the effects of branched chain amino acids (BCAAs,valine,leucine and isoleucine) on rumen microbial fermentation in vitro based on different concentrate to forage ratio (CFR) of 20:80,40:60 and 60:40,and the best combination of BCAAs on each CFR was gained after analysis of the combination effects of BCAAs.The results showed that the final pH,NDF degradability (NDFD) and acetate/propionate were not affected by BCAAs addition (P>0.05).However,DM degradability (DMD) was significantly different in various combinations.When the CFR was 20:80,the three amino acids showed a combination significant effect on the NH₃-N,VFA and TVFA (P<0.05).Along with the increase of valine levels,the concentrations of isobutyric acid and valeric acid were significantly increased (P<0.01),isovaleric acid was affected by isoleucine and leucine (P<0.05).When the CFR was 40:60,the concentration of isobutyric acid increased with the increase of valine supplementation level (P<0.01).Isovaleric acid was affected by three kinds of BCAAs (P<0.05).When the CFR was 60:40,BCAAs mainly affected the concentrations of isobutyric acid, valeric acid and isovaleric acid.Isobutyric acid was mainly affected by valine (P<0.01),isovaleric acid was affected by leucine and isoleucine (P<0.01),and valeric acid was affected by valine and isoleucine (P<0.05).Based on the concentrations of isobutyric acid,valeric acid and isovaleric acid,the three BCAAs showed obvious combination effects (P<0.05).In conclusion,taking DMD as the index,when the CFR was 20:80,the best supplementation levels of valine,leucine and isoleucine were 0.67, 2.00 and 1.33 mmol/L,respectively, when the CFR was 40:60 or 60:40,it's not necessarily to add these BCAAs.

The present study was conducted to determine the effect of different housing systems on meat flavor compound of Beijing-You chicken.A total of three hundred 49-day-old healthy Beijing-You chicken were randomly assigned to two groups (housing systems of cage-rearing and floor-rearing with outdoor access).Each group had five replicates with thirty broilers per replicate.The experiment ended at 91 days of age.The results showed that compared with the cage-rearing group,meat fatty acid of the chicken in floor-rearing with outdoor access including C16:0,C16:1,C18:1n9c,C18:2n9c,C22:0,SFA and UFA contents were significantly improved (P<0.05),C15:1,C18:0,C18:2n9t and C20:4n6 contents had a tendency to increase,but no significant difference was found between this two groups (P>0.05).The IMP content of the chicken in floor-rearing with outdoor access was significantly higher than that in the cage-rearing group (P<0.05).Volatile flavor compounds in both housing systems present different percentage rate.The percentage of ketones and alcohols of the chicken in floor-rearing with outdoor access was higher,relative to the cage-rearing group,the aldehydes content was reduced.This was the reason of odor rendering differences.The results suggested that different housing system affected the composition and relative contents of flavor compounds,the chicken in floor-rearing with outdoor access could improve meat quality and flavor of Beijing-You chicken.

The objective of the present study was to investigate the effects of dietary cation anion balance on blood biochemical parameters of heat stressed lactation dairy cows. 75 middle lactation (104.8 DIM±12 DIM) China Holstein cows were used in a randomized complete block design into five groups with similar parity, milk yield and weight, the groups Ⅰ, Ⅱ, Ⅲ, Ⅳ and control group were adjusted the DCAB value by providing with different level cation-anion salt, which were 195.76, 186.68, 177.77, 166.94 and 239.12 mEq/kg DM. The experiment was conducted for 67 days, including a 7 day adaptation period and a 60 day treatment period. The results showed that: compared with the groups Ⅲ, Ⅳ and control, the P_O2 of the groups Ⅰ, Ⅱ have increased that were significantly different (P<0.05);the effect of DCAB on dairy cows P_CO2,pH,HCO₃^-,TCO₂,SAT_O2 in blood was not significant difference (P>0.05). Compared with the groups Ⅲ, Ⅳ and control group, the groupsⅠ, Ⅱcreatine phosphokinase (CK) in serum have decreased, the effect of DCAB on creatine phosphokinase (CK)in serum was significant difference (P<0.05); the glutamic-pyruvic transaminase (GPT) in serum have increased, which increased 13.98%, 15.99%, 10.77% and 4.55%, 6.40%, 1.61%,respectively, but there were not significant difference (P>0.05). The content of T4, T3 and COR in serum were not significantly affected by change the DCAB value (P>0.05),but effect of DCBA on the content of urea nitrogen in serum was significant difference (P<0.05), the concentrations of urea nitrogen in serum have an increasing trend,with increasing DCAB values, concentrations of urea nitrogen in group Ⅰ was the highest. Therefore, comprehensive index above, under the condition of heat stress, middle lactation cows feeding DCAB value of 186.68 to 195.76 mEq/kg DM diet is more suitable to improve its health.

