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20 August 2014, Volume 41 Issue 8
Cloning and Bioinformatics Analysis of MyoG Gene in Jinghai Yellow Chicken
ZHANG Tao, ZHANG Gen-xi,WANG Jin-yu, FAN Qing-can, WANG Wen-hao, WEI Yue, CHEN Xue-sen, TANG Ying,WANG Yong-juan
2014, 41(8):  1-8. 
Abstract ( 280 )  
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Based on the published mRNA nucleotide sequence of Gallus gallus myogenin(MyoG) gene, a pair of primers were designed to clone the MyoG gene coding sequence of Jinghai Yellow chicken by RT-PCR. A variety of software and online tools were used for bioinformatic analysis of MyoG gene. Bioinformatics technology was successfully used to analyze the MyoG homology among different species, physical and chemical properties of the protein, transmembrane region of MyoG sequence, subcellular localization, hydrophilic, potential phosphorylation locus, conserved domain database, MyoG secondary structure and teriary structure. Finally, a 754 bp gene sequence was cloned, which contained CDS region, promoter region and part of the 3 ′region. Bioinformatics analysis showed that MyoG gene of Jinghai Yellow chicken was less conservative and shared 99.7%, 94.4%, 92.0%, 70.0%, 70.0%, 73.3%, 69.0%, 67.9%, 67.8% and 73.5% identity with Gallus gallus, Meleagris gallopavo, quail, horse, people, wild boar, cattle, goats, rats and sheep. The amino acid sequence analysis revealed that the MyoG gene of Jinghai Yellow chicken encoded water-soluble protein and its molecular weight was 25854 u, isoelectric point was 5.46, and did not belong to the membrane protein. Subcellular localization of MyoG was in the cytoplasm and it did not belong to the secreted protein. The MyoG protein contained six phosphorylation sites,a alkaline sequences, a HLH sequence, a low complexity sequence. The secondary structure of MyoG was mainly composed of random coil. The tertiary structure of domain area of MyoG protein showed a helix structure. The results above laid the foundation for further studies of expression regulation mechanism and function of MyoG gene in the muscle growth process.
Silico Cloning and Sequence Analysis of STAT1 Gene in Water Buffalo (Bubalus bubalis)
DENG Ting-xian, PANG Chun-ying, CHEN Ming-tang, WANG Jian, YANG Bing-zhuang, LIANG Xian-wei
2014, 41(8):  9-13. 
Abstract ( 230 )  
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The buffalo STAT1 gene was silico cloned and analyzed by bioinformatics method in this study. Using the Bos taurus STAT1 sequences to get and identify the buffalo STAT1 gene sequences by the silico cloning and RT-PCR technology. The results showed that the buffalo STAT1 gene length size was 3437 bp, contained an open reading frame (ORF) of 2244 bp that encoded 747 amino acids. The CDS of the obtained buffalo STAT1 gene showed that 99%,98%,89%,93%,86%,85% and 87% identity with that of Bos taurus, Ovi saries, Homo sapiens, Sus scrofa, Rattus norvegicus, Mus musculus and Capra hircus. Analysis of the phylogenetic tree showed that the nearest relationship existed between buffalo and Bos taurus.
Bioinformatics Analysis of Coding Regions of SCF Gene in Different Species
XI Jian-zhong, ZHOU Rong-yan, ZHANG Jun-jie,LI Xiang-long, LI Lan-hui, LI Nan, ZHANG Zhen-hong
2014, 41(8):  14-20. 
Abstract ( 199 )  
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To study the genetic variation of the SCF gene CDS in different species, the genetic diversity of the CDS of SCF gene sequences from Capra hircus, Ovis aries, Bos taurus, Ceratotherium simum, Sus scrofa, Orcinus orca, Equus caballus, Mustela putorius furo, Homo sapiens, Sorex araneus, Heterocephalus glaber, Rattus norvegicus, Chinchilla lanigera, Mus musculus and Octodon degus were analyzed using the method of bioinformatics. The characteristics of composition of nucleic acid sequences and amino acid sequences, leader peptides, signal peptide, trans-membrane topological structure and the secondary structure were also analyzed, and some of the foci of intense investigation on the function of SCF were reviewed. The results showed that a total of 294 polymorphic sites were detected from 55 sequences of 15 species, from which 36 hapolotypes were sorted. The genetic diversity of the SCF gene CDS was rich among species. The theoretical isoelectric point of the sequences was widespread, most of the peptide chains were acidic, hydrophilic,with the N-terminal signal peptide and without leading peptide. Soluble peptide had no transmembrane domain, while transmembrane type had a 23 amino acid long transmembrane domain. The mainly secondary structures of these proteins were α-helix, disorderly curly, extended chain and β-corner.
Preparation of Monoclonal Antibodies against Mycoplasma capricolum subsp. capripneumoniae and Screening of Antibody Binding Epitopes
ZHENG Ying-ying, PIAN Ya-ya, CHU Yue-feng, ZHAO Ping, HE Ying, JIAN Ying-na, JIANG Yong-qiang, LU Zhong-xin
2014, 41(8):  21-24. 
Abstract ( 245 )  
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The aim of this research was to prepare monoclonal antibodies against Mycoplasma capricolum subsp.capripneumoniae (M.capricolum subsp.capripneumoniae, Mccp), and also screen antibody binding epitopes. We inactivated Mccp with formaldehyde, and then immunized BALB/c mice. We applied conventional fusion cell technology to obtain hybridoma cell, subclones, and ascites of monoclonal antibodies. We used enzyme-linked immunoassay (ELISA) and immunoblotting (Western blotting) technologies to identify the specificity of monoclonal antibodies, and enzyme digestion method to identify antibody binding epitopes. Finally, we successfully screened five monoclonal antibodies and identified three antibody binding epitopes, which provided a preliminary and reliable experimental basis for further scientific research.
Expression of Hemagglutinin of Canine Distemper Virus in the Insect Cell and its Antigenic Analysis
QUAN Chuan-song, JIANG Qian, YU Zuo, LI Bo-tao, LI Lian-feng, QU Lian-dong
2014, 41(8):  25-29. 
Abstract ( 192 )  
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To maintain the native conformation and antigenicity of hemagglutinin (H) of canine distemper virus, the H gene which referred to HLJ2-07 strains was accurately amplified by RT-PCR and cloned into pFastBacTMHTA vector. The recombinant plasmid was successfully transformed into E.coli DH10BacTM competent cell and constructed shuttle vector, which was transfected into Sf9 cell to save baculovirus expressing destined protein. To determine the antigenicity of recombinant protein, Western blotting analysis was used to detect the interesting protein. Furthermore, the identification and characterization of B cell epitopes were predicted and analyzed by related software and online websites. The result demonstrated that the fusion protein generated by recombinant baculovirus was reacted with both anti-His tag monoclonal antibodies and canine distemper virus positive serum by Western blotting test, which showed that this protein had sufficient antigenicity. In addition, B cell epitopes were selected by between Swiss-model and online websites for consistent sequences to improve predictive accuracy, whereas the possible sites were 175 to 195, 240 to 250, 365 to 380, 480 to 508 and 526 to 555. These efforts served as a strong foundation for the development of diagnostic kits,the production of monoclonal antibodies and selection of protein epitopes.
Expression and Identification of F Protein of Peste Des Petits Ruminants Virus Strain Tibet 07 in Insect Cells
ZHAO Wen-nian, WANG Qing-hua, LIU Chun-ju, WANG Shu-juan, LEI Cheng-hong, BAO Jing-yue, WANG Zhi-liang
2014, 41(8):  29-33. 
