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Table of Content

25 December 2007, Volume 38 Issue 12
遗传繁育
Association Analysis on Myostatin 5′Regulatory Region Polymorphism and Growth Traits in Three Cattle Breeds
ZHANG Run-feng;CHEN Hong;LEI Chu-zhao;ZHANG Chun-lei;ZHANG Hai-jun;BAO Bin;CHEN Fu-ying;WANG Yi-min;NIU Hui;WANG Xin-zhuang
2007, 38(12):  1273-1278.  doi:
Abstract ( 1794 )   PDF (460KB) ( 1029 )  
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Single nucleotide polymorphisms (SNPs) of myostatin 5′regulatory region in Nanyang, Qinchuan and Jiaxian Red cattle breeds were analyzed by PCRRFLPs. The results showed that the allele T was dominant in the three breeds for the T→A mutation, and only one homozygous AA individual was detected in Jiaxian Red cattle which was at Hardy-Weinberg disequilibrium (P<0.05). The Nanyang and Qinchuan populations were at Hardy-Weinberg equilibrium (P>0.05). The effects of the T→A mutation on growth traits of the three breeds were also estimated. The results showed that six-month Nanyang cattle with genotype TT had lower heart girth and heart girth index than individuals with TA genotype, eighteen-month Nanyang cattle with TT genotype had higher withers heights than animals with TA genotype (P<0.05), but the effects of myostatin genotypes on growth traits were not significant in Qinchuan cattle and Jiaxian Red cattle. These results indicated that the growth stages and the selected traits should be comprehensively considered in the breeding practice of Nanyang cattle.
Cloning,Sequence Analysis and Molecular Evolution of the Myf-5 Gene in Gayal
CHEN Yu-hong;ZHONG Jin-cheng;MA Zhi-jie;HU Han-li;JIN Shuang;LUO Hong
2007, 38(12):  1279-1289.  doi:
Abstract ( 1125 )   PDF (745KB) ( 813 )  
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The Myf-5 gene of the gayal (Bos frontalis) was cloned and sequenced by the T-A method, including the 5′flanking ragion, three exons, two introns and partial 3′flanking region, then based on the nucleotide and amino acid sequences compared with some species (cow, chimpanzee, rhesus monkey, human, dog, house mouse, Chinese softshell, chicken and zebrafish) by DNAMAN, BioEdit and DnaSP, bio-softs, and also constructed the molecular phylogenetic tree on nucleoside acid of the coding region of Myf-5 gene with method of NJ, ME and UPGMA. The results show that: ①The numbers of exons and introns of Myf-5 gene of gayal are the same with that of cow, chimpanzee, rhesus monkey, human, dog, house mouse, Chinese softshell, chicken and zebrafish, consisted by 3 exons and 2 introns, but the size had some differences. The size of 3 exons and 2 introns was 660,76,1 226,852,407 bp,respectively. The junction sequence of exons and introns of gayal’s Myf-5 gene was accordance with gene composition of GT-AG rule. ②The homology rates of nucleotide sequences of the coding region of Myf-5 gene between the gayal and cow, chimpanzee, rhesus monkey, human, dog, house mouse, Chinese softshell, chicken and zebrafish were 99.0%,90.0%,90.5 %,90.5%,87.2%,84.8%,72.9%,64.3%,51.8%,respectively; while the homology rates of amino acid sequences were 99.6%,94.9%,94.6%,94.9%,89.9%,88.7%,81.6%,70.4%,56.6%,respectively. This result showed that the nucleotide sequence of the coding region and accordingly amino acid sequences of Myf-5 gene of gayal and cow, chimpanzee, rhesus monkey, human, dog, house mouse, Chinese softshell, chicken and zebrafish were quite conservative. ③The result was basically consistent with three methods which was NJ, ME, UPGMA constructed in molecular phylogenetic tree, the gayal was clustered together with cow firstly, chimpanzee, rhesus monkey and human clustered together, then together with each other, followed by other species. The clustering result was accordance with the results of mtDNA, other function genes and the zoological classification. This indicated that Myf-5 gene fit to construct molecular phylogenetic tree.
The SNP of CLPG Gene in Jiaxian Red Cattle and Its Effect on Growth and Development Traits
The SNP of CLPG Gene in Jiaxian Red Cattle and Its Effect on Growth and Development Traits
2007, 38(12):  1290-1293.  doi:
Abstract ( 1366 )   PDF (514KB) ( 693 )  
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The SNPs of CLPG gene in 113 Jiaxian Red cattle were detected by PCR-SSCP, and three genotypes (AA, AB and AC) were identified. The sequencing results showed that genotype AA was the wild type with C and T at the position 150 and 419, respectively, genotype AB and AC have mutations T419C, C150T, respectively. The polymorphism information contents (PIC) was 0.226, which indicated that the polymorphism of this population was relatively low. The associations of different genotypes with some of the growth and development traits were analyzed. The results showed that the individuals with genotype AA and AC had a larger body weight, body length than the individuals with genotype AB (P<0.05 or P<0.01, respectively), the individuals with genotype AA were better than those with AB in the circumference of cannon bone (P<0.05), the individuals with the genotype AB was significantly different with those of AA (P<0.05) and AC (P<0.01) in the hucklebone width, respectively.