This experiment was conducted to study the effect of pine needle powder to replace part of green-hay powder on production performance and serum biochemical indices of Jiuyishan female rabbits.Twenty four and healthy female rabbits with similar weight were randomly divided into three treatments and 8 rabbits in each treatment.Rabbits in each group were fed basal diet supplemented with pine needle powder which part of 0 (control group),7.5% (group Ⅰ),15% (group Ⅱ) green-hay powder after inseminated 12 days,respectively.The adjustment period lasted for 4 days,and the experimental period lasted for 24 days.The results showed that compared with the control group,the AFI in the groups Ⅰ and Ⅱ was significantly decreased (P<0.05),the ADG in the groups Ⅰ and Ⅱ was significantly increased (P<0.05).With the level of pine needle powder increased,the A/G in the groups Ⅰ and Ⅱ had a decreased tendency,the rabbits offspring litter weight had a increased tendency,but there were no significant differences among all groups (P>0.05).The serum content of AST in the group Ⅱ was extremely significantly higher than that in the control group (P<0.01),but there were no significant differences in the serum contents of ALT,LDH,ALP and BUN among all groups (P>0.05).In conclusion,supplementation of pine needle powder in diet to replace part of green-hay powder had different effects on production performance and serum biochemical indices of Jiuyishan female rabbits,and replaced 7.5% green-hay powder had better effect.

The roots and rhizomes of Glycyrrhiza uralensis are important Chinese traditional medicine prescription material,licorice stems and leaves could be used as medicine and forage fodder.This paper reviewed the physicochemical properties of Glycyrrhiza uralensis,and the application and the advance of the roots and rhizomeson,the stem and leaf,the residue and the by-product via extract of the roots and rhizomeson of Glycyrrhiza uralensis in animal production.

In order to study the effect of feeding patterns (extensive feeding pattern (EP),complete intensive feeding pattern (CP) and incomplete intensive feeding pattern (IP)) on serum biochemical indices and hormone content in dairy cows.30 cows with similar body weight,parity and lactation for each pattern were selected to conduct the research,blood samples were collected to detect biochemical indices and related metabolic hormones,so as to analyze theirs relationship with different feeding patterns.The results showed that the serum contents of GLU,ALB,NEFA,BHB,ALT,ALP,LPS and HIS of CP and IP patterns were higher than that of EP pattern,but the serum contents of IgG,IgA and TA of CP and IP patterns were significantly lower than that of EP pattern (P<0.05).The serum contents of INS,INS/GC,T3 and GH of CP and IP patterns were higher than that of EP pattern,but the serum content of GC of CP and IP patterns was lower than that of EP pattern.In conclusion,feeding patterns had different degree effect on serum biochemical indexes and related hormones secretion of dairy cows,and CP pattern was better than IP and EP patterns.

The experiment was conducted to study the effect of castration on weight,body measurement index and serum biochemical indices of young Xinjiang brown cattle in different periods.62 Xinjiang brown cattle (21-months-old) were chosen and divided into the experimental group (castrated) and control group (non castrated).Experimental animals were measured weight and body measurement index and detected blood samples in 22,23,24,25 and 26 months.The pre-test period lasted for 10 days and the trial period lasted for 150 days,The results showed that the daily gain of control group was significantly higher than that of experimental group in 23 and 24 months (P<0.05).The daily gain of control group was highly significantly higher than that of experimental group in 25 and 26 months (P<0.01).The whole daily gain of control group,which was significantly increased by 13.40% than that of experimental group (P<0.05).The body height of control group was significantly higher than that of experimental group in 24,25 and 26 months (P<0.05).The body length,long body straight and chest circumference of control group were significantly higher than that of experimental group in 23,24,25 and 26 months (P<0.05).The tajiri long and sciatic wide of experimental group were significantly higher than that of control group in 23,24,25 and 26 months (P<0.05).The serum levels of urea nitrogen and albumin of experimental group were significantly higher than that of control group in 23,24,25 and 26 months (P<0.05),while the serum level of globulin was opposite trend.The serum activities of alanine aminotransferase and aspertate aminotransferase of control group were significantly higher than that of experimental group in 24,25 and 26 months (P<0.05).In conclusion,castration could reduced daily gain,growth rate and protein metabolism capacity of young Xinjiang brown cattle,but that could improved the ability of fat deposition.

This experiment was conducted to study the effect of Chinese herbal formula supplementation on protein synthesis and anti-stress ability of early weaned piglets.126 crossbred piglets (Landrace×Yorkshire) weaned at (28±2) days of age were randomly assigned into 7 groups with 3 replicates per group and 6 piglets per replicate.Piglets in group 1 was control group which fed a basal diet, piglets in groups 2,3,4 were fed diets supplemented with 0.05%,0.10%,0.15% water decoctum of Chinese herbal foumula,respectively,piglets in groups 5,6,7 were fed diets supplemented with 0.05%,0.10%,0.15% powders of Chinese herbal foumula,respectively.The experiment lasted for 21 days.The results showed that compared with the control group,Chinese herbal compound additives had significantly increased the activities of serum AKP,GOT,GPT and the contents of serum TP,ALB of early weaned piglets (P<0.05),but had significantly decreased the content of serum BUN of early weaned piglets (P<0.05).Chinese herbal compound additives had significantly decreased the activity of serum CK and the content of serum GLU of early weaned piglets (P<0.05).In conclusion,under this experiment conditions,Chinese herbal compound additives could promote the protein synthesis of early weaned piglets,reduce the anti-stress of weaned piglets,and powder was better than the water decoctum of Chinese herbal compound additives form,the appropriate supplemental level was 0.15%.

