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Table of Content

20 April 2011, Volume 42 Issue 4
综述
Multidrug Efflux Systems Involved in Resistance to Antibacterial Drugs
WANG Chun-mei;HE Qi-gai;CAO Ji-yue
2011, 42(4):  455-467.  doi:
Abstract ( 942 )   PDF (118KB) ( 385 )  
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Bacteria have developed various ways to resist the toxic effects of antibiotics and other drugs, such as produce enzymes that inactivate antibiotics, target alteration by mutation or enzymatic modi?cation, inhibit drug entry into the cell and active ef?ux of drugs. Actively exporting an antibiotic from the bacterial cell by the efflux pump is essential to ensure signi?cant levels of drug resistance. The multidrug efflux pumps that affect such efflux typically export several unrelated substances, including antibiotics and molecules produced by the host organism. There are different kinds of multidrug efflux pumps in gram-negative bacteria and that in gram-positive bacteria as the different structure of cell membrane. In gram-negative bacteria, it has been reported five superfamilies, these include ATP-binding cassette family (ABC), resistance nodulation division(RND) family, major facilitator superfamily (MFS), small multidrug resistanc (SMR) family and multidrug and toxic-compound extrusion family (MATE). In gram-positive bacteria, it has reported four superfamilies, these include ABC, MFS, SMR family and MATE, except RND family. Facing the resistance of bacteria to antibiotics, it is urgent to find new molecules mechanisms against resistant bacteria. Some researchers have been searching for the efflux pumps inhibitors (e.g., MexAB-OprM specific efflux pump inhibitor) for several years, and some possible compounds have been identified from medicinal plant to restore antibiotic activity.
Multideug Efflux Systems Involved in Resistance to Antibacterial Drugs
WANG Chun-mei,HE Qi-gai,CAO Ji-yue
2011, 42(4):  455-467.  doi:
Abstract ( 121 )   PDF (1079KB) ( 1214 )  
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遗传繁育
A1116T SNP Functional Analysis of Porcine TLR3 Gene
XING Min-wei;ZHAI Chun-yuan;LI Hai-tao;YANG Xiu-qin
2011, 42(4):  468-474.  doi:
Abstract ( 644 )   PDF (1213KB) ( 376 )  
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In order to reveal the biological role of nonsynonymous SNP A1116T of porcine TLR3 Gene and to investigate the relationship of the SNP with disease resistance/susceptibility, the study was designed to construct eukaryotic expression vector of two types of alleles by splicing overlap extension PCR and site-directed mutation methods, to analyze the function of the two alleles in signal transduction in cultured PK-15 cell by dual-luciferase reporter assay and Real-time PCR methods. Two types of eukaryotic expression vectors of porcine TLR3 were successfully constructed. The dual-luciferase reporter assay demonstrated that the mutant type TLR3 allele was defective for activating TLR3-dependent reporter constructers. The real-time PCR analysis showed that the mutant type was reduced in inducing IL-6 transcription. The results indicate that the mutation influences the signal transduction ability of porcine TLR3.
A1116T SNP Functional Analysis of Porcine TLR3 Gene
XING Ming-wei,ZHAI Chun-yuan,LI Hai-tao,YANG Xiu-qin
2011, 42(4):  468-474.  doi:
Abstract ( 131 )   PDF (942KB) ( 523 )  
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遗传繁育
Expression Analysis of DECR1 Gene in Tissues of Mashen and Large White Pig
LI Bu-gao;GUO Xiao-hong;CAO Guo-qing;WANG Xiao-jing;GAO Peng-fei;LIU Hong;ZHOU Zhong-xiao
2011, 42(4):  475-480.  doi:
Abstract ( 855 )   PDF (694KB) ( 384 )  
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The purpose of this study was to analyze DECR1 mRNA and protein expression profiles of Mashen and Large White pigs in different tissues, and evaluate the potential relationships between its relative expression and lipid metabolism. DECR1 expression profiles in nine tissues (liver, kidney, spleen, lung, small intestine, subcutaneous adipose, stomach, longissimus dorsi, heart) of Mashen and Large White pigs were examined by quantitative realtime PCR and Western blot, respectively. The results indicated that, by real-time PCR, DECR1 had the tissue-and speciesspecific in mRNA expression, but highly in liver, subcutaneous adipose and heart. The DECR1 mRNA expression in subcutaneous adipose of Large White pig was significantly higher than that of Mashen pig (P<0.01). The results showed that, by Western blot, the distribution of DECR1 in tissues were significantly different, but highly in subcutaneous adipose, liver and small intestine. Moreover, the expression of DECR1 in subcutaneous adipose of Large White pig was significantly higher than that of Mashen pig (P<0.05). It was inferred that DECR1 gene may be related to regulatory mechanism of lipid metabolism.
