In order to get porcine phytase salivary glands specific express cloned embryos,this study used phytase specific expressed plasmid DNA (including the promotor and terminater sequences of parotid secretary protein (PSP) gene,Neo selected gene and EGFP report gene and high enzymatic activity phytase gene (appA)) and transfected it into porcine fetal fibroblast cells with liposome.The stably transfected cell lines were screened by G418 selection,and positive phytase transgenic embryos were constructured by somatic cell nuclear transfer.The results showed that the phytase plasmid of this experiment could be used to carry on the cell selection,and the more smaller of plasmid DNA size,the more higher transfected efficiency was got.The plasmid YM6552 with 14.89 kb size only got 7.1% transfected efficiency,while EGPF plasmid got 43.4%.In the formation of single cell colony,pYN3600 (more smaller DNA size) got 25 colony numbers,and 14 of them expressed green fluorescent protein,11 of them were phytase PCR positive.They were much higher than those of YM6552 (19,8 and 6).All the reconstructed transgenic embryoes from the positive cell lines were expressed green fluorescent protein.Though the in vitro development ability of transgenic cloned embryos were a little lower than that of control group,there were no significant differences between them (P>0.05).In conclusion,all the phytase plasmid,cell selection method and nuclear transfer techniques could be used to produce porcine phytase salivary glands specific express embryos.

This study was aimed to clone and express ORFV086 protein of Orf virus (ORFV) in prokaryocytes and prepare polyclonal antibody anti-ORFV086 which was used to detect the expression of ORFV086 protein. The ORFV086 gene was amplified from genome DNA isolated from ORFV by PCR, then cloned into prokaryotic expression vector pET-33b (+) to construct a recombinant expression vector pET33b-086. The pET33b-086 was transformed into E.coli BL21 (DE3) pLys and induced by IPTG from which the fusion protein was identified by SDS-PAGE. The purified protein was injected into New Zealand rabbits and the polyclonal antibody was prepared, which was identified by the indirect ELISA and Western blotting methods. The results showed that the recombinant protein mainly existed in the inclusion body with the expected molecular weight of about 100 ku. After purified by cutting the gel slices, target protein was used as immunogen to prepare polyclonal antibody. The titer of the polyclonal antibody was 1:128000. Western blotting method revealed that the purified polyclonal antibody had a specific affinity for the reorganizational and natural ORFV086 protein. Thus, the rabbit anti-ORFV086 polyclonal antibody was successfully prepared, providing a tool for further investigation on the role of ORFV086 protein infection.

This experiment was designed to investigate the regularity and significance of β-catenin and BMP2 mRNA expression in skin of young minks. The mRNA expressions of β-catenin and BMP2 in the skin from 0 to 6 weeks old minks were studied by Real-time quantitative PCR relative quantitative method (ΔΔCt). The dorsocentral region of skin samples were collected from three minks per week. The results showed that β-catenin significantly up-regulated at 4 weeks old (P<0.05) and reached the maximum at 5 weeks old (P<0.01), then compared to 5 weeks old,it extremely significantly decreased at 6 weeks old (P<0.01), while the expression of BMP2 mRNA kept stable with weeks increasing (P>0.05). Combined the results of hair follicle histology of young mink, we could conclude that the abundance of β-catenin mRNA was correlated with secondary follicle development. It might be involved in the development of skin and secondary hair follicle in mink, and the abundance of BMP2 did not apparently change at different developmental stages of follicles.

To get the molecular biology character of Salmonella abortus equi isolated from Xinjiang and investigate the structure and function of FliC gene, we amplified FliC gene using PCR method, cloned, sequenced and analyzed FliC gene using bioinformatic methods. The results indicated that FliC genes of 2 isolates were 626 bp (GenBank: KJ486797, KJ486798). Sequence homology analysis showed that compared with those strains obtained from GenBank, the nucleotide sequence identity of the FliC gene of these isolates in Xinjiang were 49.6% to 100%. The results showed that FliC gene was conserved during their evolution. This result would provide foundation for the analysis of molecular epidemiology of this disease and the prevention of this infection using FliC gene.

In this research,a simple and rapid extraction of microbial DNA for PCR using Chelex-100 chelating resin directly from milk was built.3 common mastitis pathogens (S.aureus,E.coli and S.agalactiae) were submitted to tenfold serial dilu-tions and artificially inoculated to milk samples.Then the sensitivities of the PCR assay using the Chelex-100 DNA extraction method were detected.In the meantime,the sensitivity of the 3 common mastitis using phenol-chloroform DNA extraction method was compared.The results showed that the PCR assay using the Chelex-100 DNA extraction method was more sensitive than that using phenol-chloroform DNA extraction method.The sensitivity of the PCR assay using the Chelex-100 DNA extraction method in detecting S.aureus, E.coli and S.agalactiae was 10³,10² and 10² CFU/mL,repectively.The sensitivity of the PCR assay using phenol-chloroform DNA extraction method in detecting S.aureus,E.coli and S.agalactiae was all 10⁴ CFU/mL.The thresholds of the sensitivities using the Chelex-100 DNA extraction method were compatible with their use as an efficient tool of diagnosis in bovine mastitis.The Chelex-100 DNA extraction method was simple,rapid,cheap,involve no organic solvents and meaningful for direct extraction of DNA of other pathogens from milk.Undoubtedly,this method will make sense for rapid diagnose of bovine mastitis by PCR.

The complete CDS of DLX3 gene was amplified from skin during Liaoning cashmere goat hair follicle growth phase by RT-PCR using one pair of primer which was designed and synthesized according to the DLX3 gene sequence in GenBank(No:XM_005694267.1),inserted into blunt-vector after DNA sequencing verification,it was then sub-cloned into eukaryotic expression vector pIRES2-EGFP.After the restriction enzyme (XhoⅠ/BamHⅠ) digestion of the eukaryotic expression vector and sequencing,the plasmid had been transfected into the goat fibroblasts by Lipofectamine 2000.We also observed the fluorescent under the microscope,examined the transcription vector pIRES2-DLX3-EGFP by RT-PCR and Western blotting.The results showed that we had successfully cloned goat DLX3 gene and constructed eukaryotic expression vector pIRES2-DLX3-EGFP,we could observe the green fluorescent after 24 h transfection of the plasmid.Then, we amplified the transcription product of 909 bp by RT-PCR.The targeted protein 32.87 ku was also detected by Western blotting.This result has paved the way for investigating the influence of DLX3 on hair follicle development and cycling as well as the research of the regulation mechanism of hair follicle growth and development.