In this study, we administrated Radix isatidis and Rhizoma coptidis oral liquid according to 1(1%), 3(3%), 5(5%) and 10(10%) times of the clinical recommended doses for 21 days, and got the data about clinical hematology, blood biochemistry, organ index and histopathology of Radix isatidis and Rhizoma coptidis oral liquid in the target animal chicken. Then we preceded t test using SPSS 17.0 software and analyzed the statistical significance of inter-group indexes to provide the relevant data for the safety of clinical application. The experimental results showed that there was no significant difference about the relevant data of Radix isatidis and Rhizoma coptidis oral liquid in the target animal chicken compared with control group (P>0.05), and Radix isatidis and Rhizoma coptidis oral liquid would be safe at 10 times of the recommended dose (10%) in the target animal chicken.

The study observed the variety of cyclic AMP phosphodiesterase (cAMP-PDE) activity in polymorphonuclear neutrophils (PMN) and contents of cAMP,IL-6,TNF-α in plasma of porcine respiratory disease complex (PRDC),the therapeutic effect and influence of self-made Chinese herbal decoction,for accumulating information of respiratory inflammation pathological mechanism and its prevention.Divided pigs with PRDC into disease group and treatment group, and normal ones as control group. Drenched treatment group with 1 g crude drug/mL Chinese herbal decoction, which was composed of reed rhizome,mongolian milkvetch root,white peony root and liquorice root,twice a day and 1.5 mL/kg body weight for ten days. Observed changes of the number of white blood cell in peripheral blood,neutrophils were isolated from whole blood,HPLC was used to test its cAMP-PDE activity and ELISA kit was used to test the contents of IL-6,TNF-α in plasma.Compared with control group,the number of white blood cell in disease group increased significantly (P<0.05),cAMP-PDE activity and contents of TNF-α,IL-6 increased extremely significantly (P<0.01);but TNF-α in treatment group increased significantly (P<0.05). Compared with the disease group,the number of white blood cell,cAMP-PDE activity and content of TNF-α in treatment group decreased significantly (P<0.05), content of IL-6 decreased extremely significantly (P<0.01).The results showed that abnormal inflammatory reaction existed in PRDC,cAMP-PDE activity,the contents of IL-6 and TNF-α,the number of white blood cell increased significantly.Self-made Chinese herbal decoction could improve the clinical symptoms of the disease,of which the function mechanism was related to the reduction of the white blood cell number,inhibition of cAMP-PDE activity,and reduction of contents of IL-6 and TNF-α in plasma.

The purpose of this study was to investigate the effects of drinking boron on liver tissue of Africa ostrich chicks,36 10-day-old Africa ostrich chicks were randomly divided into 6 groups with 6 replicates in each group. The same basic diet and different doses of drinking boron were fed,boron contents in drinking water in each group were 0,40,80,160,320,640 mg/L,respectively. Test period lasted for 80 d. After the test,weighed,carotid artery bleeding to death,removed the liver after death,liver indexes were calculated. Liver specimens were placed in 4% paraformaldehyde and then embedded in paraffin. The livers were sectioned,stained by HE,and then observed and photoed under microscope. The results confirmed that low doses of drinking boron (40,80 mg/L) were in favor of liver development,improved the organizational structure of the liver,increased the Kupffer cells in the liver sinusoid,enlarged hepatic sinusoid,the optimal dosage was 80 mg/L. High doses of drinking boron (160,320 and 640 mg/L) adversely affected the development of Africa ostrich chicks' livers and made liver tissue pathological changes obviously.

The experiment was designed to study the effect of pine needle on immune function and serum cholesterol of chickens.240 healthy Roman cocks (1 day old) were randomly divided into 4 groups,3 replicates in each group and 20 chickens per replicate.The control group was fed with basal diet and normal drinking water,groups Ⅰ and Ⅱ were fed with the adding of 0.5 and 1.0 g pine needle powder,group Ⅲ was fed with pine needle decoction of 0.5 g.Pre-trial period was 4 d,formally trial period was 70 d.The results showed that the percentage of CD3⁺CD4⁺ T lymphocytes in groups Ⅰ,Ⅱ and Ⅲ were higher than that in control group at the 15 day after the first immunization,but that percentage of groups Ⅰand Ⅱ were lower than control group at the 30,45 and 60 days,Group Ⅲ was lower than control group at the 45 day.The percentage of CD3⁺CD8⁺ T lymphocytes in groups Ⅰ,Ⅱ and Ⅲ were higher than that in control group at the 15,30 and 45 days after the first immunization,but that percentage of groups Ⅰand Ⅲ were lower than control group at the 60 day.At the 14 and 21 days after the first immunization,the level of NDV antibody in groups Ⅰand Ⅱ were lower than that in control group,but it was higher than control group at the 28 and 35 days.The level of NDV antibody in group Ⅲ was higher than that in control group at the 14 and 21 days while it was lower at the 7 and 35 days.The levels of serum total cholesterol and triglyceride in groups Ⅰ,Ⅱ and Ⅲ had lower trend compared with control group,and the level of serum triglyceride in groups Ⅰand Ⅱ was significantly lower than that in control group and group Ⅲ (P<0.05).The level of serum low density lipoprotein in groups Ⅱ and Ⅲ was lower than that in control group,while the level of serum high density lipoprotein in group Ⅱ was higher than that in control group,but there were no significant differences (P>0.05).In conclusion,the pine needle could promote the proliferation and differentiation of T lymphocytes,improve the content of CD3⁺CD4⁺ and CD3⁺CD8⁺ lymphocyte,collaborate with vaccine to enhance the specific immune,and reduce the serum cholesterol level in different degrees.

