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20 April 2013, Volume 40 Issue 4
Analysis of Genomic Characteristics of Porcine Reproductive and Respiratory Syndrome Virus GX1001 and GX1002 Strains Isolated in Guangxi Province
LIN Chang-hua, SHI Kai-chuang, TAO Chun-ai, ZHONG Cheng, MO Sheng-lan, LI Gang
2013, 40(4):  1-9. 
Abstract ( 385 )  
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To investigate the genomic characteristics of porcine reproductive and respiratory syndrome virus (PRRSV), the full-length genome of PRRSV GX1001 and GX1002 strains from Guangxi province were amplified by RT-PCR, sequenced and analyzed. The sequence data showed that, excluding the poly (A) tail, the genomic sequences of GX1001 and GX1002 were 15329 and 15318 bp in length, respectively. Full-length sequence analysis revealed that GX1001 and GX1002 shared 99.3% nucleotide identity with each other, 84.9% to 99.5% nucleotide identity with representative North American (NA) type isolates, 61.5% to 61.9% nucleotide identity with representative European (EU) type isolates, respectively. Full-length sequence comparisons demonstrated that GX1001 and GX1002 had discontinuous 30 aa deletion in 481th aa and 533th to 561th aa of NSP2 region compared with that of the NA prototype VR-2332, which was considered as the genetic marker of highly pathogenic PRRSV(HP-PRRSV), and had new multiple genomic variations such as mutation, deletion and insertion distributing in different regions. Phylogenetic analysis showed that all the NA type isolates could be divided into four subgroups, and GX1001 and GX1002 belonged to subgroup 4 represented by the highly pathogenic JXA1 strain. The results indicated that GX1001 and GX1002 strains from Guangxi province belonged to HP-PRRSV with new characteristic variations in different regions. This study provided valuable genomic data for further surveillance of HP-PRRSV.
Effect of TCA on Gsα Expression in Peritoneal Macrophage in Mouses
GUO Wen-rui, MAO Wei, GUAN Hong, LI Pei-feng
2013, 40(4):  10-13. 
Abstract ( 199 )  
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The assay was aimed to explore the effect of taurocholate acid (TCA) on Gsα expression in peritoneal macrophage in mouses, and provide clues for explaining the function mechanism of TCA on mouse peritoneal macrophages. The peritoneal macrophages of mouse were isolated and cultured. The effect of TCA on the expression of Gsα mRNA was examined quantitatively by Real-time fluorescent quantitative PCR. TCA(15,1.5 and 0.15 μg/mL) decreased significantly the expression of Gsα mRNA in peritoneal macrophage in mouses (P<0.05), compared with that in control group and LPS group. The results showed that TCA could significantly promote the expression of Gsα mRNA in peritoneal macrophage in mouses.
Cloning and Sequence Analysis of ROP17 Gene of Toxoplasma gondii
NIE Zi-xiong, ZHANG Nian-zhang, HU Qian-jiang, CHEN Rui-zhao, ZHOU Dong-hui, YUAN Zi-guo, LIN Rui-qing, ZHU Xing-quan
2013, 40(4):  13-17. 
Abstract ( 412 )  
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Toxoplasma gondii ROP17 gene was amplified from individual T.gondii strains from different geographical locations and hosts by PCR. These PCR products were subjected to be cloned and sequenced and ROP17 sequences were aligned using the ClustalX 1.81. The phylogenetic relationships among T.gondii strains were constructed using the software Mega 5.0 and Puzzle 5.2. The lengths of all ROP17 sequences were 1375 bp, and the A+T contents were 49.53% to 50.04%. The intra-specific variation among the T.gondii strains was up to 2.23%. The variation rate in amino acid sequences was 0 to 6.11%. Phylogenetic analysis revealed that ROP17 gene sequences could be used as a marker for studying genetic relationships of T.gondii isolates. The low variation in ROP17 gene sequences among T.gondii strains might indicate that ROP17 gene could be used as a potential anti-toxoplasmosis vaccine candidate molecule in further studies.
Prokaryotic Expression of Human MxA Gene and Preparation of the Rabbit Antisera
XIN Ting-ting, WANG Hong-mei, LIU Wen-hao, SUN Tao, WANG Li-qun, HE Hong-bin
2013, 40(4):  18-22. 
Abstract ( 354 )  
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This assay was aimed to express the fusion protein of the MxA in Escherichia coli and prepare the antisera of rabbit against MxA.The recombinant expression vector pET32a(+)-MxA was contrusted by combinating MxA fragment from PCR and prokaryotic expression vector pET32a(+), recombinant protein was induced to express and be purified. We immuned New Zealand rabbits with purified recombinant protein to prepare MxA antisera. Then, ELISA results showed that antibody titer was 1∶64000,results of Western blotting showed that antisera had well combination with MxA.Results indicated that we had succesfully prepared immunogenic MxA protein and antisera which laid a foundation for further study on the methods of clinical diagnosis.
Preparation and Identification of Monoclonal Antibodies against Recombinant Truncated P Protein of Canine Distemper Virus
YUAN Wei-feng, JIA Hong, YU Ping-ping, HOU Shao-hua, GUO Xiao-yu, SHI Li-jun, ZHAO Zhan-zhong, ZHU Hong-fei
2013, 40(4):  22-25. 
Abstract ( 334 )  
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The assay was aimed to prepare monoclonal antibodies (McAbs) against P1 protein of canine distemper virus (CDV). BALB/c mice were immunized with P1 protein and the immunized mice spleen myeloma cells were fused with SP2/0. Two hybridoma cell lines of E9E4C10 and E9F8D9 that stably produced antibodies against protein were identified by ELISA. The antibody titres of ascites for the hybridoma lines were both 1∶106. The subtypes of monoclonal antibodies were both identified as IgG2b. Western blotting and IFA showed that the McAbs specifically recognized certain antigenic epitopes of P1 protein of CDV. Additive index of ELISA competitive binding assay indicated that the two monoclonal antibodies were specific for different epitopes. The McAbs prepared in this study were useful tools for developing immunological diagnosis techniques of CDV and investing P protein function.
Establishment of a Duplex Real-time PCR Assay for Detection of Duck Paramyxovirus and Duck Circovirus
XU Zong-li, XIE Zhi-xun, XIE Li-ji, LIU Jia-bo, XIE Zhi-qin, PANG Yao-shan, DENG Xian-wen, FAN Qing
2013, 40(4):  26-31. 
Abstract ( 313 )  
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A duplex Real-time polymerase chain reaction(drRT-PCR) assay was developed and optimized to simultaneously detect duck paramyxovirus and duck circovirus in one reaction. Two sets of specific primers for duck paramyxovirus and duck circovirus, along with two TaqMan probes specific for each virus were used in the assay. This drRT-PCR assay was found to be specific and be able to detect and differentiate duck paramyxovirus and duck circovirus, and no positive results were observed, when nucleic acid from duck hepatitis virus, Muscovy duck parvovirus, duck plague virus and H9 subtype avian influenza virus and so on were used as drRT-PCR templates. The sensitivity of this drRT-PCR assay was 160 and 140 copies for duck paramyxovirus and duck circovirus,respectivly. The drRT-PCR assay was applied to 118 clinical samples detection. Among these samples, duck paramyxovirus positive rate was 0.85%, duck circovirus positive tate was 8.47%, no co-infection. This drRT-PCR assay was a quick, sensitive and specific test for detecting duck paramyxovirus and duck circovirus,and would be useful for the control of these viruses in ducks.
Studies on Single Nucleotide Polymorphisms at the Exon 2 of H-FABP Gene among Different Sheep Breeds
ZHAO Xue, CAO Yang, JIN Hai-guo, ZHANG Li-chun, YU Yong-sheng, LUO Xiao-tong, LI De-yun
2013, 40(4):  32-35. 