Expression Analysis of DECR1 Gene in Tissues of Mashen and Lsrge White Pig
LI Bu-gao,GUO Xiao-hong,CAO Guo-qing, WANG Xiao-jing,GAO Peng-fei,LIU Hong,ZHOU Zhong-xiao
2011, 42(4):  475-480.  doi:
Abstract ( 142 )   PDF (876KB) ( 560 )  
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遗传繁育
Studies on Population Genetic Diversity and Microsatellite Markers for Growth Traits in Luxi Cattle
BI Wei-wei;CHEN Wei-yun;WANG Hui;WANG Zhong-hua;ZHANG Yuan
2011, 42(4):  481-488.  doi:
Abstract ( 604 )   PDF (830KB) ( 473 )  
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8 pairs of polymorphic microsatellites out of 30 microsatellite loci in Luxi cattle chromosome were selected from GenBank. Genetic diversity of Luxi cattle and the relating effects between genotypes and growth traits were analyzed in 120 individuals by using least squares linear model and DNA analysis technique. A total of 34 alleles were detected. Each microsatellite loci in the number of alleles was ranging from 3.6, and the average number of alleles was 4.25. The average heterozygosity (H) was 0.561 3; the polymorphism information content (PIC) was 0.498 3. The results indicated that eight microsatellite loci had significant effect on some growth traits of Luxi cattle (P<0.05 or P<0.01). The most favorable genotypes for interrelated traits were AE at ETH185 for body height, CC at BM711 and BB at BM1824 for body length, AD at TGLA53 and CC at BM711 for cannon girth, BB at BM1824 and AB at DVGA55 for abdominal girth, AC at TGLA53, BD at ETH185, AB at BM711, AC at DVGA46 and BB at DVGA44 and DVGA55 for body weight, respectively While the disadvantageous genotypes were AD at microsatellite loci of DVGA46 and DVGA44, BC genotype at ETH185, AA genotype at DVGA55 loci The results make the genotype useful for markers associated with the growth traits for marker assisted selection.
Studies on Population Genetic Diversity and Microsatellite Markers for Growth Traits in Luxi Cattle
BI Wei-wei,CHEN Wei-yun,WANG Hui,WANG Zhong-hua,ZHANG Yuan
2011, 42(4):  481-488.  doi:
Abstract ( 133 )   PDF (1071KB) ( 477 )  
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遗传繁育
The Tissues Profile of MC3R and MC4R and the Association of Their Expressionin the Adipose with Backfat Thickness in Chinese Simmental Cattle
HUANG Meng;XU Shang-zhong;LI Jun-ya;GAO Xue;CHEN Jin-bao
2011, 42(4):  489-494.  doi:
Abstract ( 586 )   PDF (751KB) ( 333 )  
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In order to explore the expression rule of key gene and screen the candidate gene affecting fat deposition, 30 samples from 137 slaughtered 18 month Chinese simmental beef cattle were divided into thick (H) and thin (L) backfat groups, and the expression of MC3R and MC4R were analyzed in the adipose of different samples and other tissues. The result indicated that both the MC3R and MC4R expressed in the heart, liver, spleen, lung, kidney, muscle, adipose, thymus, small intestine, lymph and testis. The abundances of both the MC3R and MC4R mRNA in adipose were higher in L group than those in H group (P<0.05), and correlated with backfat thickness (r=0.784, P<0.05; r=0.836, P<0.05; respectively). The results indicate that MC3R and MC4R had significant relationship with backfat thickness and might be a candidate gene for fattening in beef cattle.
The Tissues Profile of MC3R and MC4R and the Association of Their Expression in the Adipose with Backfat Thickness in Chinese Simmental Cattle
HUANG Meng,XU Shang-zhong,LI Jun-ya,GAO Xue,CHEN Jin-bao
2011, 42(4):  489-494.  doi:
Abstract ( 131 )   PDF (1014KB) ( 596 )  
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遗传繁育
Molecular Cloning of C/EBPα in Landes Geese and the Effect of Overfeeding on Its mRNA Expression Levels
YU Sha-li;ZHANG Xiang;SU Sheng-yan;ZHOU Yang;LI Qi-fa;XIE Zhuang
2011, 42(4):  495-502.  doi:
Abstract ( 941 )   PDF (6791KB) ( 239 )  
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The experiment was conducted to clone the C/EBPα gene in Landes geese liver and to check the relative expression levels of C/EBPα mRNA of different groups. In the study, the gene sequence was obtained based on the C/EBPα gene sequence of chicken. The results showed that the C/EBPα gene has a 975 bp open reading frame (ORF) flanked by a 210 bp 5′UTR and a 397 bp 3′UTR. The deduced amino acid sequence has 324 residues, which was a nucleoprotein residing in nucleolus without signal peptide and transmembrane region.The C/EBPα protein contained a carboxyl terminal basic-leucine zipper (bZIP) domain, which was characterized by cysteinerich, doublezinc finger motifs and a distinct homeodomain. Also, the levels of C/EBPα mRNA of different groups were detected by real-time PCR. Compared to the control group, the levels of C/EBPα mRNA of all overfeeding groups increased significantly (P<0.05 or P<0.01).The level of overfeeding supplemented with T3 and betaine group was higher than that of control group (P<0.01) and other overfeeding groups (P<0.05). So it is suggested that the C/EBPα gene may be related with lipid metabolism.