In order to obtain the recombinant chicken interferon-α (ChIFN-α) and study its biological activity, the recombinant E.coli was induced to express ChIFN-α. After ChIFN-α inclusions were collected for renaturation and purification, Western blotting test for the recombinant ChIFN-α was done. By measuring the EID₅₀ of chicken embryos infected with avian influenza virus (AIV) H9 subtype, Newcastle disease virus (NDV) and infectious bronchitis virus (IBV), we investigated the antiviral activity of the recombinant ChIFN-α and determined its antiviral potency by cytopathic effect inhibition assay. The specific bands were found as expected in Western blotting test of renaturated ChIFN-α. In addition, the ChIFN-α showed significant antiviral activity against AIV H9, NDV and IBV, and antiviral potency of the interferon was about 1.5×2¹⁰ U according to the test on CEF/VSV cell line. High-concentration ChIFN-α showed cytotoxicity. The recombinant ChIFN-α had immunogenicity and antigenicity. And it could inhibit or kill viruses in chicken embryos including AIV H9, NDV and IBV, which showed broad spectrum and high antiviral activity. The study would promote our research and development of antiviral products in the future.

In order to reveal the Guangdong region of porcine reproductive and respiratory syndrome (PRRS) epidemic law, 5 strains of swine clinical samples amplified from Guangdong region of PRRSV GP5 genes of the isolates with the method of RT-PCR, cloned, sequenced and domestic and foreign strains corresponding gene nucleotide sequence comparison in this study.The results showed that PRRSV-2011-GD2, PRRSV-2011-GD3, PRRSV-2013-GD1, PRRSV-2013-GD2 and PRRSV-2013-GD3 strains with the American type reference strains (ATCC VR-2332 U87392) nucleotide sequence homology were 88.8%, 99.5%, 88.7%, 88.7% and 83.5%,respectively. The phylogenetic tree analysis indicated that PRRSV-2011-GD2, PRRSV-2011-GD3, PRRSV-2013-GD1, PRRSV-2013-GD2 and PRRSV-2013-GD3 strains belonged to the American.The results provided the basis for a better understanding of the molecular epidemiology of PRRSV features and antigenic variation law, also laid the foundation for the study of gene engineering vaccine.

For analysis melanocortin receptor 1(MC1R) genotype in swine spices with different coat color and study the gene functions on swine coat color. This research analysed the genotype of MC1R in Junmu 1 White pigs, Durocs pigs, Tibet mini-pigs and Large White pigs with PCR-SSCP and PCR-RFLP technique. The result showed that Junmu 1 White pigs and Large White pigs presented mutation on nt894insCC and G1197A site, Durocs pigs presented mutation on G668C, C1318T, and G1554A site, Tibet mini-pigs presented mutation on C1318T and G1554A site, with a different genotype on nt894insCC site. The MC1R genotype result provided a basis for studying MC1R regulation mechanism to swine coat color.

To enrich the molecular epidemic information for the newly identified porcine kubovirus, the nonstructural protein 2C of porcine kobuvirus was amplified by reverse transcription-polymerase chain reaction method from the tissues of the piglets association with diarrhea in Fujian province, and then cloned and sequenced, bioinformatics analysis were conducted to investigate the structure and function of porcine kubovirus nonstructural protein 2C. The results demonstrated that porcine kobuvirus nonstructural protein 2C was 1005 bp in length, coding an open reading frame (ORF) with 335 amino acids. The molecular weight, theoretical isoelectric point, instability index (Ⅱ), aliphatic index and the Grand average of hydropathicity (GRAVY) of porcine kubovirus nonstructural protein 2C were 36.9297 ku, 7.18, 39.41, 87.10 and -0.242, respectively. The sequenced gene shared the highest homology with CH/HZ/2011 strain (GenBank accession number was JX827598), with 91.6%, while, the lowest homology with XX strain (GenBank accession number was KC204684), with 88.8%. The porcine kubovirus nonstructural protein 2C shared three different phylogenetic branches, porcine kubovirus Hungary isolates and Thailand isolates were at the branch Ⅲ, and the porcine kubovirus Chinese isolates distributed in the other two phylogenetic branches.

In order to further study the role of Wzt protein in the synthesis of smooth lipopolysaccharide, the PCR technology was used to amplify the Wzt gene of 759 bp .Then it was ligated into pMD20-T vector.The recombinant plasmid pET-28a-Wzt was transformed into E.coli BL21(DE3) for expression under induction of IPTG. The protein product was analyzed by SDS-PAGE and Western blotting. The results showed that the size of the Wzt gene was 759 bp and the sequence homology was 99.87% compared with the sequence of Brucella melitensis 16 M strain with the access number of AF047478.1 in GenBank,proving that the Wzt gene was successfully cloned. Also, the prokarotic expression vector pET-28a-Wzt was successfully constructed and the Wzt protein with molecular weight of 30 ku was highly expressed in E.coli BL21(DE3), which all the above proved that Wzt gene was successfully expressed.

A triplex reverse transcription-polymerase chain reaction (triplex RT-PCR) was developed to detect avian influenza viruses (AIVs) and H9 N2 subtype AIVs simultaneously. Three pairs of specific primers were designed according to the conserved regions on the sequences of H9 AIV HA gene, N2 AIV NA gene and AIV M gene in GenBank. It showed that all samples containing H9N2 subtype AIV could be amplified into three specific bands, 313 bp for HA gene, 451 bp for NA gene and 667 bp for M gene by this triplex PCR. All samples containing N2 subtype AIV with different HA genes (not H9) could be amplified into two specific bands, 451 bp for N2 subtype AIV and 667 bp for M gene. All samples containing other subtypes of AIVs (not H9 or N2) could be amplified into one specific band, 667 bp for M gene. No specific bands of the same sizes were amplified from genomic materials of other avian pathogens. The detection limit of triplex PCR was 10^-2 ng/μL. The results of triplex PCR for detection of clinical samples were coincident with that of the viral isolation completely. Our results demonstrated that the optimized triplex PCR assay was quick, specific and sensitive for detection of AIVs especially H9 and N2 subtype AIV.

To establish a rapid, simple and accurate method to diagnose and detect H1 subtype swine influenza virus (SIV), the specific primers and TaqMan MGB probe were designed according to the conserved region of HA gene of H1 subtype SIV. A one-step Real-time fluorescent quantitative RT-PCR assay was developed for detection of H1 subtype SIV. A series of dilutions of recombinant plasmids pMD18-H1 were prepared and used to generate standard curves. The results showed that the one-step Real-time fluorescent quantitative RT-PCR was capable of detecting 10² copies of H1 subtype SIV per microliter. The results were negative for the detection of H3N2 and H9N1 subtypes SIV, classical swine fever virus, porcine reproductive and respiratory syndrome virus, porcine epidemic diarrhea virus, transmissible gastroenteritis virus. The coincidence rates between one-step Real-time fluorescent quantitative RT-PCR and virus isolation were 94%. The method was highly specific and sensitive, and could be used for rapid quantitative detection of H1 subtype SIV.