Abstract ( 210 )  
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In order to obtain purified peste des petits ruminants virus F protein, the F gene of peste des petits ruminants virus strain Tibet 07 was cloned into transfer vector pFastBac/CT-TOPO, plasmid pFastBacCT-PPRV-F was then transformed into DH10Bac complement cells. The recombinant bacmid DNA was isolated and transfected into sf9 insect cells to express F protein. The expressed F protein was identified by using SDS-PAGE, Western blotting and indirect immunofluorescence assay. The recombinant F protein was successfully expressed in insect cells with a relative molecular of 59 ku.The sf9 cells infected by Bacmid-PPRV-F can reacted with positive serum of peste des petits ruminants.Using the Bac-to-Bac baculovirus expression system expressed the F protein of peste des petits ruminants virus strain Tibet 07.This work provides basis for development of rapid test for detection of peste des petits ruminants antibody.
Establishment of Multiplex PCR for Detecting of Shigella flexneri Serotype 2
ZHU Zhen, WANG Jing, ZHANG Ji-yu, WEI Xiao-juan, ZHOU Xu-zheng, GUO Xiao, LIU Cui-cui
2014, 41(8):  34-39. 
Abstract ( 181 )  
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In order to build a fast, efficient and specific multiplex PCR method for detection and identification of Shigella flexneri, this study used specific genes ipaH, gtrⅡ, gtrⅩ and R002 in different serotypes of Shigella to design the corresponding primers to amplify the target fragment, and in the same PCR system validated the Shigella flexneri. At the same time through a series of exploration and adjustments to optimize the reaction conditions, we established a method to detect Shigella flexneri type 2a and 2b out of a variety of bacteria quickly. The results suggested that the multiplex PCR could identificate the Shigella flexneri quickly, efficiently and accurately, and had great significance for the prevention of laboratory contamination, clinical diagnosis and treatment of shigellosis.
Cloning and Tissues Expression Analysis of Goose Forkhead Box O1 (FoxO1) Gene cDNA Sequence
YANG Lu, WEI Shou-hai
2014, 41(8):  39-45. 
Abstract ( 202 )  
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In order to study the function of goose forkhead box O1 (FoxO1) gene, in this study, according to the highly conservative region of FoxO1 gene sequences from chicken (Gallus gallus),human (Homo sapiens),and mouse (Mus musculus)in the GenBank,the specific primers were designed to clone the CDS region of goose FoxO1 gene using the RT-PCR method,and the bioinformatics analyses of FoxO1 were conducted.The results showed that the full length of CDS region of FoxO1 gene was obtained, and the similarities of nucleotide sequence of goose with chicken, human and mouse were 97%, 83%, 82%, respectively. The expression level of FoxO1 gene in various tissues of Sichuan White goose showed: sebum> abdominal fat > crureus > pectorales > liver. In conclusion, FoxO1 gene could be expressed in several tissues, and showed an evident expression difference.
Analysis of Bm86 Gene and its Encoding Protein of Rhipicephalus microplus in South Xinjiang
LIU Yong-hong, ZHAO Li
2014, 41(8):  45-49. 
Abstract ( 167 )  
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In order to highly express the ideal Rhipicephalus microplus antigen, we analyzed genetic evolution of Bm86 gene on the laboratory acquired in this study and biological information of its coding protein. The results showed that the homologies of complete open reading frame sequences of nucleotide and amino acid sequence of Bm86 gene of Rhipicephalus microplus with all the foreign Bm86 gene logged in GenBank were 95.9% to 98.6% and 93.8% to 97.8%,respectively. The difference was very small. The evolutionary tree showed that the strain and most of logged strains were in same branch. Biology information prediction showed that the protein was unstable. Insoluble probability was 93.26%. There were 4 potential glycosylation sites. Analysis of the above information would lay the foundation for the gene expression, preparation of immune antigen and development of vaccine against Rhipicephalus microplus.
Molecular Characteristics Analysis of Two Strains of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus
LUO Cheng-bo, CUI Min, KONG Ling-ping, CHENG Zhen-tao, ZHOU Bi-jun, MAO Li-hong
2014, 41(8):  50-56. 
Abstract ( 184 )  
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In order to find out the graph variation characteristics of porcine reproductive and respiratory syndrome virus (PRRSV) of Qianxinanin Guizhou province, the Nsp2 and GP5 genes of PRRSV isolated strain GZLPS and GZSB were cloned, sequenced and analyzed. The results showed that, the Nsp2 gene of 2 isolates contained 90 nucleotides for discontinuous deletion and 36 nucleotides for mutation. Furthermore, there were different replacements of nucleotides in GP5 gene. Not only Nsp2 gene, but also GP5 gene, the nucleotide homologies were above 96% and the amino acid homologies were above 95% with strain JXA1 and isolates in Guzizhou since 2011, but there were differencex in molecular variation among these strains. It indicated that the PRRSV had already behaved diverse on variation style of Qianxinan in Guizhou province.
Cloning and Sequence Analysis of C-terminal of Toxoplasma gondii DOC2 Gene
GAO Qi, ZHANG Nian-zhang, HU Ling-ying, ZHOU Dong-hui,ZHU Xing-quan,WENG Ya-biao
2014, 41(8):  56-60. 
Abstract ( 194 )  
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In order to evaluate whether Toxoplasma gondii DOC2 gene could be used as a vaccine candidate antigen, the C-terminal of DOC2 gene was amplified by RT-PCR and then sent for sequencing. After translated the gene sequence into amino acid sequence, the basically biological characters, advanced structures and linear B cell epitopes of the C-terminal of DOC2 gene were predicted using multiple bioinformatics analysis. The results showed that the length of C-terminal of DOC2 gene was 1887 bp. The C-terminal of DOC2 protein was predicted contain 11 hydrophilic regions, 7 transmembrane domains,19 alpha regions, 1 beta turn, 17 random coils and 12 B cell epitopes, which suggested this protein was a probable vaccine candidate against Toxoplasma gondii infection.
Expression, Purification and Immunological Activity Identification of Recombinant Proteins TB27.4 of Mycobacterium bovis
GAO Xin-tao, LI Ping-jun, XIN Ting, JIA Hong, GUO Xiao-yu, ZHANG Gai-mei, LI Ming, LIU Shu-qing, ZHU Hong-fei, XU Lei
2014, 41(8):  61-65. 
Abstract ( 172 )  
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In order to evaluate the diagnostic properties of protein TB27.4 against bovine tuberculosis, the gene coding for TB27.4 was amplified from Mycobacterium bovis (strain Vallee Ⅲ) genomic DNA by PCR, then cloned into pET-32a(+). The recombinant plasmid was transformed into E.coli BL21(DE3) and parameters for protein expression and purification were optimized. Results of SDS-PAGE and Western blotting showed that the recombinant protein was expressed in soluble form, and could be recognized by serum from Mycobacterium bovis-infected cattle. During ELISA, the recombinant TB27.4 could specifically stimulate the PBMCs of Mycobacterium bovis-infected cattle to secrete high amount of IFN-γ. These results indicated that the recombinant protein TB27.4 had good reactogenicity and T cell activity.This study laid the foundation for further study of the role which the recombinant protein TB27.4 played in the diagnosis of bovine tuberculosis.
Construction and Expression of NS1 Protein Eukaryotic Expression Vector of Influenza A Virus Subtype H7N9
WU Li-wei, LI Ting, YANG Shi-mei, WANG Hui-yu, MEI Lin, HAN Xue-qing
2014, 41(8):  66-70. 
Abstract ( 216 )  
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DNA Methylation Status of ASCL2 Gene in Lungs of SCNT Cattle and Naturally Reproduced Cattle
WANG Meng-nan,ZHANG Ming-yue,LI Ning,LI Shi-jie
2014, 41(8):  71-75. 
Abstract ( 181 )  
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In an effort to determine the reprogramming of ASCL2 gene in SCNT bovines,the bisulfite sequencing method (BSP) was used to analyze the DNA methylation status in promoter region of ASCL2 gene in lung tissues of SCNT cattle and natural breeding cattle.The results showed that the methylation level of ASCL2 promoter exhibited seriously hypermethylated in SCNT bovines (28.41%) compared with naturally breed bovines (7.62%) (P<0.05). Our results suggested that the abnormal DNA methylation of ASCL2 might be associated with the defects of lung organs in died SCNT bovines,which was one of the main reasons for neonatal death of SCNT animals.
