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20 September 2013, Volume 40 Issue 9
Cloning and Expression of Buffalo Cx43 Gene
LIN Lang, GONG Yun, WANG Meng, QU Chun-feng, HUANG Shi-hai, LEI Xiao-can, SHI De-shun, LI Xiang-ping
2013, 40(9):  1-7. 
Abstract ( 257 )  
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Buffalo Cx43 gene was cloned in the present study, the Cx43 sequence was systemically analysized by bioinformatics techniques. The expression patterns of Cx43 in different tissues and different stages of follicles were also assayed with RT-PCR and immunochemistry methods. The results showed that, a buffalo Cx43 gene fragment was cloned and sequenced, including the whole CDS of 1152 bp (coding 383 amino acids). The molecular weight and isoelectric point of buffalo Cx43 protein were predicted as 43.13 ku and 8.88 respectively. The results of sequence multialigned showed that the sequence of buffalo Cx43 gene shared 99%,98%,94%,93% and 92% homology with Bos taurus,Ovis aries, Sus scrofa,horse and Homo sapiens respectively. Buffalo Cx43 protein was predicted containing special connexin protein domain. In addition, we also analyzed the mRNA expression level of Cx43 gene in buffalo tissues through Real-time fluorescence quantitative-PCR. The results showed the mRNA of Cx43 gene existed in all of the six detected buffalo tissues with the most abundant expression in ovary liver,followed by kidney,heart and skin,the minimal expression was observed in liver. The Cx43 expression was detected in buffalo follicle during various development stages by immunochemistry method. Expression of Cx43 protein increased along with the follicle development. The cloning and analysis of buffalo Cx43 gene laid an important foundation for furthur investigating the function of Cx43 gene.
Molecular Biological Identification of Dermacentor nuttalli and Dermacentor marginatus
LV Ji-zhou, WANG Zhen-bao, YUAN Xiang-fen, WU Shao-qiang
2013, 40(9):  8-15. 
Abstract ( 356 )  
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This study was to establish a method for identifying Dermacentor nuttalli (D.nuttalli) and Dermacentor marginatus (D.marginatus) with molecular markers, and to study the phylogenetic relationship of these two species of ticks. Ticks were collected from domestic animals in Xinjiang province and classified by morphology characters. Then 16S rRNA and COⅠ of ticks were amplified by PCR and subsequently sequenced. Phylogenetic trees were constructed by Mega 5.0 and Mrbayes 3.2. On the phylogenetic trees based on 16S rRNA, D.nuttalli was clustered together with their respective class D.nuttalli, and D.marginatus formed a distinct branch. Moreover, D.marginatus was clustered with their respective class D.marginatus, and D.nuttalli formed a distinct branch on the phylogenetic trees based on COⅠ. Based on morphology and combined with molecular biology identification the method was a simple, accuracy method for the identification of D.nuttalli and D.marginatus.
Development of a Double-antibody Sandwich ELISA for Detection of Subgroup J Avian Leukosis Virus
LIAO Ya-lin, LIANG Yi-yu, WANG Xiu-long, FENG Min, TAN Li-qiang, CAO Wei-sheng
2013, 40(9):  15-18. 
Abstract ( 371 )  
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A double-antibody sandwich ELISA (DAS-ELISA) was developed for the detection of subgroup J avian leukosis virus (ALV-J). Antibodies of DAS-ELISA were purified from the ALV-J gp85 mono-specific serum prepared by our laboratory earlier. Results of statistics analyses showed that DAS-ELISA had good specificity, repeatability and stability. The limitation of viral antigens for detection was 0.165 μg/mL. The coincidence between PCR and DAS-ELISA established here was 85.2%.
Identification of the microRNAome in Texel Sheep by Deep Sequencing
ZHANG Shi-fang, WEI Cai-hong, LU Jian, ZHANG Xiao-ning, ZHOU Xin-lei, ZHANG Shu-zhen, WANG Guang-kai, CAO Jia-xue, ZHAO Fu-ping, ZHANG Li, DU Li-xin
2013, 40(9):  19-22. 
Abstract ( 404 )  
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Information concerning the sheep microRNA (miRNA) had been long overdue, the discovery and identification of miRNAs would further promote the study of miRNA functions and gene regulatory mechanisms. Samples were culled that included five developmental stages: The longissimus dorsi muscles of Texel fetuses were at 70, 85, 100, 120 and 135 days. We performed a comprehensive search for Texel miRNA on the pool library by Solexa deep sequencing; then obtained 16532850 raw reads. This study used the ACGT101-miR v4.2 analysis software to explore the sequencing data. The sequencing results were analyzed using the latest mammalian mature miRNA (in miRBase v17.0), precursors (pre-miRNAs), and sheep genome (Ovis aries genome database, Feb 2010). Our results extend the repertoire of sheep microRNAome to 1529 pre-miRNAs encoding for 1999 mature miRNA. We preformed quantitative Real-time PCR experiments for selected 5 miRNA in the pool sample and found agreement between the sequencing and Real-time PCR data. These results were prelude to the advancement in sheep biology.
Establishment of a Nested PCR Technique for Detection of H1 Subtype Avian Influenza Virus
GUO Jie, XIE Zhi-xun, LUO Si-si, LIU Jia-bo, DENG Xian-wen, XIE Zhi-qin, PANG Yao-shan, FAN Qing
2013, 40(9):  23-26. 
Abstract ( 253 )  
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In order to develop a simple and quick method to detect H1 subtype avian influenza virus (AIV),two pairs of primers were designed according to the sequences of HA gene of H1 subtype AIV, and a nested polymerase chain reaction (nPCR) method was developed for detection of H1 subtype AIV. The specificity and sensitivity test showed that this nPCR was only specific to the H1 subtype AIV and could detect 13.2 fg/μL of AIV RNA. The results demonstrated that the optimized nPCR assay was rapid, specific and sensitive for detection of H1 subtype AIV.
Analysis of the Virulent Genes of One Newcastle Disease Viruses Isolated from Duck and its Pathogenicity Experiment in Duck
LI Xian-wei, SUN Min-hua, HE Jing, WANG Zhao-xiong, LIU Da-wei, YANG Feng, REN Tao
2013, 40(9):  27-32. 
Abstract ( 385 )  
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To explore the epidemiological understanding of Newcastle disease virus (NDV) in ducks, a Newcastle disease virus (NDV-104) was isolated from duck in Guangdong province in 2009. The biologically characteristic, pathogenicity and F and HN gene were explored. The results showed that MDT, ICPI and IVPI of the isolated strain were 56.4 h, 1.95 and 1.64, respectively. Combined with the analysis of the amino acid sequence of the F protein cleavage site (112 to 117), indicating it belonged to velogenic NDVs. The pathogenicity results showed that the isolated strain was infectious and pathogenic in ducks. Genetic analysis revealed that the isolate strain belonged to the gene Ⅶd subtype. The homology analysis of F and HN protein's amino acid showed that the isolated strain shared 96.6% to 99.3% and 97.0% to 99.7%, respectively, with genotype Ⅶ NDVs since 2000. However, the F and HN protein's amino acid of the isolate strains were low similarity with the common vaccine strains, such as B1, V4, Clone30, Mukteswar and LaSota, and were 88.1% and 87.6% similarity with LaSota and V4, which were different from the classical strains of NDV.
Prokaryotic Expression and Purification of the Jaagsiekte Sheep Retrovirus Receptor Hyaluronidase-2
ZHU Fu-yu, MA Xue-en, YU Li-xin, YAO Hong-qiang
2013, 40(9):  32-36. 