Abstract ( 317 )  
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PCR-SSCP approach was applied to test the single nucleotide polymorphisms (SNPs) at the exon 2 of heart fatty acid-binding protein (H-FABP) gene and four sheep populations that comprised of Northeast fine wool sheep, Small-Tailed Han sheep, Dorper sheep and Dorsets sheep were screened in this study. The results showed that four SNPs at 939(A/G), 980(G/A), 1013(A/C), 1018(T/C) and three genotypes were detected. During the Northeast fine wool sheep, Small-Tailed Han sheep and Dorper populations, the BB was the predominant genotype. While in Dorsets sheep, the AB was the predominant genotype. A chi-square analysis suggested that the allele frequencies and genotype frequencies of H-FABP gene were in Hardy-Weinberg equilibrium. Statistical analysis revealed an intermediate polymorphism information content (PIC) in the Northeast fine wool sheep, Dorper sheep and Dorsets populations (0.25<PIC<0.50), but a low PIC in the Small-Tailed Han sheep population (PIC<0.25). Further analysis showed that the different groups had different degree of genetic variation.
Prokaryotic Expression and Purification of Matrix Protein of BD-06 Strain Rabies Virus and Preparation of its Polyclonal Antibody
ZHAO Xue-chao, ZHANG Xue-wei, ZHANG Shou-feng, LIU Ye, MI Li-juan, HU Rong-liang
2013, 40(4):  36-39. 
Abstract ( 348 )  
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The objective of this study was to obtain polyclonal antibody against the matrix protein of rabies virus in order to provide materials for further study on the function of matrix protein. The full sequence of the matrix protein gene from the BD-06 strain rabies virus was successfully cloned and inserted into the expression vector pET28a. The results of restriction enzyme digesting and sequencing proved that the matrix protein gene was correctly inserted into pET28a. After the expression plasmid was transformed into E.coli Rosetta and induced by 0.5 mmol/L IPTG, 27 ku M protein was obtained. After the matrix protein was purified and vaccinated the rabbit,the polyclonal antibody was obtained. The results of Western blotting and IFA showed that the antibody could react with the rabies virus. The matrix protein in the BHK-21 cell affected by BD-06 strain rabies virus was positioned by using the polyclonal antibody. The results indicated that the polyclonal antibody against the matrix protein of rabies virus was successfully prepared, which laid a foundation for further study on matrix protein of rabies virus.
Prokaryotic Expression and Idenfication of the Preliminary Immunogenicity of the H Protein of Canine Distemper Virus
YUAN Ying-shuo, SONG Fei-fei, ZHANG Le-cui, HU Rong-liang
2013, 40(4):  40-45. 
Abstract ( 309 )  
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The assay was aimed to highly express the H gene of canine distemper virus in E.coli and analyze the immunogenicity of the products. The fragment with main epitope sequences, amplified from Ondetstepoort vaccine strain via reverse transcription-polymerase chain reaction, was cloned into vector pET28b and expressed through IPTG introduction after transformed into RosettaTM strain. The product, after purification, was identified by SDS-PAGE, western blotting and indirect enzyme linked immunosorbent assay. After that, the antibody titers of mice immunized with the purified H proteins were determined with neutralization test.The 1113 bp fragment of interest was specifically amplified by RT-PCR and expressed with the introduction of 1.0 mmol/L IPTG at 37 ℃. The protein expressed, identified by SDS-PAGE and Western blotting with a size of 42.38 ku, could be recognized by anti-CDV antibody without nonspecific reaction. In the animal experiment, the serum antibody titer was determined to be 2-3.3 by neutralization test.The target protein was expressed accurately and specifically, showing a good immunogenicity and antigenicity according to the animal experiment, and therefore could be employed as an antigen candidate for the Real-time examination of animal immune condition, which laid the foundations for the development of new-type canine distemper vaccines and ELISA kit for detecting the anti-CDV antibody.
Study on Genetic Structure of Dahe Black Pig Based on DNA Sequence Analyses of Cytochrome b Gene
TANG Li-zhou, SHI Xiao-dong, DING Wei, YU Long, WANG Jun-jie, ZHU Lei, GUO Song-chang
2013, 40(4):  45-49. 
Abstract ( 321 )  
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For revealing the level of genetic divergence and genetic structure of Dahe Black pig, this research was implemented using molecular biological methods that sequenced and analyzed the Cyt b DNA sequences based on 46 samples from 5 geographic populations of Qujing city. The AMOVA results showed that most of genetic variation (62.63%) distributed within populations. This could indicate fully that the whole population genetic structure of this species was not evident because of significant gene flow among different populations. The results of neutrality tests further validated that the instable population structure of this whole group could be affected by the population expansion from the artificial selective breeding and introduction breeding. This instable population structure or lower genetic differentiation could be induced by the large-scale introduction and expanding breeding. The significant gene flow induced the genetic convergence in various geographical populations eventually, population expanding and population genetic diversity declined significantly. This study was expected to provide important scientific basis and data for the molecular genetic breeding of Dahe Black pig and for the manager’s effective measures to protect the genetic resources of Dahe Black pig.
Effect of Echinacoside on the Expression of the OPN mRNA and Protein in Rat Osteoblasts in vitro
LI Cai-hong, LIU Zhong-jie, ZHENG Shi-jun, HAN Bo, WANG Jiu-feng
2013, 40(4):  50-55. 
Abstract ( 240 )  
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In order to investigate the effect of Echinacoside on the OPN mRNA and protein expression of rat osteoblasts cultured in vitro, the osteoblasts were derived from calvariae of neonatal rat and digested by the trypsin and collagenase. The third-generation of cultured osteoblasts were exposed to different concentrations of Echinacoside, while took oestrogen as positive control. Total RNA and protein were prepared from osteoblasts after 24, 48 and 72 h respectively. And the change from OPN mRNA was dected by fluorescent quantitation PCR,OPN protein was dected by Western blotting. The results indicated that different concentrations of Echinacoside had increased the expression of osteoblast OPN mRNA, especially 5?10-4 and 5?10-5 mg/mL Echinacoside significantly or extremly significantly up-regulated the expression of OPN mRNA at 24, 48 and 72 h. Secretion of OPN protein was more than control group after 48 h addition of Echinacoside.The conclusion was that Echinacoside significantly promoted the expression of OPN mRNA of osteoblasts, while had the trend of raised role for OPN protein.
Establishment of a Nested PCR Assay for Detection of Chicken Infectious Anemia Virus
WANG Jing-ru, YANG Xiao-jing
2013, 40(4):  55-58. 
Abstract ( 257 )  
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According to the sequence of chicken infectious anemia virus (CIAV) strain published in GenBank,two pairs of primers were designed and synthesized.The outer primers amplified a fragment of 485 bp in length, and the inner primers amplification fragment size was 297 bp in length.A nested PCR assay for rapid detection of ALV was established.A specific 297 bp fragment was amplified from DNA templates of CAV strain,but no bands were amplified with templates extracted respectively from avian influenza virus (AIV) subtype H9,Newcastle disease virus (NDV), infectious bursal disease virus(IBDV),egg drop syndrome virus (EDSV), reticuloendotheliosis (REV), avian reovirus (ARV) and Marek’s disease virus(MDV). Sensitivity of the 1st and 2nd amplifications by the nested PCR assay were 100 pg and 1 fg,respectively.The sensitivite of the 2nd amplifications increased by 105 times.The results showed that the nested PCR was specific,sensitive,rapid,and accurate,and could be used as a routine assay for the detection of CAV.This method had good reproducibility, specificity and sensitivity, and might detect low content CAV accurately and rapidly.This method could be used as a method for the diagnosis and detection of clinical cases,and molecular epidemiological investigation of CAV.
Study on Expression of Key Enzyme Genes Related to Milk Fat Synthesis in Mammary Gland of Yak
LI Yu-can, DENG Xiao-yu, JIANG Ming-feng
2013, 40(4):  59-64. 
Abstract ( 408 )  
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The Maiwa yak, which were used as milk producer in the northwest plateau of Sichuan province, were selected as experimental animal. The Real-time quantitive (RT-PCR) was applied to detect the expression level of ACSL1, ACSS1, ACSS2 and FABP3 genes in the mammary gland of yak during the lactation cycle. The results showed that ACSL1, ACSS1, ACSS2 and FABP3 gene were consistently expressed during the whole lactation period. RT-PCR results showed that the expression level of all 4 genes consistently increased from time point -15 d, and the peak appeared at time point 30 d. The expression level of all 4 genes at 30 d significantly increased (P<0.05) compared with time point -15 d. Meanwhile, the peak of milk yield was at time point 120, 90 d later than the peak of mRNA expression levels, then the milk yield consistently decreased till 240 d. These results implied that the expression pattern of milk fat synthesis gene during the lactation cycle reflected the yak’s genetic adaption to living environment in Tibetan plateau.
