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20 June 2014, Volume 41 Issue 6
Construction of Follicle-specific Expression Vector of Spider Dragline Silk Protein and Screening of the Transgenic Cell Lines
WANG Hai-tao,MENG Fan-hua,FANG Jun,ZHOU Huan-min
2014, 41(6):  1-6. 
Abstract ( 241 )  
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This study aimed at getting transgenic cell lines which could express spider dragline silk protein in the hair follicles specifically. By synthesizing spider dragline silk protein gene monomer artificially according to the published cDNA fragments of Nephila clavata spider, got the diploid (2S) by doubling, then constructed hair follicles specific expression vector pK-2S using pEGFP-N1 as original vector and transfected Xinjiang fine wool merino sheep fetal skin fibroblasts. Make use of PCR to detect positive clones at DNA and RNA level.The sequencing result showed correct sequence after gene synthesis,after enzyme digestion of the vector, we got the correct band while we could amplify target gene from the genom and cDNA of positive clones. This study established cell lines of transgenic spider dragline silk like protein gene successfully while the gene had expressed at RNA level. Our study laid the foundation of making the spider dragline silk gene specifically expressed in the hair follicles of the sheep by transgenic technology.
Effects of microRNA-152 on Milk Protein Synthesis in Bovine Mammary Epithelial Cells
WANG Jie, BIAN Yan-jie, WANG Zhuo-ran, LI Dan, GAO Xue-jun, LI Qing-zhang, WANG Chun-mei
2014, 41(6):  6-11. 
Abstract ( 205 )  
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In order to study the effect of microRNA-152 (miR-152) on milk protein synthesis in dairy cow mammary epithelial cells, liposome transfection was used to change the expression of miR-152 in bovine mammary epithelial cells. The influence of miR-152 on the cell proliferation and lactation was detected using Real-time PCR, Western blotting, cell viability analysis and other techniques. Results indicated that when miR-152 was inhibited, SOCS3 protein expression was increased, signaling pathway factors p-STAT5,p-mTOR,S6K1 and cyclinD1 protein expression were decreased, cell proliferation was decreased, β-casein secretion was decreased. The results suggested that miR-152 probably regulated bovine mammary epithelial cells by suppressing expression of SOCS3. And miR-152 could increase milk protein synthesis and cells proliferation by regulating signal transductions of p-STAT5,p-mTOR,S6K1 and cyclinD1 in mammary epithelial cells.
Regulation Effect of Lipopolysaccharide on Cyclooxygenase-2 Gene Expression in Bovine Oviduct Epithelial Cells
WANG Xiu-ming, MAO Wei, CAO Jin-shan, DONG Zhi-heng, LIU Bo
2014, 41(6):  11-17. 
Abstract ( 190 )  
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Development and Application of TaqMan Fluorescence Quantitative PCR Assay for Detecting Classical Swine Fever Virus
YUE Feng, ZHU Yan-ping, YIN Mei, WANG Xuan-nian
2014, 41(6):  18-22. 
Abstract ( 248 )  
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According to the conserved sequence of 5′nontranslated region (5′NTR)of classical swine fever virus (CSFV) in GenBank, a pair of primers and a TaqMan probe were designed. The recombinant plasmid pGEM-CSFV containing the complete gene of CSFV was as a standard control to establish fluorescence quantitative PCR standard curve. Specificity, sensitivity and reproducibility of the method were determined. CSFV load in plasma was measured at different time points after experimental infection of CSFV in pigs. The results showed that standard curve established had a good linear relationship between threshold cycle and template concentration, R2=0.999. The assay had high sensitivity and could detect 1×101copies/μL. The specificity of the method was high, there was no cross reaction between CSFV and other pathogens containing porcine circovirus 2, porcine parvovirus, porcine pseudorabies virus and porcine reproductive and respiratory syndrome virus. The method showed good reproducibility, there was a coefficient of variations less than 2% for both inter-assay and intra-assay. CSFV load in plasma increased to peak value at 7 days after experimental infection of CSFV in swine and gradually decreased later. These results indicated that the developed TaqMan fluorescence quantitative PCR assay had the advantages of good specificity, sensitivity and reproducibility. It provided a basis for quantification analysis and clinical diagnosis of CSFV.
Clone and Expression Analysis of AdipoQ Gene in Tibetan Goat
LI Xue-mei, XUE Ke, ZHONG Tao, LI Li, WANG Lin-jie, ZHANG Hong-ping
2014, 41(6):  23-28. 
Abstract ( 142 )  
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In order to study the expression pattern and functions of AdipoQ, we used the bioinformatics and comparative genomics methodologies to clone the Tibetan goat AdipoQ gene. Semi-quantative RT-PCR and fluorescence quantification PCR were performed to analyse the AdipoQ mRNA expression pattern and relative expressions in three different adipose tissues of Tibetan goat. The results showed that the AdipoQ gene in Tibetan goat had high identity with the orthologous gene in other mammals. We found that AdipoQ was highly expressed in adipose tissue, and weakly expressed in skeletal muscle and rumen, and no expression was detected in other tissues. The expression of AdipoQ in perirenal fat tissue was higher than those in subcutaneous fat tissue and mesenteric fat tissue, but the expression difference was not significant (P>0.05). In this study, we obtained the sequence and tissue expressions of AdipoQ gene which laid a foundation for further studying its functions in Tibetan goat.
Cloning and Prokaryotes Expression of Porcine Interferon-α and Interleukin-18 Fusion Gene and Detection of its Antiviral Activity
ZHANG Pan-pan, HE Jing, KANG Yin-feng, XIANG Bin, REN Tao
2014, 41(6):  29-33. 
Abstract ( 144 )  
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In order to explore the antiviral effect of porcine interferon-α and interleukin-18 fusion gene (poIFN-α-IL-18), poIFN-α-IL-18 was amplified from pMD18-T/IFN-α and pMD18-T/IL-18 by using fusion-PCR in this study. The recombinant expression plasmid pET-28b/IFNα-IL18 was constructed by inserting poIFNα-IL18 into vector pET-28b(+) separately. SDS-PAGE was used for analyzing the protein after induced by IPTG. The protein was purified by Ni2+-NTA. Antiviral activities of the purified protein were tested by cytopathogenic effect inhibition assay. Results showed that the recombinant expression plasmid pET-28b/IFNα-IL18 was successfully constructed. SDS-PAGE could detect the protein bands about 37 ku. After Ni2+-NTA affinity chromatography purification, the activity of purified fusion protein against astrovirus was better than the single protein. Antiviral activities of the purified proteins, poIFN-α-IL-18, poIFN-α and poIL-18, were about 1.03×105,1.28×104 and 0.11×104 U/mg.
Construction and Identification of Bovine Myostatin Gene Targeting Vector
ZHU He-ping, WU Kai-feng, SU Xiao-hu, WANG Yi-chao, ZHOU Huan-min, ZHAO Rui-yuan, ZHANG Yan-ru
2014, 41(6):  34-37. 
Abstract ( 155 )  
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The objective of this study was to construct a targeting vector for knocking-out bovine myostatin (MSTN) gene. Based on the bovine MSTN gene sequence, we chose third exon about 600 bp as a target site. The homologous arms of bovine MSTN gene were amplified from the genomic DNA, which were about 4.4 and 1.4 kb in length, respectively. The two fragments were cloned into pPNTⅢ vector and located at both sides of the neomycin resistance gene to construct targeting vector MSTN-KO-pPNTⅢ. By sequence analysis and restricted endonucleases digestion, the 1.4 kb short arm and 4.4 kb long arm were detected in the targeting vector. In conclusion, the replacement targeting vector for bovine MSTN gene had been successfully constructed.
Synthesis of Complete Antigen of Enrofloxacin and Preparation of Rabbit Polyclonal Antiserum
LIU Li,ZHANG Jia-he,YUAN Wei-feng,MENG Ting
2014, 41(6):  38-41. 