The optimization of primary hemocyte culture of Scylla paramamosain was successfully studied in vitro.The results showed that the hemocyte cells were morphologically kept activity better at pH 4.6 marine crustaceans anticoagulant.High concentration of fetal bovine serum (>15%) in the culture lead to development of vacuoles.The hemocyte cells survived best in L15 medium supplemented with 5% fetal bovine serum and 1.2% sodium chloride.The house keeping gene β-actin of Scylla paramamosain of cultured hemocyte maintained stably expressed through the experiment.This cell culture system could be used for in vitro experiments.

Exogenous H₂S, in many aspects, had a protective effect on the central nervous system. In preliminary experiment, the administration time and dosage of exogenous H₂S were determined, and then 48 SD rats were randomly divided into four groups, such as control group (CP group), low-dose group (LP group, 7 μmol/kg NaHS), middle-dose group (MP group, 14 μmol/kg NaHS), high-dose group (GP group, 28 μmol/kg NaHS). Animals in three dose groups were randomly administrated 7, 14, 28 μmol/kg NaHS by intraperitoneal injection, followed the administration of ketamine 20 minutes after. The body temperature, heart rate, respiratory rate, blood oxygen saturation and blood pressure were recorded. The results showed that compared to the CP group,body temperatures and blood oxygen saturations in the dose groups significantly increased (P<0.05), heart rates and blood pressures in the dose groups significantly decreased (P<0.05), respiratory rates in the dose groups slowed down, followed a tendency to increase, and these changes showed a significant dose-dependent.

5-hydroxytryptamine (5-HT), as an essential neurotransmitter and intercellular messenger, has a variety of biological functions. More than 90% of 5-HT is localized in the gastrointestinal system where 5-HT plays an important role. The occurrence of many intestinal diseases is associated with the changes of 5-HT expression. 5-HT affects proliferation and maintenance of intestinal mucosal epithelial cell, increases intestinal permeability, and promotes goblet cell to release mucus indirectly. The mechanism of those actions may be that 5-HT stimulates afferent neuron and then activates the secretion of vasoactive intestinal peptide (VIP) and acetylcholine (ACH), thus causes diarrhea. For the purpose of giving some guiding information of diagnosing and treating intestinal diseases, the article discussed the reason of 5-HT causes diarrhea and the effect of 5-HT on the barrier of intestinal mucosal epithelial cell.

Along with the extensive use of antibiotics in clinical prevention and treatment, the problem of bacterial drug resistance is on the rise, especially the numbers of pan-drug resistance strains and multi-drug resistance strains increase obviously. It is hard for drug resistance to be generated by Chinese herbs with special antimicrobial mechanism. Therefore, the research and development of antagonizing drug-resistance Chinese herbs and the study of their antimicrobial mechanism draw more and more attention. Natural flavonoids possess certain antimicrobial activity and most species of them have strong inhibiting effect but little untoward effect on gram-positive bacterium such as S.aureus, Streptococcus and Bacillus subtilis.What's more, it is difficult for them to develop drug resistance. The antimicrobial action and antimicrobial mechanism of flavonoids were summarized in order to provide theoretical basis for further study of flavonoids antagonizing drug-resistance bacteria agents in this review.

The genetic diversity of major histocompatibility complex (MHC) gets more and more attention in the area of conservation biology. MHC has been identified as a locus influencing disease resistance, mate choice, and kin recognition in mammals and fish which have been extensive and in-depth studied. However, it is unclear whether the mechanisms by which MHC genes influence behavior in mammals are applicable to birds, especially in wild populations. This article reviewed MHC polymorphism mechanism, its relationship with avian disease resistance and mate choice, and methods for MHC genotyping, and so on. It will provide references to the further study of bird conservation biology.

Circadian clocks are comprised of the central circadian clock and peripheric circadian clock. The central circadian clock mainly locates in the suprachiasmatic nuclei (SCN) of hypothalamus in the mammal, but also contains retina and pineal gland in the birds. The circadian clocks can be stimulated by environment and then modulate the physiologic activities, that many tissues, hormones and irritants are involved. This review will highlight the role of central circadian clocks in the secretions of melatonin in the pineal gland and gonadotrophin releasing hormone (GnRH) in the hypothalamus, and make a brief summary about the effect of the illumination in the procees.

