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Table of Content

20 March 2012, Volume 39 Issue 3
特约综述
Main Risk Factor Analysis for Milk Quality and Safety Ⅱ.Mycotoxins
ZHENG Nan;WANG Jia-qi;;HAN Rong-wei;XU Xiao-min;;ZHEN Yun-peng;;QU Xue-yin;;ZHAO Lian-sheng
2012, 39(3):  1-9. 
Abstract ( 520 )  
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Mycotoxin contaminant is a problem for milk safety. In the present paper, the mycotoxins might be exist in milk, maximum residue levels of mycotoxins in milk established by many government and international organizations and risk assessment of mycotoxins in milk were reviewed. These could be referred to improve surveillance system of milk safety in China.
Toxicity of Mycotoxins in Milk: A Review
ZHENG Nan;WANG Jia-qi;;HAN Rong-wei;XU Xiao-min;;ZHEN Yun-peng;;QU Xue-yin;;ZHAO Lian-sheng
2012, 39(3):  10-13. 
Abstract ( 404 )  
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Aflatoxin, ochratoxin, zearalenone, deoxynivalenol, T-2 toxin and fumonisin are main mytotoxins in milk. In the present paper, toxicity of mycotoxin in milk was reviewed, which might be as reference for risk analysis of mycotoxin in milk.
Determination of Mycotoxins in Milk: A Review
ZHENG Nan;WANG Jia-qi;;HAN Rong-wei;XU Xiao-min;;ZHEN Yun-peng;;QU Xue-yin;;ZHAO Lian-sheng
2012, 39(3):  14-18. 
Abstract ( 505 )  
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It is essential to analysis mycotoxins in milk for mycotoxin research.In the present paper the classic methods for determination of mycotoxins in milk,including TLC,HPLC,LC-MS and ELISA,were reviewed.The principle,advantage and disadvantage for each method were mentioned.Further,the trend for determination of mycotoxins in milk was proposed.The information in this paper might be as reference for research on mycotoxins in milk.
生物技术
Bac to Bac Expression and Antigenicity Identification of P72 Gene of ASFV
BAI Li-hua;HOU Shao-hua;JIA Hong;YUAN Wei-feng;GUO Xiao-yu;LIANG Qian-qian;ZHU Hong-fei
2012, 39(3):  19-23. 
Abstract ( 530 )  
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This experiment was designed to express P72 gene of African swine fever virus (ASFV) in Bac to Bac system for further research in the structure. function and immunogenicity of the P72 gene. In this study,P72 gene complete sequence (1941 bp) was amplified by PCR using pGEX-6p-1-P72 for its template. The product was cloned into pFastBac HTa granulosis virus vector and transferred into DH10Bac competent cells. Recombined plasmid transfered into the Bacmid was named rBacmid-P72 and the rBacmid-P72 was transfected into Sf9 incect cells. The recombinant baculovirus was confirmed by PCR using M13 primers. Western blotting and sandwich ELISA confirmed that the P72 protein was expressed in Sf9 cells correctly and can be recognized by rabbit anti ASFV positive serum and P72 mAb. The results indicated that P72 protein expressed in Bac to Bac system had intensive specificity,stabile activity and good antigenicity,and can be used as candidate antigen for detecting ASF.
The Propagation and Evaluation of SHIV-KB9 Adapative Virus Stock in Chinese Original Rhesus Macaques
CONG Zhe;TAO Zhen;WANG Wei;CHEN Ting;JIN Guang;YAO Nan;SU Ai-hua;WEI Qiang
2012, 39(3):  23-27. 
Abstract ( 410 )  
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To propagate SHIV-KB9 virus stock via passaged in Chinese-original monkey PBMCs and realize viral replication and immunological trauma in the monkey after inoculation intravenously with 10 fold dilution of SHIV-KB9. Infectious virus was generated by ligating pSHIV-KB9 5' and pSHIV-KB9 3', and transfecting CEMx174 cells. The peripheral blood mononuclear cells(PMBCs) from Chinese-original rhesus macaque were cocultured with the supernatant of transfection and the virus stock was collected when the viral replication reached the peak area.The variation of env gene was analized, so did the viral loads and tissue culture infective dose (TCID50)with TZM-bl cells. Three Chinese-origin rhesus monkeys were infected with this batch of SHIV-KB9 intravenously with 10 fold dilution and the viral loads in plasma and the change of CD4+ and CD8+ T lymphocytes were analysed during the whole infection. In this study, totally 95 mL virus stock was propagated in monkeys PBMCs and without any variation about the gp120 sequence of env gene. The viral loads was 2.678×105 copies/mL and had 3.16×103 TCID50 in 1 mL of cell-free SHIV-KB9 stock. 3 rhesus macaques were infected and presented severe and rapid depletion of CD4+ cells during acute phase. This panel of SHIV-KB9 adapted in Chinese-original rhesus monkeys' PBMCs were highly infectious and suitable for the animal modle.
Adiponectin and its Receptors mRNA Levels in Swine Adipose Tissue in the Condition of Cold Stress
FU Shou-peng;REN Bao-yan;LIU Ying;LI Su-nan;LI Yang;WANG Mao-peng;YANG Zhan-qing;WEI Li-bin;LI Zhi-qiang;LIU Ju-xiong;WANG Wei;GUO Bin;HU Zhong-ming
2012, 39(3):  28-32. 
Abstract ( 444 )  
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The adipose tissue samples in the neck, pars dorsalis and viscera mesenteria of pig under the temperature of -15~-10 ℃, -10~-5 ℃, -5~0 ℃, 15~18 ℃ were collected for sthdying the expression of adiponectin and adiponectin receptors mRNA by Real-time fluorescent quantification reverse transcriptase PCR to explore the effects of cold stress on lipometabolism. The results showed that the expression of adiponectin mRNA in the neck, pars dorsalis and viscera mesenteria gradually decreased with the increasing cold stimulus and the differences were significant (P<0.05). The levels of AdipoR 1 and AdipoR 2 mRNA in viscera mesenteria firstly increased and then declined (P<0.01). The expression of AdipoR 2 mRNA in pars dorsalis gradually decreased and then returned the normal level; AdipoR 1 mRNA of the different groups had no obvious difference. AdipoR 2 mRNA in the neck went up gradually and declined to the normal level(P<0.01); AdipoR 1 mRNA firstly increased then down-regulated to normal level and went up again(P<0.01). It was concluded that adiponectin and AdipoR are associated with the process of cold stress, which may play important role on redistribution of adipose tissue under animal cold stress.