Abstract ( 239 )  
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The purpose of this paper was to synthesize complete antigen of enrofloxacin (EFLX),and to prepare the rabbit polyclonalantibody.Ovalbumin (OVA) and bovine serum albumin (BSA) were used as protein carriers to synthesize small molecule drugs EFLX complete antigens by active ester method.The conjugates formed in this way was EFLX-BSA and EFLX-OVA,the serum titer was detected by use of EFLX-BSA immunogen and EFLX-OVA coating antigen by ELISA.The results showed that the serum titer detection result of three rabbits were all above 1∶51200,the second were the highest of all,its result was 1∶204800 and the sensitive was good.The result of indirect competitive ELISA indicated that this method had a linear detection range from 37.52 to 302.31 ng/mL (R2=0.9838),with IC50 values of 169.91 ng/mL.In this paper,the immunogen and coating antigen were successfully synthesized,and prepared the good sensetive of rabbit poloclonal antibody from the serum. This experiment obtained high antibody titer,which could lay the foundation for the development of the ELISA kit and colloidal gold test strip.
Sequence Variation in Mitochondrial nad5 Gene between Syphacia obvelate and Aspiculuris tetraptera
YANG Yan-chuan, LI Zuo-lei, ZHANG Nian-zhang, LIU Guang-xue, ZHU Xing-quan, SUN Xiao-lin
2014, 41(6):  42-45. 
Abstract ( 264 )  
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The objective of this study was to examine sequence variation in mitochondrial NADH dehydrogenase subunit 5 gene (nad5) between and within Aspiculuris tetraptera (A. tetraptera) and Syphacia obvelate (S. obvelate) collected from mice in Lanzhou, China. 42 pinworm samples were collected from mice in two laboratory animal centers in Lanzhou, Gansu province, and their partial nad5 sequences were amplified by PCR. Sequence analysis revealed that the nad5 sequences of A. tetraptera and S. obvelate were 865 and 753 bp, respectively. Sequence similarity in the nad5 sequences within each of the two pinworms were 99.60% and 100%, while the inter-specific sequence difference was 53.64% between A. tetraptera and S. obvelate. These findings demonstrated that nad5 sequences were quite conserved within mouse pinworms, but were significantly different among different pinworm species, indicating that nad5 sequences could be used as the genetic marker for the differentiation of different pinworms.
Establishment of SYBR Green Ⅰ Real-time Fluorescent Quantitation PCR for Detection of Gallinacin-1
LI Yan, WANG Yuan-xin, SUI Xin, ZHENG Shi-min
2014, 41(6):  46-51. 
Abstract ( 161 )  
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The chicken beta-defensin-1 (Gallinacin-1, Gal-1) was regarded as the research object. According to the chicken β-actin and Gal-1 genes sequences published in GenBank, the recombinant plasmids were constructed for the standard, the study successfully established Real-time fluorescence quantitative PCR method for detecting the expression of Gal-1 mRNA in chicken, and tested the sensitivity, repetitiveness and specificity. The results showed that this method was characterized by rapidity, high-flux, wide linear range, good specificity, high sensitivity, and so on. So this method would provide the basis for making further efforts for quantitative research on the expression of Gal-1 mRNA and laying technical foundations for research on the related diseases.
Establishment of Plaque Purification for Newcastle Disease Virus using DF-1 Cell
2014, 41(6):  51-54. 
Abstract ( 293 )  
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In order to establish a new cell culture system for the purification of Newcastle disease virus (NDV),NDV was plaque purified by using the chicken embryo fibroblast cell line (DF-1). The results showed that,compared to the common basal media (DMEM) used in cell culture, DMEM/F12 with 15% fetal bovine serum was better. The optimized purification conditions were as follows, 3×105 DF-1 cells per well were seeded into 6 well plates for 24 h, then infected with NDV for 1.5 h. After removing all virus from each plate, the first agarose overlay was added and incubated for 48 h before the operating of the second agarose overlay. After purification, the homologies of F and HN genes between the purified virus and the original virus were 100%, respectively, which indicated no mutation was found during the purification and screening. By using this method, it would take only one week to obtain the purified virus with the simply manipulability, high repeatability and efficiency.
Prokaryotic Expression and Identification of the S1 Gene Main Antigen Domain of Avian Infectious Bronchitis Virus
ZHU Yan-ping, GUO Dong-guang, YUE Feng, YANG Yuan, LI Chong, WANG Ai-guo, LI Bo-wen, RUAN Tao, KONG Ling-yun, WANG Xuan-nian
2014, 41(6):  55-59. 
Abstract ( 218 )  
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Avian infectious bronchitis virus (IBV) S1 gene was the object in this study and a pair of primers was designed for amplifying the S1 main antigen fragment of IBV, which was composed of 198 bp. And then it was sub-cloned into prokaryotic expression plasmid pGEX-6P-1. The recombinant plasmid pGEX-6P-1-S1 was transformed into E.coli Rosetta (DE3) and induced by IPTG. SDS-PAGE analysis showed a specific protein band with a molecular weight 33.0 ku, according to our study anticipation. The express production was inclusion body when it was identified. Western blotting indicated that the expressed protein could react with the chicken positive serum and be recognized by the specific monoclonal antibody of GST. The result suggested that the recombinant protein was correctly expressed and had a good antigenicity, which provided substantial antigen base for preparation of IBV monoclonal antibody.
Molecular Cloning of cDNA Sequences Encoding TGFα Gene in the SD Rats
ZHANG Ming-hao, ZHANG Jin-tao, LI Ya-ping
2014, 41(6):  59-63. 
Abstract ( 167 )  
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This study was designed to clone and analyze the cDNA encoding TGFα from SD rats. The PCR method was developed to clone the TGFα cDNA. A full-length cDNA and CDS sequences of SD rat were 970 and 483 bp, which had been accepted by GenBank (accession number: KF366251). TGFα protein encoded by this gene was composed of 160 amino acid residues. The identities of coding sequences of TGFα gene were 98.6%, 95.0%, 85.5%, 85.3%, 85.5%, 86.3%, 86.3%, 73.2%, 73.2% and 56.8% by homologous comparison among SD rat and other species, and in amino acid sequences were 98.8%, 96.9%, 90.6%, 90.6%, 91.2%, 91.9%, 91.9%, 72.6%, 72.6% and 38.8%, respectively. The results suggested a certain degree of conservation of the TGFα gene among different species, but there were some specificity in function among different species. The results derived from information searching by BLAST program revealed there were 7 SNPs sites in the sequences of TGFα gene coding sequences between SD rats and BN rats in GenBank. The results provided new data for SNPs in TGFα gene.
Expression of Porcine Epizootic Diarrhea Virus S1 Gene in Pichia pastoris
TAN Bo-min, YU Xi-yao, ZHONG Ze-min, JIANG Zhi-qiong, HUANG Yu-mao
2014, 41(6):  64-67. 
Abstract ( 231 )  
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To develop a new vaccines can efficiently express the porcine epizootic diarrhea virus S1 antigen epitope genes mainly,the recombinant plasmid pPICZαC-S1 was constructed by integrat S1 antigen epitope genes mainly into yeast expression vector pPICZαC and transformed into Pichia pastoris X33 by electroporation after linearized by Sac Ⅰ restriction enzymes. The positive transformants (pPICZαC-S1-X33) were screened by ZeocinTM. The transformants were inoculated into culture medium and induced with methanol. The result was demonstrated by SDS-PAGE and Western blotting, and showed the mainly antigen epitope of spike protein was secreted successfully expression in Pichia pastoris. Expression of the protein molecular weight for 42 ku, and the ability to combine with PEDV positive serum specificity.