Polymorphisms in 292G/A and 1168G/A locus of lipopolysaccharide binding protein (LBP) gene in 81 Meishan pigs were detected by PCR-RFLP method,the relationships between two polymorphic sites and partial important cytokines in blood serum were analyzed in this study to provide feasibility analysis of those polymorphic sites used for further disease-resistant breeding of pigs. The results showed that 292G/A and 1168G/A of LBP gene were digested by MspⅠ and Bsh1236Ⅰ, respectively, which could be divided into three kinds of genotypes (AA, AB and BB). PIC value of Meishan pigs in 1168G/A ranged from 0.25 to 0.5, which displayed moderate polymorphism. While the value of 292G/A was less than 0.25, which displayed low polymorphism. The correlation analysis indicated that IFN-α level of the BB genotype was significantly higher than that of the AA and AB genotypes in the 292G/A (P<0.05), while there had no significant differences in immune indices among the 3 kinds of genotypes based on the 1168G/A (P>0.05). To make a conclusion: The mutation in the 292G/A of LBP gene had probably some effects on general disease resistance in Meishan pigs. It is necessary to make a further analysis and verification of the hereditary effects in LBP gene 292G/A.

The objective of the present study was to assess the effects of Astragalus polysaccharides on cryopreservation of semen collected from adventive boars and provide a theorectical reference to improve the formula of diluents for boar semen cryopreservation. Semen was collected from three kinds of American-derived boars (Landrace, Large Yorkshire, Duroc) and diluted in solution supplimented with different concentrations of Astragalus polysaccharides (0, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%). After being loaded into 0.25 mL straws the extended semen was freezed. Subsequently the qualitative parameters (motility, abnormality and acrosomal integrity) of frozen-thawed spermatozoa were estimated, and comparisons of mean values among three breeds of boar at the same conoentration or among treatments at the same breed of boar were performed. The results showed that,when compared without Astragalus polysaccharides acrosomal integrity of frozen-thawed Duroc boar spermatozoa was significantly higher than that of frozen-thawed Large Yorkshire boar spermatozoa (P<0.05), but no significant differences were observed when other comparisions were carried out at the same concentration (P>0.05). Of all the three breeds of boar, 0.04% was the optimal concentration of Astragalus polysaccharides supplied to semen diluents, at which the quality of frozen-thawed semen was significantly better than that of the control (P<0.05). There weren't significant differences in the quality of frozen-thawed semen among the three breeds of boar at the optimal concentration of added Astragalus polysaccharides (P>0.05). In brief, the extender supplimented with 0.04% Astragalus polysaccharides can demonstrate desirable parameters of frozen-thawed spermatozoa in all the three breeds of boar.

In this study, Wenshang Barred chickens was selected as the research object,exon 2 of prolactin (PRL) gene were detected by PCR-SSCP and the DNA fragments were sequenced. The correlation analysis between the polymorphism of the exon 2 and egg production traits were carried out respectively. The results indicated that there were two polymorphic sites in exon 2. C→T mutation at the position of the 3838 bp was found, which resulted in 3 genotypes (AA, AB and BB). Furthermore, an amino acid changed from leucine to phenylalanine. The frequencies of genotype BB and allele B were the highest. Three genotypes of AA, AB and BB in age at first egg, living weight at first egg, egg weight of first egg, living weight at age of 300 days, egg weight at age of 300 days index of reproductive traits difference did not reach the significant level (P>0.05). The results showed that PRL gene exon 2 polymorphism and reproductive traits of Wenshang Barred chickens presented no significant correlation.

By observing the unknown sex pellets collected from Cervus elaphus songaricus wild populations of Urumqi Nanshan mountain, all pellets are presented one of two shapes of bullet-like and jujube-stone like. The bullet-like fecal pellets displayed a short thick form with a low length/width ratio. The jujube-stone like ones displayed a thin long form with high length/width ratio. Using these shapes,the 140 samples were divided into two groups of 86 bullet-like and 54 jujube-stone like feces, and determined by PCR amplification of the SRY gene. Among of them,94 samples were identified as male,and the others as female. All the samples were clustered by the ratio of averaged length and width,and then the discrimination equation was established. The statistical results showed that the consistent degree of sex determination by fecal statistic index and DNA method was 85.48%,while morphological method and real sex was 88.34%. So,the bullet-like pellets were from male wapiti,and the jujube-stone like ones from females. The results showed that wapiti sex could be directly determined by its pellet shape in the wild. At the same time,the discrimination equation could be used as associated method.