Cloning and Sequence Analysis of Interleukin 10 Genomic DNA in Common Carp (Cyprinus carpio L.)
FENG Xiang-ru, CHEN Yi-long, SUN Zhen, JIA Sheng-mei, WANG Wen-dong, ZHANG Jun-hui, YANG Zhen-guo, LU Qiang
2013, 40(4):  65-68. 
Abstract ( 231 )  
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The purpose of this study was to obtain the full-length sequence of common carp interleukin 10(IL-10) genomic DNA. The sequence of common carp IL-10 full-length cDNA(GenBank:JX 524550) was used. In this study, we designed two pairs of primers in its untranslated regions. Using the carp spleen genomic DNA as the template, we carried out PCR and obtained the IL-10 full-length genomic DNA sequence. The sequence was 2176 bp in length, and the GenBank accession number was JX524551. We compared the gained sequence with the other IL-10 and found their similarity of all harboring 5 exons and 4 introns. Moreover, all exons were comparatively conserved. The exons and introns splice sites were in accordance with "gt......ag" rules. Our results indicated that it was the sizes of introns that were crucial in the process of evolution.
Establishment and Application of PCR Method for Diagnosing Actinobacillus pleuropneumoniae
ZHANG Jian-xin, ZHU Yan-kun, MA Jin-you, ZHAO Jie, YU Yan, HAN Ke-fang
2013, 40(4):  69-72. 
Abstract ( 342 )  
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According to the APP included in GenBank serum surface antigen of D15 type 1 to 14 and 16S rRNA gene sequences were designed two pairs of primers, namely amplified two nucleotide fragments of 637, 431 bp, and built PCR detection method of APP. The results indicated that type-specific serum 1, 3, 5a, 8, 10 could amplify two expected fragments and Escherichia coli, Pasteurella, Salmonella, Streptococci and Staphylococci, and they were negative. Sensitivity checked out test results showed that the lowest concentration was 50/μL. 5 strains of clinical isolates strain suspected APP by PCR ,the results showed that in four of them were positive, one was negative. The specificity of PCR detection method of the experimental establishment was strong specificity, high sensitivity.
Rapid Detection of Vibrio alginolyticus by Using Loop-mediated Isothermal Amplification Assay
XU Yi-gang, WU Yan, LIU Zhong-mei, LI Dan-dan, CUI Li-chun, LI Su-long
2013, 40(4):  72-76. 
Abstract ( 286 )  
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Vibrio alginolyticus was one of the main pathogens that caused bacterial diseases in fish, shrimp and cowry, and seriously caused harm to the development of aquaculture. Thus, the development of a method to rapidly and accurately detect V.alginolyticus and protect the development of aquaculture was of great importance. In this work, a loop-mediated isothermal amplification (LAMP) method for the rapid detection of V.alginolyticus was developed using the collagenase gene of V.alginolyticus as target gene, and the LAMP test could be completed in 45 to 60 min. Results showed that the LAMP method showed a high specificity to V.alginolyticus and a good sensitivity that 10 CFU/mL for cultivated V.alginolyticus and 19 CFU/mL for its contaminated food. The LAMP method developed in this work was a sensitive, rapid and simple tool for the detection of V.alginolyticus.
Construction and Expression of the Recombinant Expressing Plasmids for the Tetrameric Leader Chain of Swine β2m
LIU Chang-hua, LIU Fa, ZHANG Jing, QIAO Zhen, GAO Feng-shan 
2013, 40(4):  77-81. 
Abstract ( 247 )  
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In order to construct the tetrameric leader chain of swine β2m, the light chain of swine β2m was mutated at site of 69 from phenylalanine (Phe) to cysteine (Cys) by using PCR site-directed mutagenesis technique. Then the mutant β2m gene was inserted into pET-28a and transformed into Escherichia coli BL21. After induction, the expression of target protein was detected by SDS-PAGE. The results demonstrated that the mutant β2m was about 297 bp, with coding for 98 amino acids. It was shown that the phenylalanine at site of 69 was mutated to cysteine. After induction, the mutant β2m was expressed successfully as a protein of 10.6 ku detected by SDS-PAGE. It was concluded that the recombinant expression plasmids of the mutant β2m was constructed and target protein was expressed successfully. This study would lay the foundation for construction of tetramer later.
Analysis of Sequence Variation in Mitochondrial nad4 Gene of Trichuris ovis Isolates
WANG Yan, YANG Yan-chuan, ZHANG Yue-wen, LI Jia-yuan, ZHU Xing-quan, LIU Guo-hua
2013, 40(4):  81-84. 
Abstract ( 287 )  
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The objectives of the present study were to analyze sequence variation in the NADH dehydrogenase subunit 4 (nad4) gene among Trichuris ovis isolates from Guangdong province in China, and to study its phylogenetic relationships with other whipworms using the nad4 gene sequences. The partial nad4 (pnad4) were amplified from individual T.ovis samples, and pnad4 sequences were aligned using the ClustalX 1.81. Neighbor-joining (NJ) trees were constructed using the software Mega version 5.0. The lengths of all pnad4 sequences were 827 bp. Phylogenetic analyses showed that all T.ovis isolates were clustered in the same clade.Sequence variations in pnad4 sequences within T.ovis were 0 to 1.5%, which was significantly lower than inter-species differences (37.8% to 45.6%). It was concluded that pnad4 sequence could be used as a genetic marker for population genetic studies of T.ovis, as well as for the differentiation and identification of different whipworms. The results of this study provided foundation for further studies on molecular genetics and diagnostics of T.ovis.
Research Progress of Bacillus subtilis as Drug and Immunogen Delivery Carrier
YUE Min-jie, SHI Yuan-yuan, FU Shuai-gu, WANG Yan-bin, KANG Xiang-tao
2013, 40(4):  85-89. 
Abstract ( 313 )  
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Bacillus subtilis was a Gram-positive bacteria, and also a non-pathogenic strain, which could form spores. It was a hypopus that could withstand extreme conditions and other noxious chemicals. B.subtilis, with a monolayer cells outer membrane and endotoxin-free cell wall, could efficiently secret large amounts of heterologous proteins out of the membrane directly within animal intestine. B.subtilis as a probiotic, lived in the gut of its host, had the capacity to maintain the balance and stability of the intestinal environment and stimulate the animal to release secretory immunoglobulin. All those above advanced the local immunity of the intestinal mucosa and further enhanced the immunity. Currently,B.subtilis became an ideal vaccine vector to the preparation of mucosal immune vaccine because of its unique advantages to humoral and cellular immune. Gradually, B.subtilis, as a vaccine vector that popularized easily, turn out to be a hot research object. Here, the present study was conducted to illustrate the mechanism of mucosal immune, research progress and application prospects of B.subtilis that worked as a vaccine vector.
Establishment of SYBR Green Ⅰ Fluorescent Quantitative PCR Method for Detection of Porcine Reproductive and Respiratory Syndrome Virus
HUANG Zhen, WEI Chun-ya, YAN Ming-fei, TIAN Yue-ru, ZHANG Min-ze, XIE Jie-xiong, SUN Long, ZHANG Gui-hong
2013, 40(4):  89-92. 
Abstract ( 308 )  
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A pair of specific primers conserved in the the Nsp7 genes of American porcine reproductive and respiratory syndrome virus (PRRSV) were synthesized. Optimization of reaction conditions, the detection of SYBR Green Ⅰ Real-time quantitative PCR method was compared and analyzed at the same time with the commercialization of fluorescent quantitative PCR kit. The results showed that the specificity of the method, high sensitivity, a good distinction between the American swine reproductive and respiratory syndrome virus and other viruses. 14 clinical samples were detected using this method, the positive rate was 85.7%, coincidence was 100% with commercial kits.
Effects of Two Chinese Medicinal Ingredients on Growth Performance, Digestive Utilization Ratios of Nutrients, Digestive Enzyme Activity and Intestinal Bacterial in Broilers
SUN Yao-gui, ZHANG Xiang-jie, CHENG Jia, WANG Jun-dong
2013, 40(4):  93-98. 