Amplification and Prokaryotic Expression of B23.1 Protein Gene in Vero E6 Cell
LV Mao-jie;CHEN Jian-fei;SHI Hong-yan;FENG Li
2012, 39(3):  33-36. 
Abstract ( 370 )  
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A fragment containing B23.1 protein gene of Vero E6 cell was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) using the upper/lower primers each including the recognition sequence for BamH Ⅰ and Xho Ⅰ respectively. The results showed that the ORF of B23.1 gene was composed of 901 bp, deletion and insertion were not found. By digestion with restriction enzymes BamH Ⅰ and Xho Ⅰ, the B23.1 gene were cloned into pGEX-6p-1 vector to construct recombinant plasmid named pGEX-6p-B23.1. The recombinant plasmid was transformed into E.coil BL21(DE3) and induced with IPTG. The results of SDS-PAGE assay showed that the protein was largely expressed,the GST-tagged protein was purified. Western blotting analysis showed that the target protein had a good reactivity with B23 monoclonal antibody (mouse).
Expression of Epsilon-toxin Gene of Clostridium perfringens Type D and its Primary Immunological Protective Function
WANG Guang-hua;LIN Guo-zhen;ZHENG Fu-ying;CAO Xiao-an;GONG Xiao-wei;ZHOU Ji-zhang;QIU Chang-qing
2012, 39(3):  36-41. 
Abstract ( 409 )  
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Epsilon-toxin (ε-toxin) gene was amplified from chromosomal DNA of Clostridium perfringens type D (C60-2) by polymerase chain reaction (PCR),and a 906 bp epsilon toxin gene fragment was obtained. Sequence analysis indicated that the homology of the nucleotide sequence of the strain to those other reference strains was more than 99%. The expression plasmid pET32a-ETX was constructed by inserting the epsilon toxin gene into the prokaryotic expression vector pET32a. The plasmid pET32a-ETX was transformated into E.coli BL21(DE3)plys and the recombinant strain BL21(pET32a-ETX) was obtained. Then,the transformants were induced to express with IPTG. The specific 54 ku protein was detected by SDS-PAGE and the immunogenicity of the expressed epsilon toxin was confirmed by Western blotting and ELISA. The obtained recombinant protein was transformed into epsilon toxoid vaccine by adding 0.4% formaldehyde into epsilon toxin. The protective immune response was proved after the mice was immunized with epsilon toxoid vaccine. The results showed that the recombinanted strain BL21(pET32a-ETX) could be as a candidate of epsilon toxoid vaccine to provide protective immune response against C. perfringens type D infection.
Expression of Neurotrophin-4/5 in the Reproductive Organs of the Prepubertal Gilts
ZHANG Wei;YI Kang-le;ZHOU Xu
2012, 39(3):  41-46. 
Abstract ( 398 )  
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The expression of neurotrophin-4/5 (NT-4/5) mRNA and protein in ovary, oviduct and uterus during the follicle and luteal phase of the pubertal gilts were investigated by quantitative real-time PCR and Western blotting. The results showed that the mRNA and protein of NT-4/5 were present in ovary, oviduct and uterus during the follicle and luteal phase. In addition, the expression amount of NT-4/5 mRNA in ovary, uterus during the follicle stage were significantly higher than the luteal phase (P<0.05), however, the expression level of NT-4/5 mRNA in oviduct during the follicle phase was significantly lower than the luteal phase(P<0.05). In collection, these results strongly suggest that local production of the NT-4/5 may particulate in the regulation of reproductive functions in the reproductive functions in the reproductive tract of the prepubertal gilts.
Cloning and Expression of Chicken IFN-γ Gene
LU Zhi-chuan;SHI Yao-xu;YANG Xiao-pu
2012, 39(3):  47-50. 
Abstract ( 374 )  
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The total RNA was abstracted from the concanavalin A(Con A) -activated spleen lymphocytes, a pair of specific primers was designed by Oligo software based on the sequence of chicken interferon-gamma submitted by digby in GenBank(accession No.U27465). A 500 bp DNA fragment was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and then ligated into T-easy vector for sequencing. The results showed that the fragment contained the complete open reading frame (ORF) of CHIFN-γ gene. In comparison with GenBank data, the homology of the nucleotide sequence was 100%.The sequence was inserted into pGEX-4T-1 and the recombined expressing vector was constructed, then transformed into Escherichia coli BL21, which were further induced by IPTG and cultured,and a fusion protein was obtained with about 43 ku of molecular weight in SDS-PAGE.This results showed that the cloned CHIFN-γ gene was expressed in prokaryotic cells.
Semi-quantitative RT-PCR and Bioinformatics Analysis of Sanhuang Chicken ST3Gal6 Gene
ZHOU Fei;CUI Lian-cheng;AN Yu-fu;NING Zhang-yong
2012, 39(3):  51-54. 
Abstract ( 418 )  
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This study was designed to analyse the tissue specific expression profile and bioinformatics characteristic of ST3Gal6 in Chinese yellow chicken. Based on the sequence of the ST3Gal6 of the chicken reported in NCBI,2 pairs of primers were designed and the primers were used to amplify the Chinese yellow chicken ST3Gal6 gene by PCR. Then the tissue specific expression profile was analyzed by semi quantitative RT-PCR,and the bioinformatics analysis of ST3Gal6 was summarized.The results showed that,the full length of the cloning gene of ST3Gal6 was 1169 bp,including the 1059 bp of CDS sequence of chicken ST3Gal6,which encoding a 352 amino acid residues deduced protein. This nucleotide sequence of CDS shared 62%,62%,61.9%,59%,54.4% homology with the ST3Gal6 mRNA from human,pongo pygmaeus,cattle,mouse,Xenopus laevis respectively.Semi quantitative PCR revealed that the ST3Gal6 gene was expressed in various tissues,but at different levels. The expression level of this gene was higher in the brain while lower in lung tissue. The biologic softwares were used to predict the bioinformatics of ST3Gal6,which showed that the ST3Gal6 was a kind of transmembrance protein and have 2 transmembrane domains in chicken.There were 22 phosphorylattion sites and one phosphory lation site of specific protein kinase in ST3Gal6 protein.