This experiment was to study the single nucleotide polymorphisms (SNPs) and single amino acid polymorphism (SAPs) of exon 7,exon 13 and exon 14 in Chinese Merino sheep endothelial nitric oxide synthase (eNOS) gene, which were analyzed according to the published SNPs of exons loci of endothelial nitric oxide gene of human and mouse in GenBank. 3 exons of Chinese Merino sheep eNOS gene, having the higher similarity with the human and mouse, were found through bioinformantics method, and they were exon 7,exon 13 and exon 14, respectively. Exons polymorphism in human and mouse eNOS gene were aligned and analyzed by the NCBI SNP database, found that 2 SNPs loci in the human eNOS gene exon 7 (rs1799983) and exon13 (rs148554851) had clinical significance. After that, the amplified fragments of the 3 exons in Chinese Merino sheep eNOS gene were analyzed by PCR-SSCP method, indicating that SNPs loci were not discovered 3 exons in Chinese Merino sheep, therefore, 3 exons regions of Chinese Merino sheep eNOS gene were relatively conservative.

Epigenetics modification is the heritable changes in gene activity which are not involved in changes in the DNA sequences, including DNA methylation and chromatin remodeling, histone modification and non-coding RNAs, which plays important roles in gene expression, spermatogenesis, development and maturation. During spermatogenesis, normal morphology, motility and fertility were proven to be modulated by epigenetic changes. Cryopreservation causes dramatic changes of the sperm viability, motility and fertility, and the cryoinjury might be involved in some epigenetic factors. The epigenetic modifications involved in spermatogenesis and cryopreservation and their relationship were reviewed.

In this paper,in order to find out the hybrid effect on the production performance on weight gain and wool yield of the hybrid offspring,in which the F1 generation was crossbred by French Charolais sheep (♂) with Chinese Merino sheep (Xinjiang type) (♀) and the B1 generation was crossbred by French Charolais sheep(♂) with F1 (♀),a hybrid fattening experiment was carried out. The results showed that,in the March age, the average weight of F1 generation were 2.8% (P>0.05) and 46.5% (P<0.01) higher than the B1 generation and the Chinese Merino sheep (Xinjiang type) respectively;In the June age, the average body weight of F1 generation were 4.0% (P<0.01) and 28.6% (P<0.01) higher than the B1 generation and the Chinese Merino sheep (Xinjiang type) respectively;In the March age and June age, the average weight of B1 generation were 42.5% (P<0.01) and 23.7% (P<0.01) higher than Chinese Merino sheep (Xinjiang type) respectively. From the wool,there was no significant difference (P>0.05) between of F1 generation and and B1 generation on the average wool yield, but compared with the Chinese Merino sheep (Xinjiang type),the the average wool yield of F1 generation and B1 generation were reduced by 20.4% (P<0.01) and 20.1%(P<0.01) respectively, the differences were significant. From the gross economic benefits of weight gain and wool yield, compared with Chinese Merino sheep (Xinjiang type),the gross economic benefits of F1 generation and B1 generation were increased by 14.80% and 10.88% respectively. Therefore, we can make full use of the hybrid advantages of French Charolais sheep and Chinese Merino sheep (Xinjiang type) to develop mutton production, but should not be used for the production of wool.

Some pathologic samples were collected from chickens which were preliminarily diagnosed to be infected with reticuloendotheliosis virus (REV) in Guangdong province. The samples were inoculated on DF1 cells for reproduction. With identification of polymerase chain reaction (PCR) and indirect immunofluorescence assay (IFA), one REV strain was isolated, named as GD1210. According to the published sequences of REV, six pairs of primers were designed and synthesized. The overlapping fragments were amplified and the whole genome of GD1210 strain was sequenced. Sequence analysis showed that the full genome sequence of GD1210 strain was 8443 bp in size and shared the highest nucleic acid homology with the SNV strain when it was compared with the published REV sequences. The pathogenicity experiment for one-day-old SPF chickens of GD1210 strain indicated that the infected chickens had severe immunosuppression.

In order to obtain the optimal therapeutic dosage of ethyl pyruvate (EP) in compound preparation and compare the effects of different concentrations of EP and administration time on treatment outcome,four different mixture ratios of suspensions composed by EP that prepared by Veterinary Medicine Laboratory (Qingdao Agricultural University) were applied to the experimental therapy of mice infected with E.coli. 340 Kunming mice were divided into 17 groups randomly (20 in each group) and then were injected with E.coli ATCC 25922. A ceftiofur (CE) single suspension (CE 5 mg/kg) and four kinds of EP compound suspension (EP was 40,80,120 and 160 mg/kg,respectively) were injected into the mice after 1,4 and 8 h in order to observe the mortality rates of mice and analyze statistically. The results showed that the effect of compound suspension groups of EP 40 and 80 mg/kg were much better than CE single suspension group at 1 and 4 h after vaccination (P<0.05),but there was no significant difference between the two compound suspension groups (P>0.05). At 8 h after vaccination, the therapeutic effect of all compound suspension groups and single suspension group was not significant and there was no significant difference compared to positive control group (P>0.05). The therapeutic effect of compound suspensions of EP 40 and 80 mg/kg were significantly better than CE single suspension, and could help to remarkably improve the cure rate of mice, but there was no significant difference between the two compound suspension groups. It was recommended to choose compound suspension of EP 40 mg/kg from the perspective of security and economy.