Cloning and Sequence Analysis of ORF3 Gene of Porcine Torque Teno sus Virus 1b
HU Chong-wei, CHEN Ru-jing, CHEN Qiu-yong, LIN Qun-qun, CHEN Peng-qiang, XIU Jin-sheng
2014, 41(8):  76-80. 
Abstract ( 166 )  
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In order to enrich the ORF3 target protein information of porcine torque teno sus virus 1b (PTTSuV-1b), two parts of ORF3 fragments were amplified by PCR method from sera of the piglets association with post-weaning multisystem wasting syndrome in Fujian province. Then the target PCR products were cloned and sequenced. The results demonstrated that PTTSuV-1b ORF3 gene was 600 bp in length, coding an open reading frame (ORF) with 199 amino acids. The nucleotide sequence and amino acid sequence deduced from the gene were analyzed by the bioinformatics software. Compared with the PTTSuV-1b ORF3 genes download from GenBank, the sequenced gene shared the highest homogeneity with FJ/China/2010/TTV2/2 strain at 99.7%, with the SC strain at 94.0%, with the Spain PTTSuV-1b TTV2_G43 isolates at 97.3%. However, the PTTSuV-1b ORF3 genes shared with the torque teno sus virus K2 472142 stain isolated from Germany at 60.7%, with the Spain PTTSuV-1a PTTV1_1914 isolates at 46.8%. According to the results of the phylogenetic relationship constructed with Mega, PTTSuV-1b had two distinguish branches (branchⅠ and branch Ⅱ), the Fujian isolates were at the branchⅠ.
Research Progress of Fosl2 Gene
FAN Xing-xing, LI Xin-jian, GUO Ji-li, LI Gai-ying, LV Gang, REN Guang-zhi
2014, 41(8):  80-84. 
Abstract ( 354 )  
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Fosl2 (Fos-like antigen 2) that is expressed in a wide range of tissues during animal development belongs to the activator protein-1(AP1) transcription factor family that includes the various isoforms of Fos and Jun, which play diverse and important roles in animal growing development and immunologic response. Following the indepth research of recent years, people found that Fosl2 has a very close relationship with animal fat metabolism, skeleton development and cancer growth. This article reviewed the gene structure and the tissue distribution, also generalized the function of Fosl2 in animal fat metabolism, skeleton development, cancer growth and eye development. The prospects in study on the biological function were discussed in order to show a clearly direction for the deeper study and the animal growth traits promotion.
Preparation of Anti-calves Mycoplasma pneumonia Immunity Colostrum Whey
WANG Chao-li,YAN Gen-qiang,WANG Jing-mei,GAO Jia-deng,YANG Long-long,YANG Ming-wei
2014, 41(8):  85-90. 
Abstract ( 176 )  
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In order to discuss the preparation method of anti-calves Mycoplasma pneumonia immunity colostrum whey, this experiment used the cow Mycoplasma pneumonia deactivation adjuvant vaccine to be injected into the pre-natal 2 to 4 week cow two times to prepare hyperimmune colostrums. We collected the post-natal 12 h colostrum, using centrifugal precipitation and filtration to prepare skim milk separately, using the acid treatment method, enzyme hydrolysis method, enzyme and acid union method to prepare whey, through ultraviolet dispersion of light luminosity meter and ELISA method, we detected the whey total protein content and anti-cow Mycoplasma pneumonia IgG titer. The result indicated that 4 ℃,3500 r/min centrifugal 30 min was the best to the colostrum degreasing effect, the colostrum total protein content and anti-cow Mycoplasma pneumonia IgG titer losed few; the whey separation rate of enzyme and acid union method was the highest, in the vitamin C (0.2 g/mL) processing method the protein slightly losed to the degreasing colostrum, in the hydrochloric acid processing method the anti-cow Mycoplasma pneumonia IgG loss was smallest to the degreasing colostrum. Quality synthetic evaluation result was that 0.6 mol/L hydrochloric acid, pH 4.6, the room temperature 30 min processing, 10000 r/min centrifuge for 20 min was the appropriate method to prepare hyperimmune colostrum whey.
Preparation of Monoclonal Antibodies against Ofloxacin and Preliminary Development of its Enzyme Linked Immunosorbent Assay Kit for Rapid Detection
DONG Li-xue,FENG Cai-wei,FENG Jing,ZHENG Bai-qin,JIA Fang-fang,MENG Jun-li,HAO Li-wu,ZHANG Shu-hong
2014, 41(8):  90-94. 
Abstract ( 208 )  
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Ofloxacin was coupled with BSA as an immunogen and with OVA as a coating antigen using the coupling agent EDC. Monoclonal antibodies against ofloxacin and its enzyme linked immunosorbent assay kit were prepared and established,respectively. The limit of detection was 1 μg/kg in fish, and the recovery range was 62.8% to 97.7%. The intra and inter assay RSD were both less than 10%.
Effect of Cellulase or Formic Acid on Feeding Value of Silage Pineapple Leaves
LI Meng-chu,WANG Ding-fa,ZHOU Han-lin,ZHOU Lu-li
2014, 41(8):  95-100. 
Abstract ( 168 )  
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The research was designed to study the effect of cellulose or formic acid on feeding value of silage pineapple leaves.The treatment included the silage directly (control group),silage with 0.1 g/kg cellulose group,with 0.2 g/kg cellulose group,with 3.0 g/kg formic acid group,with 6.0 g/kg formic acid group.Nutrients content and rumen degradation of nutrients were analyzed after silage for 45 days.The results showed that compared with the control group,two groups of cellulose significantly increased the content of tannins (P<0.05),while the other contents of ingredients were not significantly changed (P>0.05).Addition of 3.0 g/kg formic acid significantly decreased the contents of ammoniacal nitrogen (NH3-N),Ca and P (P<0.05),but significantly increased the content of water soluble carbohydrates (WSC) (P<0.05).Addition of 6.0 g/kg formic acid significantly increased the contents of dry matter (DM) and WSC (P<0.05),but significantly decreased the contents of crude protein (CP),acid detergent fiber (ADF),neutral detergent fiber (NDF),NH3-N, Ca and P (P<0.05).Addition of 6.0 g/kg formic acid significantly increased the degradation rate of DM within 72 h (P<0.05),but had no significant effect on the degradation rates of CP and NDF at 72 h (P>0.05).In conclusion,under this experiment condition,adding 6.0 g/kg formic acid had the best quality of silage pineapple leaves,followed by the direct silage group.
Effect of Astragalus Polysaccharin on Growth Performance,Intestinal Microflora and Immune Function of Immunosuppression Broilers
SUN Bo,CHEN Jing,WU Juan,LIU Jiang,CHEN Hong-lan,LI Wei-ping
2014, 41(8):  101-106. 
Abstract ( 332 )  
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The aim of this study was to explore the effect of different levels of Astragalus polysaccharin on growth performance,intestinal microflora and immune function of immunosuppression broilers.A total of 180 fast large yellow feather chickens with similar weight were randomly divided into 4 groups,with 3 replicates of 15 chicks in each replicate,group Ⅰ was fed with basal diet,free drinking water,groupsⅡ,Ⅲ,Ⅳ were add 200 mg/L water soluble florfenicol in the drinking water,medication for 6 days,then group Ⅱ was fed a basal diet,groups Ⅲ,Ⅳ were fed with basal diet supplemented with 500 and 1000 mg/kg Astragalus polysaccharin,respectively.The pre-trial period lasted for 7 days,and the experiment lasted for 35 days.The results showed that compared with group Ⅱ,groups Ⅲ and Ⅳ was significantly improved the growth performance of immunosuppression broilers (P<0.05),enhanced the number of beneficial bacteria (Lactobacillus and Bifidobacterium) in the gastrointestinal tract of immunosuppression broilers (P<0.05),reduced the number of harmful bacteria (Escherichia coli and Salmonella) in the gastrointestinal tract of immunosuppression broilers (P<0.05),improved the immune organs index and Newcastle disease antibody titer of immunosuppression broilers (P<0.05),decreased the serum contents of IL-10 and IFN-γ of immunosuppression broilers (P<0.05).In conclusion,adding 500 and 1000 mg/kg Astragalus polysaccharin in diet could eliminate immunosuppression effect of florfenicol test,improve the growth performance,adjust the intestinal micro ecological balance and ennhance the immune function of broilers.