Cloning and Tissue Expression of Sus Scrofa Thyroid Hormone Responsive spot14 Gene
WANG Cong-mei;GUO Yu-jie;WANG Yue-ying;HAN Li-qiang;LI Hong-ji;YANG Guo-yu;
2012, 39(3):  55-59. 
Abstract ( 380 )  
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A pair of cloning primers were designed based on the sequence information in silico and were used to clone and analysis the Sus scrofa thyroid hormone responsive spot14 (THRSP,accession: JF951726). The positive clone was identified and sequenced, at the same time the THRSP gene organization distribution rule was analyzed by RT-PCR. The length of Sus scrofa THRSP gene was 621 bp, including an open reading flame of 450 bp, encoding 150 amino acids. Identity analysis showed that the THRSP nucleotide sequence in Sus scrofa shared 83.8%, 88.5%, 80.9%, 80.7%, 85.1%, 47.1% homology with that of Homo sapiens, Bos taurus, Rattus norvegicus, Mus musculus, Oryctolagus cuniculus, Gallus gallus respectively, the predicted peptide shared 76.9%, 80.8%, 76.2%, 76.2%, 82.1%, 32.0% homology with that of Sus scrofa. The phylogenetic tree demonstrates that Sus scrofa was the closest to cattle. Semi-quantitative RT-PCR analysis showed that the expression level of THRSP was the highest in liver and adipose tissue, the lower in cerebrum, spleen, stomach, skin, caecum and rectum, the lowest in lung, duodena jejunum and ileum, without in kidney and muscle. The high expression level of THRSP in the liver, fat and other fat generation of the organization suggest that which might adjust as fat synthesis of metabolic control factor, but fat synthesis in the regulation mechanism needs further research.
Isolation, Identification and Genomic Sequencing of FS Mutation Strain of Porcine Reproductive and Respiratory Syndrome Virus
LI Jia-bin;ZHAO Meng-meng;LI Yu-gu;MA Chun-quan
2012, 39(3):  60-64. 
Abstract ( 427 )  
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A porcine reproductive and respiratory syndrome virus (PRRSV) strain isolated from visceral organs was designated as PRRSV-FS. The sequence analysis revealed that the isolated virus was a mutation strain as the Nsp2 gene existed amino acid deletion of 30 and the GP5 gene was conservative relatively. The genome of PRRSV FS strain showed 88.7%, 98.8%,58.5% of nucleotide indentity with ATCC VR-22332, JN-HS,Europena Lelystad virus respectively.
Progress of Mannose-binding Lectin-associated Serine Protease 2
WANG Xin-ju;JU Zhi-hua;WANG Chang-fa;LI Guo-rong
2012, 39(3):  65-70. 
Abstract ( 411 )  
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MBL-associated serine protease (MASP) and mannan-binding lectin (MBL) are key factors of the lectin pathway of complement activation and play an important role in the defense.MBL or ficolin can form complexes with sugars presented on pathogens through its carbohydrate recognition domain (CRD) and its collagen-like region (CLR) can recognize and binding to MASP. MBL can activity the zymogens of MASP which is a key enzyme and activate the complement cascade through lectin pathway.
Loop-Mediated Isothermal Amplification Technique and Its Application in Detection of Animal Disease
LI Jie;WANG Pei-yuan;ZHANG Ping;LI Jin-ping;LIN Zhi-bin;LI Guo-qing
2012, 39(3):  71-74. 
Abstract ( 391 )  
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Loop-mediated isothermal amplification(LAMP) technique is a sensitive,specific method which has been widely used for rapid detection of animal disease, this article reviewed LAMP amplification mechanism, technical characteristics and its application in recent years, aiming to provide reference in further research and application of LAMP.
Cloning and High-level Expression of a Core Fragment Gene of Prion Protein from Sika Deer (Cervus nippon)
LIU Dong;LU Shi-ying;LI Yan-song;LI Zhao-hui;WANG Guang-ming;ZHANG Yuan-yuan;ZHOU Yu;SONG Jie;LIU Zeng-shan;REN Hong-lin
2012, 39(3):  78-80. 
Abstract ( 426 )  
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The gene of prion protein protease resistance fragment PrPres of the Sika deer was amplified by PCR using a pair of specific primers. The product of PCR was cloned into the expression vectors of pET-Trx and pET-His. The restructuring plasmids were transformed into the host bacterium of E.coli BL21 (DE3) plys, at 37 ℃ for 4 h. SDS-PAGE result illustrated that fusion proteins of Trx-PrPres and His-PrPres were highly expressed. The expression level of fusion proteins were 38.2% and 30.1%, respectively.
Brief Introduction of Immunefluorescent Test Technology and Application Progress on Parasites Detection
WANG Xiao-huan;YANG Lian-ru;ZHAO Lin-li;WU Shao-qiang
2012, 39(3):  81-84. 
Abstract ( 396 )  
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Immunofluorescence technique is a new detection technology which is based on biochemistry,microscopy and immunology. Its basic principle is using fluorescence-labeled antibody or antigen to combine with the antigen or antibody in the samples,and the fluorescence can be recognized under a microscope. The microscopic accuracy and the immunological specificity and sensitivity can be organically combined in this method. As a rapid diagnostic method,immunofluorescence technique is widely used in medical and biological research. It is a general method in the diagnosis of virus,bacterium or parasite caused diseases,such as Toxoplasma gondii infection. With the development of labeled antigen and antibody,immunofluorescence technique will be widely used in parasitosis diagnosis.
The Influence of miRNA to Stem Cell Reprogramming
BAO A-dong;SU Zhi-fang;LU Tao-feng;MA Hui-ru;WANG Chang-shou 
2012, 39(3):  85-88. 