Abstract ( 374 )  
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In order to construct an efficient prokaryotic expression system of jaagsiekte sheep retrovirus (JSRV) receptor hyaluronidase-2 (Hyal-2),we designed a pair of specific primers for the JSRV receptor Hyal-2 gene,using PCR amplification technique to amplify the full-length of Hyal-2 gene, and directional restructuring it into the prokaryotic expression vector pGEX-4T-1.We constructed the pGEX-4T-1-Hyal-2 recombinant plasmid which was transformed into E.coli BL21(DE3) using IPTG to induce expression, optimized conditions gradually until stable expression,and then detected fusion protein by Western blotting, at last, the fusion protein was purified by the affinity chromatography methods.The results showed that Hyal-2 gene was exactly insert into the prokaryotic expression vector PGEX-4T-1,after induction, the target protein containing GST tag was efficiently expressed by expression bacteria which included recombinant plasmid pGEX-4T-1-Hyal-2,SDS-PAGE electrophoresis result showed that the molecular weight of target protein was 80 ku,consistent with the expected size,and target protein that verified by Western blotting was a fusion protein with GST tag,the target protein which purified by the glutathione affinity chromatography would lay the foundation for further preparation of Hyal-2 protein's polyclonal antibody and in-depth study of its function.
Establishment of Real-time PCR Assays for Detection of the Intramuscular Fat Deposition Related Genes in Mongolia Sheep
MIAO Hai-ming, GU Yue, YANG Jin-li, GAO Ai-wu, HUANG Ya-juan, WANG Hai-rong
2013, 40(9):  37-41. 
Abstract ( 286 )  
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This experiment was conducted to establish a detection method which was suitable for genes expression of Mongolia sheep intramuscular fat (IMF) deposition genes. According to the sequences of IMF deposition genes in GenBank, pairs of specific primers were designed and synthesized. Then the amplified fragments were detected by cloning technology to ensure the accuracy of the PCR products. The Real-time fluorescent quantitative PCR (Real-time PCR) was used to check the feasibility of method. Results showed that Real-time PCR optimum cycle parameters were 94 ℃ 4 min, 94 ℃ 30 s, 58 ℃ 45 s, 72 ℃ 1 min,40 cycles;72 ℃ 5 min.Each peak melting curve was a single gene, the target product was stable. The method of Real-time PCR was established successfully and could be specially used to detect IMF deposition genes of Mongolia sheep.
Cloning and Sequence Analysis of TLR5M Full-length cDNA in Common Carp Peripheral Blood Leukocytes
SUN Zhen, JIA Sheng-mei, FENG Xiang-ru, CHEN Yi-long, ZHAI Xin-xin, SHEN Xue-fei, ZHANG Jun-hui, WANG Wen-dong, YANG Zhen-guo, LU Qiang
2013, 40(9):  41-45. 
Abstract ( 271 )  
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In this paper, we have isolated, sequenced and characterized the full-length cDNA of TLR5M gene from common carp by the methods of rapid amplification of cDNA ends(RACE) on the basis of its EST sequences. The results showed that the TLR5M gene was 3182 bp in length,vthe gene consists of a 38 bp 5' untranslated region(UTR), 486 bp 3'UTR and a 2658 bp open reading frame(ORF), encoding a peptide of 885 amino acids. The amino acid sequence of carp TLR5 gene shared 84.46% identity with cirrhinus mrigala.
Cloning and Prokaryotic Expression of Kidney Lysozyme Gene in Lezhi Black Goat
HU Yu-fei, JIANG Ming-feng, ZHANG Peng, LUO Fan, ZHU Lian-lian, WANG Yong
2013, 40(9):  46-51. 
Abstract ( 306 )  
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The kidney lysozyme (LZLyK) gene from Lezhi Black goat was cloned, expressed and its activity was detected in this experiment. With the comparative genomics, the LZLyK gene was cloned successfully obtaining a 444 bp long cDNA which coded 147 amino acids. The phylogenetic analysis showed that the LZLyK gene shared the highest homology up to 96.9% with the sheep's lysozyme NO.2(M32498.1) in amino acids. After being transformed into E.coli BL21 (DE3), the recombinant plasmids pET-LZLyK were expressed in a prokaryotic expression system. The SDS-PAGE analysis revealed that the LZLyK protein had molecular mass of 16 and 30 ku respectively, which were consistent with the prediction. The activity of the protein mentioned above was determined by the way of nephelometry, and consequently it turned out to be 15 U/mL which indicated that the protein of LZLyK possessed a certain antibacterial activity. It was found that the content of lysozyme was more in stomach than other tissues through RF-PCR.
Cloning and Prokaryotic Expression of Outer Membrane Proteins 16 Gene of Brucella melitensis
JIA Xiao-xiao, JIAO Han-wei, GUO Shi-yu, SHI Qiao-yun, RONG Hui, ZHANG Jia-ning, ZHU Hua-pei, DU Li, CHENG Ying, WANG Feng-yang
2013, 40(9):  51-54. 
Abstract ( 269 )  
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To successfully clone the outer membrane proteins 16 (Omp16) gene and make prokaryotic expression in E.coli, one pair of primers was designed according to B. melitensis M5-90 strain Omp16 gene sequence in GenBank, and then obtained Omp16 gene which was about 507 bp by PCR from the Brucella genome. After purifying, Omp16 gene was inserted into pMD20-T vector to construct recombinant plasmid pMD-Omp16. pMD-Omp16 transformed into E.coli DH5α and identified it by sequencing, then subcloned to vector pET-28a(+). The constructed recombinant plasmid pET-Omp16 was transformed into E.coli BL21(DE3) for expression under induction of IPTG. Lastly, the expression products of recombinant protein His-Omp16 was identified by Western blotting. The results showed that the prokaryotic expression vector was successfully constructed and expressed Omp16 gene in E.coli BL21.
Selection of Reference Genes in Mice Infected with H5N1 Avian Influenza Virus
WU Qiao, ZHANG Bin, TANG Cheng, YUE Hua
2013, 40(9):  55-60. 
Abstract ( 339 )  
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This experiment was conducted to assess the stability of six candidate reference genes (Ywhaz,Hprt1,Sdha,Actb,Ppia and 18S rRNA) in spleen of mice infected with H5N1 avian influenza virus (H5N1 AIV). The mice were infected with the virulent strain A/DK/GD/378/08 of H5N1 AIV and harvested at 12,24,48,72,96,120 and 144 h post-infection. The expression levels of the six reference genes in infected and uninfected mice were determined by Real-time PCR and assessed the stability of reference genes by Genorm software. The results showed that the stability of six reference genes in normal mouse spleen tissue descended as following: Ywhaz/Hprt,Sdha,Actb,Ppia,18S rRNA, the stability of six reference genes in mouse spleen tissue infected with H5N1 AIV descended as following: Ppia/Sdha,18S rRNA,Hprt,Ywhaz,Actb. Furthermore,based on comprehensive evaluation of the normal and the infected H5N1 AIV,the stability of six reference genes in mouse spleen tissue descended as following: Sdha/Hprt1,Ywhaz,Ppia,Actb,18S rRNA. Therefore,the Ppia and Sdha were the best reference genes to use in the study of replication levels of H5N1 AIV in spleen of mice. However,for the study of organism response to H5N1 AIV infection,the Sdha and Hprt1 were the best reference genes.
Using SUMO Expression System to Efficiently Express VP0,VP1,VP3 Protein of Foot and Mouth Disease Virus Serotype O in Swine
SONG Ni, WEN Yong-jun, WANG Feng-xue, WU Hua
2013, 40(9):  61-65. 