Establishment and Application of TaqMan MGB Fluorescent-Quantitative PCR Assay for Detection of Canine Parvovirus
LIU Yi, WU Zhi-ming, YAN Ruo-qian, ZHAO Ming-jun, WANG Dong-fang,ZHAO Xue-li, LIU Mei-fen, CHENG Jun-zhen, XUE Xiao, LI Ning
2014, 41(6):  68-73. 
Abstract ( 224 )  
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In order to establish a TaqMan MGB fluorescent-quantitative PCR (FQ-PCR) assay for detecting canine parvovirus (CPV) specifically, sensitively and rapidly, a highly sensitive and specific TaqMan MGB FQ-PCR assay was developed using the specific primers and TaqMan MGB probe designed basing on the conservative sequences of VP2 gene of CPV in GenBank. The sensitivity, specificity and repetition assay of FQ-PCR assay were tested, and 46 clinic suspicious CPV infected samples were detected by the FQ-PCR assay in contrast to the routine PCR method. The results indicated that the FQ-PCR was successfully established. The developed FQ-PCR assay was able to detect as little as 1×101copies/μL of recombinant pGEX-T/CPV plasmid DNA, and the sensitivity of which was 100 times more than that of the routine PCR. The specificity assay exhibited that positive signals could be obtained from recombinant pGEM-T/CPV plasmid, but not from the genomic DNA or total cDNA of the other 5 kinds of pathogenic microorganism acting as the controls. The repetition tests were carried out by detection repeated 3 times for 3 different concentrations of recombinant pGEX-T/CPV plasmid, and the results indicated that the FQ-PCR was reproducible. Twenty-three positive results from 46 clinic suspicious CPV infected samples were obtained, which showed the better sensitivity than that of the routine PCR, with 19 positive samples from the same 46 suspected samples. The study suggested that the CPV FQ-PCR method was successfully established, and suitable for clinic rapid diagnosing of CPV and early detection of latent infection.
Development of a Rapid Test Strip for Detecting Vomitoxin Residue
LI Xiang-mei,WU Xiao-ping,LI Zhi-ping,WANG Zhen-lai,WANG Shi-en,HAN Jing-peng, SU Li-fang,YAO Lin,LIU Lei,LI Yuan
2014, 41(6):  73-76. 
Abstract ( 165 )  
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For development of a rapid test strip for detecting vomitoxin (DON) residue in milk.We used DON-OVA and secondary antibody goat anti-mouse to coat nitrocellulose membrane,and used anti-DON monoclonal antibody to prepare colloidal gold labeled antibody,then lyophilized in micro-holes of reaction plate.At last we prepared a rapid test strip that could be used for detecting DON residue in milk.The results showed that the limit of detection of the test strip was 100 μg/L,and the repeatability and the specificity were preferably,so the rapid strip could be used for detecting DON residue in milk.
Effect of Feeding Pellet Diets on Intake,Digestion and Metabolism of Pregnant Ewes
DONG Ling-yun,LUO Qiu-jiang,PAN Chao,PAN Rong,ZHOU Feng-wen,TANG Bing-hui
2014, 41(6):  77-84. 
Abstract ( 195 )  
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16 adult Small Tail Han ewes,with body weight of 53 kg,were randomly divided into 4 groups (n=4),the roughage-type ground diet (GD)(group 1) or pellet diet (PD)(group 2)(67% cornstalk),and the concentrate-type GD (group 3) or PD (group 4)(50% cornstalk) were fed from the 50th day of pregnancy.Two digestion-metabolism experiments at the 60 to 76 and 120 to 136 d of pregnancy were carried to study the effect of feeding PDs on the intake,digestion and metabolism of pregnant ewes.The results showed that under the conditions of roughage-type ration (RTR) and concentrate-type ration (CTR),the DM voluntary intake of ewes was increased by 44.8%(P<0.01) and 46.0%(P<0.01) at early pregnant stage (EPS),respectively,and 58.7%(P<0.01) and 58.5% (P<0.01) at later pregnant stage (LPS),respectively.The apparent digestibility of DM was decreased by 6.2%(P>0.05) and 6.0% (P>0.05) at EPS,respectively,and 8.0%(P>0.05) and 1.6%(P>0.05) at LPS,respectively. The apparent digestibility of CP was decreased by 13.9%(P>0.05) and 2.3%(P>0.05) at EPS,respectively,and 4.8%(P>0.05) and 3.9% (P>0.05) at LPS,respectively. The apparent digestibility of energy was decreased by 2.9%(P>0.05) and 5.7%(P<0.05) at EPS,respectively,and 0.5%(P>0.05) and 8.2%(P<0.01) at LPS,respectively.However,under the conditions of RTR and CTR,by feeding PD the digested amount of DM of ewes was increased by 19.7%(P<R0.05) and 37.0%(P<0.01) at EPS,respectively,and 65.8% (P<0.01) and 46.3%(P<0.01) at LPS,respectively.The digested CP amount of ewes was increased by 31.2%(P<0.01) and 30.4% (P<0.05) at EPS,respectively,and 60.1% (P<0.01) and 56.4% (P<0.01) at LPS,respectively. The amount of digestible energy was increased by 54.6%(P<0.01) and 36.8%(P<0.01) at EPS,respectively,and 105.2%(P<0.01) and 63.8% (P<0.01) at LPS,respectively. Under the conditions of RTR and CTR,by feeding PD the nitrogen retention of ewes was increased by 74.5%(P<0.01) and 44.7%(P<0.05) at EPS,respectively,and 101.1%(P<0.01) and 73.4%(P<0.01) at LPS,respectively. The birth weight of lamb was increased by 26.3%(P<0.05) and 34.8%(P<0.01),respectively. It was concluded that by feeding PD the voluntary intake of pregnant ewes could be increased. The digestibility of diet had the tendency to decrease,but the digested amount was increased very significantly. By feeding PD the nitrogen retention of ewes and the birth weight of lamb were increased.Therefore,feeding PD was an effective way for increasing nutrient supply to pregnant ewes.
Effect of Transgenic Bt Brown Rice as the Dietary Ingredients on Immunity in Growing Pigs
CAO Zheng-hui,WANG Zhan-bin,GU Xian-hong
2014, 41(6):  84-88. 
Abstract ( 163 )  
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The experiment aimed to study the effect of transgenic Bt brown rice as growing pig’s diet on immunity in growing pigs.30 Large White piglets were randomly allocated into 2 dietary treatments with 15 replicates per treatment.The control group diet consisted of basic diet and 70% non-transgenic Bt brown rice,and the test group consisted of basic diet and 70% transgenic Bt brown rice. This experiment lasted for 90 days.The results showed that effect of transgenic Bt brown rice on IgA,IgG,IgM contents and complements C3 and C4 levels in serum were no significant differences between the two groups (P>0.05).Submicrostructure of liver and spleen of growing pigs in treatment group had no pathological changes according to tissue slice observation of two organs and also had no significant difference compared with the control group (P>0.05).And there were no significant differences in immune organ index between the two groups (P>0.05).In conclusion,under this experimental condition,Bt brown rice as growing pig’s diet fed to growing pigs will not cause damage to immune organs and will not influence immune function.
Effect of Ensiling on Degradability of Dry Matter and Starch in Rumen about Potato Residues and Sweet Potato Residues
LI Jian-nan,LI Qiu-feng,GAO Yan-xia,CAO Yu-feng,LI Jian-guo,LI Yun-qi
2014, 41(6):  89-93. 