The purpose of this study was to identify the strain CVCC 3982 isolated from a death dairy cow's lung in Beijing and determine its pathogenicity. The suspected pathogen was identified by means of Biolog system and 16S rDNA sequence analysis. Its pathogenicity was determined by artificial inoculation in CD-1 mice, synthetic primers were used to examine the main virulence genes. The results showed that the suspected pathogen was gram positive bacillus, β hemolysis. The identification result of Biolog was Trueperella pyogenes. The homology of 16S rDNA gene sequence with the Trueperella pyogenes type strain NCTC 5224 reached 100%. By the phylogenetic analysis, the isolate was in the same branch with Trueperella pyogenes. The isolate contained hemolysin (plo) gene, neuraminidase H (nanH) gene, neuraminidase P (nanP) gene, fimbriae gene (fimA, fimC, fimE), but was lack of collagen-binding protein (cbpA) gene and fimG gene. Infection of suspected bacteria resulted in mortality to mice by intraperitoneal injection. The results indicated that the isolate was a virulent Trueperella pyogenes.

To analyze the molecular characterization and evolutionary relationship of the virulence associated genes of the BJ/YT strain which was isolated from a Yorkshire Terrier in Beijing in 2012. The results revealed that the homologies of the gE gene nucleotide sequences and amino acid sequences were 98.9% to 100.0% and 98.3% to 100.0%, respectively; meanwhile, the homologies of the TK gene nucleotide sequences and amino acid sequence were 99.2% to 99.9% and 99.0% to 100.0%, respectively. The phylogenetic tree based on gE gene showed that BJ/YT and the strains isolated from Hebei province could be grouped into a relatively independent branch, and all of them shared a high homology. Besides, a concordant mutation site was presented. Few mutation sites in gE and TK amino acid sequences were observed when compared with other referenced sequences. But those sites were not located in the main domains or antigenic epitope regions. Therefore, from the view of molecular epidemiology, it was likely that the BJ/YT isolate was the main circulating strain in Beijing and its surrounding area the last few years. However, the variations of gE and TK genes did not have obvious effects on the virulence of the epidemic strain during this period.

Guangxi Luchuan pigs and Bama miniature pigs are included in the national genetic resources list of local varieties with excellent performance.Carrying on the basic research in molecular biology of local pig has become the focus in recent years.This paper summarized the research progress on molecular biology in genetics and breeding,mycoplasmal pneumonia of Guangxi Luchuan pigs and genetics and breeding,disease model of Bama miniature pigs.

In order to establish a method to detect H3 subtype avian influenza virus (AIV), two pairs of specific primers were designed and selected according to the conserved sequences of the hemagglutinin (HA) genes of H3 subtype AIV, the reaction conditions were optimized to establish the nested PCR.The specificity and sensitivity of assays were evaluated, and 96 clinical samples were detected by nested PCR. The results showed that the nested PCR was only specific to H3 subtype AIV, without amplification of other viruses; the detection limit of nested PCR was 1×10³ copies/μL which was 100 fold higher than the routine PCR; the detection of clinical samples were consistent with the viral isolation completely. This study provided a accurate and effective method for the detection of H3 subtype AIV.

Molecular diagnostics based on nucleic acid amplification technology (NAT) were widely used in medical research, inspection and quarantine, disease diagnosis and other fields, available reference materials for NAT assays were critical to ensure the diagnostic test data and the results were accurate and consistency. This article described the development history, current situation and challenges of nucleic acid reference materials, and prospected for the future direction of development of nucleic acid standard materials.

In order to provide a more dependent and fine tool for porcine immunology research, we developed a serial of pig specific NF-κB reporter system. Firstly, 6 reporter vectors were constructed by using 6 NF-κB recognition sites. Through transfection of porcine iliac endothelial cell line (PIEC), porcine alveolar macrophages cell line 3D4/21 and porcine primary fetal fibroblasts(CPEF), the relative luciferase expression was detected. These 6 reporter vectors, constructed in this study, could effectively report the expression of p65 subunit of porcine NF-κB, and the upregulation fold-changes were all higher than that of pNF-κB-Luc. The luciferase activities of reporter vectors A to F displayed regular difference among each other, of which reporter vector C showed the highest activities, while reporter vector F was the lowest. Compared to the commercial available NF-κB reporter vetor, those all 6 reporter vectors showed higher specificity of transcription activities. So our vector series could be used as NF-κB reporter vectors for the porcine species.

4 SPF BALB/c mice were immunized with purified classical swine fever virus (CSFV) as the antigen and mouse splenic cells were fused with SP2/0 cells. Hybridoma cells were screened by ELISA and semi-solid agar cloning, 4 hybridoma cells secreting anti-CSFV monoclonal antibodies were obtained. ELISA results showed that all of these monoclonal antibodies with high antibody titer, which could specifically react to CSFV but not cross react to other pig viruses and PK-15 cell cultures. Western blotting indicated that 3 strains of monoclonal antibodies reacted with CSFV specially. Specific green fluorescence was observed in the cell membrane by IFA. This study layed a foundation for the development of new diagnostic reagents for CSFV.

The gas production procedure was conducted to determine the effect of different concentrate to forage ratio on rumen fermentation parameter and rumen microbial synthesis of niacin,niacinamide (NAM),and nicotinic acid (NA) in vitro.Rumen fluid was collected from four Holstein adult steer with permanent rumen fistula,and three dietary substrates with different concentrate to forage ratios (30:70,50:50,70:30,DM) at an equal crude protein level of 13.5% were formulated by adjusting inclusion levels of starch,avicel and casein.The results showed that with the concentrate to forage ratios increased,the in vitro rumen pH,molar percentages of acetic and acetate to propionate were extremely significantly decreased (L,P<0.01),the ammonia-N concentration was significanthy decreased (L,P<0.05),whereas the 72 h gas production,molar percentages of propionate,butyrate and total VFA were extremely significantly increased (L,P<0.01).With the dietary concentrate to forage ratios increased,the concentrations of total niacin (nicotinic acid+niacinamide),niacinamide were increased at 24 and 48 h (L,0.05<P<0.10),while the concentration of nicotinic acid had no evident changing tendency (P>0.10).In conclusion,dietary concentrate to forage ratio had a greater effect on rumen fermentation parameters and synthesis of niacin,niacinamide,and as dietary concentrate to forage ratios increased,the concentrations of total niacin and niacinamide had an increasing tendency.

This experiment was conducted to investigate the effect of lactic acid bacteria and acidic protease solid-state fermentation on crude protein,pH,acidity and antinutritional factor content of soybean meal,cottonseed meal and rapeseed meal.The total fermentation time was 72 hours,and collected samples in every 12 hours (collected samples in every 24 hours for exterminating crude protein).The results showed that the crude protein content,and acidity of soybean meal,cottonseed and rapeseed meal were increase in different extents,and the pH was declined by lactic acid bacteria and acidic protease compound solid-state fermentation.The contents of trypsin inhibitor,lupeose and raffinose in soybean meal were remarkably declined,and the contents of tannin in cottonseed meal and the contents of ITC and OZT in rapeseed meal were declined in varying degrees.But the degradation ability of phytic acid and free gossypol was very limited.It was concluded that the lactic acid bacteria and acidic protease compound solid-state fermentation could improve the crude protein content and acidity,decrease the pH of fermentation substrate,and could remarkable decline the contents of antinutritional factor contents of soybean meal,cottonseed meal and rapeseed meal,so obviously improve the nutritional value.