Abstract ( 447 )  
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Structural protein precursor P1 gene of foot and mouth disease virus (FMDV) serotype O in swine was synthesised according to published sequences in GenBank.The structural protein VP0,VP1 and VP3 gene primers were designed depending on these specific sequences,and were obtained by PCR method according to the P1 gene, respectively.The PCR products were cloned into Blunt vector,and were digested and ligated into the prokaryotic expression vector SUMO.These recombinant expression plasmids,SUMO-VP0, SUMO-VP1 and SUMO-VP3,were transformed into E.coli BL21(DE3)plysS.For detecting the efficient expression,these fusion protein products expressed by the E.coli were checked by SDS-PAGE electrophoresis,and molecular weight were 55,48 and 40 ku,respectively.The largest ratio of the bacterial protein was induced under the conditions of IPTG concentration of 1.0 mmol/L,temperature of 37 ℃ and last for 5 h. The results of Western blotting showed that the fusion proteins had a good reaction with the FMDV positive serum.
Expression of VP2 Gene of Infectious Bursal Disease Virus in Insect Cells
YANG Feng, LI Xian-wei, SUN Min-hua, YAN Fu-qiang, WEI Qing-lan, XIANG Bin, REN Tao
2013, 40(9):  65-69. 
Abstract ( 324 )  
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The VP2 gene of very virulent infectious bursal disease virus (vvIBDV) JM-1/10 strain was placed behind the CMV promoter of pcDNA-3.1(+).The CMV-VP2 gene was cloned into the baculovirus expression system pFastBacTM Dual to build pFast-CMV-VP2.Transformed it to Escherichia coli DH10Bac.Recombinant baculovirus expression plasmid Bacmid-CMV-VP2 was screened.Bacmid-CMV-VP2 was used to be transfected into Sf9 insect cells to get the recombinant baculovirus vBac-CMV-VP2 and transduced into BHK-21 cells.After 48 to 72 hours,indirect immunofluorescence assay (IFA) was conducted to detect specific fluorescence.Western blotting analysis result showed target protein was expressed.The prepared recombinant vBac-CMV-VP2 could not only be expressed in insect cells, but also expressed in mammalian cells.
Establishment of Melting Curve Analysis Assay for Molecular Identification of Oesophagostomum spp. from Pigs
SHU Li, CHENG Tian, WENG Ya-biao, LIN Rui-qing, ZOU Shang-shu
2013, 40(9):  70-74. 
Abstract ( 247 )  
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Based on the IGS rDNA sequences of O.dentatum and O.quadrispinulatum, a pair of primers (MCF/MCR) was designed for developing a melting curve analysis assay for the identification and differentiation of two Oesophagostomum species. This assay allowed the specific identification and differentiation of O.dentatum and O.quadrispinulatum by melting curve analysis, with no amplicons being amplified from heterogenerous DNA samples. DNA sequencing confirmed the identity of the amplified products. The single egg of Oesophagostomum spp. and DNA extracted using What-man FTA card could also be detected by this assay. These findings showed that the melting curve analysis assay established was accurate, sensitive, specific and rapid, allowing the detection and identification of O.dentatum and O.quadrispinulatum from pigs.
Cloning and Tissue Expression of Pig Lipoic Acid Synthase Gene
CAO Jing-jing, ZHANG Xiao-mei, LU Wei-fei, GUO Yu-jie, HAN Li-qiang, WANG Lin-feng, YANG Guo-yu
2013, 40(9):  75-79. 
Abstract ( 280 )  
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The objective of this expriment was to clone pig lipoic acid synthase (Lias) gene by using electronic extension and RT-PCR and to study its tissue distribution in pig. A pair of cloning primers was designed based on the in silico sequence information and was used to clone and analyse the gene of pig Lias (No:JN797612.1). The positive clone was identified and sequenced, at the same time the Lias gene organization distribution rule was analyzed by RT-PCR. The length of pig Lias was 1119 bp, including an ORF of 1119 bp, encoding 372 amino acids. Identity analysis showed that the Lias nucleotide sequence in pig shared 71.5%, 69.0%, 77.3%, 70.4% homology with that of Mus musculus, Rattus norvegicus, Homo sapiens, Bos taurus, respectively, the predicted peptide shared 91.5%, 79.4%, 89.4%, 88.2% homology with that of Mus musculus, Rattus norvegicus, Homo sapiens, Bos taurus, respectively. The phylogenetic tree demonstrated that pig was the closest to Mus musculus and Rattus norvegicus. Semi-quantitative RT-PCR analysis showed that Lias expressed in heart, liver, kidney, adipose tissue, spleen, skin, rectum, lung, duodena jejunum, ileum and muscle, but it was higher in the kidney and fat, and then expressed in rectun and ileum.
Optimication of Electrotransformation Conditions of Six Lactobacilli Strains
FANG Ming-jian, WANG Lei, CHEN Yu-lin
2013, 40(9):  80-85. 
Abstract ( 253 )  
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This study was aimed to establish the optimal electrotransformation conditions of six Lactobacilli strains (L.plantarum, L.casei, L.reuteri, L.brevis, L.coryniformis and L.curvatus) which isolated from Shanbei White cashmere goats' rumen, plasmid pLEM415-egfp was electrotransformed into competent cell of six Lactobacilli on different growth phase (D600 nm=0.4, 0.6, 0.8, 1.0 and 1.2), and the electrotransformation efficiency was investigated under resistance 200 Ω, capacitance 25 uF and different electric field strength (1.5, 2.0 and 2.5 kV). Results showed that the optimal D600 nm and electric field strength to obtain the maximum transformation efficiency for L.plantarum, L.casei, L.reuteri, L.brevis, L.coryniformis and L.curvatus were 0.8, 1.2, 1.0, 0.6, 0.8, 0.4 and 1.5, 1.5, 1.5, 1.5, 1.5, 2.0 kV, respectively. The establishment of optimal electrotransformation conditions for six Lactobacilli strains provided the foundation for further construction and application of transgenic Lactobacillus.
Cloning and Sequence Analysis of Lactoferrin Gene in Tibetan Sheep
LI Jian-bo, JIANG Ming-feng, WANG Yong
2013, 40(9):  86-90. 
Abstract ( 411 )  
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RT-PCR method was used to clone the full length of lactoferrin (LF) cDNA sequence in Tibetan sheep at Ruoergai county of Sichuan province. Nucleotide sequence and structure of LF amino acid sequence were deduced by bioinformatics software. The results showed that LF gene was 2127 bp in length and encoded 708 amino acids, including a signal peptide of 19 amino acids at its N terminal. Isoelectric point (pI) of this protein was 8.4, and the molecule mass of the predicted protein was 77.2 ku. Semi-quantitative RT-PCR results indicated that LF gene was expressed in mammary, trachea, lymph, liver, lachrymal and spleen, but not in lung. In conclusion,LF gene in Tibetan sheep had been successfully cloned and the nucleotide and amino acid sequence of LF and Lfc were analysed in this study, it laid a foundation for further research.
Development of a Real-time PCR Assay for Detection of Chicken IL-1β, IL-18 and Tumor Necrosis Factor α Genes
ZHANG Kun-li, XIE Zhi-xun, TENG Li-qiong, LIU Jia-bo, XIE Li-ji, PANG Yao-shan, FAN Qing, DENG Xian-wen, LUO Si-si, XIE Zhi-qin
2013, 40(9):  90-95. 