Abstract ( 389 )  
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The experiment was conducted to investigate the effects of TFHL, TSG and TFHL+TSG supplement in daily ration for broilers on growth performance, utilization rate of nutrients, digestive enzyme activity and intestinal bacterial.128 male broilers (1 day old) were divided randomly into 4 groups, the control group (fed a basal corn-soybean meal diet), groupⅠ (fed a basal corn-soybean meal diet containing 300 mg/kg TFHL), groupⅡ (fed a basal corn-soybean meal diet containing 150 mg/kg TSG) and group Ⅲ(fed a basal corn-soybean meal diet containing 300 mg/kg TFHL and 150 mg/kg TSG). Each group had 4 replacements, each replacement had 8 broilers. The results showed that:compared with the control group, the ADG in group Ⅲ significantly increased (P<0.05) during the whole experimental periods (1 to 42 days),and the F/G reduced significantly (P<0.05). The digestive utilization ratios of CP, Ca and TP in group Ⅲ were significantly higher than that in control,Ⅰ and Ⅱ groups (P<0.05); the EE digestive utilization ratio was significantly lower than that in control, control,Ⅰ and Ⅱ groups (P<0.01). Compared with the control and Ⅱ groups ,the Trypsin activity in group Ⅲ increased significantly (P<0.01); compared with the control group, the lipase activity significantly lower (P<0.01); compared with the control and Ⅰ groups, the amylase activity was significantly increased (P<0.01). Compared with the control,Ⅰand Ⅱ groups, the coliform counts in ileum and cecum significantly reduced in group Ⅲ(P<0.05). Compared with the control group, the lactic acid bacteria number of ileum in group Ⅲ significantly increased(P<0.05); compared with the control,Ⅰ and Ⅱ groups, the lactic acid bacteria number of cecum in group Ⅲ significantly increased(P<0.05). Supplement of TFHL or TSG in diet played a role in improving production trait and utility ratio of digestion, enhancing activity of intestinal trypsin and amylase, reducing activity of lipase, stimulating growth of intestinal beneficial bacterium and inhibiting reproduction of pathogen for broiler. Taking all factors into consideration, it was better to add the two ingredients to diet at the same time.
Comparison Study on Milk Amino Acids Composition of Dairy Cows from Different Farms in the Spring and the Summer
ZHANG Juan-xia, Sun Peng, YANG Yong-xin, BU Deng-pan, ZHOU Ling-yun, YANG Jin-hui, LUO Qiu-jiang
2013, 40(4):  99-104. 
Abstract ( 321 )  
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The objective of this study was to assess raw milk amino acids composition in different farms between the spring and summer to provide a basis for improving the qualitiy of milk. Milk samples were collected from 6 cities (Beijing, Anhui, Heilongjiang, Neimeng, Xinjiang and Xi’an) in the spring and summer, respectively. Amino acids were extracted by hydrolyzed with HCl, and concentrations of amino acids were determined by Hitachi L-8900 high speed amino acid analyzer. The experimental datas were analyzed by SAS 8.0 with GLM procedure. The results showed that significant variation of Cys, Val, Tyr and Phe were determined between the spring and summer (P<0.01), while others were not siginificant differences (P>0.05). Significant difference of all amino acids existed among these six regions (P<0.01). The interaction effect between seasons and locality were highly significant on the amino acids concentrations of whole milk (P<0.01). It was concluded that amino acids in raw milk were influenced by locality significantly, while Cys, Val, Tyr and Phe were also affected by seasons.
Biological Function and Application of Semen Ziziphi Spinosae Extracts in Livestock Production
PAN Long, CHENG Jian-bo, BU Deng-pan, SUN Xian-zhi, SUN Peng, ZHOU Ling-yun
2013, 40(4):  105-110. 
Abstract ( 331 )  
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Semen Ziziphi Spinosae extracts were the tranquilizing and allaying excitement of the traditional Chinese medicine, whose main components were the saponins and flavonoids compounds. The extracts were gradually paid attention by Chinese veterinarian, with broad development potential and a great market prospect in livestock production. The author reviewed that Semen Ziziphi Spinosae extracts had the effect of the central nervous system, immune system, and the application in livestock, and provided the development and application for the basis in the animal husbandry production.
Effect of Grass Particles Made from Different Proportions of Tomato Sauce Residue and Alfalfa on Rumen Degradation for Sheep
ZHANG Shu-xin, PAN Xiao-liang, WANG Zhen-guo, CUI Zheng-tao, WANG Chen-qiang, PAN Qi, CUI Yan-xia, XU Ya-nan
2013, 40(4):  110-113. 
Abstract ( 245 )  
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Three rumen-cannulated sheep were used to measure ruminal degradation, using the nylon bags method for different ratio of tomato sauce residue and alfalfa, cotton shell particles made of grass in the rumen degradation rare of 2,6,12,24,36 and 72 h,which studied to determine the tomato sauce residue and alfalfa, cotton grass system shell particles suitable ratio, mixed grass particle dynamics of nutrients in the degradation rate. The results showed that tomato sauce residue∶alfalfa∶cotton shell was 60∶35∶5,which was the highest combination of effects.Degradation rate of the group:dp(DM)= 27+40.9×(1-e-0.0139t)(n=6,R2=0.978);dp(CP)=31.9+32.6×(1-e-0.071t)(n=6,R2=0.986) ;dp(NDF)=14.3+56.1×(1-e-0.023t)(n=6,R2=0.994).
Effects of Microbial Fermented Feed on Immune Function Parameter of Beef Cattle
YU Miao, YAN Jin-xiu, PENG Zhong-li, BAI Xue, GUO Chun-hua, YU Ting-ting, AI Dan, CAO Long-kai
2013, 40(4):  114-117. 
Abstract ( 353 )  
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The aim of the study was to illustrate the effect of the microbial fermented feed on cattle by the parameters of the immune and antioxidant capability in the serum. Ninety healthy cattle with the similar age, conformation and weight (373.4 to 425.3 kg), which crossbred by Simmental bull and Native cow were randomly allotted into three groups. One group was the control group, and the others were the treatment groups. There were 30 cattles in each group with 5 replicates and 6 cattles were used in per replicate. The experiment was designed with randomized block design. The cattle in the control group were fed on the basal concentrate. Meanwhile the cattle in the treatment groups Ⅰand Ⅱ were fed on the microecological function feed instead of concentrate by the ratio of 25% and 40%, respectively. Each head of cattle was fed on the same weight of concentrate and corn straw particle, but was allowed free choice continuous access to feed on vinasse and free drinking every day. Every formal experiment period had 48 days, advanced experiment period had 10 days. The results showed that compared with the control group, the concentrations of total serum protein, albumin, IgA, IgG, IgM increased markedly(P<0.05 or P<0.01), while the blood serum concentration of as partate aminotransferase, aminotransferase, maleic dialdehyde decreased significantly (P<0.05 or P<0.01) in the treatment groups. Besides, the total antioxidant capacity and the activity of superoxide dismutase were enhanced significantly (P<0.01)in the treatment groups. This study would provide the scientific basis for improving immunity of beef cattle by microbial fermented feed.
Effects of Free Gossypol on Immunity of the HISEX Young Hens
LU Jun-xian, GAO Yu-shi, TANG Meng-jun, PU Jun-hua, ZHANG Xiao-yan, GE Qing-lian
2013, 40(4):  118-121. 
Abstract ( 260 )  
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To study the effects of free gossypol (FG) in the diets on immunity in the starter period of HISEX layers, 360 brown shell HISEX layers of 1 day of age were assigned randomly to four groups with three pens per groups and 30 chicks per pens. The chicks in control group (group Ⅰ) were fed with the basal diets and the chicks in other groups were fed the diets containing free gossypol, 70,105 and 140 mg/kg, respectively. The trial lasted for 6 weeks from 1 to 42 days old. On 14, 28 and 42 days, indices of immune organs and the level of cytokines in the sera were measured. Sera HI of Newcastle disease was detected once a week. The results showed that the organ index of bursa of fabricius, thymus and spleen were not influenced by the level of FG in the diets.Different levels of free gossypol had no significant effect on ND antibody titre of HISEX layers. Different levels of free gossypol decrease levels of sera cytokine IL-2,IL-4 and IFN-γ. It was concluded that up to 140 mg/kg of FG had no significant effect on immune organ index and titer of Newcastale disease. But it could decrease level of serum cytokine on 42 days.