Research Progress on Aflatoxin and its Antidote in Animal Nutrition
WANG Zong-wei,YANG Jian-wei,HU Chun-liang,SUI Mei-xia,LIU Xi-ping,ZHANG Ru-lei
2014, 41(8):  107-111. 
Abstract ( 229 )  
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Aflatoxin was the most serious harmful biological toxicity in livestock feed.This paper reviews the aflatoxin metabolism in the animal body,toxicity for animal growth performance,antioxidant system,immune system and liver toxicity of aflatoxin,and it introduced physical adsorption class,chemical reactions class,biodegradation class and combination class of aflatoxin antidote including its effects.It provided a theoretical basis to research how to prevent aflatoxin contamination for the animal feed industry.
Effect of Adding Different Lactic Acid Bacteria Inoculants on Nutrient Values of Alfalfa Silage
LAI Yu-jiao,LUO Hai-ling,WANG Zhen-zhen,LU Xiao-nan,QU Yang-hua,GAO Wei-min,LIU Kun,YU Zhu
2014, 41(8):  111-116. 
Abstract ( 211 )  
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This experiment was conducted to investigate the influence of adding different lactic acid bacteria inoculants on nutrient values of alfalfa silage.The test took the alfalfa as material while adding Lactobacillus casei(A-LAB),Lactobacillus plantarum(B-LAB) and Pediococcus pentosaceus (C-LAB) at dose of 106 CFU/g fresh weight to ensile and set the control group,respectively.The cornell net carbohydrate and protein systems (CNCPS) was used to evaluate nutrient values of alfalfa silage after ensiling 45 days.The results showed that adding lactic acid bacteria inoculants could decrease the content of non-protein nitrogen (PA) compared with the control group. and the effect of B-LAB reached to the significant level (P<0.05).The content of intermediately degraded protein (PB2) and slowly degraded protein (PB3) of all the lactic acid bacteria inoculant groups were higher than the control group,but the difference was not significant (P>0.05).In the aspects of carbohydrate fractions,adding lactic acid bacteria inoculant increased the content of sugars (CA) and non-structural carbohydrate (CNSC) and decreased the content of available fiber (CB2),and the effect of A-LAB reached to the significant level (P<0.05).In conclusion,adding lactic acid bacteria inoculant could improve the nutrient values of alfalfa silage to certain extent by preserving the true protein and increasing the content of rapid degradation rates and CNSC of carbohydrate fractions,and Lactobacillus plantarum and Lactobacillus casei had the better effect.
Analysis of Amino Acids in Skeletal Muscles of 5 Different Livestock Breeds
LUO Wei,YANG Zhong,MENG Li-yun,ZHANG Wen-guang,SU Rui
2014, 41(8):  117-122. 
Abstract ( 225 )  
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The aim of this study was to compare the amino acid contents of skeletal muscles among Inner Mongolia cashmere goat, sheep, cattle, pig and chicken. The results revealed that the percentages of 7 essential amino acids were abundant, which were all over 35%. The contents varied among different livestocks and the order was pig, chicken, sheep, cashmere goat and cattle. As the palatable taste amino acid, the percentage of Glu and Asn in cashmere goat and sheep were the highest in all the breeds. This might be the reason why the mutton was palatable. The coefficient of variation (CV) of amino acid revealed that the CV of Asp was lowest in all 17 amino acids. So the content of Asp could be used to calculate the contents of essential amino acid, palatable taste amino acid and total amino acid. The results of cluster analysis showed that the contents of amino acids in 17 samples from cashmere goats were clustered together with all the samples from sheep, cattle and chicken, the pig was clustered in another group. It suggested that the amino acid patterns were different between ruminant and simple stomach animals. This study supplied important evidences for the research of skeletal muscles protein and was meaningful for revealing the mechanism of the formation of meat characteristics.
Functions and Approaches of △3-△2-Enoyl-CoA Isomerase on Metabolism of Unsaturated Fatty Acid
LU Yun-feng,SHI Jian-zhou,PANG Zhen-ling,WU Ke-liang,ZHANG Hao
2014, 41(8):  123-126. 
Abstract ( 392 )  
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Study on Purple Corn Anthocyanin Inhibits Replication of Infectious Bovine Rhinotracheitis Virus on Bovine Kidney Cells
JIANG Liang, HUANG Xiu-fen, LIU Shan-shan, YAN Min-xin, ZHAO Xiao-yan, ZHANG Lin-bo, LI Yong-qing
2014, 41(8):  127-130. 
Abstract ( 176 )  
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To investigate the effect of purple corn anthocyanins (PCA) on inhibiting infectious bovine rhinotracheitis virus (IBRV) in vitro, the toxicity of PCA on madin-darby bovine kidney cells (MDBK) was measured by observation of cytopathic effect (CPE).Then the inhibition effects of PCA on replication of IBRV were investigated within safe concentrations by MTT assay. The results showed the maximum safe concentration of PCA on MDBK cell was 262.14 μg/mL, whereas PCA could apparently inhibit the replication of IBRV on MDBK cells in very low dose (7.8125 μg/mL). The median inhibitory concentration (TC50 and IC50) of PCA to MDBK and IBRV was 262.14 and 11.38 μg/mL,individually. On consequence, the treatment index (TI) was calculated as 23.04. These data demonstrated that PCA had the function of anti-IBRV, and suggested that purple corn could be as a feed material for prophylaxis and treatment of infectious bovine rhinotracheitis in dairy cows.
Effect of Astragalus Polysaccharides on Serum Biochemical Indexes of Lactating Holstein Cows
WEN Yue-ling,SHEN Yi-jun,ZHOU Jin-wei,WANG Bin,SHEN Liu-hong,YU Shu-min,CAO Sui-zhong
2014, 41(8):  131-135. 
Abstract ( 239 )  
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This experiment was aimed to explore the effect of Astragalus polysaccharides (APS) on serum biochemical indexes of lactating Holstein cows.According to the similar age,parity and weight,40 Holstein cows in lactation period were selected and randomly divided into four groups with 10 cows in each replicate in a single-factor completely randomized block design.Group Ⅰ was the control group which fed with basal diet without APS,groups Ⅱ,Ⅲ and Ⅳ were fed basal diet supplement with 10,50 and 100 g/(cow·d) APS,respectively.The experiment lasted for 20 days.The results showed that with the increase of the APS level,the serum activities of ALT,AST,γ-GGT,ALP,CK and the serum contents of TBIL,DBIL,BUN,CRE,CHO,TP,ALB and GLU in groups Ⅱ,Ⅲ,Ⅳ were no significant differences compared with the control group (P>0.05).With the increase of the APS level,the serum content of TG in groups Ⅱ,Ⅲ,Ⅳ had a decreased tendency,and 10 days after using medicine,it was significantly decreased compared with the control group (P<0.05).In conclusion,feeding the cows with recommendatory dosage of APS in the actual production condition was safe and reliable.10 times as the recommendatory dosage of APS was also safe,and APS was able to regard as a security antioxidant and immunopotentiator in cow breeding.
Separation of Active Ingredients in Grape Procyanidins and its Effects on Proliferation of Peripheral Blood Lymphocytes in Broiler
YANG Jin-yu,WANG Jing,ZHANG Hai-jun,WU Shu-geng,YUE Hong-yuan,QI Guang-hai
2014, 41(8):  136-140. 