Abstract ( 199 )  
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Four Holstein cows fitted with rumen cannulas were used for measuring degradability of dry matter and starch in potato residues samples,sweat potato residues samples and their silage samples by nylon bag technique.The results showed that the effective degradability and 72 h disappearance rate of dry matter and starch in rumen about potato residues and sweat potato residues were reduced significantly after ensiling (P<0.01).The disappearance rate of dry matter and starch in rumen about potato residues were reduced from 95.72% and 99.60% to 53.43% and 86.51% after ensiling.The disappearance rate of dry matter and starch in rumen about sweet potato residues were reduced from 97.65% and 99.81% to 66.72% and 84.69% after ensiling.The effective degradability of dry matter and starch about potato residues silage were reduced 47.34% and 35.98%, respectively, and sweat potato residues silage were reduced 51.48% and 57.39%, respectively. The effective degradability of dry matter about potato residues silage and sweat potato residues silage had no significant difference (P>0.05),but the effective degradability of starch about potato residue silage was significantly higher than that about sweat potato residue silage (P<0.05).The amount of rumen bypass starch in potato residues silage and sweet potato residues silage were increased 57.43%(42.94 g/kg) and 151.37% (122.52 g/kg),respectively.The amount of rumen bypass starch in potato residues silage was extremely significantly lower than that in sweet potato residues silage (P<0.01). Therefore,the effective degradability of dry matter and starch about the tow kinds of potato residues were reduced effectively and the amount of rumen bypass starch were increased by ensiling.The amount of rumen bypass starch in sweat potato residues were obviously higher than that in potato residues.
Research Progress on Zinc and Different Sources of Zinc in Animal Nutrition
LIU Xi-ping,SUI Mei-xia,XU Zhong-xiang,WANG Zong-wei,ZHANG Ru-lei
2014, 41(6):  94-98. 
Abstract ( 252 )  
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Zinc was one of the most important trace elements in animal body,it had important functions on the promotion of animal growth and immune function,maintenance of healthy skin.The paper summarized the biological function of zinc,the factors affected the body’s absorption of zinc and the research progress on different sources (inorganic zinc and organic zinc) of zinc in animal nutrition in recent years.It provided a theoretical basis for the zinc in the efficient and rational use in livestock food.
Study on Metabolism Levels of Serum Trace Elements in Dairy Cows with Subclinical Ketosisin
Bagedati,WANG Yin-long,LI Hong-cheng,LIU Qiang,Saiwujiafu
2014, 41(6):  98-102. 
Abstract ( 170 )  
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This study was designed to research the effects on serum trace elements of Holstein cows with subclinical ketosis.40 Holstein cows with subclinical ketosis in 1 to 15 d postpartum, through the application of atomic absorption spectrometry, we measured Cu, Fe, Mn, Zn, Se, Co contents in serum, determined GSH-Px, T-AOC, T-SOD activities and MDA content in serum by using spectrophotometer and colorimetric.Cu, Zn, Se, Co contents in serum of positive group were extremely significantly lower than that of control group in the whole testing process (P<0.01);Mn concentrations in serum of positive group in the 1st, 5th days detection were significantly lower than the control group (P<0.05), 10th, 15th day were extremely significantly lower than the control group (P<0.01);Fe contents in serum of positive group and control group were not significantly different (P>0.05), GSH-Px, T-SOD, T-AOC activities in serum of positive group were extremely significantly lower than the control group (P<0.01),MDA content in serum of positive group was extremely significantly higher than the control group (P<0.01).Subclinical ketosis had direct impact on GSH-Px, T-AOC, T-SOD activities of cows antioxidant system, indirect effect on serum trace elements Cu, Mn, Se, Zn contents;vitamin B12 had the influence on Co content in serum,so Co content in serum significantly decreased in subclinical ketosis cow;the liver and oxidases of subclinical ketosis cow were damaged, resulting in the Fe content in serum had no significant changes.
Research Progress on Detection Method,Hazard and Residual of Mycotoxins in Laying Hens
FANG Si-min,BAI Fan,BAI Xue,GUO Chun-hua,ZHANG Zheng-fan,PENG Zhong-li
2014, 41(6):  103-107. 
Abstract ( 282 )  
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Feed mycotoxins contamination in laying hens diets posed severe threat to both laying hens health and animal products safety. In this review,we concluded the effects of mycotoxins in laying hens,residual in hens tissues and eggs,as well as the detection method to promote the research on mycotoxins.
Effect of Chinese Herbal Medicine on Production Performance and Economic Benefits of Weaning Piglets
SUN Yu-hua
2014, 41(6):  108-111. 
Abstract ( 246 )  
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In order to study the effects of different doses of Chinese herbal medicine and antibiotics on production performance and economic benefits of weaning piglets, 60 three-crossbred healthy weaning piglets were selected (Duroc×Landrace×Large White), with the weight of (10±1)kg. They were randomly divided into 4 treatment groups (3 experimental groups and 1 control group) and each group had 3 replications including 5 piglets separately. In each experimental group, the basal diets were supplemented with 0.2%, 0.4% Chinese herbal medicine and antibiotics (400 mg/kg of colistin sulfate). The results showed that, in comparison to the control group, the average daily gains in 3 experimental groups increased respectively by 8.24%, 9.17% and 8.65% in turn, significant differences (P<0.05);feed to gain ratioes were decreased by 5.12%, 6.56% and 5.01%, significant differences (P<0.05);piglet diarrhea rates decreased;the gross profits per pig increased respectively by 3.78%,5.87% and 4.07%. It illustrated that Chinese herbal medicine could completely replace antibiotics to a certain extent, but there were some differences between the effects of different doses of Chinese herbal medicine.
Study on Changes in Mineral Elements Contents of Yak Milk during the Lactation Cycle
LIU Yi-li,JIANG Ming-feng,JIANG Wei-hua,LIN Zhong-li,CAI Zi-jian,ZHANG Lin,ZHU Xiao-yi,REN Hong-hui,LI Jian-bo
2014, 41(6):  112-116. 
Abstract ( 184 )  
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To clarify the changes of mineral elements of yak milk during the lactation cycle,milk samples at 12 time points from 0 to 180 d during the lactation cycle were collected from Maiwa yak at Longri livestock breeding station in Sichuan province. After milk samples were prepared using microwave digestion,fourteen elements were analyzed by the ICP-OES for four macro elements (potassium,calcium,magnesium,phosphorus) and ten micro elements (boron,barium,cadmium,cobalt,copper,molybdenum,nickel,titanium,zinc,manganese).The statistical analysis based on the statistics data from the samples on day 30 was proceeded. The results showed that the concentrations of K,Ca,Mg and P in Maiwa yak colostrum were higher.The concentrations of K on day 0 (P<0.01),120 (P<0.05),Ca on day 5 (P<0.05),Mg on day 180 (P<0.05) had significant differences as compared with that of mineral elements on day 30, but no significant difference was observed in the concentration of P (P>0.05) during the lactation cycle.The results also suggested that the concentrations of trace elements were significantly affected by the lactation cycle. The concentrations of Ba on day 0 and 1 (P<0.01),2,3,4,5 and 180 (P<0.05),Mo on day 180(P<0.05),Ni on day 180(P<0.05),Zn on day 180(P<0.05) had significant differences as compared with that of mineral elements on day 30,but there were no significant differences on B,Cd,Cu,Ti,Mn during the lactation cycle.In conclusion,the concentrations of K,Ca,Mg,Ba,Mo,Ni,Zn had significant differences during the lactation cycle (P<0.05),but no significant differences were observed in the concentrations of P,B,Cd,Cu,Ti,Mn (P>0.05).Overall,the concentrations of K,Mg,P,Ba,Zn were all firstly decreasing and then increasing,while Ca content was firstly increasing and then decreasing.
Study on the Influence of 2 Strains of Lactobacillus on the Content of IgG and IgA in Blood Serum of Mice
LV Tian-xing,YU Chen-long,CHEN Guang-ming,YANG Xiao-yu,SU Ri-na,ZHANG Qi-jin
2014, 41(6):  117-120. 