Effect of Vaccaria segetalis on Cellular Signal Transduction Pathway Associated with Milk Protein Synthesis in Dairy Cow
MENG Hai-yang, SUN Xiao-xu, WANG Li-na, LIN Ye, YUAN Xiao-han, LI Qing-zhang, GAO Xue-jun
2013, 40(4):  121-124. 
Abstract ( 349 )  
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To study the effect of Vaccaria segetalis on cellular signal transduction pathway associated with milk protein synthesis in dairy cow,water extraction of Vaccaria segetalis was prepared.Western blotting was used to study the changes of the signal molecules associated with milk protein synthesis of dairy cow mammary gland epithelial cells (DCMECs), treated with estrogen (ER), prolactin (PRL) and Vaccaria segetalis. The results showed that the Vaccaria segetalis extraction liquid, ER and PRL could promote milk protein synthesis and up-regulate the expression of p-STAT5,p100,GAS,S6K1 and p-mTOR, furthmore, methionine could enhance this action. These datas suggested that Vaccaria segetalis had a similar role with ER/PRL and up-regulated expression of the genes of DCMECs for lactation on the translational and transcriptional level.
Determination of Enramycin in Premix by TLC-UV Spectrophotometry
ZHOU Peng-fei, CHEN Min, FANG Zhong
2013, 40(4):  125-128. 
Abstract ( 605 )  
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A method of TLC-UV was set up for determining enramycin in premix. Target components were extracted by acetone aqueous solution. Using TLC plate, a mixture of N-Butanol∶glacial acetic acid∶water (2∶1∶1) was used as developer. Fixing the position under the light of a UV lamp(254 nm), detection wavelength was 271 nm. The absorbance and quantity of sample application showed a good linear relationship when the quantity of enramycin ranged from 20 to 166 μg (r=0.9989) with average recovery of 99.85% and RSD value of 3.1%(n=9). The content of enramycin in premix detected by this method was almost the same as that was detected by HPLC. This method which was simple, rapid and accurate,large amount of analysis,could be used to the quantitative analysis of enramycin in premix.
Study on Toxicology of Qishen Oral Solution
WANG Jian-fang, KONG Ling-bo, SUN Miao, WU Jie, HU Yan-jiao, GAO Hui-jun, MU Xiang
2013, 40(4):  129-133. 
Abstract ( 229 )  
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To evaluate clinical safety of Qishen oral solution, tests of acute toxicity and chronic toxicity were made according to toxicological evaluation procedures and methods. During the acute toxicity test, 40 Kunming mice were randomly divided into 2 groups. The maximum tolerable dosage of Qishen oral solution was determined by intragastric administration in mice. During the chronic toxicity test, Wistar rats were randomly divided into 4 groups, and the three experimental groups were given Qishen oral solution in doses of 3.75,7.5,15 g/kg by intragastric administration for 4 weeks, once a day, and observed after stopping administration for 2 weeks.The control group was given the same amount of physiological saline.The chronic toxicity were assessed on rats through recording daily water intake, feed intake and weekly body weight, measuring hematologic and blood biochemical indexes after administration for 4 weeks and withdrawal for 2 weeks. The results showed that the maximum tolerable dosage of Qishen oral solution administration was 40 g/kg in mice.It was equivalent to 40 times of the dosage used in clinic.Daily feed intake and water intake for some administration group showed significant (P<0.05)or extremly significant (P<0.01) differences compared with that in the control group, but average weekly gain had no significant changes (P>0.05); during 4 weeks of intragastric administration, some indexes of hematology appeared significantly (P<0.05) or extremly significantly (P<0.01) different from that in the control group, but after 2 weeks, they had no significant changes (P>0.05); blood biochemistry,viscera index and histological examination had no significant difference (P>0.05). In conclusion, the clinical medication of the oral liquid was safe under this experimental condition.
Cell Culture of Endometrial Epithelial Cells from Chinese Holstein Dairy Cows in vitro
ZHANG Shi-dong, WANG Dong-sheng, WANG Xu-rong, PAN Hu, LI Hong-sheng, YAN Zuo-ting
2013, 40(4):  134-137. 
Abstract ( 357 )  
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To establish and perfect cell culture technique of endometrial epithelial cell (EnEpC)in dairy cows, primary cell was isolated and cultured by tissue cultivation that combined with enzymolysis of collagenase. The endometrial epithelial cell was purified from primary culture cells by different time digestion of trypsase, and EnEpC was identified by immunohistochemical detection of cytokeratin expression. The results demonstrated that flagstone-like epithelial cell was obtained by the adoptive method, and cytokeratin expression was positive in the cells. The purified cells could be subcultured about 10 times, and cells from generation 2 to 8 maintained stably biological characteristics of morphology and proliferation state that were found in primary culture cells.
Influences on Apoptosis Mediated by Caspases on Spermatogenesis, Development and Cryopreservation
DING Li, ZENG Chang-jun
2013, 40(4):  138-144. 
Abstract ( 397 )  
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Caspase is a family of cysteine proteases that play an important role in regulating apoptosis. Spermatogenesis and development involve an early waves of germ cell apoptosis, which are required for maintaining the balance between germ cell proliferation and death, to ensure sufficient number and quality of well differentiation and mature spermatozoa. Main reasons probably owe to structural damage, functional lost of boar spermatozoa and apoptosis during cryopreservation, which finally lead to sperm program cell death, apoptosis. The present paper reviewed the influence mechanism of apoptosis mediated by caspase, which showed tightly relationship between apoptosis mediated by caspase and spermatozoa cryopreservation, spermatogenesis and development.
Research Progress on H-FABP Gene in Black Cattle
ZHAO Shi-quan, DONG Ya-juan, BAI Xue-jin, NIU Zhao-shan, YANG Li
2013, 40(4):  145-148. 
Abstract ( 234 )  
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H-FAPB gene, as one of the candidate genes affecting intramuscular fat content which is intimately related to meat quality traits, has become the key point of improving Black cattle meat quality by molecular breeding technology. The paper not only reviewed regulative mechanism of H-FABP influencing on fatty acid transportation and the location and structure of H-FABP gene, but also performed an analysis of scholars’ researches on different kinds of livestock and poultry, which would definitely provide theory reference for the breeding of Black cattle by means of modern molecular biotechnology.
Protection Effects of Flavonoids and Vitamin E on Diethylstilbestrol Induced Oxidative Injury
DU Juan, PAN Hong-yan, LIN Li-mei, HUANG Da-wei, GONG Zhi-yong
2013, 40(4):  149-153. 
Abstract ( 312 )  
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The purpose of this study was to investigate the oxidative damage caused by different doses of diethylstilbestrol (DES) and the protective effects for DES induced toxicity with flavonoids and vitamin E in Kunming mice. 80 Kunming male mice were randomly divided into 10 groups, the groups were normal saline control group, DES exposed groups (0.035, 0.35, 3.5 mg/(kg穌)), flavonoids interference groups(26.0, 58.0, 112.0 mg/(kg穌)) and vitamin E interference groups(56.2, 112.4, 224.8 mg/(kg穌)). The effects on liver and testis were studied in adult male mice treated for one week. Mice were killed to collect liver and testis, and then contents of malonadehyde (MDA), superoxide dismutase (SOD), glutathione perxidase (GSH-Px) and total antioxidant capacity (T-AOC) were determined. It was found that high dose DES (3.5 mg/(kg穌)) could induce mice liver oxidative injury. The protective effects of flavonoids for DES induced toxicity on liver dose dependent increased. The contents of MDA significantly decreased (P<0.01), but the contents of SOD significantly increased in the high dose group (112 mg/(kg穌)). The contents of MDA significantly decreased (P<0.01), but the contents of GSH-Px, SOD and T-AOC significantly increased in the medium and high dose groups (112.4, 224.8 mg/(kg穌)). DES appeared to cause oxidative injury in Kunming mice. The results provided preliminary experimental support for flavonoids and vitamin E as medicine for DES induces damage.