Analysis on Single Nucleotide Polymorphisms of Exon of MyoG Gene in Different Sheep Breeds
LIU Yong-bin;WANG Feng;DENG Feng;TIAN Chun-ying;LIU Mei-xia;LIANG Zhen-jun;RONG Wei-heng
2007, 38(12):  1294-1299.  doi:
Abstract ( 765 )   PDF (1080KB) ( 764 )  
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The polymorphisms of myogenin gene in Mongolian sheep, Poll Dorset sheep, German meat Merino sheep and Suffolk sheep were analyzed by PCRSSCP. The results indicated that there were three genotypes(AA,AB and BB)detected by exonⅠ, and there weren’t polymorphism in exonⅡ. To amplify fragments by exonⅠ, AA and AB genotypes were detected in four sheep breeds. BB genotype was detected in Mongolian sheep, Poll Dorset sheep and Suffolk sheep.Frequency of AA genotype in Mongolian sheep was the highest , but frequency of AB genotype in other three breeds was high. Frequency of A allele was obviously higher than frequency of B allele in four sheep breeds. The polymorphic fragments amplified by exonⅠwere cloned and sequenced, and the sequencing results showed that there was one single nucleotide mutation :T→C at cDNA 305 of myogenin gene in sheep, and this mutation resulted in an amino acid change: cysteine→arginine.
Study on Mitochondrial DNA Genetic Polymorphism of Seven Indigenous Goat Breeds(Groups) in Sichuan
ZHANG Hong-ping;LI Li;XIANG De;LIU Cheng-jian;LI Xue-wei
2007, 38(12):  1300-1305.  doi:
Abstract ( 1402 )   PDF (499KB) ( 714 )  
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Complete sequence of mitochondrial DNA(mtDNA) control region was sequenced in 43 individuals from 7 indigenous goat(Capra hircus)breeds(groups)in Sichuan province. The complete sequence of mtDNA control region was 1 212 or 1 213 base pair, and the content of A+T(59.9%)was higher than that of G+C(40.1%)significantly. 74 sites were polymorphic and all of these sequences were neutral mutation. All sequence polymorphic sites defined 27 haplotypes. High nucleotide diversity(1.868%)and haplotype diversity(0.966) were manifested. These results suggested richer genetic diversity within Sichuan indigenous goat populations and genetic divergence among them. The Kimura 2-parameter distance among breeds(groups)varied from 0.001 7 to 0.030 6. Phylogenetic unrooted trees constructed with neighbourjoining methods showed two maternal origins in Sichuan indigenous goats. More detailed studies should be continued to indicate whether the two maternal origins are Capra aegagrus and Capra falconeri.
Polymorphism of G3072A in IGF2 Gene and Its Association with Birth Weight and Early Growth Traits in Yorkshire Pigs
SUN Yi;FEI Si-qing;TANG Hui;WU Ying;FAN Xin-zhong;JIANG Yun-liang
2007, 38(12):  1306-1310.  doi:
Abstract ( 692 )   PDF (504KB) ( 799 )  
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By use of PCR-SSCP approach, the present study analyzed the single nucleotide polymorphism (SNP) at G3072A site of IGF2 intron 3 in Yorkshire, Landrace and Duroc pigs. The association of the polymorphism with birth weight and early growth traits were as well analyzed in Yorkshire pigs. The results indicated that the genotypes AA,AB and BB were found in Yorkshire, Landrace and Duroc pig populations, and the populations of both Yorkshire and Landrace pigs were in a state of Hardy-Weinberg equilibrium(P<0.01),whereas the Duroc pig population was not(P>0.05). Sequencing demonstrated that A and B alleles correspond to nucleotide A and G, respectively. In addition, for A allele, a (AG) dinucleotide was inserted at the 10-11th position downstream the mutation site. Association analysis indicated that the least square means of birth weight of AA, AB and BB pigs differ significantly (P<0.05), whereas body weight at 21, 28 and 70 days and average daily gain of body weight from birth to 21 days,from 21 days to 28 days and from 28 to 70 days of AA and AB individuals were not significantly different in Yorkshire pigs(P>0.1).
Effect and Different Expression of ER Gene in Early Embryos of Generic Hybrids of Chicken-quail
LI Yan;QIAO Ai-jun;MA Wen-xia;LIAO He-rong;ZHAO Zong-sheng;LI Da-quan
2007, 38(12):  1311-1315.  doi:
Abstract ( 1217 )   PDF (726KB) ( 618 )  
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To investigate the effect of estrogen receptor(ER) on early embryos of generic hybrids of chicken-quail.we acquired cross bred eggs of chicken(♂)-quail(♀) by artificial insemination,hatched the eggs in one batch according to the standard condition of chicken,collected early living-embryos of 2.75,3.00,3.25,3.50,3.75,4.00,4.25,4.50,4.75 and 5.00 d at random.Adopting RT- PCR and multi-ply PCR, we identified the sex of embryos with primers of Wpkci and β-actin,then selected four female and male embryos respectively from each period taken β-actin as internal standard to determined the relative quantity of ER mRNA.The patterns of ER mRNA expression of male and female were basically consistent.The expressive level were all higher in 2.75~3.00 d,declined distinctly in male(P<0.05) and very significantly in female(P<0.01) in 3.25 d,then ,all reached initial level in 3.50 d and declined great significantly in 3.75 d (P<0.01),then maintained the lower level.Compared the expression between different sex of the same periods, ER mRNA expression of female were higher than male in 2.75~3.50 d ,and the difference was very significant (P<0.01). We found 2.75~3.50 d were the gonadal differentiation period.The abnormal ER expression may induced embryos death and hybrid infertility in the periods of gonadal differentiation.Those conclusion should be confirmed after further research on ER expression and function in chicken and quail’s embryos respectively of the same period.