Abstract ( 183 )  
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In this test,the LJ2 and LZ1 Lactobacillus strains had been given to mice with different doses for 30 days continuously by the way of intragastric administration.Then we detected the content of IgG and IgA in blood serum of mice at 10,20 and 30 days. The results showed that compared with the control group,all the different groups with different doses of LJ2 or LZ1 could significantly increase the content of IgG and IgA in blood serum of mice,and the low-dose group and middle-dose group tended to exceed the high-dose group with time passing.All in all,for the increase of the content of IgG and IgA in blood serum of mice,the LZ1 strain had a better effect than the LJ2 strain.The sequence according to the effect was listed from the best to usual as following:The LZ1 low-dose group had the best effect,then came to the LZ1 middle-dose group and high-dose group,the last was the LJ2 low-dose group.
Study on Growth and Development Pattern of Chinese Holstein Calf and Heifer
YU Zhi,GAO Yan-xia,CAO Yu-feng,LI Qiu-feng,LI Jian-guo
2014, 41(6):  121-125. 
Abstract ( 249 )  
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The objective of this study was to measure the body weight,height,length,chest and pipe circumference of 150 Chinese Holstein calf and heifers from born to breeding,aiming to revealing the growth and development pattern and drawing the growth curve. The results showed that the body weight,height,length,chest and pipe circumference of heifers at 13-month age respectively were 366.26 kg,125.69 cm,168.01 cm,139.38 cm and 17.83 cm,reached the breeding requirements. The fastest growth period of heifers was from birth to 8 month old,the speeding of growth decreased after 8 month old. The estimate of body weight for calf and heifer were:Y=-284.087+4.747X1+5.664X2-3.209X3-25.352X4 and Y=96.475X22X3.
Study on Serum-free Culture of Embryonic Germ Cells from Guanzhong Dairy Goat
GE Xiu-guo, LI Ji-xia, DOU Zhong-ying
2014, 41(6):  126-131. 
Abstract ( 178 )  
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Goat embryonic germ cells (EGCs) was a kind of pluripotent stem cells derived from the primordial gonad, which isolation and culture system in vitro has an important value for the study of goat breeding. In this assay, Guanzhong dairy goat EGCs were isolated and cultured by enzyme digestion and tissue culture methods, and the effect of serum-free culture medium on cell proliferation in vitro was detected. The results were found that the medium could be used for obtaining of goat EGCs, and the colony morphology was typical, which expressed AKP, Oct4, TERT and SSEA-1. The EGCs could also differentiate into embryoid bodies, fibroblast-like cells, lipoblast-like cells and oocyte-like morphology by differentiation test in vitro. In conclusion, these data indicated that serum-free culture medium could be used in isolation and culture of goat EGCs, and our results provided a new reference to further establish the long-term culture system of goat EGCs.
Study on Jurkat Cells Apoptosis Induced by Aphidicolin Using Flow Cytometry
WAN Yu-ping,JIA Fang-fang,WU Xiao-sheng
2014, 41(6):  132-135. 
Abstract ( 216 )  
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To study cell apoptosis changes induced by different concentrations of aphidicolin, we used flow cytometry to get Jurkat cell apoptosis rates. We found that dose and time dependent effects appeared because of Jurkat cell apoptosis induced by aphidicolin. With the increasing of aphidicolin concentrations or incubation time, the early apoptosis rate, late apoptosis rate and total apoptosis rate all increased. There was an obvious increasing trend when the concentration of aphidicolin was more than or equal to 1 μg/mL, or the incubation time was more than or equal to 16 hours.
Effect of FGM Strain Passage Times on the Transformation of Astragalus Polysaccharide from Astragalus membranaceus by Fermentation
SHANG Li-ming, QIN Zhe, ZHANG Jing-yan, WANG Xu-rong, WANG Lei, KONG Xiao-jun, CHEN Jie, YANG Zhi-qiang, WANG Xue-zhi, LI Jian-xi
2014, 41(6):  136-139. 
Abstract ( 183 )  
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  The assay was aimed to further improve the stability of fermentation of Astragalus membranaceus by Streptococcus alactolyticus FGM strain. First,Astragalus membranaceus-sulfuric acid method,respectively. Finally, we made a statistical analysis. The results showed that the multiple continuous passage of FGM strain had caused the serious degeneration phenomenon of FGM strain and extremely significantly affected the yield of APS (P=0.007), so the fermentation performance was the strongest after the 4th generation of subculture. was inoculated with the successively passaged FGM strain to fermentation, then we measured the viable counts of FGM strain and the yield of Astragalus polysaccharide (APS) by bacteria plate count method and phenol The assay was aimed to further improve the stability of fermentation of Astragalus membranaceus by Streptococcus alactolyticus FGM strain. First,Astragalus membranaceus was inoculated with the successively passaged FGM strain to fermentation, then we measured the viable counts of FGM strain and the yield of Astragalus polysaccharide (APS) by bacteria plate count method and phenol-sulfuric acid method,respectively. Finally, we made a statistical analysis. The results showed that the multiple continuous passage of FGM strain had caused the serious degeneration phenomenon of FGM strain and extremely significantly affected the yield of APS (P=0.007), so the fermentation performance was the strongest after the 4th generation of subculture.
Determination of Total Flavonoids in Fengliao Decoction by UV-visible Spectrophotometry
LIU Hong-mei,CHEN Yu-ting,HE Li-mei,PENG Xin-yu,WU Jin-jie,YUAN Ming-gui,LUO Sheng-jun,WEI Guang-wei
2014, 41(6):  140-144. 
Abstract ( 202 )  
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A UV-visible spectrophotometry method was established for the determination of total flavonoids in Fengliao decoction. Rutin was the control, NaNO2-Al(NO3)3-NaOH was the color developing reagent and the detection wave length was 510 nm. The standard curve was linear in the range of 0.008 to 0.048 mg/mL for total flavonoids, the average recovery of total flavonoids was 100.97% with RSD of 1.66%. The method established was fast and reliable, and could be used to determine the content of total flavonoids in half finished product and finished product of Polygonum hydropiper L. and Daphniphyllum calycinum Benth..
Research Progress on Mechanisms of Oxidative Stress Generation in Water Body
WANG Qiu-ju,JU Xue,WANG Qing-bin,YANG Yi-yu,LUO Sha,LIU Hong-jian,ZHANG Dong-ming
2014, 41(6):  144-149. 
Abstract ( 183 )  
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Reactive oxygen species (ROS) were an unenviable part of aerobic animal,but they could caused cell damage of aquatic animals if accumulated too much,then caused diseases.In order to provide theoretical basis for works of against oxidative stress in China intensive aquaculture,these were reviewed that mechanisms of reactive oxygen species generation and elimination in aquatic animals,what were the mechanisms that the factors could lead to oxidative stress in aquatic environments,such as temperature,oxygen level,salinity,transition metal ions and pesticides.
Study on Fed Batch of BHK-21 Suspension Cells
WU Fa-ju, LIU Ping, AN Fang-lan, DONG Wen-jiao, SONG Yu-xia, DONG Jin-jie, MU Ke-bin, WANG Chao-ying, LIU Xue-rong
2014, 41(6):  150-153. 
Abstract ( 269 )  
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The research was aimed to study the impacts of different feeding nutrients, start time of feeding and cell inoculation density on the growth of BHK-21 suspension cells.The result showed that amino acid concentrated liquid had a apparent influence on promoting the growth of BHK-21 suspension cells. But serum components played a more important role in the promotions of cell growth, maintenance and cell activity. When initial inoculation density was 3×105 to 6×105 cells/mL, adding a certain amount of concentrated liquid at 48 h, BHK-21 suspension cells grew well. In addition, BHK-21 suspension cells did not obviously occur apoptosis in the optimized conditions.
Protective Effect of Thioredoxin on H2O2-induced Injury of BRL-3A Cells
2014, 41(6):  154-158. 