Abstract ( 192 )  
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This experiment was conducted to study the effect of grape procyandins (GPC) and its ingredients on the proliferation of peripheral blood lymphocytes (PBLs)of broiler,and to develop the method to screen active ingredients of GPC.Six concentrations (0,0.24,1.2,6.0,30,150 μg/mL) of GPC were chosen to study the best concentration on the proliferation of PBLs of chickens.RT-HPLC was used to gotten chromatograms of GPC,different ingredients were gotten by Real-time fractionation and different ingredients in the best concentration were used to get the best one.The result showed that the best concentration of GPC was 30 μg/mL and the best ingredient fraction was 16 to 25 min.
Research Progress on Insulin-mimetic Effects of Selenium
JI Feng,HUANG Jian-guo,WANG Ya-lei,MA Chun-jian,SU Qi
2014, 41(8):  140-144. 
Abstract ( 159 )  
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During recent years,it has been found that selenium (Se) has insulin-mimetic effects on the promotion of glucose uptake into cells,glucose metabolism and signal transduction based on the relationship of Se and diabetes, but the effect dose of Se can only be obtained with nearly toxic doses and are not suitable for humans.This paper gives a brief review of research progress on insulin-mimetic effects of Se,in favor of an extensive study on the regulation mechanism of Se in glucose metabolism,so as to provide a reference for applying Se for regulation of disorders of glucose metabolism in stressed animals.
Isolation and Identification of Lactic Acid from Bovine Rumen
YANG Long-long,JIANG Jian-jun,YAN Gen-qiang,WANG Jing-mei,GAO Jia-deng,WANG Chao-li,YANG Ming-wei
2014, 41(8):  145-148. 
Abstract ( 191 )  
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To screen and separate viable preparations for bovine strains of lactic acid bacteria,this separated lactic acid bacteria from bovine rumen were identified by API 50CHL.For biochemical identification of strains by resistance for oxygen,acid and alkali,determination of the growth temperature, drug resistance and antibacterial property.The results showed that the three strains of lactic acid bacteria have been identified,which were L. acidophilus (L1), L. fermentum (L2) and L. holzapfelii (L3), respectively. L1 had strongest acid,L2 had strongest alkali,L1 was strong in 37 to 40 ℃ were grown between good and facultative anaerobic,the drug resistance rate of L1,L2 and L3 was 60% to 100%.The three strains compare with the Escherichia coli and Staphylococcus showed their inhibition,the L1 had the best bacteriostatic effect,the antibacterial diameter of 18.30 and 16.40 mm,respectively.In conclusion,L1 and L2 will be a candidate viable lactic acid bacteria strains resistant formulations.
Study on Hyperkalemia and Hypercalcemia in Dogs and their Therapeutic Mechanism
ZHANG Yi
2014, 41(8):  149-154. 
Abstract ( 305 )  
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The assay was aimed to explore the effects of Ca2+ or K+ on dog cardiac function, and the role of Ca2+ and/or K+ on treating canine electrolyte disorders. 20 Chinese rural dogs at the age of 24 to 48 months were divided into two groups by age, gender and weight. Hyperkalemia (group Ⅰ) model was created by intravenous injection with 5% KCl at flow velocity of 1.5 mL/min, and then relieved the symptoms of hyperkalemia by intravenous injection with CaCl2 at flow velocity of 1 mL/min; hypercalcemia (group Ⅱ) model was created by intravenous injection with 5% CaCl2 at flow velocity of 1 mL/min, and then relieved the symptoms of hypercalcemia by intravenous injection with KCl at flow velocity of 1.5 mL/min.The blood was collected before and after the models built, and at the time of CaCl2 (KCl) was injected to the models.The sera were separated and used to detect the concentrations of K+, Ca2+, Na+, Cl-. Simultaneously, blood pressure, heart rates and electrocardiogram (ECG) were monitored at the same collecting points. The results showed that hyperkalemia model was established by intravenous injection with 5% KCl to the dog at flow velocity of 1.5 mL/min.At this time the serum concentration of K+ was significantly increased (P<0.05),heart rate extremely significantly decreased (P<0.01),SBP significantly decreased (P<0.05). The ECG showed that the model dogs were arrhythmia, P wave became flat or even disappeared, T wave was tall as mantle.While CaCl2 was injected to the hyperkalemia model by intravenous injection, the concentration of K+ was significantly decreased (P<0.05),the heart rate significantly increased (P<0.05), The ECG showed that P wave reappeared,mantle-like T wave disappeared.Hypercalcemia model was established by intravenous injection with 5% CaCl2 to the dog at flow velocity of 1 mL/min.At this time the serum concentration of Ca2+ was extremely significantly increased (P<0.01), the concentration of Na+ decreased (P<0.05),the heart rate significantly increased (P<0.05),pulse pressure extremely significantly decreased(P<0.01). The ECG showed that T wave was widened,ST segment was shortened or even disappeared,QT interval was shortened,U wave was appeared.While KCl was injected to the model dogs by intravenous injection, the serum concentration of Ca2+ decreased (P>0.05), the heart rate significantly decreased (P<0.05) and returned to normal, but pulse pressure continued to decline.The ECG showed that T wave was apparently elevated.The time limit of ST segment increased.U wave was disappeared. The conclusions were that hyperkalemia (hypercalcemia) model could established after 5% KCl (5% CaCl2) injected to the healthy dogs. The heart fuction recovered obviously when CaCl2 was injected to the hyperkalemia dogs, therefore, CaCl2 had a better therapeutic effect for hyperkalemia. However, it had no good therapeutic effect when KCl injected to the hypercalcemia dogs.
Expression Changes of Relative Factors in Mice with Aristolochic Acid Nephropathy
TIAN Ya-hui, CHANG Di, YANG Yao-hui, WU Guo-ying, ZHAO Hai-jiao, WU Jin-dong, WU Guo-juan, ZHANG Zhong-wen
2014, 41(8):  155-160. 
Abstract ( 159 )  
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The study was aimed to observe the expression changes of relative factors in mice with aristolochic acid nephropathy (AAN). 30 mice were given aristolochic acid Ⅰ (AAⅠ) at 8 mg/(kg·d), and mice were killed at a different time. Then observe the renal pathological changes by hematoxylin-eosin (H&E), Real-time PCR and Western blotting technology were used to detect the expression changes of α-SMA, vimentin and CK-18. The results showed that with the extension of treatment time, we could found the kidney structure was damaged more seriously, and confirmed that AAⅠcould induce epithelial-mesenchymal transition (EMT) in renal, promoted the expression of α-SMA and vimentin, inhibited the expression of CK-18. The study laid a foundation for a deeper mechanism study of AAN.
Research Progress on Toll Like Receptors Mediated Signaling Transduction Pathway
MA Si-hui,YANG Huan,WU Tian-cheng,CUI Huan-zhong,ZHANG Hui,LI Yu-meng,HONG Pan,ZHENG Xin
2014, 41(8):  160-164. 
Abstract ( 177 )  
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Toll like receptors (TLRs) can recognize pathogen-associated molecular patterns and raise the adaptor protein molecules containing toll IL-1 receptor (TIR) domain.Moreover,TLRs can initiate specific immune response through activating signal transduction by the MyD88-dependent pathway or TRIF-dependent pathways.This article summarizes the latest research review on structure features,distribution,ligand recognition,adaptor protein molecules participating in signal transduction and signal transduction pathway of TLRs.
Effects of Oral Echinacea purpurea on Heart Rate,ECG, Blood Pressure and Respiration of Awake Beagle Dogs
PANG Yun-lu,ZHONG Ying-jie,FU Hai-ning,ZHONG Yan,JIANG Yi-hai
2014, 41(8):  165-169. 
Abstract ( 180 )  
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The experiment studied the effects of oral Echinacea purpurea on heart rate, ECG, blood pressure and respiration of awake Beagle dogs. Experimental results showed that oral Echinacea purpurea 0.125 to 1.25 g/kg could apparently reduced the heart rate of awake Beagle dogs, and had no significant effects on blood pressure, respiration and ECG between P wave, T wave, R wave, QT interval, PR interval as well as QRS duration (P>0.05). It showed that Echinacea purpurea had no apparent effects on the dog’s respiratory system and cardiovascular system, which was safe and reliable in application doses.