Abstract ( 421 )  
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The varied roles played by miRNA in the maintenance of embryonic stem (ES) cell self-renewal and pluripotency. Several recent studies that have begun to probe the roles of miRNAs during the reprogramming of somatic cells into induced pluripotent stem (IPS) cells. Subsequently, the function of numerous miRNAs has been shown to control the fate of ES cells and to directly influence critical gene regulatory networks controlled by pluripotency factors Sox2, Oct4, and Nanog. miRNAs are likely to influence the process of reprogramming. In addition, we posit that more than a dozen additional miRNAs are excellent candidates for influencing the generation of IPS cells.
Research Advance on Microneme Protein 3 of Toxoplasma gondii
CHEN Jin-ling
2012, 39(3):  88-91. 
Abstract ( 360 )  
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Microneme protein 3(MIC3) of Toxoplasma gondii is a secreted protein. MIC3 has high immunoreactivity,and plays an important role in recognization, adhesion and invasion for T.gondii to host cells. Studies on MIC3 of T.gondii will promote the development of diagnostics agents and vaccines to prevent T.gondii infection.Here,the research advances on molecular characterization of MIC3 and its potential applications on developing diagnosis mathods and vaccine were reviewed.
Cloning and Sequence Analysis of 16S rRNA Gene for Haemophilus parasuis in Swine
WANG Zhu-rong;LU Ji-jun;ZHANG Jin
2012, 39(3):  92-94. 
Abstract ( 519 )  
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Haemophilus parasuis were identified by PCR from Hunan province isolates,and used 16S rRNA sequences to reconstruct their phylogenetic relationship. The partial 16S rRNA was amplified from each H.parasuis, and sequences were aligned using the ClustalX 1.81. MP and NJ trees of 16S rRNA were constructed using the software Phylip 3.67 version 4.0 and Mega version 4.0, and ML tree was also constructed using Puzzle version 5.2. Sequence homology analysis was performed using the Megalign program of the software DNAStar version 5.0. The results showed that the lengths of 16S rRNA sequences were 783 bp. The constructed phylogenetic tree revealed that the Hunan isolates and the H. parasuis serovar 05 available in GenBank were clustered in the same clade. The results of the present study provided foundation for further studies of molecular epidemiology of H.parasuis and for diagnosis of the resultant disease.
动物营养与饲料科学
Effect of Grape Procyanidins on Growth Performance and Immune Function in Broilers
ZHANG Hai-jun;XU Lei;YUE Hong-yuan;WU Shu-geng;PAN Ying-zi;WEI Shu-min;WANG Jing
2012, 39(3):  99-103. 
Abstract ( 525 )  
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This experiment was conducted to investigate the effects of dietary grape procyanidins on growth performance and immune function in broiler chickens. Two hundred and forty day-old Arbor Acre male broiler chickens were randomly allotted into four dietary treatments with different inclusion ( 0,7.5,15 or 22.5 mg/kg) of grape procyanidins. The experiment period lasted for 6 weeks. The results showed that chicks fed higher dose of grape procyanidins intake more feed and grew faster during the first 3 weeks. During the second 3 weeks and the whole 6 weeks,chicks fed 7.5 mg/kg grape procyanidins had higher body weight gain and superior feed efficiency,and chicks fed 15 mg/kg grape procyanidins diet had inferior growth performance compared to the controls. Dietary grape procyanidins did not affect the growth of the immune organs. The percentages of CD3+,CD4+,CD8+,αβ and γδ T lymphocytes in peripheral blood had the increased tendency in chicks fed 7.5 mg/kg grape procyanidins. Overall,optimal dose of grape procyanidins in the diet would stimulate late growth and modulate immune function in broiler chickens.
Effects of Different Dietary Non-fiber Carbohydrate to Neutral Detergent Fiber Ratios on Ruminal and Blood Endotoxin and Histamine Concentrations
HU Hong-lian;LIU Da-cheng;LU De-xun;GAO Min;SHAN Dan;REN Xiao-ping;NIU Wen-yi
2012, 39(3):  104-109. 
Abstract ( 489 )  
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The objectives of this study were to explore the effect of different dietary NFC/NDF ratios on ruminal and blood endotoxin and histamine concentrations,3 Guanzhong dairy goats with permanent ruminal cannulae were divided into 4 groups and each time lasted for 10 d according to self-contrast design experiment. The goats were fed diets with different NFC/NDF ratios,which the ratios were 1.02(Ⅰ group),1.24(Ⅱ group),1.63(Ⅲ group)and 2.58(Ⅳ group),respectively. The results showed that with dietary NFC/NDF ratios increased,ruminal endotoxin and histamine increased. Resulting in blood endotoxin and histamine increasing. When NFC/NDF ratio was 2.58,concentrations of histamine and endoxin in blood and rumen increased significantly with the development of SARA,indicating that increased release of histamine and endotoxin in blood and rumen played an important role in the development of SARA.
The Production and Applications of Taurine in Livestock and Poultry Aquaculture
HAN Zheng-qiang
2012, 39(3):  110-113. 
Abstract ( 410 )  
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Taurine as a kind of semi-essential amino acid, has comprehensive biological functions and is important in the regulation of normal physiological function. The article introduces taurine's metabolic in animal's body and the production of taurine. Show the biological function such as enhance immunity, antioxidation and summarize the applications of taurine's biological functions. Give the outlook of taurine's application and proposed the direction of research in future.
生理生化
Preparation of Arsanilic Acid-TMP and its in vitro Antibacterial Activity
CAI Hao-shuai;HAN Jian-zhong;BI Li-jun;QU Dao-feng
2012, 39(3):  119-124. 
Abstract ( 402 )  
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Arsanilic acid and TMP were utilized as the raw materials and then arsanilic acid-TMP was prepared. The new compound was characterized by IR and melting point. Orthogonal test showed that the best reaction condition was that being heated at 80 ℃ for 3 hours under reflux, and the molar ratio of arsanilic acid and TMP was 1.1∶1. The in vitro antibacterial indicated that the inhibitory zone diameter of arsanilic acid-TMP against the five kinds of tested bacteria is longer than 20 mm and arsanilic acid-TMP had greater bacteria-inhibiting effect than the physical mixture of arsanilic acid and TMP (P<0.05); the minimum inhibitory concentration (MIC) of arsanilic acid-TMP against Escherichia coli, Staphylococus aureus,Salmonella pullorum,Shigella flexneri and Micrococcus luteus were 32,64,256,32,128 μg/mL, respectively, which were less than the MIC of the physical mixture of arsanilic acid and TMP. According to this study, arsanilic acid-TMP has greater antibacterial effect than arsanilic acid, TMP and their physical mixture.