The experiment was carried out to compare diarrhea status between Min pig and Landrace pig in a week after weaning, detect two varieties piglets jejunum, ileum and colon tight proteins Zonula occludens-1(ZO-1)and Occludin mRNA expression before and after weaning. The intention was to discuss the relationship between tight proteins expression and piglets weaning diarrhea. The results showed that Min piglets diarrhea rate, diarrhea frequency were very significantly lower than Landrace pig within one week after weaning (P<0.01), diarrhea days was significantly lower than Landrace pig within one week after weaning (P<0.05);Min piglets not weaning group ileum and colon ZO-1 and Occludin mRNA expression was very significantly higher than Landrace pig not weaning group (P<0.01);Min piglets weaning diarrhea group ileum and colon ZO-1 and Occludin mRNA expression was significantly higher than weaning health group (P<0.05). The results indicated that Min piglets intestinal tight junction proteins mRNA high expression before weaning might be had relation with guarantee piglets intestinal health and decrease occurrence diarrhea;Min pig fast recover intestinal mucosa function and rehabilitation by increased tight junction proteins mRNA expression after weaning. So tight junction proteins mRNA high expression before and after weaning had an important significance for piglets to resist weaning diarrhea.

In order to simplify theseparation steps of a single protein in the outer membrane protein P2 (OmpP2) of Haemophilus parasuis (Hps) and reduce the equipment requirements of membrane protein separation,Hps standard type 3 (SW114), 5 (Nagasaki) strains were used as experimental materials in this experiment.First, lysozyme, nucleic acid were used to enzyme digest enzymatic hydrolysis the Hps for its complete outer membrane. And then, using buffer containing 2% triton-X-100 for further disintegrate. At last, used the appropriate concentration of pancreatic enzymes to digest the strong hydrophobic protein. Finally, a rapid extraction of the Hps high-purity OmpP2 in vitro was found. The result of sodium dodecyl sulfate-poly propylene amide gel electrophoresis method (SDS-PAGE) showed that it was very consistent with our expection: The two strains of Hps OmpP2 was about 43 and 38 ku, respectively.The protease off peptides matched with Hps OmpP2 perfectly in mass spectrum (MALDI-TOF-TOF). This experiment had established a rapid extraction method of Hps OmpP2. It reduced the equipmental requirement and simplified the separation step of a single protein OmpP2 from the membrane protein. Both type 3 (SW114) and 5 (Nagasaki) strains OmpP2 had been found hybridization chromogenic reaction with serum collected from sick pigs in Dot-blot. It simplified that OmpP2 was an immunological antigens. This study would be worthy of the function and immune protective research for Hps OmpP2 and offered a new reference for Hps immunological diagnosis.

The effects of carbon, nitrogen, serum and coenzyme on culture of Haemophilus parasuis were investigated by single factor test, and on the basis of single factor test, the orthogonal design of four factors and three levels was employed to test the concentrations of sugar, yeast powder, ammonium sulfate and horse serum.The results showed that the optimized concentrations were that sucrose was 2 g/L, yeast powder was 2 g/L, ammonium sulfate was 2 g/L, and horse serum was 5%. Comparing the impact of different supplement methods on Haemophilus parasuis fermentation in 10 L fermentor, a good fermentation result was obtained when the residual sugar concentration was maintained at 1 g/L by supplementing constant sugar.The result showed that bacterial density and the number of viable bacteria were 16.8 and 3.6×10⁹ CFU/mL,respectively.

Strain SK-1 was isolated from naturally infected Tincaeus. Its morphological, physiological and biochemical experiments, 16S rDNA sequence were performed, and its antibiotic sensitivity was analyzed by Kriby-Bauer disc diffusion method. The results showed that strain SK-1 was gram negative bacteria,with short rod shape and oxidase-positive.Its physiological and biochemical characteristics were consistent with the physiological and biochemical characteristics of Vibrio anguillarum. The length of this 16S rDNA sequence was 1447 bp, which showed 99% homology with Vibrio anguillarum, GenBank database number was KF978124. Phylogenetic analysis indicated it grouped together with Vibrio anguillarum. So it was confirmed that strain SK-1 was Vibrio anguillarum. The antibiotic sensitivity presented that strain SK-1 was highly sensitive to 13 antibiotics such as doxycycline,tetracycline,minocycline, and middle sensitive to 9 antibiotics such as streptomycin, ceftriaxone, trimethoprim, and resistant to bacitracin and lincomycin. These results will be beneficial to the identification of Vibrio anguillarum and prevention and control of bacterial disease in Tincaeus.

Antimicrobial peptide (AMP) could kill bacteria, which had been used as a natural preservative and potential alternative to antibiotics. To understand whether the CecropinXJ could effectively inhibit bacteria from palm skin, we used sterile water to swab hands and treated with swabbed liquor by different concentrations of CecropinXJ in this study. The antimicrobial activity was evaluated by plate count method. The results showed that the growth of bacteria derived from palm skin could be effectively inhibited via adding different concentrations of CecropinXJ, and the antimicrobial activity became stronger with increasing the concentrations of CecropinXJ. When the concentration of CecropinXJ was up to 0.4%, its inhibitory capacity could be comparable to commercial hand sanitizer (P>0.05). Further study elucidated that swabbed liquor included most of gram-negative bacteria through 16S rDNA sequence analysis. These findings suggested that CecropinXJ could be used as a novel disinfector or cleanser for application potentially.