Molecular Cloning of C/EBPα in Landes Geese and the Effect of Overfeeding on Its mRNA Ecpresssion Levels
YU Sha-li,ZHANG Xiang,SU Sheng-yan,ZHOU Yang,LI Qi-fa,XIE Zhuang
2011, 42(4):  495-502.  doi:
Abstract ( 94 )   PDF (2241KB) ( 457 )  
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遗传繁育
Effect of Royal Jelly on Growth and Development before PubertyFemale Japanese White Rabbits
XU Bao-hua;;ZENG Zhi-jiang
2011, 42(4):  503-507.  doi:
Abstract ( 1006 )   PDF (470KB) ( 343 )  
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This experiment was conducted to explore the effect of royal jelly(RJ) on growth and development of Japanese White rabbits before puberty. Thirty six 50-day-old female rabbits were randomly allocated to three groups of 12 rabbits each: high dose group(H-RJ), low dose group(L-RJ) and control group(C). Rabbits were administrated daily with RJ weighted 0.2% and 01% of body weight in the high dose group and low dose group via gastric, respectively; saline water weighted 0.1% of body weight was administrated in the control group via gastric. After 40 days, plasma sample was collected from each rabbit and the levels of E2, FSH, LH, P4, PRL and T were analyzed by radioimmunoassay. The heart, left uterus and left ovary were collected from six slaughted rabbits in each group and the expression of the estrogen receptor alpha (ERα) in right ovary and estrogen receptor beta (ERβ) in right uterus were detected by real time RT-PCR. The results showed that: (1) The molting rate of rabbits in C, L-RJ and H-RJ was 0, 75% and 67% after 12 days, respectively; (2) There was no influence of RJ on the body weight, heart, uterus and ovary coefficients of rabbit (P>0.05); (3) The concentration of E2 was significantly higher in RJ treated groups than that in control group (P<0.01), but no significant difference was observed in the other sex hormones (P>0.05); (4) The expression of ERα in the uterus and ERβ in the ovary were lower in RJ treated groups than that in control. The results indicated that royal jelly can greatly improve the molting rate of the rabbits, but it did not influence the growth of the heart, uterus and ovary, it increased the serum estrogen level and inhibited the expression of estrogen receptor in uterus and ovary of Japanese White rabbits before puberty.
Effect of Royal Jelly on Growth and Development before Puberty Female Japanese Rabbits
XU Bao-hua,ZENG Zhi-jiang
2011, 42(4):  503-507.  doi:
Abstract ( 76 )   PDF (1021KB) ( 474 )  
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动物营养
Effects on Lactation Ability of Dairy Cow Mammary Gland Epithelial Cellsby stat5 High Expression
LIU Xiao-fei;GAO Xue-jun;LI Qing-zhang;TONG Hui-li
2011, 42(4):  508-512.  doi:
Abstract ( 588 )   PDF (1066KB) ( 299 )  
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The expression vector Stat5 was analysed for the change of effects on lactation ability of dairy cow mammary gland epithelial cells (DCMECs). The expression vector was stably transfected by G418. The proliferations and viabilities of DCMECs, expression of β-casein, lactose, stat5 gene and STAT5 protein were detected by cell counter analyser, HPLC, Western Blotting and Real-time PCR. Compared with the control group, the proliferations and viabilities of DCMECs, expression of stat5 gene and STAT5 protein were both increased (P<0.01);β-casein, lactose and pSTAT5 protein were increased (P<0.05). The results showed that, eukaryotic expression vector pcDNA3.1+- stat5-αS1 could be highly expressed in DCMECs, and could raise lactation ability of DCMECs.
Effects on Lactation Ability of Dairy Cow Mammary Gland Epithelial Cells by stat5 High Ecpression
LIU Xiao-fei,GAO Xue-jun,LI Qing-zhang,TONG Hui-li
2011, 42(4):  508-512.  doi:
Abstract ( 115 )   PDF (1186KB) ( 524 )  
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动物营养
Developmental Characteristics of Gastrointestinal Tract in Confined Lambsat the Age 0-56 Days
GUO Jiang-peng;ZHANG Yuan-xing;LI Fa-di;HAO Zheng-li;MA You-ji;PAN Jian-zhong
2011, 42(4):  513-520.  doi:
Abstract ( 962 )   PDF (763KB) ( 349 )  
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The objective of this study was to investigate the developmental characteristics of gastrointestinal tract in confined raising lambs with supplementary feed from the age of 7 days. Forty-five male lambs (Gansu modern breeding sheep group) were divided into 9 treatments (5 animals in every treatment), which were slaughtered and sampled at the age of 0, 7, 14, 21, 28, 35, 42, 49, 56 days, respectively. The relative weight of compartments in stomach (%live weight, %stomach weight), relative capacity of compartments in stomach (%gastrointestinal tract capacity, %stomach capacity), and relative weight of compartments in intestine (%live weight, %intestinal weight) were measured. The results showed that the relative weight of stomach (% live weight, RWS-L) at 0 d was about 1% of the live weight. RWS-L at 56 d was 2.33 times as large as that at 0 d. At the same time, declines were observed in RWA-L of lambs, and RWA-L at 56 d was 79.86% as large as that at 0 d. RCF-G and RCF-S were notably increasing after 7 d, and it could be appeared that RCF-S was exceed than RWF-S at 28 d. RCA-G and RCA-S were significantly decline during 7 to 28 d. With the age in days increasing, there were some increasing in relative intestinal weight (%live weight), and it was 1.43 times at 56 d as large as that at 0 d. The growing speed of large intestine (% intestinal weight) were more quickly than that of small intestine, and the large intestine and small intestine were 1.71 and 1.36 times at 56 d as large as that at 0 d, respectively. There was more growing speed in ileum than other compartments in small intestine, but almost similar growing speeds were found in every compartment (cecum, colon, rectum) of large intestine. These results indicated that different growing speeds were appeared in gastrointestinal organs at the age of 056 d in lambs. The growing speeds of relative weight in stomach were more than that in intestine. And 7 to 14 days would be ahead on the development of weight and capacity in stomach for confined raising lambs and providing enough supplementary feed from 7 d after birth. With confined raising and supplying the suited supplementary diet, the feasible weaned age in days was 35 or 28 d after birth in lambs.