动物营养
Effects of β-1,3/1,6-glucan from Saccharomyces cerevisiae on Cellular Immune and Specific Humoral Immune in Weanling Piglets
WANG Zhong;GUO Yu-ming;YUAN Jian-min;YANG Ying;NIE Wei
2007, 38(12):  1316-1322.  doi:
Abstract ( 901 )   PDF (405KB) ( 938 )  
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The present study with weanling piglets rearing in common environment was conducted during an experimental period of 4 weeks to evaluate the effects of dietary supplementation with 50, 100, 200 mg/kg of β-1,3/1,6-glucan extracted from Saccharomyces cerevisiae on peripheral blood lymphocyte subgroups, lymphocyte proliferation and production of classical swine fever (CSF) antibody. Some humoral and cellular immune parameters were measured after 2 and 4 weeks.Lymphocyte proliferation of peripheral blood was determined indirectly by the MTT colorimetric assay method, peripheral blood lymphocyte subgroups were determined by Flow Cytometry using mouse-anti-pig monoclonal antibody, CSF antibody was determined by ELISA Test kit from IDEXX Inc. A total of 160 (Landrace×Large White) weanling piglets at (28±3)d of age were randomly allocated into four treatments according to initial bodyweight and gender, five replicates per treatment, 8 pigs per replicate (per pen). All pigs were vaccinated against CSF at 25 days of age. The results indicated that oral supplementation with β-1,3/1,6-glucan can significantly enchance lymphocyte proliferation of peripheral blood in weanling piglets after 14 days of administration (P<0.01) but no significant effect was observed on d 28 (P>0.05).We also observed that oral supplementation with β-1,3/1,6-glucan can enchance significantly CSF antibody titer on d 21 after inoculation(P<0.05).In the present study, we also found that low dose β-1,3/1,6-glucan (50 mg/kg)has a numerical increase in Th cells, Tc cells, CD4+ T cells, CD8+ T cells and CD4+ /CD8+(P>0.05), but high dose β-1,3/1,6-glucan (100 and 200 mg/kg)can significantly increase the number of CD8+ T cells (P < 0.05), significantly decrease CD4+ /CD8+(P<0.05). Based on above results, we concluded that oral supplementation with β-1,3/1,6-glucan might have a transient and slight influence on immune function of weanling piglets, might be used as immune adjuvant but its actions were influenced by the dose level and times of administration. Low dose and short term of β-1,3/1,6-glucan supplemenation are beneficial to immune but high dose or long period supplemenation might lead to immune maladjustment.
Alleviation Effect of L-Arginine on the Intestinal Damages of Weanling Piglets Induced by Lipopolysaccharide Challenge
HUANG Jing-jing;LIU Yu-lan;ZHU Hui-ling;HOU Yong-qing;DING Bin-ying;FAN Wei;GUO Guang-lun
2007, 38(12):  1323-1328.  doi:
Abstract ( 1338 )   PDF (418KB) ( 888 )  
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The experiment was conducted to study the effect of L-arginine on the content of protein,DNA, RNA and disaccharidase activities in small intestinal mucosa and the content of D-xylose in plasma of weanling piglets after lipopolysaccharide (LPS) challenge.Pigs were allotted into four treatments (6 replicates/treatment and 1 pig/replicate) including: (1) Nonchallenged control (NONC); (2) Challenged control (CHAC); (3) 0.5% arginine (0.5% Arg); (4) 1.0% arginine (1.0% Arg). On d 16 after weanling, pigs in the CHAC, 0.5% and 1.0% Arg groups were injected intraperitoneally with 100 μg/kg BW LPS, whereas pigs in the NONC group were injected with an equivalent amount of sterile saline. D-xylose was orally administrated 2 h at the dose of 1 mL/kg BW after LPS challenge, and blood samples were collected 3 h after LPS challenge. On 48 h after LPS challenge, the pigs were slaughtered to collect intestinal mucosa for analysis. The results showed that: (1) Compared with NONC, the protein content of duodenal mucosa, DNA content of jejunal and ileal mucosa, suctase activities of jejunal mucosa, lactase, suctase, maltase activities of ileal mucosa and D-xylose content of plasma for CHAC were decreased significantly (P<0.05), which indicates that LPS induces intestinal damages. (2) Compared with CHAC, the protein content of duodenal mucosa, DNA content of jejunal mucosa, lactase, suctase, maltase activities of ileal mucosa, D-xylose content of plasma for 0.5% Arg group, and DNA content of jejunal mucosa, suctase activities of jejunal mucosa, lactase, suctase, maltase activities of ileal mucosa, D-xylose content of plasma for 1.0% Arg group were numerically higher, and had no significant difference from those of NONC (P>0.05), which indicates that L-arginine can alleviate the intestinal damage induced by LPS to some degree.