This study aimed to prove up the effects of fermented green juice and xylanase on quality of rice straw silage.A control group (CON) and three treatment groups which added 2 mL/kg fermented green juice (FGJ),20 mg/kg xylanase (XYL),compound them (2 mL/kg FGJ+20 mg/kg XYL,MIX) respectively and were both set up with conventional moisture (MC1) and low moisture (MC2) of rice straw silage.Each treatment was repeated 3 times.The materials were ensiled at room temperature and opened 60 days later.The results showed that in the MC1 silages,compared with the CON group,the contents of moisture and ADF in the MIX group were significantly decreased (P<0.05),the content of AN in the FGJ、XYL and MIX groups was extremely significantly decreased (P<0.01),the content of WSC in the FGJ、XYL and MIX groups was significantly or extremely significantly increased (P<0.05;P<0.01),the content of NDF in the XYL and MIX groups was significantly decreased (P<0.05),the pH in the FGJ group was extremely significantly decreased (P<0.01).In the MC2 silages,compared with the CON group,the content of moisture in the FGJ group was significantly decreased (P<0.05),the content of WSC and DMR in the FGJ group were significantly increased (P<0.05),the contents of AN and NDF in the FGJ and MIX groups were significantly or extremely significantly decreased (P<0.05;P<0.01),the pH in the FGJ,XYL and MIX groups was extremely significantly decreased (P<0.01),the content of ADF in the MIX group was significantly decreased (P<0.05).Compared with the MC1,the pH and DMR in the MC2 were extremely significantly increased (P<0.01),the content of WSC in the MC2 was significantly increased (P<0.05),the content of AN was extremely significantly decreased (P<0.01).In conclusion,three additive had significant effect in the MC1 silages and MC2 silages,especially the effect of the mixed addition of FGJ and XYL was the best,and the MC2 silages had higher quality than the MC1 silages.

The development of buffalo milk industry in China come across the problems of small high productive herds population and not enough of high quality breed source,therefore,it is necessary to carry out dairy herd improvement (DHI) to increase dairy buffalo herd productivity.This paper reviewed the situation and problems of DHI in dairy buffalo were analyzed,and the opinion and suggestions was came up with to carry out dairy buffalo DHI in the future.

In order to investigate the effect of layer housing range model with perches on production performance,egg quality and immune function of laying hens,basing on the layer's biological characteristics and behavior habits,the study initially constructed a feeding model which consists of perches,bottom web,feeding,drinking,and laying nest.The results showed that compared with control group,the age of producing first egg of the perches model could be delayed by 3 weeks and the feed-egg ratio of laying period reduced by 9.05%.At the meantime,the average egg weight and laying rate were increased by 2.62% and 3.45%,respectively. The protein height,hunt unit and egg shape index of experiment group were significantly higher than that of control group (P<0.05),higher above 24.13%,8.03% and 6.35%,respectively.The indexes of thymus,spleen,bursa and the antibody titer of Newcastle were increased by 5.44%,3.48%,6.57% and 16.74% of experiment group than that of control group,respectively.In conclusion, the housing range model with perches could not only reduce the feed-egg ratio but also improve the egg quality and immune function of laying hens.

This experiment was conducted to study the effects of dihydromyricetin (DMY) on immune organ indexes, serum biochemical parameters and breast meat quality of Yaoshan chickens. 96 healthy Yaoshan chickens (40 days old) were selected and divided into 4 treatments for 70 d. Each treatment contained 3 replicates with 8 chickens per replicate. Chickens in group Ⅰ were fed with basal diet as control group, treatment groups Ⅱ, Ⅲ and Ⅳ received basal diets supplemented with 0.025%, 0.05% and 0.1% DMY, respectively. The results were as follows,thymus index was extremely significantly increased with supplementation of 0.05% DMY (P<0.01); spleen index and bursa index of three treatment groups were not significantly affected (P>0.05); supplementation of different doses of DMY had no significant effects on the content of albumin and total protein (P>0.05), but the 0.05% DMY could significantly reduce the GOT activity and improve the LZM activity in serum (P<0.05); DMY could significantly reduce the shear force and drip loss of breast muscle (P<0.01), but the pH, meat color and cooked meat rate were not significantly affected (P>0.05). These results indicated that supplementation of DMY could improve the development of immune organs of Yaoshan chickens. The immune function and liver cell metabolism function could also be improved, and DMY could improve the breast meat quality.

The aim of this study was to determine the adsorption and desorption of aflatoxin B₁ (AFB₁),zearalenone,T-2 toxin to montmorillonite and attapulgite in the artificial gastric juice and intestinal juice. The results showed that the adsorption equilibrium of AFB₁,zearalenone,T-2 toxin to montmorillonite and attapulgite were established within 1 h. The adsorption rates of AFB₁ and T-2 toxin to sodium montmorillonite were 96.8%,38%,respectively. Zearalenone adsorbed by montmorillonite and attapulgite were less than 10%.The pH of artificial intestinal fluid had a significant influence on adsorbing mycotoxins (P<0.05).The adsorption rate of AFB₁ to sodium montmorillonite was larger at pH 5 than pH 3 and pH 7 (P<0.05),90.1%.High adsorption efficiency of zearalenone to montmorillonite and attapulgite was achieved at pH 3.The adsorption rate of T-2 toxin to montmorillonite and attapulgite was larger at pH 7 than pH 3 and pH 5 (P<0.05).The complex compound formed by montmorillonite or attapulgite with mycotoxins has the desorption phenomena,expecially AFB₁ and zearalenone.Because of the adsorption of AFB₁,zearalenone,T-2 toxin to montmorillonite and attapulgite,they are important adsorbent.However,the desorption phenomena should not be neglected.Selection of montmorillonite or attapulgite as a feed additive should be considered their adsorption and desorption in practice.

Blood gas index in venous blood of 12 Yili horses in different stages of 1000 m trot training (motionless, after the warming up, after the game instantly, 15 min, 30 min, 60 min) were detected by i-STAT blood gas analyzer, and the differences and correlation of the blood gas index were analyzed. The results showed that before and after exercise PvCO₂, PvO₂, HCO₃, TCO₂, SvO₂ and Glu of the venous blood did not change significantly (P>0.05). Blood pH, BE and Ca²⁺ were lower immediately after the game, but after 15 min rest basic rebounded to motionless level. K⁺, Hct and Hb immediately after the game were significantly increased (P<0.05), but after 15 min rest basic back down to motionless levels. Blood pH value was highly significant positive correlation with BE and HCO₃ (P<0.01), and significantly positive correlation with the TCO₂ concentration (P<0.05), Hct, Hb were highly significant negative correlation with pH, HCO₃(P<0.01),Hct, Hb were highly significant positive correlation with K⁺(P<0.01). The analysis showed that BE, HCO₃, TCO₂, Hct and Hb on acid-base balance all had certain adjustment, Na⁺, K⁺ and Ca²⁺ on the body movement had certain adjustment.