Abstract ( 316 )  
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In order to develop a SYBR GreenⅠ Real-time PCR assay for detection of chicken IL-1β,IL-18 and tumor necrosis factor-α (TNF-α) genes, four specific primer pairs were designed according to the chicken's IL-1β,IL-18, TNF-α and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene sequences in GenBank. The four fragments were amplified by RT-PCR from chicken embryo fibroblasts, cloned and sequenced. The recombinant plasmids containing the target gene were constructed and used as the Real-time PCR standard templates. Real-time PCR assays based on SYBR GreenⅠfor detection of chicken IL-1β,IL-18, TNF-α and GAPDH were established. The results showed that each gene's melting curve also had a single peak,each gene's amplification efficiency was 101.2%, 95.6%, 100.1% and 98.2%, R2 was 0.9996,0.9998,0.9957 and 0.9989. Moreover,the assays were highly sensitive,the detection limit of 100 copies in 35 Ct and each gene's coefficient of variation less than 1.4 percent for intra-assay. This reliable Real-time PCR assay might be used for decting chicken's IL-1β,IL-18 and TNF-α mRNA expressing and provided the basis for quantitative analysis of cytokine expression in host cell after virus infect which cause immunosuppressive diseases.
Study on the Construction and Expression Characteristics of Canine Distemper DNA Vaccine Based on SFV-replicon
SUN Jian-tao, HOU Shao-hua, YU Ping, ZHU Hong-fei, LIANG Qian, HAO Zhi-ming
2013, 40(9):  96-100. 
Abstract ( 360 )  
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The clone vector pEASY-Blunt-F with fusion gene of canine distemper virus(CDV) isolated from vaccine strain was successfully constructed by T-A clone technique and was digested with restriction enzyme of BamHⅠ. The F gene was cloned into eukaryotic expression vector pSCA1. Finally a recombinant plasmid named pSCA1-F was constructed and identified by PCR, restriction enzyme analysis and sequencing. pSCA1-F was transfected into BHK-21 cells by using X-tremeGENE 9 DNA Transfection Reagent,IFA and Western blotting were performed to detect F expressions,the results showed that F gene had expressed in the transfected cells.Furthermore,apoptosis assay confirmed that pSCA1-F could induce the transfected cells apoptosis.
Research Progress on Application of Baculovirus Expression System in Veterinary Subunit Vaccine Field
LIU Chun-ju, REN Wei-jie, WANG Zhi-liang
2013, 40(9):  101-105. 
Abstract ( 346 )  
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Baculovirus expression system, based on baculovirus as the protein expression vector and insect cells or larvas as the host, is an eukaryotic expression system. In comparison with prokaryotic cells, the insect cells permit post-translational modfications of foreign proteins, such as folding, glycosylation, acylation, phosphorylation, disulfide bond formation, proteolytic cleavage and so on. Moreover, the baculoviruses cannot infect livestock. These advantages make baculovirus expression system be a safe and efficient platform for the production of veterinary subunit vaccines. Here, we critically review the baculovirus expression system as a platform to produce veterinary subunit vaccines.
Research Progress on Yeast Two-hybrid Technology and Application
ZHENG Li-shuang, LI Xiang-nan, SUN Cheng-tao, LIU Hong-yu, LIU Xun, HE Ming, LV Wen-fa
2013, 40(9):  105-108. 
Abstract ( 330 )  
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Yeast two-hybrid technology is an effective molecular biology method which is used to study the interaction of proteins. It is relatively simple, fast, efficient, sensitive, extensive and can directly detect the interaction of intracellular proteins. This paper mainly introduces the basic principle, applications, advantages and disadvantages of yeast two-hybrid technology, now the research on yeast two-hybrid system technology and application will be reviewed.
Real-time Quantitative RT-PCR Analysis of Animal miRNA Expression
SONG Jia-zhe, SUN Fu-bo, ZHUANG Kai-ge, XUE Kai
2013, 40(9):  109-113. 
Abstract ( 418 )  
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About 60% of human protein-coding genes are regulated by miRNA. These genes produce different proteins, which participant in many biological processes. The functions of these proteins are involved in cell cycle,development,differentiation and metabolism. Nowadays, researchers can detect the expressions of miRNA by various approaches. However, the RT-PCR-based amplifying techniques are the most flexible methods, which are applied in extensive fields. The RT-PCR-based amplifying techniques mainly include miR-Q RT-PCR for miRNA quantification, end-point stem-loop Real-time RT-PCR for miRNA quantification, Poly(A)-tailed universal reverse transcription, multiplexing RT-PCR for high-throughput miRNA profiling and miRNA amplification profiling (mRAP). Here we introduce these important Real-time PCR methods for detecting miRNA, respectively. And the theory, strategy, application and limitation of each method are summarized synoptically.
Effect of Different Lactobacillus Forms on Quality of Fermentation Liquid Feed
QIN Chun-fu, FAN Yong-hui, ZHANG Pei-hua, LI Jia-ling, CHAI Yu-guang, ZHU Dan
2013, 40(9):  114-118. 
Abstract ( 278 )  
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The objective of this research was to investigate the effect of different Lactobacillus sources on sensorial evaluation, pH and bacterial colony forming unit (CFU) in fermentation liquid feed (FLF). Fermentation liquid Lactobacillus enzymes (LE), fermentation liquid Lactobacillus enzymes mixture (EM) and 5 mg/kg of fermentation liquid Lactobacillus spawn mixture (SM) were designed as trial groups, respectively, and fermentation liquid substrate as a control (CON). All groups above maintained a fermentation substrate to water ratio as 1:3, and kept at (36±1)℃ thermostatically for 192 hours. Sensorial evaluation and pH measurement were conducted daily, and samples were obtained after 72 h of fermentation and then cultured. CFU was counted at 48 and 72 h after culturing, respectively. The results showed that EM and SM had superior sensorial characteristics, longer shelf lives, and had extremly significantly more colony counts than LE and CON (P<0.01), whereas there was no significant differences in CFU between EM and SM (P>0.05). The pH of all treatments showed a U curve and pH of LE was significantly (P<0.05) or extremly signifrcantly (P<0.01) low compared to other treatments. The results proved that LE could not improve fermentative quality of FLF, both EM and SM improved quality of FLF.
Effects of Nutrition Restriction and Compensation on Meat Quality of Lambs
LAN Ru-bing, GAO Ai-wu, YANG Jin-li, ZHANG Chong-zhi, HOU Xian-zhi, KAO Gui-lan
2013, 40(9):  118-123. 
Abstract ( 334 )  
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This experiment was conducted to investigate the effects of the nutrition restriction period (0 to 60 d) with different energy (ME) and protein (CP) levels and compensation period (61 to 150 d) on meat quality of lambs. Eighty Mongolian lambs were randomly divided into four treatment groups, the control group (CG), the moderate restriction groups (RG1 and RG2), and the severe restriction group (RG3).Dietary nutrition level were ME 10.88 MJ/kg and CP 15.0%, ME 10.88 MJ/kg and CP 10.0%, ME 9.41 MJ/kg and CP 10.0%, ME 8.62 MJ/kg and CP 5.7% during the nutrition restriction period for CG, RG1, RG2 and RG3, respectively. In nutritional compensation period all groups were offered the same levels diet (ME 9.75 MJ/kg, CP 12%). Four lambs of each group were slaughtered at the end of the restriction period and compensation period. The pH1value, pH24 h value, the rate of water loss and shear force values of lamb the longissimus dorsi (LD) muscle were measured, and the chemical compositions of carcass were also detected after the nutrition restriction period and compensation. The results showed that at the end of the restriction period, the pH1 value of LD and the content of carcass crude protein, crude ash of RG3 group were significantly higher than the CG group (P<0.05), the shear force value of LD was significantly lower than the CG group (P<0.05), and the content of carcass dry matter was extremely significantly lower than the CG group (P<0.01). At the end of the compensation, the pH1 value of LD of RG2 group was significantly higher than the CG group (P<0.05),the shear force value of LD and the content of carcass crude protein of RG1 and RG2 groups were significantly higher than the CG group (P<0.05). The content of carcass crude ash of RG3 group was significantly higher than the CG group (P<0.05). The results suggested that the pH value,tenderness and the carcass composition were all affected by the nutrition restriction. However, there were no differences between the control group and treatment groups except tenderness. The compensatory rates decreased with the increasing of nutrition restriction.