Study on Quality Standard of Jin Gen Qin Lian Powder
REN Li-hua, XIN Rui-hua, CHENG Long,LUO Yong-jiang, WANG Gui-bo, LUO Chao-ying, XIE Jia-sheng, LI Jin-yu, ZHENG Ji-fang
2014, 41(8):  169-172. 
Abstract ( 157 )  
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The assay was aimed to establish the thin layer chromatography (TLC) quality standard for Jin Gen Qin Lian powder. In this experiment TLC was used for qualitative identification of Chinese medicinal herbs in the components,and ultimately determined the TLC for Flos Lonicerae, Radix Scutellariae, Fructus Forsythiae and Radix Bupleuri.Under this selected chromatographic conditions, good chromatographic spots were separated. This method could effectively identify Flos Lonicerae, Radix Scutellariae, Fructus Forsythiae and Radix Bupleuri in the compound. The method established in this experiment was simple and focused with good reproducibility, so it could be used to provide a reliable basis for control the quality of compound.
Effects of Freezing Methods for Dorper Sheep Frozen Semen Quality
WANG Xin-qi, SU He, DA Lai, YANG Yan-yan, CAO Gui-fang
2014, 41(8):  173-178. 
Abstract ( 152 )  
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The sheep semen were freezing by liquid nitrogen vapour method and apparatus freezing frozen method respectively, measuring thawed semen by sperm motility, acrosome integrity, membrane integrity and aspartate aminotransferase and lactate dehydrogenase activity to compare effect of different methods of freezing semen quality after thawing;measuring enzyme activity after semen dilution, equilibration, apparatus freezing frozen method to compare the release amount of aspartate aminotransferase and lactate dehydrogenase at different stages. The results showed that the sperm motility and membrane integrity after apparatus freezing frozen method were highly significant higher than the liquid nitrogen fumigation freezing method (P<0.01);aspartate aminotransferase and lactate dehydrogenase activity after apparatus freezing frozen method were highly significant lower than that of liquid nitrogen fumigation method (P<0.01). Aspartate aminotransferase and lactate dehydrogenase activity of semen diluted were highly significant lower than the activity of frozen semen (P<0.01). The results showed that the sheep semen quality by apparatus freezing frozen method was better than the liquid nitrogen fumigation freezing method. For sperm, the release of aspartate aminotransferase was in the equilibrium phase, and the release of lactate dehydrogenase was in the freezer stage.
Expression Analysis of Prolificacy-related ESR Gene during Synchronized Estrus Treatment for Small Tail Han Sheep
GONG Ping, ZANG Chang-jiang,WU Wei-wei, HANIKEZI, YU Li-juan, XU Xin-ming, TIAN Ke-chuan
2014, 41(8):  178-182. 
Abstract ( 221 )  
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This study was aimed to explore the underlying reason of low implantation rate after embryo transplantation. Exogenous hormones and CIDR were used to treat Small Tail Han sheep, the estrogen receptor (ESR) gene expression in ovarian cells derived from estrus synchronization treatment (EST),natural estrus control (NEC) and anestrus (ANE) Small Tail Han sheep were compared, and the methylation levels of CpG island was analyzed. The results showed that there was no significantly difference in estrus performance and time between EST and NET sheep, and the transcript levels of ESR were also not significant difference (P>0.05), but they were significantly higher than that in ANE sheep ovarian (P<0.05). Furthermore, though the expression amount of ESR at protein level was greatly higher in EST sheep ovarian compared to that ANE sheep (P<0.05), it was significantly lower than that in NEC sheep (P<0.05). Methylation levels at the CpG island of ESR promoter was 0 and 10.77% in NEC and EST sheep, respectively; while the promoter was hypermethylation in ANE sheep(92.30%).These results suggested that EST sheep showed similar estrus performance and time to that of NEC sheep after treated with exogenous hormones, but ESR gene expression at protein level was significantly different,and the methylation level of promoter was also significant difference. Perhaps it was one of the main causes resulting in the low implantation rate after embryo transplantation of Small Tail Han sheep.
Study on FABP Transgenic Mice about the Effects of Growth and Development
FAN Han-lu,DING Xiang-bin,LIU Xin-feng,LIU Hai-xue,GUO Hong
2014, 41(8):  183-188. 
Abstract ( 153 )  
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In the research, tissue biopsies, fourier infrared spectrum, and the apparent growth index were used to test and analyse hearts,livers, skeletal muscle, uterus, ovarians of the first and the second filial generation FABP transgenic mice. The results showed that there was no significant difference between FABP transgenic mice and wild-type mice and genetic stability.
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YUAN Yu-guo,PENG Qiu-ling
2014, 41(8):  188-192. 
Abstract ( 184 )  
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Zinc-finger nucleases (ZFNs) are engineered site-specific DNA cleavage enzymes that may be designed to recognize long target sites and thus cut DNA with high specificity. ZFNs mediate permanent and targeted genetic alteration via induction of a double-strand break at a specific genomic site. Compared to conventional homology-based gene targeting, ZFNs can increase the targeting rate by up to 100000-fold; gene disruption via mutagenic DNA repair is similarly efficient. The utility of ZFNs has been shown in many organisms, including insects, amphibians, plants, nematodes, and several mammals, including humans. This broad range of tractable species renders ZFNs a useful tool for improving the understanding of complex physiological systems, to produce transgenic animals, cell lines, and plants, and to treat human disease. This review summarized recent advances in the understanding of role and function of ZFNs and described the utility of ZFNs for mammalian transgenesis.
Determination and Correlation Analysis on Body Size Traits and Slaughter Performance of Kirin Chicken (Frizzle Chicken)
LI Nai-bin, YANG Fen-xia, DU Bing-wang, TAO Lin, CHEN Jie-bo, LI Dong-hua,YE Wei-qing,ZHU Zi-jing
2014, 41(8):  193-197. 
Abstract ( 301 )  
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In order to study the meat development value of anti-heat stress species-Kirin chicken (Frizzle chicken), 12-week-old Frizzle chicken was used to measure their body size traits and slaughter performance indexes and then studied these data with correlation and regression analysis. The results showed that keel length (12.05 cm) and shank length (8.70 cm) of male 12-week-old Frizzle chickens were significantly higher than those of female chickens (10.17, 7.47 cm)(P<0.05). The live weight, carcass weight, semi-eviscerated weight, eviscerated weight and the leg muscle of male 12-week-old Frizzle chickens were significantly higher than those of female chickens (P<0.05), abdominal fat weight (27.22 g) and sebum thick (4.38 mm) were significantly lower than those of female chickens (35.33 g, 5.12 mm)(P<0.05),other indicators were no significant differences (P>0.05). There were highly significant or very highly significant correlations between the slaughter performance and body size in Frizzle chicken. The regression equation of related indicators was identified. This conclusion provided a certain theoretical reference to predict slaughter performance indicators through a living body size measurements and contributed to the genetic breeding process of Frizzle chicken.
Effects of Different Parities on the Reproductive Performance of Duroc, Landrace and Yorkshire Sows
ZHU Shi-ping, SUN Li, YIN Xue-mei, HOU Xiao-liang, BAO Wen-bin, WU Sheng-long
2014, 41(8):  197-200. 