Screening,Identification of a Strain with High Keratinase Production and the Primary Study of its Characterization of Fermentation
GUO Gang;CHU Jie;WANG Jun-gao;LIU Ke-chun
2012, 39(3):  125-129. 
Abstract ( 397 )  
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A strain with high keratinolytic activity was obatained by casein plate firstly and feather as the sole carbon source and nitrogen source in submerged medium secondly.The strain S6 was identified as Bacillus licheniformis based on its morphologic and physicochemical characteristics.After fermenting 72 h in submerged culture,the Bacillus licheniformis S6 demonstrated highest keratinolytic activity and maximum of soluble protein, 186.8 U/mLand 160.87 μg/mL,respectively,which expresses that Bacillus subtilis S6 has a promising prospect in the degradation process of feather.
Separation Culture and Characterization Analysis of Mongolian Sheep Fetal Skin Fibroblasts
MENG Fan-hua;XIAO Hong-mei;ZHANG Dong;ZHOU Huan-min `
2012, 39(3):  129-133. 
Abstract ( 427 )  
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In this study, selecting Mongolian sheep fetus as materials, determining the fetas sex, the results showed that they were male. The skin tissues were cultuered in vitro and the cells obtained were purified and fibroblasts were screened. Through observing, the cell modality was better in 10 generations. The spindle-shape cells showed a elongated trend with the generations increasing and the spindle-shape cells stretched obviously in 20th generation and the number of cells per unit area reduced significantly. In the meantime, the growth force of cell s declined slightly with the increasing generations by the growth curve of F5 and F10 cells. But the trend was not obvious. Through the karyotype analysis of F5 and F10 cells, we found that diploid rates were 85.71% and 76.67%, which met the reques of nuclear transfer somatic cell. Recovery of adherent cells after cryopreservation was in good conditions.
Interaction between Virus Infection and NF-kappaB Signal Transduction
HUANG Hui;YAN Yu-lin;GAO Hong;GAO Li-bo;ZHAO Ru;CHEN Ling;JIANG Huan;CHEN Gang
2012, 39(3):  134-136. 
Abstract ( 450 )  
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Since we found NF-kappaB,it has been studied widely about its molecular biology characteristics,function mechanism and the relationships with diseases. Recently many studies reveal that the NF-kappaB plays an important role in the process of virus infection and disease. The translocation of NF-kappaB from the cytoplasm to the nucleus can be induced when virus infect eukaryotic cells,consequently or inducing transcription of some inflammatory gene to produce a lot of inflammatory factors,or blocking apoptosis and prolonging survival of the host cell to gain time for replication and increase viral progeny production,or promoting the expression of oncogene to make cells cancerization,and so on. Once virus infection,body properties will be changed through NF-kappaB signal transduction and diseases will appear soon. Many fators,which has been found at present,such as stress stimulator,LPS,virus,oxygen free radical and so on,are all able to activate the NF-kappaB and then affect the body's metabolism through NF-kappaB signal pathway. In this paper,the interaction between virus infection and NF-kappaB signal transduction is reviewed.
Advances of Telomerase Activation Regulation in Cells Cultured in Vitro
SUN Wu-sheng;WANG Nan-nan;XU Li-jie;FANG Nan-zhu;LI Zhong-shu;LI Fu-jun
2012, 39(3):  137-140. 
Abstract ( 370 )  
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Telomerase is capable of maintaining telomere lengths by balancing out the gradual loss of telomeric sequences that occurs during cell division. It plays an important role in regulating life span of animal cells.In order to improve cell culture efficiency in vitro,a great deal of research has been attempted. And the way of increasing the activity of telomerase is considered to be one of the most effective approaches to improve cell culture efficiency. This article will make a brief summary of these recent achievements.
遗传繁育
Association of CA Repeat Polymorphism at Intron 1 of Insulin-like Growth Factor Ⅰ Gene with Body Weight and Body Size in Shanxi White Pig
WANG Xiao-jing;WANG Song-bai;SHI Jian-zhong;YANG Xiao-fen;JIA Jing-min;GAO Peng-fei;GUO Xiao-hong;LI Bu-gao;CAO Guo-qing
2012, 39(3):  140-144. 
Abstract ( 429 )  
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A total of 174 pigs from Shanxi White pig were used to investigate associations between the polymorphic (CA)n sequence repeat located at the first intron of the insulin-like growth factor (IGF-Ⅰ) gene and the traits of body weights at different growth stages and body sizes at six-month age by unvariate animal model. There were six alleles,which were 198,202,204,206,208 and 210 bp,were detected in Shanxi White population. The frequencies of alleles of 198 and 202 bp,with less CA repeat number,were 0.2759 and 0.2385,respectively,which were greater than those of alleles with more CA repeat number,such as the allele of 208 bp,whose frequency was 0.0747. Twelve genotypes were found in Shanxi White population,the frequencies of homozygotes were greater than 0.010 apart from 208/208 bp,and the frequencies of heterozygotes were 0.017. The associations of IGF-Ⅰ genotype with the traits of birth weight,weaning weight,body weight,body height,body length,chest girth,back fat thickness alive at six-month age were significant (P<0.05). Individuals with genotype of 204/204 bp has greater body weight and thinner back fat thickness than those of others,which was in accordance with the pig breeding direction in China,should be selected as boars or sows in the selection and breeding programme.
Research Progress of the DNA Molecular Genetic Diversity of China Native Chicken Breeds
LI Xiao-qing;LI Chun-feng;CHU Xiao-hui;XU Zhi-qiang;CHEN Xiao-bo;JIA Jun-jing
2012, 39(3):  145-147. 
