Acta Veterinaria et Zootechnica Sinica

Melanocortin-1 Receptor (MC1R) Gene Variation in 4 Cattle Populations

TANG He;GAO Ying-kai;MIAO Yong-wang;

2010, 41(6): 639-643. doi:

Abstract ( 738 )

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In order to evaluate the influence of the extension (E) locus on the color of cattle′s coat, the genetic characteristic and variations of the encoding region for melanocortin-1 receptor (MC1R) gene in four cattle populations sampled in Yunnan Province, including three indigenous populations and an imported breed, were assayed and analyzed by using DNA sequencing and PCRRFLP techniques. The alleles E+, e and genotypes E+/E+, E+/e, e/e were identified in Wenshan Yellow cattle, Zhaotong Yellow cattle and Shorthorn cattle. The frequencies of E+ and e alleles were 0.54 and 0.46 in Wenshan Yellow cattle, 0.70 and 0.30 in Zhaotong Yellow cattle. The alleles E+, ED and e were identified in Diqing Yellow cattle with the frequencies of 0.76, 0.14 and 0.10, respectively. The genotypes E+/E+，ED/ED, E+/ED, E+/e and ED/e were identified in Diqing Yellow cattle, with the only exception of e/e. In contact with the coat color of the selected samples, it was found that E locus had an important influence on determinating the color of the cattle′s coat. E+ and ED alleles were associated with black coat, and e allele was connected to red coat. Whereas yellow, brown and red fur were also influenced by other loci.

Genetic Polymorphisms within 5′Flanking Region of Bovine A-FABP Gene and Its Relationships with Beef Performance Traits in Qinchuan Cattle

LIU Yan-yan;ZAN Lin-sen;WANG Hong-bao;JI Shu-han;FAN Yue-yuan

2010, 41(6): 644-650. doi:

Abstract ( 748 )

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The sequence of 1 441 bp from 5′flanking region of A-FABP gene was sequenced. The sequencing results showed only one SNP, namely -389（T/C）, then the genotypes of five native cattle breeds(Qinchuan cattle, Nanyang cattle, Luxi cattle, Jiaxian Red cattle and Xia′nan cattle) in China were examined by PCRSSCP. The SNP was detected in each group and devided into three genotypes: AA, AB and BB. χ² test indicated that this polymorphism site fit Hardy-Weinberg equilibrium（P>0.05）in five native cattle breeds. The study of relationships between some beef performance traits and genotypes in Qinchuan cattle showed that slaughter weight, carcass weight, moisture content of individuals with genotype AB were significantly lower than that of individuals with genotype AA and BB (P>0.05), but no significant difference between AA and BB genotypes. The marbling score of individuals with genotype BB was very significantly better than that of individuals with genotype AB（P<0.01）, and significantly better than that of individuals with genotype AA（P<0.05）, but no significant difference between AB and AA genotypes. The results indicate that this locus is a probable major QTL affecting marbling score and moisture content or tightly links to those genes affecting beef performance traits, it could be a candidate molecular marker for the beef cattle breeding.

Cloning and Activity Analysis of Promoter of Chicken Lipoprotein Lipase Gene

BI Jing;DING Ning;WANG Ning;LI Hui

2010, 41(6): 651-656. doi:

Abstract ( 663 )

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The objective of this study was to analyse the promoter structure and activity of the chicken lipoprotein lipase (LPL) gene. 2 kb 5′ flanking region of chicken LPL gene was amplified by PCR, cloned and sequenced. Subsequently, the luciferase reporter gene plasmids containing LPL gene promoter and its serial promoter deletions were constructed and introduced into chicken fibroblast cell line DF-1. The expression of luciferase was measured in transient expression assays. Bioinformatics analysis showed that the promoter fragment contained binding sites of transcription factors Oct1, GC box, CCAAT, GATA, AP1 and it also had a CpG island from －575 to +137 bp. Luciferase reporter assays demonstrated that the promoter region from －359 to +163 bp confered basal transcriptional activity and the region from －601 to +163 bp confered the most powerful transcriptional activity of the chicken LPL gene. These results indicated that chicken lipoprotein lipase gene could be regulated by variety of transcription factors and upstream elements. This study pays the way for further research on the regulation mechanism of chicken LPL gene.