In order to further study the distribution and adhesion of Lactobacillus plantarum in animal gastrointestinal tract,biological characteristics of Lactobacillus plantarum (LP) and recombinant Lactobacillus plantarum with GFP marker gene (Lp-GFP) had been systematically compared.The results showed that the growth curve of Lp-GFP was basically the same as Lp and Lp-GFP growth speed slightly lagging behind.The optimum growth temperature of Lp and Lp-GFP were all 37 ℃ and the optimum initial pH were all 6.5.The high temperature resistance,tolerance to artificial gastric fluid and artificial intestinal fluid,tolerance to bile salt of Lp and Lp-GFP had the same tendency.There was no significant difference on anti-bacteria character of Lp and Lp-GFP (P>0.05).The sensitivity to drugs of Lp and Lp-GFP had no differences except erythromycin.In conclusion,recombination has no significant effect on biological characteristics of Lactobacillus plantarum.These results laid an important foundation for cultivation and mechanism study of reconstructed Lactobacillus plantarum.

This experiment was aimed to study the inhibition effects of nitric oxide (NO) on Brucella in the process of Brucella infecting mice macrophage, and the interaction relations between NO and asymmetry dimethylarginine (ADMA). Brucella vaccine strain M5 was used to infect mice macrophags, then NO production and ADMA levels were determined by the Griess reagent method and ELISA. CFU were counted for each infection period. Results suggest that the NO production of macrophages rose when they were infected by Brucella, and the extracellular release quantity was significanty higher than the intracellular (P<0.05). Compared with the control group, the differences were significant (P<0.05). But the ADMA levels showed the opposite trend with NO content. There were extremely significant differences (P<0.01) compared with the control group. Combined with CFU count results, it showed that the inhibition effects of NO on Brucella only occurred at the early stage of the infection (the former 12 h), but not at the later period (the last 12 h). The ADMA had certain inhibitory effect on NO generated in the process of Brucella infecting mice macrophage.

The objective of this experiment was to study the effect of Astragalus polysaccharin (APS) on adhesion of peripheral lymphocyte (PLC) and jugular vein endothelial cells (JVEC) in broiler. The JVEC and PLC of broiler were treated independently with four doses of APS (0,200,600 and 1000 μg/mL) and the adhesion changes of them were observed subsequently. Also the two types of cells were incubated with interleukin-1 (IL-1,1000 U/mL) or tumor necrosis factor-α (TNF-α,1000 U/mL) to investigate the effect of APS on intercellular adhesion and the regulatory mechanism of APS to intercellular cell adhesion molecule-1 (ICAM-1) and T-lymphocyte subsets CD4⁺.The results showed that the PLC-JVEC adhesion increased significantly when the JVEC was incubated with APS (600 μg/mL) (P<0.05).The same significant increment of intercellular adhesion was observed when the JVEC was incubated with the combination of IL-1 and APS (1000 μg/mL),or the PLC was incubated with the combination of IL-1 and APS (200,600 and 1000 μg/mL) (P<0.05).Compared with the control group,the PLC-JVEC adhesion increased significantly when the JVEC was incubated with the combination of TNF-α (P<0.05).Compared with the control group,the values of ICAM-1 and percentage of positive cell (CD4⁺) increased significantly with the combined treatment of IL-1 and high dose of APS (1000 μg/mL) (P<0.05).In conclusion,the dose of APS affected its regulation on PLC-JVEC adhesion. Under this experimental conditions,both the medium and high doses of APS (600 and 1000 μg/mL)could increase the PLC-JVEC adhesion alone or with the combination of IL-1 and TNF-α.At the same time,the high dose of APS (1000 μg/mL) could exert the immunomodulatory by changing the secretion (expression) of ICAM-1 and CD4⁺.

This paper studied the character of N-acetyl-β-D-glucosaminidase (NAGase) from the sperm of Putian Black pig. The purification steps involved the following procedures: Ammonium sulfate precipitation, anion-exchange chromatography on DEAE Sepharose Fast Flow, gel filtration chromatography on Sephadex G100. The purified enzyme preparation was homogeneous judged by polyacrylamide gel electrophoresis. It was found that the specific activity of the enzyme was 1561.42 U/mg. The enzyme molecular weight was estimated as 58 ku. The optimal pH value was 5.6 and the optimal temperature was 45 ℃. The enzyme was stable in the pH ranges of 3.6 to 7.8. The enzyme followed typical Michaelis-Menten kinetics for the hydrolysis of pNP-GlcNAc and the K_m and V_m values were determined to be 0.82 mmol/L and 39.23 μmol/(L·min), respectively. The activation energy of the enzyme for hydrolysis of pNP-GlcNAc was 27.30 kJ/mol. The effects of some metal ions on the enzyme activity were determined, Na⁺, K⁺, Mg²⁺, Ca²⁺ had no effects on the enzyme activity, while Zn²⁺, Cu²⁺, Pb²⁺ had inhibitory effect on the enzyme activity.

Cytochrome P450 2E1 (CYP2E1) is one of the body's metabolite of drugs and low molecular weight compounds mainly enzymes, and the increasing of activity or expression of CYP2E1 is closely related with the clinical variety of diseases. This paper reviews the research progress of protein structure, function, positioning mechanism and other aspects of genetic polymorphisms of CYP2E1 at home and abroad. Thereby the paper could lay the foundation for further exploration of the biological effects of CYP2E1 and its relationship with clinical disease.

The aim of this investigation was to study the effect of brilliant cresyl blue (BCB) staining on IVM apoptosis in cow, COCs without BCB staining was considered as control group, and COCs stained with BCB in 26 μmol/L for 90 min as treatment group before mature culture. Treated oocytes were divided into BCB^-(colourless cytoplasm) and BCB⁺(coloured cytoplasm). After IVM, to assess the developmental potential of the oocytes. The results showed that, the BCB⁺ oocytes had a significantly higher proportion of mature rate (80.92%) than control group (60.00%) and BCB^-oocytes (51.61%)(P<0.05);The BCB⁺ oocytes yielded a significantly lower apoptosis rate (6.50%) than that in control (28.21%) and BCB^- oocytes (39.06%)(P<0.05). These results suggested that the staining of cow cumulus-oocyte complexes with BCB before in vitro maturation could be used to select developmentally competent oocytes.