Abstract ( 433 )  
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The main gentic diversity have morphological level,chromosomal level,protein level and DNA levels, etc. The kinds of native chicken in china,study the genentic diversity not only can enhance biodiversity conservation, but the great significance in the origin, evolution, clasification, identification and genetic breeding. Since the 1980s, the rapid development of molecular biology techniques for the deterction genetic diversity provides a more direct and accurate method that directly analyzing DNA sequence changes in the level of detection of animal genetic diversity. DNA analysis method to be the most effective methods of genetic analysis, to avoid the inference based on genotype phenotype of error may arise. Research of the DNA molecular genetic diversity of china native chicken breeds in this paper, excavation the good genes for the native chicken breeds, and provied the reference for the native chicken breeds in molecular breeding and improving.
Isolation and Identification of Embryo and Ear Marginal Fibroblasts in Zaosheng Cattle
CHEN Shi-en;TONG Wei-jian;GUO Peng-hui
2012, 39(3):  148-152. 
Abstract ( 395 )  
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The embryo and ear marginal fibroblasts in Zaosheng cattle were isolated and identified in vitro using enzyme digestion and explant culture. The survival rate of ear marginal fibroblasts cultured with smaller explant were higher than that with bigger tissue. The growth curves of the embryo and ear marginal fibroblasts cultured with enzyme digestion were sigmoid type. The doubling time of embryo fibroblasts (29.7 h) was shorter than ear marginal bibroblast (31.2 h). The survival rate of defrozen embryo fibroblasts (96.8%) was higher than that of ear marginal fibroblasts (94.7%). The immunohistochemical staining was used to identify that intermediate filament proteins of isolated fibroblasts were stained tan, showing vimentin reaction is positive.
Study on Growth Development Law and Slaughter Performance in Different Crossbreed Geese
ZHANG Ling;DUAN Xiu-jun;DAI Yan-ping;YUAN Xu-hong;DONG Biao;LI Xiao-fen;SUN Guo-bo;WANG Jian;BIAN You-qing
2012, 39(3):  153-158. 
Abstract ( 425 )  
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Three crossbreed geese (LYLTB,LYB,LLYB) were selected to systematically study the growth development law and slaughter performance with Logistic model,Gompertz model and Von Bertallanffy model. All the geese were provided by National Waterfowl Germplasm Resource Pool (Jiangsu Taizhou).The results showed that the three models fitted well the growth-development of Sumu white geese. The fitting rates were over 0.99,the rate of Logistic model was the highest,but no significant difference between the models. According to the actual raising materials,the Logistic model was the best model to fit the growth of Sumu white geese,and could be used to establish the growth standard during the production. The growth rate of LLYB geese was the fastest,the body weight and measurement were faster than LYLYB and LYB geese according Logistic model estimated values. The meat performance of three hybrid combinations were all good,the slaughtered rates were over 83%,the eviscerated were rates over 68%. So LLYB geese was the best hybrid combination among three combinations,and growth rate was fastest(temporarily name: Sumu white geese).
SNPs Detection and Genetic Analysis on H-FABP Gene of Jiangsu Local Goat Breeds
WANG Lan-ping;GENG Rong-qing;JI De-jun;CHANG Hong
2012, 39(3):  159-162. 
Abstract ( 400 )  
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PCR-SSCP and DNA sequencing approaches were applied to assess the single nucleotide polymorphisms (SNPs) and analyze the population genetics of heart fatty acid binding protein (H-FABP) gene of two goat breeds in Jiangsu province. The results showed that only the 132 G/C mutation was detected at the H-FABP gene exon 2 of the amplified fragments, with the formation of GG and GC genotypes. Homozygous GG was the dominant genotype of two goat groups, and the locus was at Hardy-Weinberg equilibrium. Heterozygosity, effective number of alleles and polymorphism information content of Huanghuai goat population was higher than the Yangtze River Delta white goat populations, indicating a relatively large degree of genetic variation and more abundant genetic diversity within population.
Advances in Function of Melanopsin
HU Liang;HE Hui;DAI Xue-long;HE Bo;LI Yin-xia;KANG Bo;JIANG Dong-mei
2012, 39(3):  162-164. 
Abstract ( 368 )  
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Melanopsin are the photoceptor and photoactive substance in the non-image-forming system. It plays an important role in mediating biological rhythm and reproduction. The study on the functions of melanopsin has been the focal point of opsin family functional research recently. In order to promote the clarity of molecular regulation mechanism in the non image-forming system, the advances in the functions of melanopsin was reviewed in the paper.
疾病防治
Development of Nested Real-time RT-PCR for Detection of Influenza A Viruse and Subtype H5
YANG Su;LIAO Xiu-yun;LIAO Ming;SHA Cai-hua;XU Hai-nie;CAO Wei-sheng;CHEN Wei-sheng 
2012, 39(3):  165-171. 
Abstract ( 501 )  
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Combining nested PCR (NP-PCR) with real-time RT-PCR,superimposing advantages of sensitivity and specificity of these two methods,establwhed nested real-time RT-PCR for detecting AIV and subtype H5,which was more sensitive than conventional real-time RT-PCR. Basing on the conserved AIV NP and HA gene downloaded from GenBank,designed nested real-time RT-PCR primers and TaqMan probes for detecting AIV and subtype H5. Target segments recombined plasmids were constructed,and positive controls were prepared with technology of in vitro transcription.Nested real-time RT-PCR was established after reaction mixtures and condition screened out. Test results indicated that this established method can specific detect AIV and H5 subtype,19 of 30 fish aquaculture water samples were presented positive results,multitudinous samples can be detected simultaneously within 5 h. It is a good method for detecting trace AIV from samples of the natural environment,with distinguishing feature of high sensitivity,good specificity,high throughout and rapidity.
Isolation, Identification and Biological Characteristics of GPV XKY10 Strain
MAO Wen-zhi;SHAO Hong-ze;YAO Xin-hua;GAO Hong-wei;SUN Jian;XU Zhi-lin
2012, 39(3):  171-173. 
Abstract ( 400 )  
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Using electron microscopy, PCR testing, blood agglutination and agar method to identify a virus isolated from the liver, spleen and intestinal tissue having typical symptoms in the 10-day-old goslings goose from a goose farm in Baicheng, and then measured the minimum death dose of goose embryo lethal average time (MDT) and the half goose embryo lethality (ELD50) to analyze the virulence. The identification results indicate that the virus is goose parvovirus, and the virulence analysis shows that this strain is a virulent strain.