Studies on Differentially Expressed Genes in the Ovary of Zi Geese

KANG Bo;JIANG Dong-mei;GUO Jing-ru;YANG Huan-min

2010, 41(6): 657-663. doi:

Abstract ( 687 )

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To elucidate the molecular mechanism of the laying process, the differentially expressed genes in the ovary of the laying Zi geese compared with the prelaying geese were identified, and their functions were classified. Suppression subtractive hybridization (SSH) was employed to isolate the differentially expressed genes, and their functional categorizations were predicted by the tool GOTM in search of the Gene Ontology database. The results showed that the subtractive efficiency of the SSH-cDNA library was approximately 20 folds, and the clones were ranged from 300 bp to 750 bp. Eighty-six positive clones were subjected to sequencing and further analysis. Sequence analysis showed that 11 known (including a mitochondrial gene) and 4 unknown ESTs were highly expressed in the ovaries of laying geese compared with prelaying geese. Eleven known ESTs belong to the functional groups of binding, catalytic activity, enzyme regulator activity, transporter activity and other activity. These results indicated that the SSH-cDNA library of the Zi geese ovary was successfully constructed and the expression levels of 15 differentially expressed ESTs were up-regulated in the laying Zi geese ovary compared with the prelaying Zi geese ovary, these ESTs could play an important role in the process of egg-laying.

Bottleneck Effect Analysis of Chinese Goat Breeds Using Microsatellites

WANG Zhi-gang;WU Jian-ping;LIU Chou-sheng;QIU Xiao-tian

2010, 41(6): 664-670. doi:

Abstract ( 695 )

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The present study was conducted to evaluate different Chinese goat breeds for mutation drift equilibrium and occurrence of any recent genetic bottleneck. A total of 2 027 animals from forty different breeds were analyzed with a set of 9 heterologous microsatellite markers. Under the two-phase model, all the goat breeds except Jining Qing goat were found to be in mutation drift equilibrium when evaluated by Wilcoxon signed-rank test. The result revealed significant heterozygosity excess suggesting possible cryptic demographic bottleneck in Jining Qing goat.

Expression of Growth Differentiation Factor 9 in Goat COCs during in vitro Maturation

ZHENG Ai-yan;WU Man;SHANG Li-qing;DING Jia-tong

2010, 41(6): 671-677. doi:

Abstract ( 776 )

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To analyze the relationship between GDF9 gene and expansion of cumulusoocyte complex (COCs), the expression of GDF9 in goat COCs during oocytes in vitro maturation (IVM) (0, 6, 12, 18, 24 and 27 h) was detected by fluorescent quantitative RTPCR method. The result showed that GDF9 mRNA were expressed at low levels in immature oocytes and increased to the highest level at 12 h of IVM, which coincides with the time of cumulus cell expansion, then declined slowly during IVM, with a slight increase at the end of IVM. The expression pattern of GDF9 suggested that it might be important in cumulus expansion and oocyte maturation of goat.

Apoptotic Law of Porcine Cloned Embryos during Development in vitro

JU Shi-qiang;GUO Hui-li;WANG Li-qin;LIN Peng-fei;LU Qing;RUI Rong

2010, 41(6): 678-684. doi:

Abstract ( 732 )

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The purpose of this experiment was to determine the relationship between embryonic development and apoptosis in porcine nuclear transfer (SCNT) embryos during development in vitro. The apoptotic incidence was evaluated via Comet assay, and the mRNA expression of gene implicated in antiapoptosis (Bcl-2) was also examined using Real-time RT-PCR. The results showed that the SCNT embryos had a similar cleavage rate (77.6% vs 80.3%, P>0.05) but a lower developmental rate of blastocysts (18.9% vs 24.5%，P>0.05) compared to in vitro fertilization (IVF) embryos. The results of Comet assay demonstrated that the apoptotic rate both of the two types of embryos maintained increased tends along embryonic developmental stages, but the SCNT embryos exhibited higher apoptotic rate from 16-cell to morula stage compared to IVF embryos. In the whole developmental process, the relative abundance of Bcl-2 mRNA was gradually reduced both in SCNT and IVF embryos, and the expression of Bcl-2 mRNA in SCNT embryos was significantly lower than that of IVF embryos at 16-cell and 32-cell stages. It was concluded that the higher degree of apoptotic incidence may be associated with the low efficiency of porcine SCNT in vitro production.