Abstract ( 143 )  
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BRL-3A cells were injured by different doses of hydrogen peroxide (H2O2) and the cell survival rate was assessed by methyl thiazolyl tetrazolium (MTT) assay to select the optimal concentration of H2O2. The BRL-3A cells were divided into normal control,H2O2-induced and thioredoxin (Trx) pre-treatment groups with different concentrations. The assay kits of malondialdehyde (MDA),superoxide dismutase (SOD) and catalase (CAT) were used to detect the alterations of oxidative stress.Cell proliferation of BRL-3A was inhibited significantly by 1000 μmol/L H2O2 (P<0.05). 2.5 μmol/L Trx had the protective effect on H2O2-induced injury of BRL-3A cells by significantly decreasing MDA contents which were induced by H2O2 and increasing the activities of SOD and CAT (P<0.05), thus protecting liver cells.Trx had protective effects on BRL-3A cells induced by H2O2.The mechanism might be related to its antioxidant activity.
Isolation and Comparison of Endometrial Stromal Cells from Pyometra Bitches and Healthy Bitches
ZHOU Peng-yang, JIN Tao, DU Miao, MA Wen-zhi
2014, 41(6):  158-161. 
Abstract ( 152 )  
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Through separating endometrial stromal cells (ESCs) in pyometra bitches and healthy bitches, the paper analyzed the differences of cells from the two resources, observed their growth and identified them by immunofluorescence assay. The results showed that the morphologies and characteristics of ESCs from pyometra bitches were basically consistent with the healthy ones. The former cells were stronger and grew faster. After inoculated 4 days, they integrated into the film. The majority of cells were spindle, while a few showed multiple angular. By immunofluorescence, they expressed vimentin with positive rate of 98% or more. It could be concluded that the isolated cells in the experiment were canine ESCs and the cells were not much different between mild pyometra bitches and healthy bitches.
Analysis of Genetic Diversity and Phylogeny of the Mitochondrial Cytochrome b Gene of Mongolian Horses and Mongolian Wild Horse
HAI Tang, HUANG Jin-long, ZHAO Yi-ping, HAN Hai-ge, SA Ru-la, MANG Lai
2014, 41(6):  162-167. 
Abstract ( 199 )  
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Genetic relationship and genetic diversity among Mongolian horses, Mongolian wild horse and donkeys were investigated by genetic variation analysis of the mitochondrial cytochrome b (Cytb) gene sequence. mtDNA Cytb gene of all samples were detected by PCR amplification and then sequenced. The date was analyzed by modern molecular biology software and the analysis showed that 90 polymorphic loci formed 15 haplotypes and the haplotype diversity (Hd), nucleotide diversity (Pi) and average number of nucleotide differences(K) were 0.9356, 0.01022 and 8.966, respectively. Genetic differences between the groups were not obvious. By construction of phylogenetic tree and NETwork, the results indicated that genetic diversity of 4 kinds of Mongolia horse were higher, Mongolian horse and Mongolian wild horse had close relationship and appeared several branches, which explained that Mongolian horses originated multiply or had been domesticated many times.
Heritability Estimates and Non-genetic Factors Analysis of Birth Weight of Chinese Merino Sheep (Xinjiang Type)
XU Xin-ming, LI Yan-fei, FU Xue-feng, YU Li-juan, ZHANG Yan-hua,HUANG Xi-xia,TIAN Ke-chuan
2014, 41(6):  168-171. 
Abstract ( 150 )  
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The article estimate the heritability and analysis the influence of non-genetic factors on birth weight of the number of 2125 Chinese Merino sheep (Xinjiang type) by using SAS 8.1 program of least-squares analysis and MTDFREML software, the data collected from Nanshan sheep breeding farm. The results showed that the birth weight heritability of Chinese Merino sheep was 0.30, it belonged to the moderate heritability (0.1<h2<0.3). The maternal age, year of birth, lambing type and group were highly significantly effect on birth weight (P<0.01), but the gender was no significantly effect on birth weight (P>0.05).
Genetic Diversity of Mitochondrial DNA D-loop Sequences in Huili Black Goat
ZHANG Wen-li
2014, 41(6):  172-176. 
Abstract ( 186 )  
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To explore the genetic diversity and origin for genetic resource protection of Huili Black goat, the mitochondrial DNA (mtDNA) D-loop was investigated. mtDNA D-loop sequences of 41 goats were analyzed by PCR, sequencing techniques, and biological information and the phylogenetic trees were constructed. The mtDNA sequences of the Huili Black goat ranged from 1211 to 1213 bp, and 2 sequences were 1211 bp, 29 sequences 1212 bp, and 10 sequences 1213 bp. The content of A+T (60.1%) was higher than one of G+C (39.9%). There were 9 haplotypes, and the haplotype diversity was 0.842+0.00368. The nucleotide diversity was 0.01542+0.00034. The phylogenetic analysis showed that Huili Black goat was distributed in a branch, and were closed to Jianchang Black goat, Chengdu Ma goat, Jintang Black goat, Guizhou White goat, Guizhou Black goat, but they were less related to Capra falconeri. Huili Black goats had rather abundant genetic diversity, and were greatly affected by other goat breeds in history.
Association of Calpastatin Gene Polymorphisms with Meat Tenderness in Altay Sheep from Xinjiang
TIAN Jia, ZHOU Wei-dong, MA Xiao-yan, ZHENG Wen-xin2, GAO Wei-ming, DI La-re?SHA Ta-er, ZUO Xiao-jia, SHI Shuai, ZHANG Rong-yin, LIU Wu-jun
2014, 41(6):  177-181. 
Abstract ( 234 )  
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In this study, Altay sheep was used for investigating the association between sheep tenderness and single nucleotide polymorphism markers on the calpastatin (CAST) gene. Its genetic polymorphism was detected using SSCP and DNA sequencing technology in exon 1, exon 2 and exon 21. However, only three genotypes AA, AB and BB were detected in exon 21, the sequencing result showed that the mutation of A into G, association analysis indicated that the mutation in exon 21 affected the meat tenderness markedly of Altay sheep (P<0.05). In summary, AA genotype might have certain improvement effect on meat tenderness.
Resource Investigation Report of Bashang Long-tail Chicken
2014, 41(6):  182-187. 
Abstract ( 247 )  
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Bashang Long-tail chicken is a good indigenous breed with high productivity and dual-purpose of egg and meat. It distributed in northern of Hebei province, mainly in the bashang region of Chengde and Zhangjiakou. The main characteristic of Bashang Long-tail chicken was its longer tail feather, The cock had 2 or 3 big warped plumes and the longest was 52 cm among the measured data. The hen was tail up and with the long tail feathers of 15 to 20 cm. The big egg yolk inside the transparent egg white was golden and had an elastic delicate taste and rich in various nutrients. The Bashang Long-tail chicken with succulent tender and delicious quality had strong ability to look for food in fields and to resist diseases. From this research we put the Changliang and Pingdingbao township of Guyuan county, Danqinghe township of Kangbao county as the key protective region to conserve their larger number of Bashang Long-tail chicken. To further establish a core group of field and storage to consolidate and enhance its original excellent characters and features by pedigree selection.
Study on Influence of Sows’ Backfat Thickness at Farrowing and Weaning on Reproductive Performance
LIU Zhu, YANG Zhi-yuan, LI Xiao-yu, YANG Xue, HUANG Tao
2014, 41(6):  187-190. 