Determination of Slaughter Performance and Meat Quality in Different Sex of Leizhou Black Ducks
MA Meng, TANG Qi-ming, SU Ying, CHEN Qi, ZHANG Hui-teng
2013, 40(9):  124-127. 
Abstract ( 293 )  
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Randomly selected healthy disease-free sixty 300 days of age Leizhou Black ducks, male and female half-and-half, carried on a slaughter test. The results showed that the whole net carcass rate and lean percentage of male duck were significantly higher than female(P<0.05), the percentage of abdominal fat and subcutaneous fat percentage of female duck were extremely significantly higher than male(P<0.01), but its breast muscle percentage was extremely significantly lower than male(P<0.01), the meat color (L, a, b), cooking lose rate of thigh muscle and tenderness of breast muscle in male and female were all extremely significantly different(P<0.01), the crude protein content of thigh and breast muscle in female duck were extremely significantly higher than male(P<0.01), the fat of breast in female was significantly higher than male(P<0.05). The results showed that Leizhou Black ducks had higher slaughter rate and whole net carcass rate. The meat quality and nutrient content of muscle in female were higher than male, the breast muscle was better than thigh.
Comparative Study on Slaughter Performance between Two Hybrid Combinations
HOU Ming-feng, MENG Fan-ming, CAI Geng-yuan, WU Zhen-fang, LIU De-wu
2013, 40(9):  127-130. 
Abstract ( 522 )  
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Carcass and meat quality traits were determined by comparing 64 heads of (Pietrain×Duroc) (PD)× (Langdrance×Largewhite) (LY) crosses with 18 heads of (Pietrain×Duroc) (PD)×(Langdrance×Lantang) (LN) crosses in order to provide reference data on producing high quality pork. The results indicated that the PD×LN corsses were significantly higher than PD×LY crosses in the content of IMF, shoulder thickest backfat and the last rib backfat (P<0.01) in live testing performance, while slaughter weight and daily gain were extremly significantly lower than the latter (P<0.01). In the carcass traits, the shoulder thickest backfat of PD×LN crosses were extremly significantly higher than PD×LY crosses (P<0.01), but lower in carcass length and longissimus muscle area (P<0.01). In meat quality traits among IMF, pH1, marbling and tenderness, the PD×LN crosses were better than PD×LY crosses (P<0.01), but worse in fleshcolor b (P<0.01), pH24 h and drop lose (P<0.05), at last both line crosses showed normal in meat color with score 3 to 4, and meat quality traits with pH1>5.9 and pH24 h<6, the IMF of the PD×LY was detected between 1.5% to 2.0%, the IMF value of the PD×LN crosses had exceed 2%, shear force of two breed was less than 4 kg. In conclusion, meat quality of the two crosses catered to consumer taste.
Establishment of a ICR Mice Model Infected by Listeria monocytogenes
YIN Hai-chang, LV Xing-feng, LIU Si-guo, WANG Wei, WANG Jian-chao, MENG Qing-wen
2013, 40(9):  131-135. 
Abstract ( 445 )  
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Listeria monocytogenes (LM) was a zoonotic foodborne pathogen that could cause severe symptoms of infection in human and animals.Measuring LD50 of mice models which had immune activities was widely used for evaluation of pathogenicity of LM strains. In order to create a ICR mice model infected by LM, the N21 LM strains of the 1/2a serotype were inoculated orally into 6-week-old ICR mice groups (each group containing 10 mice) with the dose of perfusion 106,107,108,109 and 1010 CFU per mouse, respectively, the other 10 mice were inoculated by PBS as a control group to measure LD50 of LM infecting ICR mice. Another 40 mice were divided into two groups by gender, and inoculated with the dose of calculated LD50 of LM.Clinical symptoms, changes of histopathological, changes of body weight and tissue bacterial load were evaluated in each group, respectively.The results found that the LD50 of N21 LM was 109.25 CFU. The cycle of infection was about 10 days, the infected mice showed the clinical symptoms of rough coat, listlessness, penis hanging out, weight loss,etc.The histopathology analysis suggested that liver had appeared bacterial clumps in parenchyma, thrombosis and necrosis in sequence; spleen mainly showed white intramedullary lymphopenia decreased;lung mainly showed fiber simply pneumonia. Colony counting and Real-time PCR found that the highest bacterial load was in the liver, spleen followed. The experimental results showed that the ICR mice could be utilized as a LM infection model. This study laid the foundation for research on LM pathogenic mechanisms, vaccine development and evaluation of antimicrobial peptides transgenic mouse's resistance to Listeria monocytogenes.
The Effect of Colchicine and Nocodazole in Different Concentrations on Cycle Synchronization of Fibroblast in Cashmere Goat
YANG Hui-ru, XIAO Hong-mei, CAI Ting, JU Hong-li, YU Xin-lei, LIU Zhi-hong, LI Jin-quan, WANG Zhi-xin
2013, 40(9):  136-139. 
Abstract ( 739 )  
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This experiment was conducted to explore optimized conditions to make more Cashmere goat fibroblasts which had been treated in G2/M. In the experiment, the colchicine and nocodazole reagents were used in the processing of Cashmere goat fibroblast cell cycle synchronization, and the number of the cell in G2/M was measured by flow cytometry to observe the optimal concentration and action time. The results showed that the colchicine concentration should not be too large;furthermore, the time should not be too long in treatment of colchicines. In addition, under the same conditions, the treatment of nocodazole worked better than colchicine in synchronization. The optimum processing conditions included the concentration was 300 ng/mL and the action time was 24 h. Then the percentage of fibroblast in G2/M accounted for 15.10%. The results suggested that the effect of nocodazole reagents was more pronounced for Cashmere goats fibroblasts, when conducting the cell cycle synchronization processing.
Study on Subchronic Toxicity Test of Jizhikang Oral Liquid
HONG Wei-ming, ZUO Wei-yong, SONG Liang, HUANG Wen-qiang, YU Jie
2013, 40(9):  140-143. 
Abstract ( 275 )  
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In order to study the subchronic toxicity of the Jizhikang oral liquid, the test was conducted in accordance with the animal drug technological specification compile of Ministry of Agriculture. Experiment mice were divided into 4 groups, the 3 experiment groups were given Jizhikang oral liquid of 0.01,0.02,0.04 mL/(g·BW) for 28 days. The control group was given the same amount of naomal saline. The results showed that the body weight, organ coefficient, index of hematology and blood biochemistry indices in 3 experiment groups were not significantly different from that in the control group(P>0.05). Results of histopathology examination showed that it had no adverse reaction in experiment groups. The results showed that the Jizhikang oral liquid had no subchronic toxicity,and could be safely used for experimental animals.
Effects of env Gene on Subgroup J Avian Leukosis Virus Infection and Replication
WU Xiao-chan, LI Jiao, CAO Wei-sheng, LIAO Ming
2013, 40(9):  143-146. 