Antibacterial Activity and Stability Study of Antimicrobial Peptides [L6,K11]-IsCT
ZHENG Wen-guan, ZHANG Jie, YANG Gang-qi, YANG Min, PAN Yun, LI Yong-xin
2013, 40(4):  153-158. 
Abstract ( 441 )  
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IsCT was a non-cell-selective antimicrobial peptide isolated from the scorpion.In order to be used in animal feed,[HT5] [HT5"]it was necessary to study the effects on inhibiting and killing common pathogenic bacteria. Agar gel diffusion and dilution methods were used in this experiment to study the antibacterial activities and stabilities of [L6,K11]-IsCT which was the derivative of IsCT. The results indicated that [L6,K11]-IsCT had better antibacterial activities to Escherichia coli than Staphylococcus aureus. The minimal inhibitory concentration (MIC) of Escherichia coli was 50 μg/mL. When treated with 121 ℃ 0.12 MPa for 30 minutes or 2.0≤pH≤11.0 for 30 minutes and incubated in artificial gastric juice for 2 h,[L6,K11]-IsCT remained excellent antibacterial activities. Therefore,[L6,K11]-IsCT presented excellent heat stablility and could resist to extreme pH environment as well as artificial gastric juice.
Study on Inhibition Effect of Broussonetial papyrifera Leaf Extract on Staphylococcus aureus and its Mechanism
LIU Xiao-jun, LIU You, CHEN Shao-hong, LIU Yan-fen
2013, 40(4):  159-162. 
Abstract ( 328 )  
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To investigate the inhibition effect of Broussonetia papyrifera leaf extract and its mechanism, the Staphylococcus aureus was used as test bacteria, the inhibition zone diameters were observed by the Oxford cup method, the growth curve, protein synthesis, and the membrane permeability of the the Staphylococcus aureus were detected to study the antibacterial mechanism of the Broussonetial papyrifera leaf extract. The results indicated that the antibacterial circle diameter of the Broussonetial papyrifera leaf extract and 75% ethanol extract were 17.5 and 18.0 mm respectively, the minimal inhibitory concentration (MIC) was 6.8 mg/mL, and the minimum bactericidal concentration (MBC) was 12.5 mg/mL. The results also revealed that the growth of the Staphylococcus aureus significantly decreased owing to the inhibition of the bacterial protein synthesis caused by the Broussonetial papyrifera leaf extract, but the membrane permeability was not affected. The Broussonetial papyrifera leaf extract showed obvious antibacterial activity on Staphylococcus aureus in vitro, and the antibacterial effect was probably associated with protein synthesis inhibition of the bacteria.
Research Progress on Application of Ultra Performance Liquid Chromatography Technology in the Field of Veterinary Drug Analysis
ZHOU Chun-feng, GUO Shi-jin, MEI Jian-guo, ZHANG Ying, YANG LI-mei, SHEN Zhi-qiang
2013, 40(4):  162-166. 
Abstract ( 245 )  
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Ultra performance liquid chromatography technology is a rapid development technique which based on small particle packing in recent years. The technique, which has many advantages including fast, sensitive, accurate, efficient and so on, has become one of the research focus in the field of drug testing and analysis. As the development of the technology, it has been widely used in veterinary medicine research development, and detection of veterinary drug residues. In this paper, the technical characteristics of ultra performance liquid chromatography and its practical applications were reviewed to show the wide application prospect of the technology in the future of veterinary analysis field.
Polymorphism of CETP Gene and Association Analysis with Production Performance-related in Dahe Blank Pig
LI Fu-quan, YANG Wen, GUO Bin, HUO Jin-long, SHI Xian-wei
2013, 40(4):  167-173. 
Abstract ( 375 )  
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The associations between SNPs in the cholesteryl ester transfer protein (CETP) gene and production performance of pigs were reported in this study, the polymorphism of CETP gene was detected by a AvaⅡ RFLP in 334 Dahe Black pigs across F4 to F6 generations. These results showed that b alleles was excellent gene with frequencies of 0.5821, 0.6042 and 0.6885 in F4, F5 and F6 generations, respectively. Seven SNPs (19, 69, 132, 175, 203, 224 and 234 bp, respectively) were found by the homozygous samples sequencing. Associations between the founded AvaⅡ RFLP and birth weight, body weight at 45 days, body weight at 4 months, body weight at 6 months, body height at 6 months, body length at 6 months and cannon circumference at 6 months and back-fat thickness at 6 months were tested by least square analysis. The results showed that in F4 generation, body length at 6 months with Bb and bb genotype was higher than that with BB genotype (P<0.05);cannon circumference at 6 months with bb genotype was higher than that with BB genotype (P<0.05), that with Bb genotype was not significant (P>0.05). In F5 generation, the birth weight with Bb and bb genotype was higher than that with BB genotype(P<0.05), no significant difference between them (P>0.05).In F6 generation, the birth weightand body weight at 45 dayswith bb genotype was higher than that with BB genotype (P<0.05), that with Bb genotype was not significant (P>0.05). In addition,the birth weight showed the higher significant difference between bb genetype and female/male (P<0.05). These results suggested that the CETP gene had some effects on the growth traits of porcine,and the selection of b allect was favored with regard to the growth traits of porcine, this SNP might be considered as a potential genetic marker for improving growth.
Effects of IGF2 Gene on Growth Traits in Luchuan and Large White Pigs
ZHANG Bing, MA Qing-yan, QIN Qian, SUN Jun-li
2013, 40(4):  173-176. 
Abstract ( 306 )  
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To reveal the SNPs of insulin-like growth factor-2 (IGF2) gene and evaluate its relationship with growth traits, we used PCR-SSCP to identify one SNP of IGF2 gene, and analyzed the effects of the SNP on birth weight(BW), weaning weight(WW), average daily gain(ADG) in Luchuan and Large White pigs. The results showed that IGF2 gene had significant effects on BW, WW, ADG in Large White pigs(P<0.05), and had significant effects on BW, ADG in Luchuan pigs (P<0.05). It could be inferred that IGF2 gene might be one of candidate genes affecting the growth traits in Luchuan pig. The polymorphic site could be used to marker assisted selection program for growth traits in Luchuan pig.
The Effect of Melatonin on Semen Quality of Bovine
LI Xiao-xia, CAO Ping-hua, YU Xue-li, LI Ying-hua
2013, 40(4):  177-179. 
Abstract ( 262 )  
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The antioxidative effect of melatonin (MLT) was evaluated on preserved bovine semen. Progressive motility (using visual observation), plasma membrane integrity (using the hypoosmotic swelling test, HOST) and acrosome integrity (using coomassie blue) of bovine spermatozoa were examined at 27 ℃ with 0, 10-3, 10-4, 10-5 mol/L concentration and for 1, 4, 8 h incubation time. The results showed that progressive motility, the percentage of plasma membrane integrity and acrosome integrity of the bovine spermatozoa were higher for 10-3 and 10-4 mol/L MLT than that of the control group (P<0.05). And progressive motility, the percentage of spermatozoa with plasma membrane integrity and acrosome integrity in melatonin (10-3 mol/L ) groups at 1, 4 h were higher than that in the control group (P<0.05). In conclusion, addition of optimal concentration of MLT helped to improve semen quality and life of preserved bovine semen.
Research Progress on Effect of Maternal Immunity during Pregnancy in Stress
LIU Xue-yin, YANG Qing-chun, YU Zhong-jia, LIU Guan-hui, CHEN Yao-xing, MA Bao-chen, WANG Zi-xu, DONG Yu-lan
2013, 40(4):  180-184. 
Abstract ( 242 )  
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Environmental stress on animal growth and reproduction of influence were more and more serious with the development of intensive aquaculture. Pregnancy maternal stress would induce maternal emotional tension, affect fetal development, and even cause abortion. Mechanism of stress influence might be caused maternal immunity and the number and activity of uterus lymphocytes, NK disharmony, and then induced the disorder of maternal immunity through the activation of the nerve-endocrine, immune network, which would initiate the embryonic immune rejection, affect fetal dysplasia and even death. The research progress of stress on pregnancy, and the influences role path, especially stress on maternal immune during pregnancy were summarized.
Cytogenetic Research on Dehong Buffalo,Nili-Ravi Buffalo and Their F1 Hybrid Bulls
LI Si-yin, YANG Liang-yu, YANG Yu-ai, BAI Wen-shun
2013, 40(4):  184-189. 