Histopathology Observation of Cecum and Rectal in the Cold Stress Quail
LIAO Wang;FU Jing;BAI Xiu-juan
2012, 39(3):  174-181. 
Abstract ( 414 )  
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In this study, intestinal tissues were obtained from quail and were kept under the temperature of (12±1)℃ for acute(0 h,30 min,3 h,12 h) and chronic(5,10,20 d) cold stress. Our aim is to explore the effects of cold stress on gut mucosal immune function of quail,through the observation of histopathology changes,in order to provide the scientific data for studying the mechanism of the effect of cold stress on gut mucosal immune system in quail. The results show that it is proved that the cold stress can cause morphological changes of the cecum and rectal in electron microscope levels.
Pathology of Chicken Experimentally Infected with a Avian Influenza Virus, A/goose/Guangdong/2/1996(H5N1)
YE Yuan-lan;LIU Qi-hong;LI Yu-gu
2012, 39(3):  181-186. 
Abstract ( 370 )  
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Chickens intramuscularly inoculated with a avian influenza virus,A/goose/Guangdong/2/1996(H5N1),produced approximately 70% of morbidity and 60% of mortality, and had micropathological and ultrapathological lessions in some degree in multiple organs,such as, systemic hyperemia, congestion, haemorrhage, thrombosis, and cell degeneration,necrosis,and inflammation etc. As well as apoptosis was detected in pancreas, kidney, liver, heart, lung, cerebrum, intestines, thymus, spleen and bursa of fabricius etc.
Development of RT-PCR Assay for Rapid Detection and Subtyping of Swine Influenza H1,H3,N1,N2 Viruses
QI Hai-tao;KONG Wei-li;ZHANG Gui-hong
2012, 39(3):  187-191. 
Abstract ( 407 )  
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According to the genomic sequence of H1N1,H3N2 subtype swine influenza virus (SIV) published in GenBank, the specific primers were designed according to the conserved region of H1,H3 HA,N1,N2 NA and M genes of swine influenza virus (SIV) H1N1,H3N2 subtype. RT-PCR assay was developed for detection of swine influenza H1,H3,N1,N2 subtype. The results showed that the RT-PCR was capable of detecting 104 EID50 of M and 104 EID50 of HA or NA. The results were negative for the detection of porcine reproductive and respiratory syndrome virus,CSFV and other subtypes except the corresponding subtype. The coincidence rates between RT-PCR and virus isolation were 100%. This assay provides a rapid,sensitive,and cost-effective laboratory diagnosis for detecting and subtyping of SIV in pigs.
Survey on the Prevalence of Intestinal Parasites in Yaoshan White Goat
LI Meng-jie;ZHU Dan;QI Meng;SONG Dan;SHI Ya-dong;ZHAO Zi-fang;DU Hai-li;ZHANG Yun;ZHANG Long-xian;NING Chang-shen
2012, 39(3):  191-194. 
Abstract ( 635 )  
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To understand the infection status of intestinal parasites in Yaoshan white goat, Henan province, a total of 63 fresh fecal specimens from a farm were collected and examined the presence of oocysts or eggs using the centrifugation method, the Lugol's iodine-solution staining method and the Sheather's sugar flotation. Seventeen intestinal parasite species were discovered and the overall infection rate was 98.4%, there were Eimeria spp.(n=9), Cryptosporidium, Giardia, Entamoeba, Strongylus spp., Nematodirus, Trichuris, Moniezia and Trematoda. Eimeria spp.was found with the highest infection rate of 95.2%. The Coccidia oocysts obtained were identified and nine Eimeria species were discovered, with most of which being mixed infections (the highest number of coinfection reaching to five). The infection of intestinal parasites in Yaoshan white goat was very common, thus, necessary measures of prevention and curation are needed to strengthen.
Investigation on the Infection and Prevention of Nematodes in Digestive Tract
XIAO Fang-ping;ZHOU Si-xuan;LIU Hui;XU Jian;XU Hong-zhong;DONG Feng-xian;ZHONG Hu
2012, 39(3):  195-197. 
Abstract ( 461 )  
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Nematodes in digestive tract of one breeding goat farm were examined, the results showed that the infection rate of nematodes in digestive tract was 100% and nematodes, trematode, tapeworm and coccidia were included.There were Haemonchus contortus,Trichostrongylus, Chabertia, Strongyloides papillosus, Bunostomum, Oesophagostomum, Marshallagia, Nematodirus spathiner, Fasciola hepatica, Eurytrema pancreaticum, Ogmocoty, Moniezia, Helictometra and Eimeria, etc. The integrated measures to prevent and control nematodes in digestive tract were made according to the survey results.
Histopathological Observation and PCR Diagnosis of Avian Leukosis in Qingyuan MA Chicken
HUANG Xing-guo;WANG Zheng-fu;GAO Ming-chao;CAI Li-li;WANG Jun-feng;HUANG Shu-jian
2012, 39(3):  197-201. 
Abstract ( 385 )  
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15 Qingyuan MA chickens which appeared obviously clinical symptom and were tested to be avian leukosis positive were selected. The tumor organs, such as liver, spleen, kidney and so on, were carried on histopathology and PCR diagnosis. The pathology tissue slices from organs of liver, kidney, spleen, marrow, gland stomach, heart, lung, pancreatic gland ovary and so on, were observed, results showed that the proliferation of massive lymphoblasts or scatter and of small lymphomyelocytes were observed. The PCR identification indicated that all 15 samples can amplify a fragment about a length of 545 bp, which consisted with the expected size. The results suggested that all 15 isolates were the ALV-J avian Leukosis viruses.
Research Progress and Outlook of Hemsleya and its Preparation
SU Hong;BAI Hua;QI Jing;HU Ming;ZHU Xiao-ling;LIU Yu-qing;LUO Yan-bo;
2012, 39(3):  202-205. 
Abstract ( 377 )  
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Because Hemsleya has comprehensively medical and economical worth and in order to make full use of Hemsleya source comprehensively, the article review ingredients, pharmacological action, clinical using, preparation and quality control et al of Hemsleya, and provide some advice of studies and clinical practice of Hemsleya.