To explore the influence of recombinant chicken interferon-α/chicken interleukin-2 fusion protein (rChIFN-α-Linker-ChIL-2 protein, recombinant fusion protein) on the percentage content of peripheral T lymphocyte subsets (PBLC) of SPF chicken,the flow cytometry was used to detected the percentage contents of CD3⁺CD4⁺, CD3⁺CD8⁺ and the CD4⁺/CD8⁺ ratio in PBLC at different days post injection in the 14-day-old SPF chicken injected with rChIFN-α-Linker-ChIL-2 protein (group 2) and recombinant chicken interferon α (rChIFN-α) protein (group 3). The results indicated that the recombinant fusion protein and rChIFN-α protein could increase the percentage content of CD3⁺CD4⁺ T lymphocyte, decrease the percentage content of CD3⁺CD8⁺ T lymphocyte, improve CD4⁺ /CD8⁺ T lymphocyte ratio during 3 to 14 d post injection. The percentage contents of CD3⁺CD4⁺, CD3⁺CD8⁺ and CD4⁺/CD8⁺ ratio in chickens of group 2 were significantly different from the PBS control group (group 1) (P<0.01), and were notably different from group 3 during 3 to 7 d post injection (P<0.05). These results showed that both the rChIFN-α-Linker-ChIL-2 protein and rChIFN-α protein could affect the percentage contents of lymphocyte subsets, improve CD4⁺/CD8⁺ ratio, enhance the cellular immune function in chicken. The influences of rChIFN-α-Linker-ChIL-2 protein on the percentage contents of CD3⁺CD4⁺ and CD3⁺CD8⁺ T lymphocyte, and CD4⁺/CD8⁺ value were notable higher than rChIFN-α, which suggested that recombinant fusion protein play a synergistic role of interferon alpha and interleukin-2 on the cellular immune function by this pathway in chicken.

Objective of the study was to develop a preparation treatment for salbutamol in feed by enzyme-linked immunosorbent assay (ELISA).The assay optimized the influence factor of the preparation and used to extract salbutamol residues in feed.The results showed that the inhibition concentration (IC₅₀) for the developed ELISA was 0.606 ng/mL with the detection range of 0.221 to 1.658 ng/mL and R² was 0.9998.After optimization,pH 7.5,0.06 mol/L PBS with the dilution ratio of 10 was selected to extract salbutamol residues in feed and the limit of detection (LOD) was 5.0 ng/mL in feed.The recoveries ranged from 77% to 110% for the spiked samples (5 and 10 μg/kg),and RSD was less than 8%.

This experiment was conducted to use four defined transcription factors that induced sertoli cells of transgenic mice to induced pluripotent stem (iPS) cells. Sertoli cells of transgenic mice were infected with retrovirus vectors that lead to the expression of Oct4, Sox2, c-Myc and Klf4, green fluorescence protein (GFP) was also transduced to monitor the infection efficiency as well as an indicator of exogenous gene in part of induction experiments. As early as 3 days post-infection, the cells started to express GFP and morphologically differ from uninfected sertoli cells. As the culture prolonged, the single cell was getting smaller and more cells aggregating to form colonies. At 16 days, the GFP were silenced and compact and round-shaped colonies formed. Alkaline phosphatase staining showed that iPS cells in the undifferentiated condition, reverse transcription-polymerase chain reaction (RT-PCR) showed that iPS cells could express the specific genes of embryonic stem cells, and could form embryoid body (EBs) which could differentiate into myriad cell types of three layer.

The aim of this study was to investigate the correlativity between expression of microRNA-21 (miR-21) and transforming growth factor beta-induced (TGFBI) in skeletal muscles of Landrace pigs. Comparative analysis was conducted between miR-21 and TGFBI in different tissues and in skeletal muscles at 28 developmental stages by quantitative PCR (qPCR). The results showed that the miR-21 was expressed without specificity in different tissues and fluctuated in skeletal muscles at developmental stages; TGFBI was expressed highly in skeletal muscles at embryonic periods and has different expression levels during developmental periods of skeletal muscles. The correlativity analysis showed that there was significantly negative correlativity between miR-21 and TGFBI in embryonic period of skeletal muscle development, which indicated that TGFBI was regulated by miR-21 in pig skeletal muscle development.

In this study, Guizhou White goat, Qianbei Ma goat, Guizhou Black goat from Guizhou native goat breeds were selected for testing subjects, the technology of DNA pools and the direct sequencing of PCR products were used to detect the single nucleotide polymorphism (SNP) of follicle stimulating hormone receptor (FSHR) gene in goat. Moreover, through bioinformatics software to analyze the impact of secondary structure of RNA. The results showed that three SNPs (C126T,C1246A and A1412C) were found, among these SNPs, C126T and C1246A lay in the exon 1 and exon 10, which were sense mutation and the A1412C lay in intron. Bioinformatics analysis that the two SNPs in the exon 1 and exon 10 lead to the alteration of secondary structure of mRNA.

One pair of special primer was designed according to the conservative region of tyrosinase (TRY) gene in Mustela putorius furo, Ailuropoda melanoleuca, Canis lups familiaris and Felis catus deposited in GenBank database. The partial sequence of exon 1 from Neovison vison(Black), Neovison vison(Regal White) and Neovison vison (Palomino) mink were obtained by PCR amplification and cloning sequencing. The results showed that the length of TYR gene in mink with three kinds of coat color was 796 bp, comprising partial exon 1(795 bp) and 5'UTR (1 bp). Five singleton variable sites were detected at position 34 (A/G), 138 (T/A), 248(T/C), 545(A/G) and 765(C/A), including four missense mutations and one frameshift mutation. The accession numbers were KJ198847(Black), KJ658343(Regal White) and KJ152769 (Palomino), respectively. The nucleotide similarity between mink and ferret, giant panda, dog, cat, rabbit, pig, alpaca, goat, sheep, cattle, human, rhesus monkey, chimpanzee, chicken and japanese quail was 72.8% to 98.6%, and the amino acid similarity was 75.0% to 98.1%. The phylogenetic tree constructed by TYR amino acid indicated that the evolution distance of mink was the most homogeneous to ferret beloning to Mustelidae. This study would lay the bioinformatics foundation for further investigating correlations between polymorphisms of TYR gene and coat color phenotype in mink.