Decelopmental Characteristics of Gastrointeseinal Tract in Confined Lambs at the Age 0-56 Days
GUO Jiang-peng,ZHANG Yuan-xing,LI Fa-di,HAO Zheng-li,MA You-ji,PAN Jian-zhong
2011, 42(4):  513-520.  doi:
Abstract ( 100 )   PDF (1256KB) ( 534 )  
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预防兽医
Construction of Recombinant Vector of VP2 Gene of Foot-and-mouthDisease Virus and Establishment of Stable Cell Line for Its Stable Expression
XIE Jin-lu;;WANG Hong-mei;LIU Guo-yi;WU Jian-ming;LIU Xiao;GAO Yun-dong;YU Li;ZHONG Ji-feng;HE Hong-bin
2011, 42(4):  521-526.  doi:
Abstract ( 676 )   PDF (1545KB) ( 328 )  
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The aim of the study was to construct a recombinant vector, which including VP2 gene of Asia I strain of foot-and-mouth disease virus (FMDV), and to establish the BHK-21 cell line, which stably expressing VP2 gene. VP2 gene of FMDV was amplified from Asia I strain by RT-PCR, and its complete cDNA was cloned into pIRES2-EGFP vector. The recombinant pIRES2-EGFP-VP2 vector, which could express VP2 and EGFP proteins, was confirmed by sequencing analysis, the transient expression of VP2 in 293T cells transfected with the recombinant plasmid via LipofectamineTM 2000 was then determined by Western blot. Furthermore, BHK-21 cells were transfected with pIRES2-EGFP-VP2, and anti-G418 cell clones were screened using G418, both G418 and GFP positive cell clones were continually cultivated after GFP detection, their expressions of VP2 gene were confirmed by Western blot. The results showed that the recombinant plasmid pIRES2-EGFP-VP2 correctly encoded VP2 gene and transiently expressed in 293T cells- BHK-21 cells were transfected with recombinant plasmid, then BHK-21 cell clones expressing VP2 were obtained by G418 pressure selection, after continuous passage for 60 days, BHK-21 cell lines stably expressing VP2 gene of FMDV were established. The above results indicated that BHK-21 cell lines stably expressing VP2 gene of FMDV were established, and it provides a good platform to study the role of VP2 gene during FMDV induces apoptosis.
Construction of Recombinant Vector of VP2 Gene of Foot-and-mouth Disease Virus and Eatablishment of Stable Cell Line for Its Stable Expression
XIE Jin-lu,WANG Hong-mei,LIU Guo-yi,WU Jian-ming,LIU Xiao,GAO Yun-dong,YU Li,ZHONG Ji-feng,HE Hong-bin
2011, 42(4):  521-526.  doi:
Abstract ( 125 )   PDF (1438KB) ( 370 )  
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预防兽医
Stable Expression of T7 RNA Polymerase Gene Mediated by RetroviralVector in PK15 and SK6 Cells
WU Jin-yan;TIAN Hong;ZHENG Hai-xue;CHEN Yan;JIN Ye;SHANG You-jun;LIU Xiang-tao﹡
2011, 42(4):  527-532.  doi:
Abstract ( 556 )   PDF (1057KB) ( 275 )  
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The experiment was conducted to insert T7 RNA polymerase (RNAP) gene into cells of swine by retroviral vector system, then to analyze its hereditary stability. The bacteriophage T7 RNAP gene was amplified via PCR from lysogen DE3, and the gene was cloned into pBABEpuro retrovial vector to construct a recombinant plasmid that named as pBABEpuro/T7. Then the pT7BABEpuro and pVSV-G plasmids were cotransfected into GP2-293 packaging cells by Lipfection 2000, some pseudotype viruses were ingathered and transfected into PK15 and SK6 cells under polybrene, respectively. The T7-PK15 and T7-SK6 cells were propagated in DMEM with puromycin respectively. The genome extraction from the cells transfected different times, and the T7 RNAP gene was amplified from the genome by PCR- Expression and activity of T7 RNAP gene in T7-PK15 and T7-SK6 cells were analyzed by immediate fluorescence and SDS-PAGE assay. Results showed that the T7 RNAP gene had been integrated into the chromosome of T7-PK15 and T7-SK6 cells, and expressed stably at high level. After 40 times passage, transcriptional activity of T7-PK15 and T7-SK6 cell lines were not weaked. T7 RNAP gene had been stably integrated into the chromosome of T7-PK15 and T7-SK6 cells, construction of cell lines provided favorable tools for virus rescue in vivo.