Abstract ( 260 )  
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To investigate the effect of parities on the reproductive performance of sows, the production records of Duroc (128), Landrace (681) and Yorkshire (610) sows were collected in a national breeding farm in Jiangsu province from 2009 to 2011, then analyzed the total number born, number born alive, litter size at weaning, newborn litter weight and corrected litter weight at 21 days of different parities. Besides, the sows were divided into three stages (1st to 2nd parities, 3rd to 6th parities and 7th to 8th parities) to explore the effects of different breeds on the reproductive performance according to the parities. The results showed that the parities have significant effect on the reproductive performance (P<0.05). The total number born, number born alive, litter size at weaning and newborn litter weight of sows were improved gradually from 1st to 4th parity, the peak of which was at the 4th parity, and the reproductive performance of the 4th parity was significantly higher than other parities (P<0.05). The reproductive performance of sows from 3rd to 6th was preferable, and then it decreased rapidly after the 7th parity. The corrected litter weight at 21 days of each parity (except for the 4th parity) presented an increasing trend. The breeds also had significant effect on the reproductive performance (P<0.01). The reproductive performance of Landrace and Yorkshire sows was significantly higher than that of Duroc sows (P<0.01). Landrace sows was significantly higher than Yorkshire sows in total number born and corrected litter weight at 21 days (P<0.05). In summary, the reproductive performance of 3rd to 6th parities was the best in all three breeds; The reproductive performance of Landrace was superior to Yorkshire in general, so Landrace sows might be more suitable for female parent in the production of two-breed cross sows to this breeding farm.
Histological Observation of Development of Skin and Hair Follicle in Neonatal Mink
WANG Zhuo, LI Guang-yu, YANG Ya-han, ZHANG Ting, HUANG Jian, ZHOU Ning, SUN Wei-li
2014, 41(8):  201-205. 
Abstract ( 185 )  
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The study investigated the structure and development traits of skin and hair follicle of 0-week-old to 6-week-old neonatal male minks. Skin samples were obtained from central back of minks. Tissue sections were prepared by using HE staining method. The results showed that the guard hair and underfur of minks had hair medulla; mink was born without the secondary follicles, but the structure of the primary follicles had formed. The density of primary follicles firstly increased and decreased at 4-week-old with the increase of week age. Primary hair follicles no longer developed or rarely developed after 4-week-old. At 4-week-old, some of the secondary follicles had formed. The density of the secondary follicles increased with the increase of week age. Secondary hair follicles and primary hair follicles average ratio (S/P) was 2.95±0.23. Secondary hair follicles still developed after 6-week-old. The epidermises of neonatal minks were firstly thicken, and then thinning. The dermal of the minks thickened and the primary and secondary follicles deepen with the increase of week age.
Effects of Lipopolysaccharide on Antiviral Immune Response Induced by Porcine Reproductive and Respiratory Syndrome Virus
WANG Dong-mei, FENG Ya-qiong, CUI Chun-xiao, LI Na-na, DU Dong-ying, QIN Yun-jie, FU Peng-fei, XIA Ping-an
2014, 41(8):  206-210. 
Abstract ( 181 )  
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To study the effects of lipopolyscaccharide on antiviral immune response induced by porcine reproductive and respiratory syndrome virus (PRRSV), porcine alveolar macrophage (PAM) cell was prepared and cultured with PRRSV solution 100 TCID50. After inoculated 12 h, cells were treated with 100 ng/mL LPS. Then collected cells and supernatants after treated 12, 24, 36 and 48 h, respectively. The level of the virus, IFN-α and TNF-α mRNA in the PAM cells were quantitated by Real-time qPCR including lipopolysaccharide (LPS) stimulation of PRRSV infected PAM cells group, PRRSV infection control group, LPS stimulated cells control group and cell control group. The results showed that compared with unstimulated with LPS, the replication of PRRSV in PAM cells infected with PRRSV stimulated with LPS group was less than that of PRRSV infection control group from 12 to 48 h. The transcription level of IFN-α mRNA improved significantly (P<0.05) and the transcription level of TNF-α mRNA improved and reduced at 48 h. Compared with LPS stimulation control group, LPS stimulation of PRRSV infected PAM cells group, IFN-α mRNA transcription level improved at 12 h and reduced at 24, 36 and 48 h. So PRRSV infected PAM cells stimulated with LPS, the transcription of IFN-α, TNF-α mRNA was significantly improved (P<0.05), thereby inhibiting the replication of PRRSV in PAM cells.
Research Progress on Avian Coccidiosis Disease
LIN Yu-xin,SUN Ming-ming,DAI Guo-jun,ZHANG Jing-jing
2014, 41(8):  210-214. 
Abstract ( 185 )  
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It had became a trend that the research of disease-resistant breeding which were to reduce disease losses, improve the quality of livestock and poultry products, implement pollution-free production with the development of molecular biology and genetic engineering. The article summarized on the current status of the prevention of avian coccidiosis, the research and development of coccidiosis prevention of drugs and vaccines and especially on the progress of coccidiosis-resistant breeding. Although the major prevention of coccidiosis was still drugs and vaccines currently, in the long run, nurturing the new disease-resistant varieties or new lines of chicken by coccidiosis-resistant breeding would become the new way for disease prevention and control.
Isolation, Identification and Phylogenetic Analysis of Proteus mirabilis from Goats with Diarrhea
CHEN Xi, WANG Yuan-wei, WANG Yong, TANG Cheng
2014, 41(8):  215-218. 
Abstract ( 269 )  
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Proteus mirabilis were identified from goat manure samples with diarrhea from a goat farm in Chengdu of Sichuan province (14/20). 5 isolates (5×108 CFU each) were randomly selected from 17 Proteus mirabilis and infected into mice by intraperitoneal injection. The results showed that all of the injected mice were dead within 12 h. 16S rRNA genes of Proteus mirabilis isolates were sequenced and analyzed. The genes shared 99% to 100% homology with the sequence of Proteus mirabilis available in GenBank, and shared a homology of 99.8% to 100% within isolates in this study. Phylogenetic analysis showed that isolates clustered into two branches and presented some diversity, but no significant correlation of evolution between different areas and hosts. In conclusion, Proteus mirabilis was closely involved in goat diarrhea. Its harm to the sheep industry and public health significance should be seriously concerned.
Antibacterial Activity of Endophytic Fungi from Podocarpus macrophyllus
RAN Xue-qin, HE Ying-xia, SUN Duan-fang, YE Jing, WANG Jia-fu
2014, 41(8):  219-223. 
Abstract ( 206 )  
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In order to study whether the secondary metabolites of endophytic fungi had antibacterial activity or not, 96 endophytes were isolated from P.macrophyllus in Guizhou province. The antibacterial activity of chloroform/methanol extracts of fermentation broth for 96 fungi was determined by cylinder plate method. 49% endophytic fungi from P.macrophyllus exhibited antibacterial activities against at least one bacterium among three pathogenic bacteria, Escherichia coli, Bacillus subtilis or Staphylococcus aureus. Out of the fungal endophytes isolated from P.macrophyllus, the extracts with 100 folds dilution of strain LHS18, LHS42, LHS67, LHS92 and LHS93 still displayed strong inhibitory activities against S.aureus and strain LHS30 against E.coli. Two strains, LHS18 and LHS42, were further identified to be Aspergillus fumigatus, LHS92 to be Neurospora tetrasperma, LHS30 to be Aphanocladium sp., LHS67 to be Phomopsis liquidambari, respectively. LHS93 was not identified for the low identity of internal transcribed spacer (ITS) sequence between rRNA genes with the known sequences of fungi in Nucleotide database. It suggested that 49% of the endophytic fungi isolated from P.macrophyllus showed potent antibacterial activity, which could be a resource for new antibiotics in future.
Isolation, Identification and Antibiotics Susceptibility of Mannheimia haemolytica from Sheep Lungs
FENG Xu-fei, DAO Xiao-fang, WANG Zhi-min, TANG Cheng, LI Ding-fei,YANG Fa-long
2014, 41(8):  224-228. 
Abstract ( 274 )  
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The aim of this study was to isolate the etiological bacteria of sheep pneumoniae and evaluate the antibiotic susceptibility of the isolates. Bacteria was isolated from lungs of sick sheep, and further identified with bio-chemical tests and PCR method, the antibiotic susceptibility of the isolates was determined. The results showed that the isolates were gram negative, organisms with haemolytica. Bio-chemical and PCR method proved that the isolates were Mannheimia haemolytica. The isolates were susceptible to most of the tested antibiotics such as enrofloxacin, gentamicin and tetracycline. The results obtained in this study provided useful information for treatment of Mannheimia haemolytica infection.