Abstract ( 228 )  
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In order to rescue the env element-substitution virus, we constructed the recombinant plasmid pNX-HNenv that contained the whole myelomas strain ALV-J NX0101 genome, but with env element of hemangiomas strain ALV-J HN06. The plasmids were transfected into DF-1 cells and the rescued viruses were identified, respectively. The recombinant virus was stable in cell culture during nine passages, and could be identified by ALV-J specific monoclonal antibody JE9 and virus NX-HNenv was rescued. The results showed that the env substitution had no apparent influence on ALV-J virus rescue, virus entry and replication.
Study on the Bacteriostasis of Aloe Vera Extracts against Aeromonas veronii
YANG Yong-li, WANG Li, WU Man-lin
2013, 40(9):  147-150. 
Abstract ( 254 )  
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To discuss the bacteriostasis of extracts from aloe vera on Aeromonas veronii by juicing method, water extraction and alcohol extraction method, the inhibition activities, the minimal inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the extracts from three kinds of methods which had significantly antibacterial effect against Aeromonas veronii were determined with K-B programe and the double broth dilution method. The results showed that the extracts from aloe vera with three kinds of methods all displayed sterilization ability on Aeromonas veronii inordinately. MIC of the extracts from aloe vera with juicing method, water extraction and alcohol extraction method were 1, 0.5, 0.125 g/mL, respectively. The MBC were 1, 1, 0.25 g/mL, respectively. Alcohol extract method showed higher activities than the others. But water extraction was considered as the most suitable method for extracting based on both of the antifungal activity and extraction efficiency.
Effects of the Steep of Seriphidium on Immunologic Function of Chicks
DUAN Hai-feng, YAN Gen-qiang, WANG Jing-mei, WANG Rong, XING Wei-yuan, WAN Peng
2013, 40(9):  151-154. 
Abstract ( 280 )  
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To investigate the effects of Seriphidium in the Nanshan area, Shihezi, Xinjiang province on immunologic function of chicks,the influence of the weight was studied by weighing method,the proliferation of T lymphocyte was studied by MTT method,the phagocytic function of peritoneal macrophages was studied by Colony counting method,the number of T-lymphocyte of peripheral blood was studied by ANAE method,the death rate in chicks was studied by Escherichia coli tapping poison. The steep of Seriphidium could levelly improve the chickens'weight, proliferation abilities of T lymphocyte and positive rates of T lymphocytes ANAE, and decrease phagoeytosis ability to bacterium and the incidence of chicks to E.coli. The results showed that the steep of Seriphidium depended on its dose had a two-way effect on the immunologic function of chicks.
Study on Extraction and Purification of Baicalin from Hairy Roots of Scutellaria baicalensis Georgi
SHI Chun-yang, QI Xiang-jun, QIAN Wei-dong
2013, 40(9):  154-157. 
Abstract ( 278 )  
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Using the content of Baicalin as the standard, the optimum macroporous resin was selected, and the factors of Baicalin's extraction and purification were investigated. LX-68 macroporous resin was the suitable one, Baicalin was extracted under the optimum conditions of a sample concentration of 8.69 g/L, a adsorption speed of 6 BV/h, a sample volume of 16 BV, a diameter-high ratio of 1:14, a elution reagent of 70% alcohol, a elution speed of 6 BV/h, a elution volume of 10 BV, and then purificated by silica gel column chromatography, the condition was chloroform-ethyl acetate-methanol-formic acid(7:3:1.5:0.5). The LX-68 macroporous resin could be used to extract Baicalin from the hairy roots of Scutellaria baicalensis Georgi, the adsorption capacity of total flavone was 131.2 mg/g, the returns-ratio was above 69.1%, and the purity of Baicalin was 98.45%.
Research Progress on Meat Quality Traits Candidate Genes in Cattle
FU Shi-jun, GUO Shi-jin, KAN De-ying, LI Jin-lin, LI Feng, TANG Shao-wei, SHEN Zhi-qiang
2013, 40(9):  158-165. 
Abstract ( 319 )  
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Calpain system played an important role in muscle protein hydrolysis and muscle tenderization after animal slaughter. Fatty acid binding proteins affected muscle tenderness, flavor and juiciness, and had high relevance to backfat thickness, marbling, tenderness and water capacity of meat traits through taking part in the transport and metabolism of fatty acids. Myogenic regulatory factors controled the fusion of muscle cell and formation of muscle fiber during the muscle differentiation process. Myostatin, a negative regulator for muscle growth, could affect lean meat percentage of animal and better meat quality traits. Several meat quality candidate genes in cattle including calpain system family, fatty acid binding protein gene, myogenic regulatory factor, and myostatin were summarized in order to find the way to improve the quantity and quality of meat and further improve the meat quality traits through marker-assisted selection.
Study on the Effect of Fagopyrum dibotorys Superfines Powder on Immune Function of Mice
WANG Hang, TANG Cheng, YUE Hua, WANG Hong-ge, WANG Cheng-long
2013, 40(9):  165-168. 
Abstract ( 304 )  
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To study the effect of Fagopyrum dibotrys superfines powder on immune function and weight of mice,60 Kunming mice were divided into 4 groups,high, middle and low dose groups of mice were gavaged with the drug at a dose of 1.75, 1.25 and 0.75 g/(kg·BW) once a day for 7 consecutive days, repectively, the mice of control group were administered with the same volume of saline. 12 h after the last once, mice were injected into the muscle with Newcastle disease vaccine,then we got the peripheral blood on day 7, 14 and 21 after the medicine administrate. The enhancement of innate immune responses was evaluated by using of CCK-8 method, hemagglutination and hemagglutination inhibition test, macrophage phagocytic function test and organ coefficient method, and record the data of weight. In middle dose Fagopyrum dibotrys superfines powder group, the level of lymphocytes proliferative capacity, serum antibody, peritoneal macrophage phagocytosis and the index of immune organs were extremly significantly enhanced(P<0.01), and the weight was also extremly significantly increased(P<0.01). Fagopyrum dibotrys superfines powder could significantly regulate the immunity function and improve the weight in mice.
Study on the Variation of Cashmere Yields of Inner Mongolia White Cashmere Goats
ZHOU Juan-juan, TANG Pei-rong, WANG Zhi-xin, WEI Yong-long, MENG Rui-qiang, WANG Zhi-ying, ZHANG Wen-guang, ZHANG Yan-jun, SU Rui, LIU Zhi-hong, WANG Rui-jun, LI Jin-quan
2013, 40(9):  169-175. 
Abstract ( 317 )  
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In this paper,data from 2003 to 2011 production performance records of Inner Mongolia White Cashmere goats were used to analysis and discuss the relationship between cashmere yield and weight after fleecing (WT). Excel and SAS were used to organize data in the study. Cashmere yield decreased in 2004,2008,2009,2011 years,but WT increased. As the growthof the age, the cashmere yield gradually increased before the fourth years old, and declined but there were a few still increasing phenomenon of varying degrees after the fourth years old,WT showed an increasing trend. Ignoring year's environmental factors the cashmere yield and WT had a same trend as the growth of the age before the fourth years old, but with the increasing of cashmere yield, the increasing of WT presented decrescent trend.Otherwise, with the increasing of WT, the added of cashmere yield slowdown. While the cashmere yield increased whereas WT decreased, and cashmere yield decreased but the fleece weight increased in the same wool production season. This research showed that there were some competition between the cashmere yield and WT grew together at the same time at the post-growing period.
Study on the Milk Somatic Cell Count and the Changes of Milk Composition in Sanne-goat
LI Wei-juan, HU Zhong-ren, LIU Qin, ZHOU Ya-ping, LI Dong-jiang, DU Wen-hua, ZHAO De-hong, ZHAO Fu, WANG Lian-mei
2013, 40(9):  175-179. 