Abstract ( 363 )  
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In order to make clear the hybrid F1 (Dehong buffalo譔ili-Ravi buffalo) bulls karyotype and fertility and the relationship between them and the parents, the study cultrued lymphocyte in the hybrid F1 (Dehong buffalo, Nili-Ravi buffalo, Nili-Ravi譊ehong buffalo)males by peripheral blood lymphocyte culturing, made chromosomal sample and G-band dyeing. Then chromosome karyotypes in them were analysed and made sure by BEION Karyotyping system. The results showed that the karyotype of Dehong buffalo was 2n=50, XY, Nili-Ravi buffalo was 48,-4,-4,-9,-9,+t(4;9),+t(4;9),XY and hybrid F1(Nili-Ravi譊ehong buffalo) 49,-4,-9,+t(4;9),XY.
Isolation and Identification of Duck Flavivirus SDbz Strain
LIN Chu-wen, ZHUANG Jin-qiu, CHEN Jin-long, WANG Wen-xiu, LI Feng, WANG Jin-liang, SHEN Zhi-qiang
2013, 40(4):  190-194. 
Abstract ( 318 )  
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Duck flavivirus(DFV)disease was a kind of new duck disease. It had brought great economic losses to the duck industry in recent years. In order to study thoroughly and prevent the disease,it was isolated and identified as follows.The bacterial infection was initially ruled out from the samples by bacteria isolation. The samples showed DFV positive by RT-PCR detection. Then they were inoculated into DEF cells and healthy duck embryo for virus isolation and passages. Cytopathic effect (CPE) in DEF cells were observed in the first generation in the 48 h after inoculation, CPE was more evident with the extension of the time, and the typical CPE were observed as from 72 to 96 h after inoculation. Each generation of inoculated duck embryo appeared to be death, and the time of death were more concentrated in 60 to 72 h after inoculation. The dead duck embryo body were observed as edema, hemorrhage, dysphasia, severe hepatic hemorrhage, swelling or necrosis mottled. The virus isolated in DEF cells and duck embryo were detected and identified by hemagglutination test, titer determination, virus neutralization test, RT-PCR and artificial infection test. The results showed that the virus isolated was DFV and named as DFV SDbz strain.
Isolation and Identification of Inclusion Body Hepatitis Virus in Shanxi
ZHANG Tan, ZHENG Jie, ZHANG Fa-ming, SUN Feng-ting, ZHANG Hong, YANG Guo-ling, LONG Jin-xue, ZHANG Liang, WANG Wen-quan
2013, 40(4):  194-197. 
Abstract ( 216 )  
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The purpose of this study was to evaluate a suspected chicken inclusion body hepatitis virus. A case of suspected chicken inclusion body hepatitis occurred on a chicken farm in Shanxi province. A live suspension culture of deadly suspected chickens was made and inoculated into the yolk cavity of a 6-day old SPF embryo. The virus was then isolated. After further analysis of the infected lesion using RT-PCR, gene sequencing, BLAST search and animal regression testing, it was determined that the suspected culprit was the inclusion body hepatitis virus.
Epidemiological Investigation of Canine Distemper in Minks
WANG Cong, BING Qi-zheng, ZHANG Ying, HUANG Juan, SHAN Hu 
2013, 40(4):  198-201. 
Abstract ( 343 )  
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In order to investigate the epidemiological situation of canine distemper in minks of Shandong province, 996 cases of illed minks from Zhucheng,Weifang and other areas of Shandong were subjected for clinical diagnosis and pathogen detection during 2010 to 2012.The results showed that 223 positive samples were detected,among of which, 60.99% cases were the coinfection of canine distemper virus and other pathagens.Most positive samples were from Zhucheng. 3-month old minks were deadly infected with the highest morbidity. The disease was typically characterized by manifestations of conjunctivitis and diarrhea. Skin pustules, hyperkeratosis, and central nervous system signs were also seen in some animals. It indicated that canine distemper became a serious impact on the mink-raising development,and mixed infection was becoming more serious,the susceptible mink should be vaccinated before 3-month old.
Survey on the Prevalence of Intestinal Parasites in Sheep
LI Meng-jie, PENG Yong-shuai, ZHU Dan, JIAN Fu-chun, WANG Guo-chun, MA Hui-hai, ZHU Wen-guang, NING Chang-shen
2013, 40(4):  201-205. 
Abstract ( 752 )  
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A total of 1109 fresh fecal samples from six cities of Henan, Shuangliao of Jilin, Chaoyang of Liaoning and Dongying of Shandong 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 technique. Nine kinds of intestinal parasite species were discovered and the overall infection rate was 97.2%. Eimeria spp. was the most common parasite species, with the infection rate of 93.5% and the highest OPG reaching to 652000. The infection rates of Strongylus spp.and Entamoeba were 51.2% and 71.3%, respectively, and the highest EPG of Strongylus spp. was 69400. The infection rates of Cryptosporidium, Giardia, Trichuris, Nematodirus, Moniezia and Trematoda were 1.6%, 5.7%, 9.7%, 0.3%, 5.1% and 2.6%, respectively. 80.7% of positive samples were mixed infections, with the highest number of mixed-infections being five. The infection rate was 98.5% in no more than 1-year-old sheep and 87.5% in more than 1-year-old sheep, respectively. The infection rate of parasites was 97.5% in drylot feeding sheep and 96.8% in grazing sheep. Summer and autumn had a higher prevalence of parasites, compared to winter and spring. The present results showed intestinal parasites were commonly found in sheep. Thus, the general measures for the control and prevention were needed to strengthen.
Research Progress of Anti-babesiosis Drugs
WANG Jing, ZHOU Xu-zheng, LI Bing, WEI Xiao-juan, FAN Ying, LIU Cui-cui, ZHANG Ji-yu
2013, 40(4):  206-211. 
Abstract ( 367 )  
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Babesiosis were exsiting in the tropical and subtropical country generally, and often resulted in great ecomomic losses in the livestock industry worldwide, and there was no sepcial drugs for this disease at present. This reviewed fouced on the progress on anti-babesiosis drugs in recent years. The effect drug imidocarb had deleterious effect on the liver function. The parasties genes had to be influenced by various mutations after treatment with atovaquone and azithromycin combination, particularly the cytochrome b gene, furthermore, the relapses after treatment by some anti-babesiosis drugs and the presence of drug resistant were still very serious. To develop compounds of anti-babesia had been focus on the heterocyclic compounds and their deribatives, such as thiazole and imidazole etc. To find the appropriate target genes in the parasite and its antagonist using molecular biology methods had achieved certain results, moreover, some experiments had showed that some natural medicines had effect on anti-babesia.
Research Progress of Porcine Reproductive and Respiratory Syndrome
GUO Min, LANG Guang-ping, FENG Jiao, LI Bin, FU Yun-yun, SHAN Yue, ZHAO Jian-zeng, GAO Ying-jie
2013, 40(4):  211-215. 
Abstract ( 335 )  
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Porcine reproductive and respiratory syndrome (PRRS) is presently one of the most important viral infectious disease of pigs. This article reviews the latest research progress on PRRS in recent years, particularly in the immunology and molecular biology. It will provide guidances and directions for the prevention and treatment of PRRS.
Research Advance on Cefquinome
LIU Mei-duan, ZHAO Di-shun, LIU Bao-shu, LIU Lin-ying, WANG Xue-liang, LI Jun-pan, GE Jing-jing
2013, 40(4):  216-221. 
Abstract ( 395 )  
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As the fourth generation of aminothiazolyl cephalosporin,cefquinome was developed especially for using in animals, and a new drug for animal severe infection, which had antibacterial mechanism and antibacterial activity. Cefquinome was known as broad antibacterial spectrum, strong antibacterial activity, superior pharmacokinetics, and no carcinogenicity, mutagenicity, teratogenicity, reproductive toxicity. The blood concentration of cefquinome reached its peak in short time, and had high concentration. The paper summarized the cefquinome’s antibacterial activity, physicochemical properties and pharmacological properties, synthetic routes, a number of problems in synthesizing cefquinome, such as low yield and post treatment difficult were also analyzed, to lead pharmaceutical scientific worker with thorough knowledge and guide clinical application.