Stable Expression of T7 RNA Polymerase Gene Mediated by Retroviral Vector in PK15 Cells
WU Jin-yan,TIAN Hong,ZHENG Hai-xue,CHEN Yan,JIN Ye,SHANG You-jun,LIU Xiang-tao
2011, 42(4):  527-532.  doi:
Abstract ( 120 )   PDF (1336KB) ( 599 )  
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预防兽医
The Comparison of Serologic Relativity and CPE Types of 3 Avian ReovirusStrains Induced Different Diseases
CHEN Shi-long;CHEN Shao-ying;CHENG Xiao-xia;JIANG Bin;LIN Feng-qiang;WANG Shao;ZHU Xiao-li;LI Zhao-long;ZHANG Shi-zhong;
2011, 42(4):  533-537.  doi:
Abstract ( 917 )   PDF (2272KB) ( 275 )  
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This study was conducted to compare the antigenic relativity and cytopathic effect (CPE) characteristics of 3 avian reovirus isolates induced different diseases. The serologic relatedness of avian reovirus (ARV) S1133 strain, muscovyduck reovirus (MDRV) MW9710 strain, novel duck reovirus (NDRV) NP01 strain were determined using cross virus-neutralization (VN) tests Two avian and five mammalian cell lines were evaluated for their application in propagating these avian reovirus strains. Relatedness (R) values, determined by cross VN, revealed that three isolates’ antigen had major differences. These strains could be cultivated successfully in Vero, MDEK, CEF, MDEF, ST, AD293 T cell lines and make some different types of CPE to different cell lines. These results indicated that ARV, MDRV and NDRV’ antigen had major differences and did not share same antigenic relationship with each other. Avain reovirus of different disease-type could be distinguished by the types of CPE.
The Comparison if Serologic Relativity and CPE Types of 3 Avian Reovirus Strains Induced Different Diseases
CHEN Shi-long,CHEN Shao-ying,CHENG Xiao-xia,JIANG Bin,LIN Feng-qiang,WANG Shao,ZHU Xiao-li,LI Zhao-long,ZHANG Shi-zhong
2011, 42(4):  533-537.  doi:
Abstract ( 117 )   PDF (1451KB) ( 560 )  
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预防兽医
Multiple-locus Variable-number Tandem Repeat Analysis of Polymorphismand Genetic Relationships of S. aureus Isolated from Bovine Mastitis
GAO Jian;LIU Xiu-quan;WANG Zhi;SU Jing-liang;HAN Bo
2011, 42(4):  538-543.  doi:
Abstract ( 554 )   PDF (1237KB) ( 274 )  
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Molecular typing of Staphylococcus aureus is valuable for presenting the strain spreading, detecting the strain geographical distribution and developing specific vaccine. We aimed at developing a MLVA method to type S. aureus isolates coming from Chinese herds,and then to perform a molecular epidemiological study. Another aim is to evaluate the discriminatory power of MLVA method by comparing with RS-PCR typing method. MLVA method (multiplex PCR system) was constructed to type 27 S. aureus isolated from different regions and herds. Meanwhile, all of the isolates were typed using RS-PCR method. Both of the methods can type all 27 isolates obtained from 17 herds located in eight regions. Nineteen distinct types were identified by MLVA method, all of which belong to individual herds. And the strains with the same source were typed identical other than those from two herds in Shanghai and Beijing. The discriminatory power of MLVA is 0.969. As for RS-PCR method, it can identify the isolates into 10 types, of which type Ⅰ, Ⅱ and Ⅴ were distributed in different regions. The discriminatory power of RS-PCR is 0.829. From the above results, we conclude that MLVA is a rapid and convenient method which shows good discriminatory power to type bovine S. aureus isolates. Meanwhile we infer that types of S. aureus are more likely to be restricted to a single herd, than found in multiple herds in China.
Multiple-locus Variable-number Tandem Repeat Analysis of Polymorphism and Genetic Relationships of S.aureus Isolated from Bovine Mastitis
GAO Jian,LIU Xiu-quan,WANG Zhi,SU Jing-liang,HAN Bo
2011, 42(4):  538-543.  doi:
Abstract ( 127 )   PDF (1430KB) ( 523 )  
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预防兽医
Cloning, Sequencing, Secondary Structure and B-cell Epitopes Prediction of Bordetella avium Outer Membrane Protein OmpA
TAN Yan-ling;ZHU Rui-liang;WANG Hui;WANG Xin-jian;WEI Kai;SUN Zhen-hong;SHENG Peng-cheng
2011, 42(4):  544-550.  doi:
Abstract ( 549 )   PDF (3095KB) ( 411 )  
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The aims of this study was to clone and sequence the ompA gene of Bordetella avium, and predict its secondary structure and B-cell epitopes, which provides a theoretical basis for the study of immune mechanism and B. avium subunit vaccine. The B. avium ompA gene was amplified, cloned, and sequenced. By using bioinformatics softwares and methods, the secondary structure and B-cell preponderant epitope of B. avium OmpA were predicted The results indicated that B. avium ompA gene is 597 bp, encoding 199 amino acids. Using three methods, prediction of the secondary structure of B. avium OmpA indicated that random coils were the main structural type of the flexible region in secondary structure, and contain a small amount of α-helix, β-sheet and a rare β-turn. The OmpA protein was supposed contain 5 potential antigen epitopes and 2 potential glycosylated sites. These results provide a theoretical basis for the further study of immune mechanisms, monoclonal antibodies preparation and epitope vaccine design.