Abstract ( 381 )  
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In this study, in order to study the effect of the somatic cell count (SCC) on the main composition and content of goat milk, somatic cell were collected and analyzed of Sanne-goat by using DHI method, which could provide the reference for raw milk product quality evaluation. The results showed that the SCC had no significantly positively related with milk fat percentages (P=0.141), and were significantly positively related with milk protein percentages (P<0.01). The content of lactose was significantly negatively related with SCC (P<0.01).The content of total solids was highly significantly positively related with SCC (P<0.05).The content of urea nitrogen was highly significantly negatively related with SCC (P<0.01). Rising of SCC could greatly affect the quality of goat milk, induce the increase of milk protein percentage, milk fat percentage and the content of total solids and the decrease of the contents of lactose and urea nitrogen. In summary, the somatic cell count and the portion of milk components in June could be used as reference to evaluate the quality of Sanne-goat milk.
Analysis of Growth and Development Models of the Gansu Meat Sheep New Breed Population
MA You-ji, HU Xiang, LI Fa-di, JIANG Zhong-wen
2013, 40(9):  179-182. 
Abstract ( 286 )  
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The Gompertz and Logistic models were used to study the production potential of Gansu meat sheep new breed population, so as to provide theoretical basis for meat sheep breeding. The results showed that the fitnesses (R2) of two kinds of models were above 0.91, the Gompertz model had higher R2 value and better predicting effect towards bodyweight. The Gompertz model was constructed for the Gansu meat sheep new breed population.The body weight at inflection age were 24.06 and 16.16 kg, inflection age were 2.73 and 1.80 months, the maximum monthly weight were 7.92 and 7.64 kg, the instantaneous growth rate were 7.27 and 5.18, the relative growth rate were 0.21 and 0.17, respectively. The results proved that the Gompertz model could be used to guide the production of Gansu meat sheep new breed.
Detecting Copy Number of Foreign Gene in Human Alpha-lactalbumin Transgenic Cloned Cattle
WANG Jie-wen, WANG Jing-jiao, ZHAO Shu-jun, LI Ning, LI Shi-jie, ZHANG Cui
2013, 40(9):  183-187. 
Abstract ( 378 )  
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To explore the changes of the exogenous gene copy number in transgenic cloned animals during subculture, we detected the copy number of human alpha-lactalbumin gene in five transgenic cloned cattles, including a parental cattle(K060923) and four offspring cattles(172,081,189 and Longwa), with Real-time fluorescent quantitative PCR. Among the five cows, K060923 was parental and the rest are offspring. The copy number of K060923 was 1.145, and that of the four offspring (172, 081, 189 and Longwa) was 0.998, 1.107, 0.891 and 0.842, respectively. The obtained results indicated that the copy numbers of all the five cows were about one. The results showed that exogenous gene was integrated into the host genome as a single copy and was stably inherited.
Signal Regulation for Melanoblast Cytopoiesis Process:A Review
YANG Chun-ling, HAO Tian-tian, LI Qiang-fei, CHEN Ting, MAO Hua-ming, DENG Wei-dong
2013, 40(9):  187-194. 
Abstract ( 371 )  
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Melanocytes synthetise melanin which determine skin and coat color for animals. There are two generate ways about melanoma cells throughout the body. One way is through the retinal pigment epithelial cells which are divided from medullary epithelium cells. The other way is through melanoblasts, in the early embryonic development, neural crest cells differentiate into melanoblast firstly, these cells continue to develop into mature melanocytes. After reviewing on the development of mouse and human genetics, we had proved that it was a complex process of melanin derived from neural crest cells. The development and differentiation of melanocytes were regulated by multiple signaling factors. Among the rest there were some transcription factors SOX10, MITF and PAX3, and three signaling pathways, KIT and KITL, WNT/β-catenin, EDN3 and EDNRB. The MITF was considered as the most important regulatory factor during the development of melanocytes.Three pathways were also considered closing to the development of melanocytes and regulating the function and activity of MITF. This article summarized the research progress on the development of melanocytes and indicated that early developmental regulation of neural crest cells might be the fundamental reason which caused the formation of hyper-pigmentation traits in silky fowl and Black-boned sheep.
Study on Mix Infection of Pathogenic Proteus mirabillis and E.coli Causing Joint Abscess and Diarrhea of Dairy Cows
WANG Xu-rong, WANG Guo-qing, ZHANG Jing-yan, YANG Zhi-qiang, MENG Jia-ren, LI Jian-xi
2013, 40(9):  195-198. 
Abstract ( 416 )  
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In order to identify the cause of joint abscess and diarrhea of dairy cows in Gansu province and choose sensitive drugs for treatment,clinical samples were isolated and identified by BVDV and BRV detection, bacteria isolation and culture, mouse pathogenicity, antibiotic sensitivity tests and content detection of Ca and P in the sera which were collected from diary cows suffering from joint abscess and diarrhea. The results showed that mix infection of pathogenic Proteus mirabilis and E.coli causing joint abscess and diarrhea of dairy cows. The two isolates were medium sensitive to florfenicol,resistant to penicillin,amoxicillin, gentamicin, kanamycin neomycin,tetracycline, norfloxacin and cotrimoxazole. This disease was controlled by the treatment of florfenicol and enhancing feeding management.
Isolation and Identification of a Resistant K.peneumoniae Strain from Rabbits
ZHANG Zhi, LI Zai-xin, ZENG Guang-ju, LI Bing-chao, HU Wei-dong, ZHAO Zhi-ping, LIU Zhang-qin, CHEN Yu-yun
2013, 40(9):  199-203. 
Abstract ( 301 )  
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A suspected pathogen was newly isolated from rabbits with diarrhea and respiratory tract infection by isolation and culture. To further understand some features of this suspected bacteria, microscopic examination, biochemical detection including capsular stain, drug sensitive test and 16S rDNA sequence analysis were performed. The results showed that the isolated bacteria was in rod containing rich capsular polysaccharide, and demonstrated a significant multi-drug resistance to streptomycin, gentamicin, ampicillin, gentamicin sulfate, kanamycin sulfate and chloramphenicol at different levels. Furthermore 16S rDNA sequence of this suspected bacteria was cloned by PCR and sequenced, the result showed that 16S rDNA sequence shared about 99% similarity to that of K.pneumonia standard strains provided by GenBank. In addition, infection of suspected bacteria (109 CFU/mL) resulted in mortality to rabbits by intraperitoneal injection, but not by oral administration. Altogether, it could be concluded that the isolated bacteria was a member of K.pneumonia and contributed to the disease of rabbits.
Separation and Identification of Multi-drug Resistant and High Pathogenic Escherichia coli Causing Piglets Watery Diarrhea
XUE Yu-jia, YU Da-yong, CUI Min, LV Yan, YAN Guang-mou
2013, 40(9):  204-208. 
Abstract ( 301 )  
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Piglets watery diarrhea with high pathogenity and high mortality rate happened in many farms in Changchun area, the diarrhea piglets repeatedly tried a variety of medications,but no effect came out. From the onset of piglets intestinal wall lining,we had separated and identified some drugresistant and highly pathogenic E.coli strains.After the biochemical test,and cloning and sequencing of 16S rRNA of the strains,we compared them with the sequences in GenBank,the isolated strains had 99% homology with the 16S rRNA of E.coli sequence. After animal testing, we determined that this kind of E.coli could cause piglets watery diarrhea. Drug sensitivity test results showed that drugs commonly used in E.coli disease as treatments such as gentamicin,ofloxacin and ciprofloxacin were invalid.