流行病防治
Research of Synthesis Prevention and Control Technology of Porcine Reproductive and Respiratory Syndrome
CUI Ya-lan;ZHOU Bi-jun;WEN Ming;WANG Kai-gong;CHENG Zhen-tao;LI Jing-dan;LONG Hua;YUE Jun
2012, 39(3):  206-210. 
Abstract ( 453 )  
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In order to explore the synthesis prevention and control technology of PRRS, field investigation, serology monitoring and etiology diagnosis were used to analysis the PRRS sample which occurred in Guizhou province, March, 2007 to October, 2009. Different vaccine immunity effect and newborn pig's female source immune body increase and decrease rule also be researched. The study of serology monitoring and etiology diagnosis indicated that PRRS easy transmit in July to October; the newborn pig and sow's disease infected rate are high, achieves 96.15% and 74.73%; the modified live virus vaccine are superior to killed preparations; the immunity PRRSV sow produces newborn pig's head to exempt the date age to decide as on 17 to 20 d; once infected the PRRSV sow to produce newborn pig to exempt the date age to decide as on 30 to 34 d. Investigated the possibility of the mosquito and fly of carrying the PRRSV in the pig farm and proved that mosquito and fly portable carry PRRSV.
A Review of the Molecular Biology Techniques in Detection of Avian Influenza Virus
DAN Xiao-ya;DONG Ying;ZOU Ming-qiang;XUE Qiang
2012, 39(3):  213-216. 
Abstract ( 460 )  
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Avian influenza (AI) is a poultry infectious disease and a highly infectious disease between human and animal caused by influenza A virus.In recent years,the molecular biology techniques in detection of avian influenza virus(AIV) had rapidly been developing.And these were reviewed in this study.
兽医临床
Development of New Animal Models of Arthritis
LIU Ming-qiang;LI Pei-feng;GUAN Hong;
2012, 39(3):  217-219. 
Abstract ( 463 )  
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This review summarizes the major developments of new animal models of arthritis in the past. It concretely focuses on three aspects: the categories and etiopathogenesis of novel transgenic animal models of arthritis, the categories and etiopathogenesis of immune complex animal models of arthritis, the action of related cytokine and synovial cell in the development of arthritis.
经验交流
Study on the Synthesis of Paracetamol Succinate and its Characteristics
JIN Chao;JIN Xiao-you;TANG Wen;ZHOU Tao;FU Hua-lin;
2012, 39(3):  220-223. 
Abstract ( 467 )  
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Paracetamol succinate(AP-S) was synthesized from paracetamol and succinic anhydride by using 4-dimethylaminopyridine (DMAP) as catalyst and acetone as solvent. AP-S yield was selected as the main index, the synthetic technology was optimized by an orthogonal design including factors such as raw material monomer ratio, solvent dosage, reaction temperature, reaction time and so on. Characteristics of AP-S were identified by chemical identification, pH test, melting point test, IR, UV and HPLC. In result, the features of resultants was consistant with AP-S. The optimized synthetic technology was succinic anhydride and paracetamol monomer ratio 2∶1, catalyst 0.15 g, acetone 30 mL, temperature 60 ℃ and time 6 h, and the yield was 81.7%.
Determination of 3 β-agonists in Pork by Liquid Chromatography-tandem Mass Spectrometriy
ZHENG Bai-qin;LI Ai-jun;ZHANG Jian-min;DONG Li-xue;MENG Jun-li;XIAO Jin 
2012, 39(3):  224-227. 
Abstract ( 428 )  
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To established a method to detect the residues of clenbuterol,salbutamol and ractopamine in pork by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and optimized it. The samples was treated by 0.2 mol/L ammonium acetate buffer and added the beta-glucuronidase to it,purified the extracting solution with PCX column,determined by LC-MS/MS and quantitated with internal standard method. The results showed that the residual concentration of the three kinds of β-agonists in pork was 0.2 to 10.0 μg/kg and the linear correlation was well. The linear correlation coefficient were 0.9997,0.9999 and 0.9993,respectively. We detected the limitative concentration was 0. 5 μg/kg and the coefficient of recovery were 93.57% to 96.63%,92.35% to 96.01% and 91.66% to 96.54%,respectively. This method has the advantages of good sensitive,accuracy and easy repetition and it is suitable for the detection of clenbuterol,salbutamol and ractopamine in pork.
Effect of Fluoride on Growth and Fluoride Content of Gonads in Male Mice
SUN Zi-long;NIU Rui-yan;WANG Jun-dong
2012, 39(3):  227-229. 
Abstract ( 454 )  
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To explore the effect of fluoride on growth and fluoride content of gonads in male mice with sexual maturation, 40 healthy male Kunming mice were distributed into 4 groups randomly including the control group (distilled water), low dose fluoride (30 mg/L NaF in their drinking water), middle dose fluoride (70 mg/L) and high dose fluoride (150 mg/L) group. During the exposure, the healthy condition of mice was observed and the body weight of mice was recorded every week. After 49 days, blood, femur, testis, epididymis, and seminal vesicle were collected and fluoride contents were deternined. Compared with the controls, there was no significant difference in all fluoride groups (P>0.05). In high group, body weight gain was significantly decreased (P<0.05). The fluoride content of femur in all fluoride groups were significantly increased compared to control group (P<0.05). Testis and epididymis fluoride contents increased in a dose-dependent manner, and were significantly higher in 150 mg/L group than those in controls (P<0.05). There was no significant difference in blood and seminal vesicle between all fluoride groups and the control group (P>0.05). The results showed that high fluoride inhibited the growth of male mice with sexual maturation, and can permeate the blood barriers of testis and epididymis to damage reproductive system.
Advances in the Research on Gene Detection Technology of Main Pathogenic Bacteria of Cow Mastitis
HAN Xu;WU Rui;SUN Dong-bo
2012, 39(3):  230-232. 
Abstract ( 437 )  
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Cow mastitis is one of three major disease in cow,bringing serious economic loss,at the same time,indirectly impact on human health. It is very important to pathogens rapid,accurate diagnosis for cow mastitis treatment and prevention. In this paper,the technology of PCR,real-time PCR,gene chip and loop-mediated isothermal amplification (LAMP) technology were comprehensive reviewed in the cow mastitis,in order to provide reference to integrated control of mastitis in cows.