Study on Flow and Digestibility of Amino Acid in Intestine with Different Protected-lysine Levels Medicate into the Rumen of Steers
JIANG Shu-zhen;YANG Zai-bin;LUAN Yu-jing;YANG Wei-ren;XU Teng
2007, 38(12):  1329-1334.  doi:
Abstract ( 796 )   PDF (382KB) ( 680 )  
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The method of basal diets plus amino acid was applied to study the flow and digestibility of amino acid in intestine with different protectedlysine levels of steers. The basal diet was composed of wheat bran, corn and soybean, the nutrition level of energy and protein was supplied according to the nutrition requirement and feed standard of steers. Six healthy crossbred(Limousin×Luxi) steers were selected in the study, which were surgically fitted with permanent fistula in the rumen, the proximal of duodenum and the dismal of ileum. 6×6 Ladin square was adopted in this experiment to study the amino acid flow in duodenum and ileum. The treatments were basal diet plus 0,6,9,12,15 and 18 g/d protectedlysine,respectively.The results showed that the flow of EAA,TAA,Met,Ile,Leu,Lys,Glu in duodenum were increased(P< 0.05), and digestibility of EAA,TAA,Thr,Cys,Val,Met,Ile,Lys,Ala,Tyr,Pro in intestine were increased(P< 0.05). This showed that the optimal accession of lysine level is 15 g/d.
预防兽医
Study on Avian Influenza Virus T Cell Epitopes Binding with Reconstructed Chicken MHC Ⅰ Molecules in vitro
LI Xin-sheng;CHEN Hong-ying;YAN Ruo-qian;GAO Feng-shan;FANG Qin-mei;XIA Chun
2007, 38(12):  1335-1340.  doi:
Abstract ( 1434 )   PDF (1041KB) ( 718 )  
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In order to identify the peptides binding with different type of chicken MHC Ⅰ molecules, the constructed BF2*04sh-linker-β2m,BF2*09sh-linker-β2m,BF2*12sh-linker-β2m,BF2*05wj-linker-β2m proteins were mixed with seven designed and synthesized T cell epitopes (three derived from avian influenza virus, two derived from swine foot-and-mouth diseases virus and two derived from grass carp reovirus) at a respective mole ratio of 1∶10, incubated at 37℃ for 12 h. The mixtures were centrifuged in Amicon Ultra-15 protein Centrifugal Filter (3000) at 4 500 rpm for 45 min to remove unbound nonameric peptides. Then, the retained solution was mixed with 5 mL citrate-phosphate buffer, incubated at 37℃ for 5 min, then centrifuged as above to obtain the filtered solution.The peptides in acid elution solution were concentrated and desalted using Sep-Pak C18 cartridges. The sample plate which obtained the lyophilized peptides dissolved in 10 μL of matrix solution was put into a 4700 Proteomic Analyzer, and the primary mass spectra (MS) and secondary spectra (MS/MS) were conducted.The results revealed that peptides KILTIYSTV and LLLAIVSLV which derived from avian influenza viruses (AIV) could bind with three different types of chicken MHC Ⅰ. The results confirmed that the binding ablity of different MHC Ⅰ with peptides are different, the T cell epitopes derived from different virus are MHC Ⅰ restricted. KILTIYSTV and LLLAIVSLV are avian influenza’s candidate T cell epitopes.
Detection and Localization of the Viral RNA in Bovine Tissues Experimentally Infected with Foot-and-mouth Disease Virus with in situ Hybridization
SHAO Jun-jun;CHANG Hui-yun;LIN Tong;CONG Guo-zheng;DU Jun-zheng;XIE Qing-ge
2007, 38(12):  1341-1344.  doi:
Abstract ( 737 )   PDF (1457KB) ( 709 )  
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RNAs of Foot-and-mouth disease virus in bovine tissues from six experimentally infected cattle was localized and detected by in situ hybridization. Results showed that strong positive staining was viewed in four bovine tongue epithelial tissues 7-28 days post-infection. The strong positive staining particles were observed in the epithelial tissues of larynx and pharynx 7-21 days postinfection. The positive staining was not observed in the tongue epithelial tissues from 35 days post-infection cattle and the normal control cattle. No positive staining was viewed in the tissues from tonsils, lymph nodes and cuneus ungulae of all six experimental cattles. Results indicated that epithelium of tongue, larynx and pharynx were infected by foot-and-mouth disease virus. The results are very valuable and helpful to further elucidate persistent mechanisms of foot-and-mouth disease virus in cattle.