Abstract ( 193 )  
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This experiment was aimed to reveal the effects of sows’ backfat thickness at farrowing and at weaning on reproductive performance. The backfat thickness at farrowing and at weaning was determined by type-A ultrasonic device. According to the backfat thickness at farrowing, sows were divided into group 1 (<10 mm), group 2 (10~14 mm),group 3 (>14 mm);According to the backfat thickness at weaning, sows were divided into group A (<7 mm), group B (7~10 mm), group C (10~13 mm),group D (13~16 mm),group E (>16 mm);According to the loss of backfat thickness during the lactation period, sows were divided into group Ⅰ(<3 mm),group Ⅱ(3~6 mm) and group Ⅲ(>6 mm). The results showed that the rate of sows producing stillbirth was 10.5% in group 2, which was the least in the three groups, and the average weaning-estrus interval of group C was significantly shorter than other groups (P<0.05). During the lactation period, the loss of back fat thickness in group Ⅲ was higher than groupⅠand groupⅡevidently, and the average estrus interval of group C was also markedly higher than groupⅠand groupⅡ (P<0.05). The study indicated that sows with medium backfat thickness at farrowing and weaning shown the best reproduction performance. And an regression equation was established to forecast estrus date based on backfat thickness at weaning: Y=31.787-6.169X+0.440X2-0.009X3.
Expression Changes of Dab2 in Kidney Tissue of the Aristolochic Acid Nephropathy Mice
WU Jin-dong, CHANG Di, YANG Yao-hui, TIAN Ya-hui, ZHAO Hai-jiao, WU Guo-juan
2014, 41(6):  191-196. 
Abstract ( 154 )  
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The study was aimed to observe the expression changes of Dab2 in kidney tissue of the aristolochic acid nephropathy (ANN) mice. In the foundation of ANN mice model, utilizing the immunohistochemistry, Real-time fluorescence quantitative PCR (qRT-PCR) and Western blotting technology to detect the expression changes of Dab2 in kidney tissues. With the extension of treatment time, the kidney structure was more seriously damaged, the expression change of Dab2 decreased on the 6th day (P>0.05), suddenly increased on the 12th day and extremely significantly decreased (P<0.01), but the overall trend of Dab2 expression change decreased, and the change of mRNA was consistent with this;at the same time, the location of Dab2 proteins changed, transferred from the brush border to the cytoplasm, and transferred into the nucleus of tubular epithelial cells. This study showed that Dab2 involved in the progress of ANN, this study layed the foundation for the function research of Dab2 in the progress of AAN.
Reliability Evaluation of ELISA for Antibodies to ALV-A/B in Chicken Serum Samples
DUAN Lun-tao, ZHAO Peng, DONG Xuan, WANG Yi-xin, CUI Zhi-zhong
2014, 41(6):  197-203. 
Abstract ( 153 )  
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To evaluate the reliability of the positive rate of ALV-A/B specific antibodies detection in chicken serum and illustrate the stability and specificity problems existing in the commercialized Avian Leukosis Virus Subgroup A/B Antibody Test Kit, two batches of serum samples submitted by the company A which had no positive samples detected previously and the company B which had positive samples detected yet were tested using the same lot of ELISA kit by the professional operators. Then, among the positive samples detected for the first time, eight samples from company A and forty samples from company B whose S/P ratio all ranked higher were re-detected triply, and IFA were tested using the forty-eight samples above as first antibodies. The consistence of the results between ELISA and IFA assay was compared. The results demonstrated that, eight samples from company A turned into negative in the re-detection by ELISA kit, and IFA results were negative as well, which indicated stability problems existing in some batches of commercialized kit. Four samples (4/40) form company B turned into negative in the re-detection by ELISA kit, so as IFA results;thirty-six samples (36/40) form company B were positive in the re-detection by ELISA kit, however, twenty of them were IFA positive and sixteen of them were IFA negative, the complicated results indicated non-specificity detection were existed in some batches of commercialized kit except for the stability problem. Therefore, to achieve higher veracity, samples should be determined by both of ELISA and IFA in the massive detection.
Isolation and Effect of Antagonistic Bacteria against Vibrio in Shrimp Aquaculture
WANG Jiao,QU Peng,SUN Fei,TIAN Xiang-li,LIANG Jing-jing,CAI Yu-yong
2014, 41(6):  203-206. 
Abstract ( 151 )  
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A total of 22 strains were isolated from aquatic environment.The potential probiotic strains were selected by dot inoculating method to screen the bacteria with antagonism property. The isolate A4 was identified as Bacillus subtilis based on conventional biochemical tests and 16S rRNA sequence homology, which exhibited significantly inhibitory activity against Vibrio parahaemolyticus,Vibrio alginolyticus and Vibrio brilliant.The probiotic effect of Bacillus subtilis showed that it could reduce the levels of the pathogenic bacteria and affect shrimp health after two days of culture (P<0.05).The Bacillus subtilis had potential applications for controlling pathogenic bacteria,which could be used as biocontrol agents in the aquaculture industry.
Drug-resistance Analysis of Coagulase-negative Staphylococcus Isolated from Dairy Cow Mastitis
WANG Xin-yu,PEI Lin,YU En-qi,LIN Wei-dong,WEN Xin-tong,ZAI Xu-sheng,QIN Ai-jian,JIN Wen-jie,ZHANG Jun,LIANG Xiang-huan,WANG Jin-rong,HANG Bo-lin
2014, 41(6):  207-210. 
Abstract ( 259 )  
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In this study, pathogenic epidemiology of subclinical mastitis in part of Jiangsu province dairy farms was investigated. It included sampling, bacteria cultivation, gram staining, microscopy observation and biochemical identification. 35 strains of coagulase-negative Staphylococcus (CNS) were isolated. Results of drug sensitivity test showed that 60% to 80% of CNS isolates were highly sensitive to clinical commonly used antibiotics like vancomycin, clarithromycin, benzene azole pennicillin, norfloxacin, chloromycetin, ciprofloxacin, cefotaxime, fosfomycin and nitrofurantoin. While for penicillin G, SMZ and polymyxin B, these isolates showed high drug resistance, the resistance ratio was 70% to 100%. All these CNS isolates showed multi-drug resistance, 3 CNS isolates were resistant to 20 drugs, 1 was resistant to 15 drugs,1 was resistant to 14 drugs,62.9% of these CNS isolates were resistant to more than 5 tested drugs. All these results indicated that the resistant status of CNS caused subclinical mastitis was very serious.
Comparison of Three ELISA Kits for the Detection of Porcine Reproductive and Respiratory Syndrome Virus Antibody and its Application
HE Hai-jian, XING Gang, MA Tao, QIU Xiao-huo, JIANG Chun-yan, ZHOU Cai-qin
2014, 41(6):  211-215. 
Abstract ( 183 )  
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To comparatively analyzed piglets maternal antibodies and the antibody dynamics in immunized pigs using three different of antibody detection Kits of porcine reproductive and respiratory syndrome virus (PRRSV),in order to provide selection refereace for Kits of monitoring PRRSV antibodies of pigs. The results showed that the Kappa value =0.784>0.6 between Kit A and Kit C,the compared results were highly-consistent;the Kappa value was 0.422 between Kit B and Kit A,the Kappa value was 0.374 between Kit B and Kit C.Maternal antibody monitoring showed that PRRSV maternal antibodies of three Kits had become negative at 42 days old. PRRSV associated antibodies of using Kit A and Kit C were detected after immuned one week, three weeks to reach the peak, while the PRRSV antibodies of using Kit B were detected three weeks after immunization, and reached the peak in five weeks. This study provided the experimental basis for the three Kits were used to antibody surveillance and immune assessment in the clinical PRRSV.
Resistance Analysis of Florfenicol to Aeromonas hydrophila from Freshwater Fish
WANG Xiao-liang,XU Li-pu,WANG Shu,WANG Jing-bo,CAO Huan
2014, 41(6):  216-220. 
Abstract ( 183 )  
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To evaluate Aeromonas hydrophila in the status of drug resistance to florfenicol which was the major pathogen from freshwater fish, we tested resistance to florfenicol of 26 strains of Aeromonas hydrophila collected from freshwater fish in Beijing area nearly three years using disk diffusion method and concentration dilution method, respectively. Then, we detected the florfenicol resistance gene of strains using PCR method. The results showed that 15.38% of stains were resistant to florfenicol. The minimum inhibitory concentration of florfenicol to strains was mainly 2 to 4 μg/mL presently, and 5 strains were more than 8 μg/mL. Florfenicol resistance gene floR was detected in 1 strain. Combined with previous data and the relative data of other provinces, it was obvious that the florfenicol resistance rate and resistance concentration of Aeromonas hydrophila were both higher than before, and the strains carried floR gene located in plasmid on the risk of transferring the gene to other species of fish pathogens. Therefore, it was necessary to use florfenicol scientifically in aquaculture.