Cloning, Sequencing, Secondary Structure and B-cell Epitopes Prediction of Bordetella avium Outer
TAN Yan-ling,ZHU Rui-liang,WANG Hui,WANG Xin-jian,WEI Kai,SUN Zhen-hong,SHENG Peng-cheng
2011, 42(4):  544-550.  doi:
Abstract ( 92 )   PDF (1876KB) ( 595 )  
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预防兽医
Expression and Bioactivity Analysis of Porcine IL-2 in Pichia pastoris
CHEN Guo-hua;JIA Huai-jie;ZENG Shuang;FANG Yong-xiang;LI Xiao-qing;JING Zhi-zhong;CAI Xue-peng
2011, 42(4):  551-556.  doi:
Abstract ( 843 )   PDF (862KB) ( 232 )  
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A pair of primers was designed to clone IL-2 gene according to the codon bias principle and the recombinant expressed vector pPIC9k-IL-2 was constructed. The recombinant plasmid was transformed into P. pastoris GS115 by electroporation, the positive recombinant strains was screened with different G418 concentration and induced with methanol. The result showed that IL-2 gene was integrated with chromosome of P. pastoris by PCR identification. The expressed product had a molecular weight of 16 and 20 kD bands by SDS-PAGE analysis. The result of deglycosylation analysis indicated that the protein was glycosylated moderately. The protein was purified with sephadex column and had a bioactivity of lymphocyte proliferation. The Western blotting analysis showed that target protein had immunological activity. P. pastoris expressed system is an efficient way of production of pIL-2, the expressed product had a good bioactivity.
Expression and Bioactivity Analysis of Porcine IL-2 in Pichia pastoris
CHEN Guo-hua,JIA Huai-jie,ZENG Shuang,FANG Yong-xiang,LI Xiao-qing,JING Zhi-zhong,CAI Xue-peng
2011, 42(4):  551-556.  doi:
Abstract ( 127 )   PDF (1446KB) ( 653 )  
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基础兽医
Determination of Tulathromycin Residues in Swine Tissues
NI Heng-jia;HUANG Xian-hui;HE Li-min;FANG Bing-hu;ZHAO Yong-da
2011, 42(4):  557-561.  doi:
Abstract ( 612 )   PDF (385KB) ( 233 )  
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This experiment was conducted to study the tissue residues of tulathromycin in swine, preparing for the determination of the withdrawal time. Pigs received 2.5 mg·kg-1 of tulathromycin by intramuscular(i. m.) route. Animals were sacrificed at 12 hours and 5, 12, 18, 25, 36, 48 days after administration. Tulathromycin was extracted from tissues by adding acetonitrile. The supernatant was defatted by adding N-hexane, then was applied to the C18 SPE cartridge.The identification was carried out by LC-MS/MS. The concentration of tulathromycin in tissues was calculated by Analyst 4.1.5 software. The results indicated that injection site held the highest residue level at 12 hours, while the concentration of tulathromycin in all of the tissues at 36 days was lower than the Maximum Residue Limit(MRL).The pharmacokinetic parameters were estimated using WinNolin software package. The results showed that the elimination rate was injection site>liver>skin+fat>muscle>lung>kidney, with the elimination half-life of 117.06, 193.14, 197.60, 207.64, 228.99, 232.61 h respectively. Compared with other tissues, the area under the plasma concentration-time curve(AUC) in lung (1 220.59 μg·h·g-1)was greater than in muscle, liver and skin+fat, lower than injection site and kidney. Kidney was considered as the metabolically active organ, so it suggested that lung was the target organ According to the MRLs set by EU and USA, etc, the longest withdrawal period was suggested in injection site, 33days. The results demonstrated that the absorption and distribution of tulathromyxin was wide. Besides, the active time is long. It is proposed that the withdrawal time should be 33 days after administration.
Determination of Tulathromycin Residues in Swine Tissues
NI Heng-jia,HUANG Xian-hui,HE Li-min,FANG Bing-hu,ZHAO Yong-da
2011, 42(4):  557-561.  doi:
Abstract ( 183 )   PDF (1314KB) ( 484 )  
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基础兽医
The Effects of Bacillus subtilis on Local Innate Immune and Expressionof TLR of Pigs
LI Yun-feng;DENG Jun;ZHANG Jin-hua;YANG Qian
2011, 42(4):  562-566.  doi:
Abstract ( 561 )   PDF (1402KB) ( 451 )  
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This experiment was conducted to study the effects of Bacillus subtilis on small intestine local mucosal immune responses and the expression of TLRs of pigs. The newborn piglets were orally administrated with Bacillus subtilis at 0, 7, 14, and 26 d after birth.The results showed that Bacillus subtilis could significantly promote the expression of TLR 9 and cytokine IL-6 in duodenum (P<0.05) and increase the quantity of IgA excreting cells (P<0.01), as well as increasing the expression of IL-1 in ileum (P<0.01)The results implied that Bacillus subtilis may increase the expression of IL-6 and the quantity of IgA excreting cells through interact with TLR 9. Orally administrated with Bacillus subtilis to prevent the newborn piglet diarrhea is promising.