Correlation between Antimicrobial Resistance and Integrons of Escherichia coli Strains Isolated from Bovine Mastitis Milk Samples
WANG Gui-qin;WU Cong-ming;CAO Xing-yuan;SHEN Jian-zhong
2007, 38(12):  1345-1350.  doi:
Abstract ( 762 )   PDF (586KB) ( 728 )  
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This study investigated the correlation between antimicrobial resistance of Escherichia coli strains isolated from bovine mastitis milk samples and the integrons. After acquirement of the antimicrobial resistance patterns, integrase gene was amplified by PCR using degenerate primers. The integrons were confirmed by restriction fragment length polymorphism (RFLP) analysis of positive PCR products, then integrated gene cassettes were examined. The incidence of class Ⅰ integron in the 80 strains of E. coli was 46.25% (37/80). All integrons were revealed as class Ⅰ integron. Neither class Ⅱ nor class Ⅲ integron was detected. Thirty-one strains carried the integrated gene cassttes were detected in thirty-seven integron-positive srtains. The class Ⅰ integron was prevalent in strains which were resistant to more than seven antimicrobials. ClassⅠintegrons harbored dfr and aadA gene cassettes confered resistance to trimethoprim and streptomycin, respectively. The order of main inserted gene cassettes was dfrA17-aadA5. There was a good correlation between integron carried and the multi-resistance, but the antimicrobial resistance patterns did not consist with the gene cassettes.
Cloning,Expression of the Mature Interleukin 15 Gene from Rongchang Porcine and Biological Activities Detection of the Expressed Protein
LUO Ai-fang;GUO Wan-zhu;HAN Guo-quan;SONG Zhen-hui
2007, 38(12):  1351-1356.  doi:
Abstract ( 1249 )   PDF (637KB) ( 770 )  
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The complete open reading frame (ORF)of interleukin-15 gene was amplified from Rongchang porcine peripheral blood lymphocyte stimulated by ConA, which includes 489bp and encodes 162aa. Sequence analysis indicated that the nucleotide shared 99.4% homology with two other porcine IL-15 genes reported previously. Phylogenetic analysis showed that the evolution relationship is comparatively close to the human and mammal IL-15 gene while it is fairly away from poultry. The gene encoding mature IL-15 protein was amplified by PCR from the recombinant plasmid containing IL-15, which includes 345 bp in all. Then it was cloned directionaly into prokaryotic expression vector pET-32a(+) and the fusion expression was induced in E.coli BL21. SDS-PAGE demonstrated that the fusion protein expressed in form of inclusion body is approximately 34 ku, the product of expression account 38.7% of the total bacterium proteins. Western-blotting analysis indicated that there is a specific electrophoresis strip at the place where the relative molecular mass is about 34 ku. A 356 bp specific fragment strip was observed by detection after the recombinant bacteria was transcribed. After the protein was purified roughly and renatured, MTT essay confirmed that it can enhance lymphocyte proliferation obviously. Rongchang porcine interleukin15 mature peptide was expressed successfully in vitro and it possesses certain biological activities.
基础兽医
Effects of Different Ratios of Calcium to Phosphorus on the Formation and Activation of Duck’s Osteoclasts Cultured in vitro
GU Jian-hong;LIU Jun-dong;ZHAO Rui-ying;WANG Fu-min;LI Hui-min;LIU Zong-ping
2007, 38(12):  1357-1361.  doi:
Abstract ( 785 )   PDF (1482KB) ( 793 )  
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Osteoclasts (OC) were isolated from long bones of 1 to 7 days old ducks. In the culture process, different concentration ratios of calcium (Ca) to phosphorus (P) (CCa : CP = 2∶1,1∶1,1∶2) were added to the culture system. The formation and activation of these OC were detected by phasecontrast microscopy, histochemical staining for tartrate-resistant acid phosphatase (TRAP) staining at the 1st and the 3rd day, and detection of lacunar resorption through scanning electron microscopy at the 7thday. For three days culture, the number of OC in experimental group was more than control group very significant. There was no difference between CCa∶ CP = 1∶1 group and CCa∶ CP = 2∶1 group.The difference among other groups was very significant at the 3rd day. The results demonstrated that CCa∶ CP = 2∶1 inhibits bone absorbing activity by inhibitng the formation and activation of OC.
Changes of Number of Mucosal Immunity-associated Cells in Small Intestine of Goats at Different Developing Stages
CHEN Fu-jü;CHEN Yao-xing;WANG Zi-xü;LIU Yun-fang;XIE Dian
2007, 38(12):  1362-1367.  doi:
Abstract ( 756 )   PDF (2022KB) ( 955 )  
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In order to reveal the underlying rules of small intestinal mucosal immunity in goat at different developing stages, 0.5, 2 and 12 months, respectively, the distribution and number changes of intraepithelial lymphocytes (IELs), goblet cells (GCs) and mast cells (MCs) were investigated by histological and histochemical methods.The results showed that the number of IELs and MCs were significantly increased (P<0.05) with age, while the number of GCs decreased substantially (P<0.05).On the other hand, the distribution of IELs was decreased from duodenum to ileum, and the number of IELs in duodenum and jejunum was 84.45% and 98.14% of ileum respectively at 0.5 month old. The distribution of MCs was similar to that of IELs. The number of GCs, however, increased from duodenum to ileum.The results suggested that GCs played a crucial defensive barrier role in the early postnatal period, while the effect of IELs and MCs enhanced concomitant with postnatal development of the gut mucosal immune system.