Study on Bacteriostatic Action of Sodium Humate on Escherichia coli
Narengaowa,AN Xiao-ping,QI Jing-wei,LUO Xu-guang,WU Lan,YU Chang-qing,TONG Bao-sheng
2014, 41(8):  228-231. 
Abstract ( 181 )  
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The role of sodium humate on Escherichia coli was investigated to provide potential novel antibacterial drugs for treating disease which caused by enteropathogenic bacteria.Antimicrobial activity for sodium humate on Escherichia coli was determined using dilution plate counting method.Ultrastructure of sodium humate against Escherichia coli was observed under transmission electron microscope.The results showed that adding sodium humate (5%,10%,15%) decreased the number of Escherichia coliP<0.05).The more obvious inhibition effect (P<0.05) as the level of sodium humate content increased.Different strains in the same processing conditions,with the extension of time,the more obvious inhibitory effect.Observation under transmission electron microscope was carried out that adding 5% sodium humate and adding 5% sodium humate with pH regulating changed the permeability of cell membrane, made cell excrescent and inhibited cell division. The results provided a theoretical basis to further understand sterilization mechanism of sodium humate.
Resistance Survey of Escherichia coli Isolates to Antimicrobial Drugs from a Cattle Farm in Ili
NAN Hai-chen, GUO Qing-yong, XIA Li-ning, WANG Jin, DI Li-na, GAO Juan
2014, 41(8):  232-236. 
Abstract ( 218 )  
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In order to investigate Escherichia coli isolates from a cattle farm to commonly used antimicrobial drugs in Ili, Xinjiang, minimal inhibitory concentrations (MICs) of the antimicrobial drugs to these isolates from drinking water, feed and fecal samples were determined by broth micro-dilution method. The results showed that 25 Escherichia coli isolates were confirmed from 25 drinking water samples (100.0%), the resistance rates to amoxicillin/clavulanic acid, ampicillin, norfloxacin, enrofloxacin and apramycin were 12.0%, 4.0%, 4.0%, 8.0% and 8.0%, respectively. 47 Escherichia coli isolates were confirmed from 72 feed samples (65.3%), the resistance rates to amoxicillin/clavulanic acid, ampicillin, norfloxacin and apramycin were 36.2%, 19.1%, 4.3% and 4.3%, respectively. 80 Escherichia coli isolates were confirmed from 80 fecal samples (100.0%), the resistance rates to amikacin, ampicillin, enrofloxacin, gentamicin, norfloxacin, ciprofloxacin, amoxicillin/clavulanic acid and ceftiofur were 12.5%, 7.5%, 7.5%, 5.0%, 2.5%, 2.5%, 1.3% and 1.3%, respectively, only to apramycin sensitive. Resistances of Escherichia coli isolates from the cattle were general to the antimicrobial drugs, but were higher rate of intermediary to some antimicrobial drugs. So treatment of the diseases caused by bacteria should avoid using antimicrobial drugs of non-sensitivity and high intermediary rate. Drinking water and feed in the cattle farm existed the risk of being contaminated by resistant Escherichia coli.
Research Progress on Avian Influenza A Virus Subtype H10N8
TAN Wei, XIE Zhi-xun
2014, 41(8):  236-241. 
Abstract ( 213 )  
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It was found for the first time at the end of 2013 in China that avian influenza A virus subtype H10N8 could infect human beings and result in death. No such a case has been reported in other parts of the world. Similarly to H7N9, H10N8 AIV is less pathogenic in poultry. However, when an H10N8 virus infects a human being, the virus can become highly pathogenic. A person infected with an H10N8 AIV deteriorated quickly and the mortality rate is high, and H10N8 infection poses a great threat to human health. At present there are no effective means to prevent and control H10N8 AIV infection. It is currently not clear how an H10N8 AIV is transmitted to humans. There are great concerns in public whether an H10N8 AIV will cause another outbreak in humans. This article will briefly describe the characteristics of H10N8 subtype viruses and the current research on these viruses.
Comparative Study on Effect of Bacteriostasis and Antivirus of Lactobacillus amylovorus Metabolites
ZHAO Yi-bo,TIAN Zong-min,WANG Ying-ying,WEI Ping
2014, 41(8):  242-246. 
Abstract ( 174 )  
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The aim of this research was to study the mechanisms of Lactobacillus amylovorus metabolites on bacteriostasis and antiviral activity,and identify the active ingredients of bacterial metabolites.The supernatants were treated by acids,hydrogen peroxide and protease.The agar well diffusion system was performed for detecting the bacteriostasis activity of the supernatants of Lactobacillus amylovorus against Escherichia coliSalmonella and Staphylococcus aureus.Then evaluated the anti-TGEV activity on the IPEC-J2 cells with MTT.The results showed that the bacteriostasis of the supernatants of Lactobacillus amylovorus reduced significantly by treating with hydrogen peroxide,trypsin and pepsin (P<0.05),but antiviral activity was not reduced significantly (P>0.05).However,the bacteriostasis and antiviral activity of the supernatants of Lactobacillus amylovorus reduced significantly by treating with acids and protease K (P<0.05),it was found that the bacteriostasis and antiviral activity of Lactobacillus amylovorus metabolites possess consistency and the component was acids or protease K-sensitive proteins.
Study on Isolation and Identification of Mountain Tai Bacillus subtilis and its Degradation of Aflatoxin B1
SUN Ling-yu,LI Chao,HAO Hai-yu,CHAI Tong-jie
2014, 41(8):  246-250. 
Abstract ( 229 )  
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The aim of this experiment was to select the Bacillus subtilis which was useful for the degradation of aflatoxin B1 from the soil of Mountain Tai.Through the 16S rRNA gene sequencing,we identified one Bacillus subtilis which had high degradation effect of aflatoxin B1,named as Mountain Tai Bacillus subtilis,the determination of degradation efficiency and active components of aflatoxin B1 and the effect of degradation aflatoxin in feed were researched in this experiment used Mountain Tai Bacillus subtilis.The results showed that the degradation rate of aflatoxin B1 of Mountain Tai Bacillus subtilis was 90.28%,the degrading efficiencies of aflatoxin B1 by exocellular coarse extraction liquid was highest,the activity of aflatoxins degradation mainly happened in the culture supernatant of Mountain Tai Bacillus subtilis.Characteristic research of active component including heating and degeneration of proteinase K,the detoxification decreased significantly,which suggested that the effect of supernatant from Mountain Tai Bacillus subtilis on aflatoxin B1 might be due to the enzymatic reactions.Further application tests proved that Mountain Tai Bacillus subtilis could degrade aflatoxin B1 in mouldy feed and it could be applied in the feed or food industry.
Comparative Proteomic Analysis of Escherichia coli Protein during Growth Phase
ZHAO Xiao-wei,YANG Yong-xin,CHENG Guang-long,HUANG Dong-wei,ZHAO Hui-ling
2014, 41(8):  251-254. 
Abstract ( 215 )  
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To investigate the differentially expressed proteins of Escherichia coli from mastitic cow during growth phase,two-dimensional electrophoresis coupled with mass spectrometry method were used to separate and identify the changes of proteins in the lag phase,exponential phase and stationary phase of Escherichia coli.The results showed that some protein compositions of Escherichia coli were changed in lag phase,exponential phase and stationary phase.Relative abundance of outer membrane protein W and phosphoenolpyruvate carboxykinase in Escherichia coli were increased,while periplasmic protein,outer membrane protein Ⅱ and phosphotransacetylase were decreased in the lag phase comparative with exponential phase and stationary phase.Results of this study help provide the relationships between proteins expression and the growth of this pathogen,and were used to control and protect the host against the pathogen.