Isolation,Identification and Drug Resistance Analysis of Staphylococcus aureus from Sheep Lungs
PEI Zhi-hua, SUN Xiao-ning, WANG Kai
2014, 41(6):  220-223. 
Abstract ( 202 )  
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One strain of Staphylococcus aureus was isolated and identified by 16S rRNA sequence analysis and biochemical analysis in a large-scale sheep farms, it was named as JLY-1. The drug resistance analysis of the strain was conducted. The result showed that JLY-1 was highly resistant to erythromycin, ampicillin, chemitrim (MIC≥512 μg/mL);resistant to doxycycline, amikacin (MIC≥64 μg/mL) and ciprofloxacin, enrofloxacin (MIC≥4 μg/mL);and sensitive to florfenicol and cefatriaxone (MIC≤0.25 μg/mL).
Study on Identification and Pathogenicity of Salmonella Isolates from Dairy Cattles in Inner Mongolia
GUAN Hong, ZHAO Jun-li,ZHAO Hong-xia,LI Zhi-fang,DING Yue-xia,SHI Rui-li
2014, 41(6):  224-229. 
Abstract ( 215 )  
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The objective of the present study was to observe the distribution of the serotypes and pathogenicity to mice of Salmonella isolated from dairy cattles with mastitis or endometritis. Salmonella strains were isolated by selective medium. The serotype of the isolates were determined by A to F polyvalent and O and H monovalent diagnostic serum agglutination test;Mice infection model was induced by intraperitoneal injecting of the Salmonella isolates. The morbidity and mortality of the mice were observed. Tissue lesions were also observed by using tissue sectioning. A total of 38 of Salmonella strains were isolated and identified from 460 samples of bovine mastitis or endometritis, the isolating rate of pathogenic Salmonella was 8.3%. 38 strains all belonged to A to F group, and 8 groups and 14 serotypes were observed. S.typhimurium (39.5%)was the most commonly isolated Salmonella. Mortality of five selected strains ranged from 50% to 80% respectively. Meanwhile, Salmonella strains were recovered from each experimental mouse’s hearts and livers. There were necrosis in heart, liver and kidney of mice. Bacterial embolus was observed in lung of infected mice. Salmonella isolated from dairy cattles had complex serotypes and were strongly pathogentic to mice, but there were differences in pathogenicity of different Salmonella isolates.
Research Progress on Subgroup J Avian Leukosis
YANG Xiao,ZHANG Xun-hai
2014, 41(6):  230-235. 
Abstract ( 318 )  
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Subgroup J avian leukosis is a name of tumorigenicity and immunosuppressive disease in avian caused by extrinsic subgroup J avian leukosis virus (ALV-J). ALV-J can infect chicks by the vertical and horizontal transmission. ALV-J is widespread through the flock and becomes more complicated in China. It has influnenced the development of poultry industry seriously. We have no vaccine and effective treatment to overcome up to now. Purifying breeder flocks is regarded the main control measure. The author reviewed the etiology, epizoology, isolation along with identification and controlling of subgroup J avian leukosis.
Prevention and Treatment Effect of Ben Lamge Granules on Chicken Infectious Bursal Disease
DU Ji-hong, ZHU Hua, CUI Xiao-qi, WANG Xiu-min, QIN Jun-jie
2014, 41(6):  236-240. 
Abstract ( 155 )  
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In order to probe into prevention and treatment effect of Ben Lamge granules on chicken infectious bursal disease, 26-day-old SPF chickens were infected with IBDV virulent strain BC-6/85 artificially, and Ben Lamge granules with different doses were joined in drinking water before and after challenge. Clinical symptoms, pathological changes, statistics lesion score, immune organ index of chicken were observed. The results showed that Ben Lamge granules with middle and high doses groups could improve the infectious bursal disease state of mind, alleviate the damage of the disease of chicken immune organs, increase blood leukocyte phagocytosis rate, increase the food intake and body weight gain administered via drinking water. The results showed that 3 g/L Ben Lamge granules could effectively prevent and control the chicken infectious bursal disease administered via drinking, continuous administration of 5 d.
Research Progress on Porcine Reproductive and Respiratory Syndrome Virus and its Vaccines
TAN Wei, DENG Xian-wen, XIE Zhi-xun
2014, 41(6):  240-247. 
Abstract ( 239 )  
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Porcine reproductive and respiratory syndrome (PRRS) severely affects prosperity of swine industry, and results in enormous economic losses every year. Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of PRRS. Pregnant sows infected with PRRSV may suffer from early farrow, abortion or stillbirth, and newborn piglets may develop respiratory symptoms. Although at present inactivated and live-attenuated PRRS vaccines are commercially available, their protective effects are not ideal. Therefore, study on PRRSV gene structure, protein functions, biological characteristics, and pathogenesis will facilitate research and development of safe and effective PRRS vaccines. This article will introduce research progress on PRRSV and its vaccines.
Effects of Different Doses of Florfenicol Added in Drinking Water on Immune Effects of Newcastle Disease Live Vaccine in Chicken
ZHANG Jing, DIAO Fu-hua
2014, 41(6):  248-251. 
Abstract ( 248 )  
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The assay was aimed to explore the effects of different doses of florfenicol on immunity effects of Newcastle disease live vaccine, determine the florfenicol optimal dosage, and provide scientific basis for the clinical application of florfenicol. 150 broilers of 14-day-old were divided into 5 groups (control group, groupsⅠ, Ⅱ, Ⅲ and Ⅳ). The control group normally drank water, groupsⅠ, Ⅱ, Ⅲ, Ⅳ respectively added water-soluble florfenicol 50,100,200,400 mg/L in the drinking water, through the determination of serum antibody titer of Newcastle disease, IL-10 and INF-γ contents, we analyzed the immune effect of different doses of florfenicol on Newcastle disease live vaccine. The test results showed, compared with control group, serum antibody titers of Newcastle disease, IL-10 and INF-γ contents of groups Ⅰ and Ⅱ had no significant differences (P>0.05), but groups Ⅲ and Ⅳ had significant differences (P<0.05). Low doses of florfenicol added in drinking water (50,100 mg/L) had no effect on immune effect of Newcastle disease live vaccine, but high doses of florfenicol (200,400 mg/L) had immunosuppression on immune effect of Newcastle disease live vaccine.
Study on Antibacterial Activity in vitro and Anti-inflammatory Abilities of Yinhuang Zhike Granules
DUAN Kai-wen,OUYANG Qing-fang,CHEN Xiu-qin,LIN Xu-hui,SHI Da-you,GUO Shi-ning
2014, 41(6):  251-254. 
Abstract ( 187 )  
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The aim of the study was to prove whether or not Yinhuang Zhike granules had effect on the antibacterial activity in vitro and anti-inflammatory abilities. The antibacterial activity in vitro was carried out by broth two-fold agar dilution method, animal model of early inflammatory was induced by injection with 0.6% acetic acid, the peritoneal fluid was collected to detect D590 nm. The results showed that MIC of Yinhuang Zhike granules to Escherichia coli,Staphylococcus aureus,Pseudomonas aeruginosa,Streptococcus pneumoniae were 7.8,7.8,3.9,125 mg/mL, while MBC were 15.6,15.6,7.8,250 mg/mL, respectively. 30,16,8 g/kg Yinhuang Zhike granules all had obvious inhibition effects on increasement of capillary permeability in mice induced by acetic acid.