In order to determine the antibiotic resistance of pathogenic Escherichia coli (E.coli) in Guangdong province,a total of 62 E.coli isolates were collected from the diseased geese of four breeding goose farms in 2012.The susceptibilities of 62 isolates to 17 antimicrobials were determined by agar dilution method.The results showed that the resistance of the isolates was serious and the resistance rates to most of the antimicrobials were over 60%.Only cefoxitin, ceftazidime and colistin remained effective to the isolates. Combined antimicrobial susceptibility testing,it showed that doxycline and florfenicol had remarkable synergy activity against multi-drug resistant E.coli strains, with FIC varied from 0.375 to 0.75.Twenty-four strains were divided into 17 types by the test of PFGE analysis.43.5%,34.8% and 21.7% isolates belonged to phylogenetic group D,A and B1,respectively.This study might provide theoretical basis and reference basis for clinical medication.

To understand the Mycoplasma pneumonia pathogen of Mashan Black goat and provide a basis for prevention and control of Mycoplasma pneumonia of Mashan Black goat, morphological observation, biochemical characteristics, growth inhibition, PCR amplification and sequence, animal pathogenicity test were used to identify the pathogen of Mycoplasma pneumonia. The results showed that M. ovipneumoniae was the pathogen of Mycoplasma pneumonia in Mashan Black goat. Drug susceptibility test showed that M. ovipneumoniae were sensitivity to spectinomycin, kanamycin, gentamycin, amikacin, lincomycin, ciprofloxacin, levofloxacin, SMZ-TMP and florfenicol.

Sheep scrapie is a central nervous system progressive degeneration disease which is caused by scrapie infection factor in sheep and goats.The main clinical symptom features of this disease are long incubation period, itching, central nervous system degeneration and high mortality. Sheep scrapie is harmful for the sheep industry, and it is the earliest known animal prion diseases.The disease is rare in China but is very popular in the world, and the potential threat to China is growing. Therefore, strengthening of the awareness of disease etiology and epidemiology is very important to comprehensive prevention measures of sheep scrapie.In order to provide certain theoretical basis for the prevention of the disease,the author mainly introduced the pathogen, epidemiology and comprehensive prevention measures of sheep scrapie at home and abroad.

Lactic acid bacteria have been widely used as the vehicle to deliver various pathogen antigens for their advantage, such as probiotic, adjuvant effect, simple preparation, and low production cost. Live vaccine with lactic acid bacteria as the delivery vector is now one hot vaccine research field, for they can stimulate the humoral, mucosal, and cellular immune responses at the same time. In this paper, the mechanism and the way to deliver heterogenous antigen of this kind of live vaccine were generalized, the application of them in preventing viral, bacterial, and parasitic diseases was discussed, and the existed problem and prospect in application were finally analyzed and forecasted.

As a part of the congenital immune, autophagy plays a "double-edged sword" role in the process of intracellular pathogen infection. On the one hand, autophagy could suppress and eliminate the infection of intracellular pathogens, protect the body from the invader; on the other hand, some intracellular pathogens could inhibit, block the occurrence or mature of autophagy, or autophagy is even used to escape the immune mechanism, it is conducive for the proliferation and survival of intracellular pathogens in the body.The author reviewed the interactions between autophagy and intracellular pathogen, and the function of autophagy played in intracellular pathogen infection.

Escherichia coli disease is a common bacterial infectious disease of poultry industry, is also a kind of zoonosis. Emergence of multi-drug-resistant strains caused great difficulties to the prevention and treatment of the disease, caused a great economic loss to our country aquaculture, but also posed a potential threat to human health in recent years. Factors of chicken Escherichia coli disease model infected by artificial and Chinese traditional veterinary diagnosis and treatment were discussed in this article, summarizes the research advancement in recent years, and provide new ideas for prevention and control of chicken colibacillosis in drug screening and the development of new drugs.

In order to analyze the causes of calf diarrhea in Nanjing, 27 strains bacteria were isolated from the material of diarrhea calves. The cultured characteristics, staining characteristics, biochemical characteristics, serotype characteristics, drug susceptibility and TEM, SHV, CTX-M-1, CTX-M-9 type extended-spectrum β-lactamase (ESBLs) gene of 27 strains bacteria were detected. The results showed that 27 strains bacteria were all E.coli, and only one bacteria was resistant for cephalothin,cefazolin and cefuroxime,and this strain was detected in CTX-M-1 type extended-spectrum β-lactamase gene. The results showed that calf diarrhea were caused by E.coli, the calf diarrhea E.coli could produce CTX-M-1-type extended-spectrum β-lactamase to resolve cephalosporin antibiotic that led to drug resistance.