临床兽医
Pharmacokinetic Study of the Baicalin from Ultramicro-pulverised Powder of Huanglian jiedusan(HLJDS) in Rabbits
MA Yu-fang;LIN Xue-ling;YU Dao-jin;YAO Jin-shui;HUANG Xiao-hong;HUANG Yi-fan
2007, 38(12):  1368-1372.  doi:
Abstract ( 753 )   PDF (391KB) ( 851 )  
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High performance liquid chromatography(HPLC) was applied to determine the concentrations of plamsa baicalin in two groups of rabbits after administering the ultramicro-pulverised powder and the ordinary powder of HLJDS (in water suspension) by gastrogavage respectively. The plasma concentrationtime data of baicalin were dealt with Pharmaceutical Kinetics Software (PKS). Pharmacokinetics characteristics of both the ultramicro-pulverised powder and ordinary powder of HLJDS were compared. The results showed that the best pharmacokinetic model of baicalin in either group was two-compartment open model.The main pharmacokinetic parameters of baicalin of ultramicropulverised powder were as follows: Ka=0.497 h-1, t1/2α=2.556 h,t1/2β=8.46 h,AUC0→∞=3.401 μg·h/mL,Vd=6.752 L/kg, Tpeak=3.005 h, Cmax=0.311 μg/mL;The main pharmacokinetic parameters of baicalin of the ordinary powder were as follows:Ka=0.419 h-1, t1/2α=2.74 h,t1/2β=5.83 h,AUC0→∞=2.611 μg·h/mL,Vd =12.890 L/kg,Tpeak=3.645 h,Cmax=0.215 μg/mL. As compared to the baicalin from ordinary powder, the relative bioavailability of baicalin from ultramicro-pulverised powder was increased by 30.26%. These data presented above indicated that the bioavailbility of baicalin from HLJDS could be greatly improved after processing and preparing by the technique of ultromicro-pulverization.
The Effect of Selenium Compounds on Oxidative Damage in Mice
WANG Jin-qiu;LIN De-gui;WANG Lei;SONG Xiao-yu;HAN Chun-yang
2007, 38(12):  1373-1376.  doi:
Abstract ( 1225 )   PDF (333KB) ( 646 )  
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To study effects of selenium compounds on the oxidative damage, sixty ICR mice were randomly divided into six groups: control group, radiation group, high and low dose Na2SeO3 groups, high and low dose MSeC groups, 10 mice per group. Intragastric administration of 0.3 mL solutions was performed once per day and lasted for 14 days,high and low dose Na2SeO3 groups were given Na2SeO3 (20 and 10 μg/mL, respectively), high and low dose of MSeC groups were given MSeC (20 and 10 μg/mL, respectively), same volume water was given to control group and radiation group instead. 7 days later all groups accepted one time radiation except the control group, at the dosage of 4.5 Gy, and the oxidative damage model was established. During the experiment, immunity and antioxidation functions were measured respectively. 3 days after radiation,white blood cells (WBC) of groups accepted radiation decreased significantly; 6 days after radiation, WBC of radiation group and the two Na2SeO3 groups continued declining, and the two MSeC groups increased steadily. In radiation group the GSHPx, SOD activities, GSH decreased significantly and MDA increased significantly compared with normal control group. There was no difference between normal control group and the groups accepting selenium compounds. The Spleen Index (SI) and Thymus Index (TI) of those groups accepting selenium compounds had significant difference with radiation group. Radiation results in oxidative damage and selenium compounds ,especially methylselenocysteine (MSeC)has protect effects on anti-oxidative damage, and the protect effect of high dosage within the range of safe dosage is obvious.
研究简报
SNP Detection of Intron 1 of PRL Gene and Its Associations with Egg-laying Traits in Gaoyou Duck
ZHANG Tang-jie;LI Hui-fang;CHEN Kuan-wei;ZHAO Yong-gao;CHANG hong;XUE Min-kai;ZHANG Jing-xin
2007, 38(12):  1377-1382.  doi:
Abstract ( 732 )   PDF (1006KB) ( 717 )  
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In present study, SNP of prolactin(PRL) gene intron 1 of Gaoyou duck were detected by PCR-RFLP and DNA sequencing. The correlation analysis between the polymorphism of PRL gene intron 1 and egg-laying traits was carried out. The results indicated that there were two Dra I digestion sites in intron 1 of PRL gene, but only one was polymorphic. A T/C mutation at the position of the 1 326 bp of the PRL gene was found by DNA sequencing, and resulted in 3 genotypes (AA,AB and BB). The frequencies of genotype BB and allele B were the highest. The least square analysis showed that the BB ducks had higher egg weight at the age of 30 weeks than AB ducks (P<0.01), the proportion of double-yolk of the AB ducks was higher than that of BB ducks (P<0.05), but there was not significant difference in egg number, the longest days of continual laying and the body weight at first laying egg among 3 genotypes.