The Effects of Bacillus subtilis on Local Immune and Expression of TLR of Pigs
LI Yun-feng,DENG Jun,ZHANG Jin-hua,YANG Qian
2011, 42(4):  562-566.  doi:
Abstract ( 97 )   PDF (1508KB) ( 676 )  
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基础兽医
Comparative Study on Small Intestinal Mucosal Immunity-associatedCells between Tibetan Goats and Tibetan Sheep
CHEN Fu-ju;CHANG Lan;WEI Fang
2011, 42(4):  567-571.  doi:
Abstract ( 561 )   PDF (2558KB) ( 316 )  
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In order to reveal adaptive characteristics of intestinal mucosal barrier structures in Tibetan goats and Tibetan sheep. The numbers of intraepithelial lymphocytes (IELs), goblet cells (GCs) and mast cells (MCs) in different parts of small intestine of 4 adult Tibetan goats and 4 adult Tibetan sheep were measured by histological method and the Scion image software analysis techniques method. Results showed that the number of GCs and MCs in different parts of the small intestine of Tibetan goats were significantly(P<0.05)higher compared to those in Tibetan sheep The number of GCs in duodenum, jejunum and ileum in Tibetan goats was higher than that of Tibetan sheep by 37.87%(P<0.05), 21.43%(P>0.05)and 31.68%(P<0.05), respectively. While the number of MCs in duodenum, jejunum and ileum in Tibetan goats was higher than that of Tibetan sheep by 85.20%(P<0.05), 50.73%(P<0.05)and 22.52%(P>0.05), respectively However, the number of IELs in different parts of the small intestine in Tibetan goats (36.35±0.98)was obviously lower(P<0.05)than that of Tibetan goats(48.84±2.12). The results suggested that the intestinal mucosal barrier function of Tibetan goats was stronger than that of Tibetan sheep. The IELs played a crucial defensive barrier role in Tibetan sheep, while the GCs and MCs play an important mucosal immune defenses in Tibetan goats.
Comparative Study on Small Intestinal Mucosal Immunity-associated Cells between Tibetan Goats and Tibetan Sheep
CHEN Fu-ju,CHANG Lan,WEI Fang
2011, 42(4):  567-571.  doi:
Abstract ( 116 )   PDF (1740KB) ( 645 )  
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基础兽医
The Study of Mouse Kidney Injury by Aristolochic AcidⅠ
DONG Xiao-kai;ZHANG Zhong-wen;PENG Xiao-lan;LIAO Fang-fang;GAO Lei;WU Guo-juan
2011, 42(4):  572-577.  doi:
Abstract ( 860 )   PDF (3135KB) ( 403 )  
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To provide theoretical basis for investigating pathologic mechanism, we observed the effects of aristolochic acidⅠ(AAⅠ)on kidney fine structures and physiological index of mice to preliminary interpret the damage of AAⅠon kidney. Male Kunming mice(20 g±0.2 g)were divided into four groups, randomly. The experimental groups were intragastricly administrated with low, moderate, and high concentration of AAⅠ solution, and the control group with equal-volume water. After 30 days, we examined pathological changes of kidney, and measured correlated physiological index in blood and urine The results showed that with the increasing doses, mice body weights lowered obviously, urinary volumes increased in the initial stage, and then hypourocrinia, anuria, renal glycosuria occurred. While the TGF-β1 contents in serum went up gradually, and erythrocytes, hemoglobins, monocytes decreased. Compared with the control group, in cortex parts and the boundary between cortex and medulla parts, epithelial cells of proximal convoluted tubules damaged, and fibration penetrated deeply with the increasing doses from cortex to medulla parts. At the same time, we found that cell organs harmed and nuclei varied in electron microscope. The results showed that AAⅠ could damage kidney dose-dependently, lead mostly to pathological changes of renal tubule mesenchymal, and affect its normal fine structures.
The Study of Mouse Kidney Injury by Aristolochic Acid Ⅰ
DONG Xiao-kai,ZHANG Zhong-wen,PENG Xiao-lan,LIAO Fang-fang,GAO Lei,WU Guo-juan
2011, 42(4):  572-577.  doi:
Abstract ( 138 )   PDF (1831KB) ( 707 )  
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研究简报
Screening and Analyzing of Genes on Prolificacy in Goat
QIAN Li-li;JIA Qing;LI Xue-mei;LI Xun-ye;ZHENG Hui-qin;FENG An-xue
2011, 42(4):  578-584.  doi:
Abstract ( 578 )   PDF (1360KB) ( 394 )  
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To search the genes associated with prolificacy of goats, and research the genetic mechanism of fertility regulation, twelve Jizhong goats were divided into 2 groups according to their reproductive records. Differential display reverse transcript PCR (DDRT-PCR) method was used in the analysis of the discrepancy of gene expression in ovarian tissue of goats in estrual period 4 segments differentially expressed in quantity were finally obtained via second PCR amplification, reverse Northern blot and semi-quantitative PCR After sequencing and cloning, these segments were submitted to GenBank, and analysised by BLAST. Two of the 4 were new segments and high homologous sequences in NCBI was not found. Another segment shared 81% similarity with one clone segment (THY010003E05) of wild boar (Sus scrofa), but with unknown function. The other segment was 94% similar to the mRNA of PCNP gene in cattle. It indicate that the segment is a homolog of PCNP gene and it could affect goat prolificacy via involving in cell cycle regulation of oocyte according to the analysis of its function.