The objective of this study was to clone the sequence of duck type Ⅱ gonadotropin releasing hormone receptor (GnRHR-Ⅱ) gene, and to analyze the association of its expression levels with reproduction trait heterosis. RT-PCR method was used to clone GnRHR-Ⅱ gene fragments, and the expression levels during early laying days and laying fastigium period in Gaoyou duck, Jinding duck and crossed populations were tested by Real-time PCR method. Sequencing and homology analysis showed that the cloned duck GnRHR cDNA was about 500 bp in length, and it belonged to the avian type Ⅱ GnRHR isoforms. It was 96% identical to the cGnRHR-Ⅱ sequence reported in domestic chicken. From the early laying days to laying fastigium period, expression levels of GnRHR-Ⅱ gene in pituitary increased for Jinding duck, Gaoyou duck and crossed populations, with the highest expression levels in Jinding×Gaoyou hybrid group (P<0.01) . The expression levels of GnRHR-Ⅱ gene in hypothalamus also increased for Jinding duck, Gaoyou duck, however, it decreased in two crossed populations. The expression levels of GnRHR-Ⅱ gene in ovary were significantly higher for Jinding duck and two crossed populations than Gaoyou duck (P<0.01), but it decreased during laying fastigium period. Compared to Jinding duck and Gaoyou duck breeds, the two crossed populations had positive heterosis in 42-week production and 42-week egg fertilization percent, and negative heterosis in the first laying age and 42-week fertilized egg hatching rate traits. It suggested that the short-lived higher expression of ovary GnRHR-Ⅱ gene before early laying days be correlated with heterosis of the first laying age and 42-week production traits, and its higher expression levels in pituitary could aid to keep laying fastigium.

In this study, the relationship of MyoD gene polymorphisms and meat yield was studied by molecular biology methods. The aim was to provide scientific basis for improving meat performance of Altay sheep by molecular marker assisted breeding methods. One year old ram of Altay (n=120) were chosen in representative area to jugular vein blood, MyoD gene 5' control region and the part of the exon 1 pieces were selected to detect genetic polymorphism by PCR-SSCP, and the effects on weight traits were analyzed. It was used to research the relationship between MyoD genetic polymorphisms and meat performance. There was no polymorphism in MyoD-P1 gene locus (5' flanking region );There was polymorphism in MyoD-P2 gene locus (326-542 bp in exon 1), the primer MyoD-P2 gene locus existence three kinds of genotypes of AA, AB and BB. Sequencing results showed that there was a base mutation A→G in location 408 bp in MyoD gene exon 1. The mutation causesd its coding changing from histidine into alanine. By correlation analysis it was found that the individual weight of AA genotype were much higher than BB genotype (P<0.05). Mutational site exon 1 of MyoD gene might be the important site of Altay sheep's meat performance, MyoD gene might be the candidate genes of Altay sheep's meat performance.

microRNAs (miRNAs) play a key role in cell proliferation,differentiation and apoptosis. miRNA involved in the regulation of mammary gland development in cell proliferation and differentiation. In this experiment, we used mice as an animal model to study the role of miR-138 in mouse mammary gland development and lactation. Maternal tail vein injection of miR-138 inhibitors, pups weighing method applied to detect the changes of milk yield;electron microscopy techniques was used to observe ultrastructure changes in mammary gland tissue. The results showed that after intravenous injection of miR-138 inhibitor, female mouse increased maternal mammary epithelial cells, increasing the amount of milk secretion,ultrastructure of cells found in the metabolic activity,collect female mouse milk after miR-138 inhibitor, detect lactose and casein of the milk were all increased. The results suggested that the inhibition of miR-138 could stimulate mammary epithelial cell proliferation, increase the secretion of milk, and regulation of the ingredient of the milk.

This experiment was conducted to study the slaughter of the relationship between meat traits and genotypes. This study selected Red cattles as the experimental group, using PCR-SSCP method of detection of bovine IGFBP6 gene exon 2 polymorphism analysis with the slaughter of the relationship between meat traits by the GLM model of the SPSS 13.0 statistical software to analysis. The results showed that IGFBP6 gene had a polymorphic site A180G in exon 2, at the same time had three genotypes of AA, AB and BB. Between the three genetypes,AB and BB genetypes were very significant higher than AA genetype in net meat weight (P<0.01),and were significant higher than AA genetype in eye muscle area and bone weigh (P<0.05), while in the ante-mortem live weight, carcass weight, kidney fat weight and rib fat weight in the three genotypes were no significant differences (P>0.05). These results indicated that the slaughter of meat was associated with certain traits, accessibility for the future protection of breeding species of Red cattle to provide a theoretical basis, and Red cattle of beef with traits improve the delivery methods and theoretical on the guidance.

Long noncoding RNA (lncRNA) is a class of RNA being more than 200 nucleotides in length, which also has no protein coding function. The researches suggest that lncRNA can mediate genomic imprinting, transcriptional activation and interference, post transcriptional regulation, chromosome dosage compensation effect, developmental regulation and many other biological processes. The detection and classification, biological function of lncRNAs, and the research progress of lncRNAs associated with muscle development would be reviewed in this paper.

China is the largest country in rabbit industry in the world, but basic research in rabbit genetics and breeding is the serious lag actually, which makes progress a little only in recent years. The latest research progress on domestic rabbit genetics and breeding in 2013 were reviewed in the present paper so as to provide references for rabbit breeders. Firstly in aspect of traditional breeding, there were some introduction or investigation about genetic resources e.g. Jiuyishan rabbit, and researches on screening the best hybridized combination in meat or Rex rabbits. Secondly as for molecular marker assisted breeding, it focused on the polymorphisms and expressions of candidate genes related to rabbit hair (hair color, hair follicle development), production (growth, slaughter and meat quality) and disease-resistant traits, furthermore several functional genes and potential molecular markers have been found. Finally, there were certain studies on techniques for inducing oestrus in females and improvement of semen dilution and preservation ways in males. Additionally,research of rabbit muscle proteomics was a bright spot in 2013. These research findings will favour the development of rabbit industry in China.

The present study was aimed to determine the efficacy of Rhizoma Dryopteris Crassirhizomae (RDC) soluble powder in artificial infection with infectious bursal disease virus (IBDV). Firstly, SPF chicken were inoculated with the IBDV LX strain in order to induce virus infectious model, and then the chicken of high,middle,low 3 groups were administered orally with 198, 99 and 50 mg/kg RDC soluble powder while the chicken were administered with Korean RDC oral solution or Astragalus polysaccharide for 5 days as the control group. Meanwhile, chicken inoculated with IBDV and without IBDV were regarded as the infectious control and healthy control groups. Body weight gain, cure rate, survival rate and immune organ index were employed to determine the efficacies. After experimental infection with IBDV, an extremely significant increase of body weight gain was found in the RDC soluble powder middle and low dose groups compared to Korean RDC group (P<0.01).RDC soluble powder low dose group was significantly higher than RDC soluber powder middle dose group and Astragalus polysacharide group (P<0.05), however, no significant difference was found between RDC soluble powder middle dose group and Astragalus polysaccharide group (P>0.05). With respect to the survival rate, 80% chicken recovered from IBDV infection in the RDC soluble powder high,middle,low dose groups, which were significantly higher compared to Korean RDC group (66.7%) and Astragalus polysaccharide group (53.3%) (P<0.05). In terms of immune organ index, the significant increase of thymus and spleens indexes was found in RDC soluble powder middle and low does groups compared to that of the infectious control group (P<0.05). However, no significant difference of bursal index was found among RDC soluble powder high,middle and low dose groups, Korean RDC group and Astragalus polysaccharide group (P>0.05).In summary, chicken administered orally with 50 mg/kg RDC soluble powder for 5 days would contribute to the immune activation by improvement of thymus and spleen indexes, leading to high survival rate against the artificially challenge with IBDV. With respect to survival rate and recovery protection, it was higher than that of Astragalus polysaccharide.