Diversity of Immune Traits between Chinese BeijingYou Chicken and White Leghorn
Diversity of Immune Traits between Chinese Beijing-You Chicken and White Leghorn
2007, 38(12):  1383-1388.  doi:
Abstract ( 1358 )   PDF (379KB) ( 811 )  
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The objective of this project was to examine the diversity of immune traits between two chicken populations which is the Beijing-You Chicken (BJY), a Chinese indigenous breed, and the other is White Leghorn (WL), a laying type breed. 250 healthy female chickens were hatched during the same period and were reared under the same farming system from 0-170 days of age. Heterophil/lymphocyte ratio (H/L), the blood white cell count(BWCC), the sheep red blood cell (SRBC) antibody titer and Avian Influenza (AI) and Newcastle Disease (ND) antibody titer were determined on bird from each group. The results showed that: (1)There were significant differences in the H/L ratio between two breeds (P<0.001). Mean of H/L ratios were 0.36±0.21 and 0.52±0.51 for BJY and WL, respectively. The total number of Blood White Cells of BJY were higher than that of WL distinctly (P<0.001) .(2)No differences were found in SRBC antibody titers between two breeds (P>0.05); (3) H5N1 AI antibody titers were measured at 50, 71,100,134 days of age after vaccine were injected at 14 and 28 days of age. The AI antibody titers of 100 days of age are highest among the titers of four ages for both groups. There were significant differences between two breeds, titers of 50,71,100,134 days of age for Beijing-You Chicken are higher than that of White Leghorn chicken ,respectively(P<0.001); (4) At the same age like AI, LaSota ND antibody titer were measured at 50, 71,100, 134 days of age after vaccine were injected at 14 and 28 days of age. The antibody titer of 71 days of age is highest among the titers of three ages for WL, while for BJY chickens the highest one is the titer of 50 days of age. Titers of 50, 100 ,134 days of age of BJY is significantly higher in BYC than that in WL ,respectively(P<0.001, P<0.05, P<0.001), while there was no difference for titers of 71 days of age between two groups. The results demonstrate that the two breeds differed markedly in immune traits. The general immune performance of BJY chickens, the resistance to stress and disease those, as some other local breeds, were superior to those of WL chickens.
Production of Transgenic Embryos Expressing Green Fluorescent Protein by Nuclear Transfer of Preadipocytes in Pig
PAN Deng-ke;WAN Rong;LIU Ji;MU Yu-lian;ZHANG Li;LI Kui;FENG Shu-tang
2007, 38(12):  1389-1393.  doi:
Abstract ( 1416 )   PDF (866KB) ( 639 )  
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Primary culture of porcine preadipocytes was established using fat tissue from an adult pig. The preadipocytes to re-differentiate induction into mature adipocytes was confirmed by oil red O staining. A porcine preadipocyte (PreA) lines were transfected with pEGFP-N1 plasmid DNA by lipofectinmediated way. GFPPreA clones were obtained by G418 selection. PreA, fetal fibroblast (FF), and GFP-PreA were used as nuclear donors for somatic cell nuclear transfer in pig. The results were as follows: For the blastocyst rate of cloned embryos, no significant difference was observed between PreA and FF (8.8% vs 8.3%, P > 0.05). Blastocyst rate of reconstructed with GFP transfected PreA decreased in comparison to those derived from nontransgenic PreA.(3.7% vs. 8.8%,P > 0.05)。The results indicate that GFP transfected PreA can produce procine transgenic blastocytes. In spite of low blastocyst rate, cloning pigs from preadipocyte is possible.
Effects of Biological Additive and Moisture Content on Fermentation Quality of Alfalfa and Timothy Silages
ZHUANG Yi-fen;ATAKU Kazuo;ZHANG Wen-chang
2007, 38(12):  1394-1400.  doi:
Abstract ( 1453 )   PDF (484KB) ( 743 )  
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Two kinds of alfalfa and timothy with high and low moisture content were made silages to study the effects of alfalfa fermented with green juice (AFGJ),timothy fermented green juice (TFGJ),lactic acid bacteria(LAB),cellulose (CEL) and lactic acid bacteria(LAB)+cellulose (MIX) on fermentation quality of alfalfa and timothy silages. Each treatment had two repeats. Silages were fermented for 50 days. The fermentation quality of silages were observed. The fermentation quality of alfalfa and timothy silages were significantly affected by additives(P<0.05), all the additives improved the fermentation quality of alfalfa and timothy silages with high and low moisture content, but there were no significant role in high moisture content alfalfa silages with AFGJ and TFGJ. The fermentation quality of alfalfa and timothy silages were significantly affected by the moisture content(P<0.05),the fermentation quality of both alfalfa and timothy silages with low moisture content were better than that with high moisture content silages. There were significant interaction between the additives and moisture content(P<0.05). The fermentation quality of alfalfa silage with high and low moisture content with MIX were the best; the fermentation quality of timothy silage with high moisture content with FGJ and TFGJ were similar,but the quality were better than other additives. The fermentation quality of timothy silage with low moisture content with AFGJ,TFGJ,LAB and MIX were similar ,but the quality were better than CEL. The results showed that the treatment by reducing the moisture content and choicing the additives could improve silage quality.