Screening and Analyzing of Genes on Prolificacy in Goat
QIAN Li-li,JIA Qing,LI Xue-mei,LI Xun-ye,ZHENG Hui-qin,FENG An-xue
2011, 42(4):  578-584.  doi:
Abstract ( 84 )   PDF (1595KB) ( 595 )  
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研究简报
Culture of Bovine Fetal Skin Fibroblasts in vitro and Ipr1 Gene Transfection
SONG Yong-li;HE Xiao-ning;HUA Song;LAN Jie;ZHENG Yue-mao;HE Xiao-ying;LI Ji-xia;QUAN Fu-sheng;ZHANG Yong
2011, 42(4):  585-592.  doi:
Abstract ( 579 )   PDF (2443KB) ( 282 )  
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In order to provide the donor cells for nuclear transfer, in this study, the eukaryotic expression vector PEGFP-SRA-Ipr1 was constructed, the bovine fetal fibroblast cells were isolated in vitro and the cells with PEGFP-SRA-Ipr1 were transfected. Ipr1 gene and SR-A promoter were successfully cloned and then used to construct the macrophage-specific eutherin expression vector- Bovine fetal fibroblast cells from ear were cultured in vitro by tissue explantation, passaged and purified. Then the eutherin expression vector PEGFP-SRA-Ipr1 was transfected into the cells at the passages of 4th to 10th by using electroporation. The fluorescence was observed after 24 h. 600 μg·mL-1 G418 was added into the culture medium after 48 h, which lasted 1 week. After that, the concentration of G418 decreased to 300 μg·mL-1 for screening. Then the monoclones were picked and cultured for amplification The positive cells were determined via PCR and flow cytometry. GFP was observed 24 h after transfection. The bovine fetal fibroblast cells permanently carrying PEGFP-SRA-Ipr1 were obtained after the screening of G418. The interested fragment was confirmed to be correct by PCR and the positive cells had the normal karyotype, suggesting that the interested gene was successfully integrated into the genome and it failed to disrupt the stability of the genome. In conclusion, the obtained cells could be utilized as the donor cells for research of transgenic cloned cattle by nuclear transfer.
Culture of Bovine Fetal Skin Fibroblasts in vitro and Ipr1 Gene Transfection
SONG Yong-li,HE Xiao-ning,HUA Song,LAN Jie,ZHENG Yue-mao, HE Xiao-ying,LI Ji-xia,QUAN Fu-sheng,ZHANG Yong
2011, 42(4):  585-592.  doi:
Abstract ( 119 )   PDF (1792KB) ( 542 )  
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研究简报
Identification of Mycoplasma capricolum subsp. capripneumoniae Hubei Strainin China and the Immunogenicity Study of Its Outer Membrane Proteins
ZHAO Ping;GUO Han;HE Ying;GAO Peng-cheng;ZHANG Nian-zhang;LU Zhong-xin
2011, 42(4):  593-599.  doi:
Abstract ( 527 )   PDF (1348KB) ( 333 )  
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A strain of Mycoplasma named GH2 was isolated successfully from lung tissues of sick goats in Hubei province. This isolate was purified several times and were identified by biochemistry test, microscope observation, PCR, REA, genome analysis and animal pathogenicity tests. The immunogenicity of outer membrane proteins and whole cell proteins of GH2 were analyzed by western blotting. The results were compared with antiserum of Mycoplasma mycoides subsp. capri and Mycoplasma ovipneumoniae. Result showed that the strain GH2 belongs to the Mycoplasma capricolum subsp. capripneumoniae (Mccp) and the proteins about 60 and 49.7 kD possess its capital immunogenicity and they were also the specific antigen of GH2 which distinguished with Mycoplasma mycoides subsp. capri and Mycoplasma ovipneumoniae. Result of this study lay good theoretical foundation for diagnosis and vaccine development of Mycoplasma capricolum subsp. capripneumoniae.
Identification of Mycoplasma capriolum subsp. capripneumoniae Hubei Strain in China and Immunogenicity Study of Its Outer Membrane Proteins
ZHAO Ping,GUO Han,HE Ying,GAO Peng-cheng,ZHANG Nian-zhang,LU Zhong-xin
2011, 42(4):  593-599.  doi:
Abstract ( 99 )   PDF (1626KB) ( 512 )  
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研究简报
Relationship between Fluoroquinolone Resistance and GeneMutation in Streptococcus suis
RUI Ping;MA Zeng-jun;DUAN Ling-xin;LIU Xie-rong;NI Jing;SHEN Jian-zhong
2011, 42(4):  600-604.  doi:
Abstract ( 524 )   PDF (1724KB) ( 317 )  
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To detect the relationship between fluoroquinolone resistance and gyrA and parC gene mutation in Streptococcus suis. The MIC of 34 strains of Streptococcus suis were determined by microdilution. The genes encoding the quinolone-resistance determining region (QRDRs) of parC and gyrA in fluoroquinolone-susceptible and -resistant Streptococcus suis clinical isolates were identified and sequenced. Ser to Phe and Arg to Leu mutation at position 79 and 87 of the parC gene was detected in fluoroquinolone resistance S.suis, and Arg to Ser, or Ser to Arg mutations at the position 66 or 81 of gyrA gene were detected in 4 highly resistance strains; no amino acid changes in gyrA or parC were detected for 9 fluoroquinolonesusceptible strains,the mutations in both genes was found in the strains with MIC of fluoroquinolone higher than 32μg·L-1. Mutations in parC gene result in low level resistance against fluoroquinolone,and high level resistance is resulted from the mutations in gyrA and parC both genes.
Relationship between Fluoroquinolone Resistance and Gene Mutation in Streptococcus suis
RUI Ping,MA Zeng-jun,DUAN Ling-xin,LIU Xie-rong,NI Jing,SHEN Jian-zhong
2011, 42(4):  600-604.  doi:
Abstract ( 104 )   PDF (1672KB) ( 608 )  
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