Screening of Porcine Circovirus Type 2 Inactivated Vaccine Candidate Strain
GAO Jin-liang, YANG Xia, CHEN Lu, LI Wei-hao, CHANG Hong-tao, ZHAO Jun, WANG Xin-wei, LIU Hong-ying, YAO Hui-xia, WANG Chuan-qing
2013, 40(4):  221-224. 
Abstract ( 801 )  
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To select porcine circovirus type 2 strain to meet the requirements for producing porcine circovirus type 2 inactivated vaccine, the efficacy and immunogenicity of 8 porcine circovirus type 2 strains from 8 regions of Henan province were assessed by measurement the antibody level of mice immunized with inactivated oil adjuvant vaccine which was manufactured with virus stock solution and diluted solution of same titer (4.233×104 copies/μL). The antibody levels were induced by vaccines manufactured with 8# and 12# strains stock solution were higher and lasted longer than other strains and reference vaccines. The antibody titers at 1 week after the secondary immunization were above required minimum protective level of 1∶800 according to national quality standards of porcine circovirus type 2 inactivated vaccine. These results indicated that porcine circovirus type 2 of 8# and 12# strains could propagate well and had good immunogenicity and therefore could be chosen as candidate vaccine strains. This study enriched the resources of porcine circovirus vaccine strains and provided basis for developing high-quality porcine circovirus vaccine.
Research Progress on the Coccidiostat of Arprinocide
XIE Jin-fu, LI Mei-di, WU Li
2013, 40(4):  225-228. 
Abstract ( 308 )  
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Arprinocide was one of a class of ribonucleotide possessing broad-spectrum anticoccidial activity in chickens, and there were no relative products in China. The author reviewed the mechanism of action and the current study of arprinocid at home and abroad, and analysed the application prospect of prevention and treatment coccidiosis in chickens,which could provide the reference evidence to study, spread and use for chickens in the farms.
Diagonosis and Treatment of Tibia and Fibula Fracture of Kunming Canine
YU Ze-zhong, WANG Ping
2013, 40(4):  229-231. 
Abstract ( 324 )  
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Through recalling analysis of the clinical use of the external fixatOR treatment of 11 cases of Kunming canine tibia and fibula fracture cases, we explored the clinical effect of external fixator for the treatment of Kunming dog canine tibia and fibula fractures. 100 days after the surgery, by a return visit, all fractures were healed, the shortest and longest fracture healing time were 30 and 90 days, respectively, in addition to the few pinholes infection and pins loosening outside, no other significant complications. Results showed that external fixaor was an ideal method for the treatment of the Kunming canine tibia and fibula fracture.
Epidemiology, Diagnosis, Prevention and Control of Peste des Ptits Ruminants
SHI Yan, WENG Shan-gang
2013, 40(4):  231-234. 
Abstract ( 423 )  
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Peste des petits ruminants (PPR) is a highly contagious, infectious, viral disease of domestic and wild small ruminants characterized by fever, stomatitis, conjunctivitis, gastroenteritis and pneumonia. It is one of the major notifiable diseases of the World Organization for Animal Health (OIE).This review presents comprehensive information on this dangerous disease including its history, spread, and diagnosis of the disease. It also deals with relevant preventive and control measures to be implemented in areas with PPR outbreaks.
Research Progress on the Bokhi of Camel
GUO Chun-yan, MING Liang, LIU Wei, ZHANG Min, HE Jun-xia, HA-Si-su-rong, JI-Ri-mu-tu
2013, 40(4):  235-238. 
Abstract ( 318 )  
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Bokhi is a kind of liquid which is secreted from the poll gland of male camel. The main components of Bokhi are steroid hormone and acids, and it has a positive effect on tumor,kidney yang deficiency and immune suppression. The author had a systematic review on the latest research progress on chemical constituents and pharmacological effects of Bokhi,in order to provide a scientific basis for the development and utilization of Bokhi.
Comparative of Propofol Intravenous Anesthesia and Enflurane Inhalation Anesthesia in Cardiopulmonary Bypass of Bama Minipigs
CHEN Ke-yan, DIAO Yu-gang, JIN Qiang, WANG Hong-qian, YAO Guo-quan, ZHANG Tie-zheng
2013, 40(4):  238-241. 
Abstract ( 278 )  
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The assay was aimed to compare of propofol intravenous anesthesia and enflurane inhalation anesthesia in cardiopulmonary bypass of Bama Minipigs. 10 Bama Minipigs, the average into two groups were propofol intravenous anesthesia and enflurane inhalation anesthesia, and in preoperative, operative and postoperative the anesthesia monitoring. After openning breast, whole blood were treated with heparin, then the ascending aorta, superior vena cava and inferior vena cava were intubated respectively, and connected to a cardiopulmonary bypass machine line the CPB. The vital signs, nose temperature (NT) , heart rate (HR), mean arterial pressure (MAP), pH and blood oxygen saturation(SPO2) of each experimental pigs were observe and recorded within before introdution of anesthesia (T0), after induction of anesthesia (T1), immediately before CPB (T2), cooling to 30 ℃ (T3), blocking the aorta former immediate (T4), blocking the aorta 4 min (T5), start thawing immediate (T6), stop CPB immediate (T7), closed chest immediately after (T8) and drawing tube operation over 10 min (T9). Two groups of weight number of pigs, operation time and anesthesia time all had no significant difference (P>0.05). Different fluorine ether group restore spontaneous breathing a short time in propofol group (P<0.05). Compared with the basic value(T0 and T1), the NT, HR and MAP value of the experiment pigs were significantly lower (P<0.05). But each rooms and in the group SPO2 value and pH were not significant different (P>0.05). Propofol intravenous anesthesia during the operation should be based on the experimental animals of the reaction conditions appropriate adjustments propofol pump into the concentration; and different fluorine ether inhaled anesthesia during the procedure of smooth, anaeesthetic effect was good, postoperative came quickly, suitable for complicated cases, and the time was long operation.
Research Progress on Inulin in Raising Pigs Production
SHANG GUAN Ming-Jun, WANG Fang, ZHANG Hong-gang, ZHANG Yuan-qing, ZHANG Bian-ying
2013, 40(4):  242-245. 
Abstract ( 424 )  
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Inulin is a kind of special natural fructan of carbohydrate,usually extracted from plants. It had a probiotic effect, and could selectively improve the beneficial intestinal microflora and inhibit enteric pathogens, furthermore. It could enhance production performance and immune function of animals. This article expounded the research progress on inulin in raising pigs as a new feed additive, and whose effect on the intestinal flora, the growth performance, the immune function, the mineral absorption and utilization, and meat quality of pigs and so on were analyzed in this paper.
Effect of Grape Seed Powder Replacing for Corn on the Weight and Economic Benefit of Adult Ewes
ZHANG Peng, SUN Zhan-peng, ZHANG Xue-shan, AN Huai-yong, LI Hui-ju, WU Zhi-guang, YANG Xin-jun
2013, 40(4):  245-247. 
Abstract ( 307 )  
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The research was conducted to study the influence of different doses of grape seed powder replacing for corn on weight and economic benefi of adult ewes. 32 adult sheep with similar age (15 to 16 months) and in good health were randomly divided into 4 groups according to the principle of similar weight. The control group was fed the basal diet, the testⅠ,Ⅱ and Ⅲ groups added different doses of grape seed powder replacing for 7%, 14% and 21% corn in the basal diet. The results showed that the test Ⅰ,Ⅱ and Ⅲ groups were no significantly difference compared with the control group in weight gain (P>0.05), the average weight of test Ⅰ group increased 14.50%, and the economic benefit of test Ⅰ group increased 37.74%. In short, adding grape seed powder replacing for 7% corn in the basal diet was the best for weight gain and economic efficiency of adult sheep in this research.
Advance on Digital Development Mode of Science and Technology Journals
ZHAO Lian-sheng, LI Li
2013, 40(4):  248-250. 
Abstract ( 219 )  
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Developing digital journals which provides a good extended platform for journal development has become a strategic choice for the development of the publishing industry in China. The purpose of this article was to discuss the advantages and disadvantages of the network development modes of science and technology journals and analyse 5 publishing models of scientific and technology journals,Which might offer reference for digital development of science and technology ournals.