Effect of Inulin on Microflora of Culture Medium with Fresh Feces of Growing Pigs
GU Xian-hong;ZHANG Ming-tao
2007, 38(12):  1401-1404.  doi:
Abstract ( 625 )   PDF (324KB) ( 615 )  
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In order to determine the ferment properties and bifidogenic action of inulin, an in vitro fermentation experiment was conducted to anaerobically incubate fresh feces of growing pigs with inulin (0, 1, 2, 4, 6, 8 g/L, respectively) for 24 h at 37℃, and then determine pH and the populations of Escherichia coli, Lactobacillus, Bifidobacterium and total bacteria in the culture medium. The results showed that pH and the number of Escherichia coli linearly decreased (P<0.001),and the number of Lactobacillus, Bifidobacterium and total bacteria in the culture medium linearly increased (P<0.01 or P=0.059). It was concluded that there were dosageresponse relationships between inulin additive level and bacteria (Escherichia coli, Lactobacillus, Bifidobacterium and total bacteria) amount or pH in the culture medium.
Analysis of Immune Responses Induced by Recombinant Secretion and Non-secretion Type Lactobacillus casei Expressing Porcine Parvovirus VP2 Protein via Oral Immunizations
. College of Veterinary Medicine;Northeast Agricultural University;Harbin 000;China; . Northeast Forestry University;Harbin 000;China
2007, 38(12):  1405-1409.  doi:
Abstract ( 705 )   PDF (590KB) ( 734 )  
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The objective of this research was to compare the immune responses induced by mucosal immunizations with recombinant secretion and nonsecretion type Lactobacillus casei expressing porcine parvovirus (PPV) protective antigen VP2 protein. The mice of vaccine groups received recombinant L. casei strains constructed in this work harboring cell surface expression system pPG-VP2 or secretion expression system pPGS-VP2, respectively. The control mice were immunized with equivalent doses of PBS. All the mice were immunized via orogastricroute. The immune protocol was administered on three consecutive days at day 0, 1 and 2. A booster immunization was given at day 14, 15 and 16 and a second booster was given at day 28, 29 and 30. Specific anti-PPV-VP2 secret immunoglobulin A (sIgA) antibody in the feces and intestines mucus and anti-PPV-VP2 IgG antibody in the serum of immunized mice was detected by indirect enzyme linked immunosorbent assay (ELISA) after orogastric immunization. The results showed that the levels of mucosal and systemic immune responses induced by secretion type recombinant lactobacilli were higher than that induced by cell surface expression type recombinant lactobacilli.
Immune Efficacy of the Recombinant Plasmids Expressing HA Gene of H1N1 and H3N2 Subtype SIV in Mice
HU Yong-hao;BAO Shi-jun;ZHAO You-shu;WEN Feng-qin;YAO Xue-ping;YANG Huan-liang;CHEN Hua-lan
2007, 38(12):  1410-1413.  doi:
Abstract ( 867 )   PDF (339KB) ( 685 )  
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BALB/c mice were inoculated with the recombinant plasmids(pCI-H1-HA,pCAGGS-H1-HA,pCI-H3-HA and pCAGGS-H3-HA) which included the HA gene of H1N1 and H3N2 subtype SIV to screen valuable vaccine candidate. The immune efficacy was evaluated by detecting HI antibody titers,analyzing weight changes of mice and examining virus titers in lungs. The results indicated that the constructed DNA vaccines induced immune responses in BALB/c mice Meanwhile, by the statistical analysis of the weight changes of BALB/c mice, there were high significant differences between the immunization groups and the control(P<0.01), and the DNA vaccines constructed with pCAGGS as the expression vector had better immune efficacy than that of DNA vaccines constructed with pCI(P<0.05).
Analyisis of Classical MHC Class I Alleles Expressed by PeripheralBlood Mononuclear Cells from 5 Horses in Heilongjiang Province
MA Jian;GUO Wei;SHEN Nan;KONG Xian-gang;SHEN Rong-xian;ZHOU Jian-hua
2007, 38(12):  1414-1420.  doi:
Abstract ( 1572 )   PDF (2329KB) ( 583 )  
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To study CTL epitopes in EIA attenuated vaccine, cDNA sequences encoding majority region of classical MHC class I from 5 horses were amplified by RT-PCR and analyzed. The amplified cDNA fragments included the polymorphic peptide binding region, the more conserved regions of alpha 3, transmembrane domain and cytopasmic domain. The RT-PCR products were synthesized from total RNA of PBMC and then cloned into pMD-18T vector.21-23 clones from each horse were randomly picked up for DNA sequencing. These sequences were further analyzed for their alleles. Our data revealed that each clone contained 3-5 alleles, which indicated there were at least 2-3 loci in MHC class I in horse genome.This results were consistent with the published data of other researches. In addition, the expression variation of classical MHC I alleles between each individuals were different. There was no identical or highly similar allele among horses No.6, No.7 and No.10. The other animals had at least one allele that shared all or more than 98% of nucleotide sequence when compared each other(data not show). These results not only provide fundamental information for our on-going study on CTL epitopes and EIAV-induced immune responses, but also served as important data for polymorphic analysis of horse classic MHC class I in China.