Research Status and Application Prospects of H5N1 Subtype Avian Influenza Virus Vaccines
FENG Shu-ying, DONG Shi-zhen, ZHANG Shu-xiao
2013, 40(9):  208-213. 
Abstract ( 454 )  
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The H5N1 subtype avian influenza virus (AIV) had been known as a pathogen with specialties of the strongest infection, the highest mortality and the widest epidemic. H5N1 subtype AIV was endemic in many countries around the world and had caused considerable economic losses to the poultry industry. However, owing to no effective treatment measures, vaccine inoculation had become the most-effective prevention measure and the key loop. Therefore,the development of the safe, high-efficiency and low-cost vaccine was one of the current hotspots for the control of avian influenza. Based on the large numbers of research achievements of avian influenza vaccine, we comprehensively described the research status of avian influenza vaccine from the following aspects,the virus inactivated vaccine, attenuated live vaccine, genetic engineering vaccine and other technology application etc. Moreover, we summarized the difficulties and drawbacks of studies and gave the future prospects of vaccine application with the purpose of providing a comprehensive background and reference for the control of avian influenza.
Dynamics Research of Serum and Egg Yolk Antibody of Laying Hens Immuned by Canine Distemper Virus
ZHUANG Jin-qiu, MEI Jian-guo, YAO Chun-yang, SHEN Zhi-qiang, DING Zhuang
2013, 40(9):  214-217. 
Abstract ( 256 )  
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In order to provide the basis for the preparation of egg yolk antibody,the dynamics of the serum antibody and egg yolk antibody in laying hens against canine distemper virus (CDV) antigen was studied,Vero cell and DF1 cell cultures were inoculated with CDV to detect the virus titer. Select the Vero cytotoxic strain with earlier cytopathic and high virus titers as inoculated virus antigen to immunize laying hens. The immune method was to immunize laying hens once every 10 days, for four times successively, then to strengthen the immune every 30 days. Blood sera were separated every 10 days and egg yolk antibodies were collected every 5 days before and after immunization,the antibody levels were determined by agar diffusion and indirect ELISA. The results showed that serum antibody titer was higher than egg yolk antibody, and both were positively correlated,the appearance of egg yolk antibody was later for 3 to 5 days than serum antibody. Egg yolk antibody had reached the highest level in 3 to 5 days after the third immunity, and then you could start to collect eggs to prepare egg yolk antibody.
Research Progress on Mastitis Treatment of Microecologics in Dairy Cows
ZHU Dan-dan, LIU Yu, XU Ming, LI Dan, WANG Yan, WU Bo, WANG Shuang, SHI Tong-rui
2013, 40(9):  218-221. 
Abstract ( 409 )  
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Mastitis was one of the common diseases in dairy cows, it produced a great impact on dairy farming business and affected milk quality. Microecologics were proposed as a valid alternative to antibiotic therapies and were wildly applied because of their safety. This paper reviewed the mastitis pathogenesis and the present situation of microecologics controling mastitis in dairy cows, including its mechanism of action.
Determination of Proantho Cyanidins B2 of Fagopyrum dibotryis Superfine Powder by HPLC
WANG Hang, TANG Cheng, YUE Hua
2013, 40(9):  222-224. 
Abstract ( 245 )  
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To determine the content of proantho cyanidins B2 in Fagopyrum dibotryis superfine powder, the Fagopyrum dibotryis crushed less than 0.75 nm superfine powder extracted by 50% ethanol,an HPLC method was used. Epicatechin showed a good linear relationship in 0.0352 to 1.4080 μg/mL and the regression equation was Y=59797X+592.3,r=0.996. Proantho cyanidins B2 of Fagopyrum dibotryis superfine powder content was between 1.305 to 1.541 mg/g.The method was simple and suitable for determing the content of proantho cyanidins B2 in Fagopyrum dibotryis superfine powder. It provided the scientific basis for Fagopyrum dibotryis preparation quality control.
Research Progress on the Regulation of Epididymal Specific Glutathione Peroxidase (GPx5) Gene
ZHANG Guo-lin, LI Zhen, GAO Jin-sheng, CAO Ning-xian, ZHANG Chun-xiang
2013, 40(9):  225-228. 
Abstract ( 248 )  
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Epididymal specific glutathione peroxidase (GPx5) protected sperms from oxidative stress, maintained the integrity of sperm genetic information and promoted sperms to develop, its physiological functions played an irreplaceable role in improving animal reproduction. But the mechanism of its gene expression regulation of transcription was not yet clear. Based on the achievements the concerning literature had been made, the study had carried on the summary of characteristics, expression and regulation of gene expression and transcription regulation etc.
Study on Selection of Growth and Development Performance of Jinfen White Pig
GUO Xiao-hong, WANG Xiao-jing, ZHANG Min, LIU Hong, SHI Jian-zhong, XU Kai, LI Wei, ZHANG Yan-qing, GAO Peng-fei, ZHOU Zhong-xiao, CAO Guo-qing, LI Bu-gao
2013, 40(9):  228-231. 
Abstract ( 607 )  
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Jinfen White pig was a new lean-type variety by crossing Mashen pig, Taihu pig, Landrace, Yorkshire pig and selecting by means of combining the marker assisted selection with systematic breeding. The selection of growth and development of six generations showed that compared with the base population, body weight at birth, body weight at weaning, body weight at 70-day-old, body weight at 180-day-old, and average daily gain during the period of 70 to 180 d of boars at the sixth generation were remarkably increased by 0.13 kg, 0.56 kg, 1.69 kg, 12.46 kg and 107.00 g, respectively (P<0.05). Compared with the base population, body weight at 180-day-old of gilts at the sixth generation was significantly increased by 10.84 kg (P<0.05). Body weight at birth, body weight at weaning, body weight at 180-day-old and average daily gain during the period of 70 to 180 d of gilts and boars at the fourth and fifth generation were significantly higher than these traits of the base population, respectively (P<0.05). It indicated that growth and development performances of Jinfen White pig had made significant genetic progress.
Effect of Different Proportions of Bamboo Powder in the Basic Diet on Rabbit Growth and Slaughter Performance
TAN Qian-hong, WANG Yong-kang, LIANG Huan, ZUO Guo-jiang, HU Yuan
2013, 40(9):  232-234. 
Abstract ( 337 )  
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176 thirty-day-old California weaned meat rabbits with consistent weight were randomly divided into four groups,Ⅰ, Ⅱ, Ⅲ groups were the test groups, adding 10% bamboo powder and 17% alfalfa meal, 15% bamboo powder and 12% alfalfa meal, 18% bamboo powder and 9% alfalfa meal in the basic diet, respectively. Ⅳ group was control group, adding 27% alfalfa meal in the basic diet.The aim was to study the different proportions of bamboo powder instead of alfalfa meal on growth performance and slaughter performance in meat rabbit. The results showed that feed to gain ratio of each test group was extremly significantly lower than control group (P<0.01), reduced by 16.27%, 10.62%, 3.47%, respectively. Daily gain of each test group was extremly significantly higher than the control group (P<0.01), increased by 19.29%,9.19%,3.67%, respectively. Half net slaughter rate of each test group was extremly significantly higher than control group (P<0.01), increased by 2.61%, 1.35%, 0.26%, respectively. Net whole slaughter rate of each test group was extremly significantly higher than control group (P<0.01), increased by 2.98%, 2.55%, 0.80%, respectively.The results revealed that adding 10% bamboo powder and 17% alfalfa meal had the best effect.