Ninty-four milk samples of clinical mastitis and seventy milk samples of subclinical mastitis were collected from part of the dairy farms (pastures,farming community) in Ningxia. Four Leclercia adcarboxylata (L.adcarboxylata) were isolated and identified, including clinical mastitis 1 strain and subclinical mastitis 3 strains. Drug sensitive test results showed that four strains of L.adcarboxylata were all susceptible to streptomycin, aminoglycosides, quinolones and cephalosporins, but they had different degrees of resistance to other drugs such as sulfonamides and lincosamides.

To investigate the effect of Tonifying Qi-Nourishing Spleen herbs on small intestinal mucosa in diarrheal piglets, antibiotics and Chinese herbal medicine were adopted to heal diarrheal piglets. Piglets were sacrificed by bloodletting in veins, dissected peritoneal cavity, and gathered small intestinal mucosa abacterially. Then, the changes of histopathology and cellular structures in small intestinal mucosa were observed by using pathological and electron microscopic sections. The results showed that small intestinal mucosa had no obvious pathological changes except slight hemorrhage in Chinese herbal medicine group. Nevertheless, there were more pathological changes in antibiotics group, for instance, small intestinal mucosa had desquamated with extensive hyperemia and hemorrhage concomitantly; epithelial cells were degenerated and necrotic;eosinophilic granulocyte displayed cellular infiltration, which included inflammatory cell infiltrate principally as lymphocyte;microvillus had desquamated and arranged indiscriminately; mucous membrane cells were damaged seriously and mitochondria presented structural abnormalities. We concluded that the therapy of using antibiotics in diarrheal piglets might cause pathological changes and functional impairment in small intestinal mucosa, and using Chinese herbal medicine might be beneficial for plerosis of small intestinal mucosa, which would provide the theoretical foundation to cure diarrhea and improve growth performance of piglets.

Foot-and-mouth disease virus (FMDV) virus-like particles (VLPs) are particles lack virus-specific genetic materials (RNA), which are self-assembled by four structural proteins and are similar to natural virus particles in morphology. Also, VLPs have better biological activity and immunogenicity. This review focuses on the assembly of FMDV capsid, advances on VLPs, the influencing factors of forming VLPs, and the prospect of the development and application of FMDV VLPs vaccines.

The purpose of this study was to understand the pollution condition,popular serotype and drug resistance of Salmonella in farmer's market and supermarket, in Shanghai. A total of 240 chicken samples and 80 chickens' cloaca swabs were obtained from market in 2012. Among them, 70 (21.9%) were positive for Salmonella. Of the 11 serotypes in 70 Salmonella isolates, S.enteritidis (47.1%), S.indiana (17.1%), S.typhimurium (8.6%) and S.derby (8.6%) were prevalent.Drug susceptibility test was conducted among the Salmonella isolates using the Kirby-Bauer disk diffusion method. The highest level of resistance was found for nalidixie acid (75.7%), followed by sulfisoxazole (60.0%), streptomycin (60.0%) and ampicillin (51.4%). No resistance to imipenem was observed in any isolate examined.The isolated chicken Salmonella strains had severe resistance to most drugs. 40 strains of Salmonella were millti-drug resistant,the most resistart strain was resistant to 14 drugs.This study had important practical significance for prevention and control and rational drug use of salmonellosis.

To provide the references for application of Lactobacillus, this review summarizes the regulation of Lactobacillus on the intestine from four aspects including role on surface of intestine, role on the immune system, role of balancing microbial flora in the intestine and regulation on metabolism. Application of Lactobacillus in animals are also reviewed in this paper. The issue described in this paper will focus on mechanism of action and application effect of Lactobacillus.

The study was aimed to observe the allergic reaction of ivermectin microemulsion injection on animals. Active systemic allergy test and passive cutaneous anaphylaxis test were carried out to study the allergic reaction of ivermectin microemulsion injection on Wistar rat and guinea pig, respectively, according to . The results showed that the allergic reaction of ivermectin microemulsion injection on Wistar rat and guinea pig were both negative. The study provided experiment evidences for the safety evaluation of ivermectin microemulsion.

The aim of the experiment was to research the safety of cefquinome sulfate intramammary infusion for lactating cows.12 healthy lactating cows (6 were primiparous and 6 were multiparous) were selected to inject 1 peice of cefquinome sulfate intramammary infusion (dose:8 g:75 mg/piece) into four dairy regions of each cow for 3 times,and its interval of the administration was 12 h,and the evaluation indicators included the differences of clinical manifestation grade,daily yielding,somatic cells number and the changes of pathogen in dairy regions in milk before and after administration.The results showed that there were no significant differences in indicators all above before or after the treatment (P>0.05),which suggested that cefquinome sulfate intramammary infusion was safe to lactating cows when administrated according to the dosing regimen above.

To analyze the virulence genes distribution of Listeria monocytogenes(L.monocytogenes, LM) isolated from commercial duck wings in Luoyang. 8 virulence genes (inlA, inlB, virR, mprF, dltA, dltB, dltC, dltD) from 11 LM isolated strains were detected using PCR method. The results showed that there existed 3 isolated strains with virulence gene-deleted. Detected ratioes for dltA, dltC and mprF were 90.9% (10/11), 81.8% (9/11) and 81.8% (9/11),respectively. The data indicated that these virulence genes were common for LM isolated from commercial duck wings in Luoyang, and some measures were taken to strengthen the surveillance of LM for the potential threat to commercial duck wings safety.

Canine third eyelid hyperplasia was an ophthalmopathy, because of gland hypertrophy and other reasons with the nictitating membrane together from the eye turned outwards and induced inflammation, most of which were treated by excision method and embedding method. In order to research the advantages and disadvantages of excision method and embedding method for canine third eyelid hyperplasia, we used 20 dogs which both eyes had third eyelid hyperplasia and not suffer from other diseases, and divided into excision group and embedding group to do operation and track observation. The results showed that the excision method had simple operation,recovery time was short and other advantages, but the dogs would have the sequelae, dry eye disease after curement, there was great harm to their health. While embedding method's operation was complex, recovery time was longer, but as long as the operators mastered skilled operation, the cure effect could also be achieved, and would not have other bad effect on the dog. Although the two methods had their advantages and disadvantages, the disadvantages of excision method outweighed its advantages, sequelae would bring dogs more inconvenience, and embedding method had more advantages than disadvantages, as long as they could master the operation method,do well postoperative nurse, and overcome the drawbacks. In future clinical practice, we should try to persuade the dog's owner to use embedding method to treat the third eyelid hyperplasia.