Effect of Yeast β-glucan and Antibiotics on Growth and IntestinalMicroflora in Early-weaning Calves

ZHOU Yi;DIAO Qi-yu;TU Yan;YUN Qiang;GUO Xu-dong

2010, 41(6): 685-691. doi:

Abstract ( 683 )

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This experiment was conducted to investigate the effect of yeast β-glucan and bacitracin zinc on growth performance and intestinal microflora in earlyweaning calves. Twenty healthy Holstein male neonatal calves were randomly allotted to four treatments with five replicates each. The calves were fed with the diets supplemented with 0 (Treatment A), 75 mg·kg^-1 yeast β-glucan (Treatment B and C), 60 mg·kg^-1 bacitracin zinc (Treatment D). The experiment lasted for 28 days. On d 21, the calves of Treatments A, B and D were challenged orally with Escherichia coli (O141：K99), and calves of Treatment C were fed normally. Feed intake (FI) was recorded daily, body weight was measured and average daily gain (ADG) was calculated every two weeks, and fecal samples were collected from superior ampullar mucosa of the rectum to determine the intestinal microflora. The results showed as follows: Compared with Treatment A, the ADG of calves in Treatment B increased by 26.18% and 24.93% in the two phases before the Escherichia coli challenged (P<0.05), the ADG of calves in Treatment B and D increased by 30.38% and 30.82% after the Escherichia coli challenged (P<0.05). As for the F/G, which in Treatment B and D were significantly lower than that in Treatment A (P<0.05). The amount of Escherichia coli in rectum was rapidly increased after the challenged, compared with Treatment A, the amount of Escherichia coli in rectum at 12 and 24 h in Treatment B and D were significantly decreased (P<0.05), and the amount of Lactobacillus was significantly decreased in the Treatment D (P<0.05). PCR-DGGE of 16S rDNA was used to investigate the similarity index, the band number in Treatment C was significant higher than that in Treatment A and D (P<0.05).The degree of similarities of treatments were range from 50% to 75%. According to the results, β-glucan could improve the growth of claves and adjust the structure of intestinal microflora, thus using β-glucan in calves feed may decrease the usage of antibiotics.

Isolation and Identification of Urease from the Rumen Content of Holstein Cows by a Culture-independent Strategy

ZHAO Sheng-guo;WANG Jia-qi;LIU Kai-lang;LI Dan;YU Ping;BU Deng-pan

2010, 41(6): 692-696. doi:

Abstract ( 1074 )

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This experiment was conducted to isolate urease active protein from cow rumen. Rumen contents were collected from Holstein cows via fistula. Cell free rumen fluid and ruminal cellular protein fluid were collected by centrifugation and ultrasonication. After precipitated by the addition of solid (NH4)2SO4 to 85% saturation and dialyzed, the fluid was applied to a HiTrap Capto Q ion exchange column. The fractions with urease activities were pooled, and concentrated by lyophilization. Then urease was separated by native-PAGE, and identified by modified Fishbein staining. The urease strap was excised from the gels, digested by trypsin and then analyzed by LCMS/MS. Searches of the peptide mass fingerprint data against NCBI databases were performed with the SEQUEST software. The results showed that higher activity urease was isolated and purified from cell free rumen fluid and ruminal cellular protein fluid. After staining, one brownish red strap containing urease from ruminal cellular protein fluid was observed. Three kinds of urease from Streptococcus thermophilus, Streptococcus salivarius and Bacillus halodurans were detected by mass spectrum. These results demonstrated the feasibility of isolating urease protein directly from rumen mixture without microorganism cultivation and this strategy could be expected to facilitate the research on uncultured microorganisms in rumen.

Comparisons of the Quality and Nutritional Value of Whole-plant CornSilage at Various Depths under the Top Surface of Silo

YAN Gui-long;CAO Chun-mei;DIAO Qi-yu;HONG Mei;WANG Jian-hong

2010, 41(6): 697-704. doi:

Abstract ( 633 )

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The effect of simple covering of silo on the quality and nutritional value of whole-plant corn silage was evaluated in this study. The silage samples were taken from 10, 20, 40, 80, 160 or 320 cm under the top surface of the aboveground silo and analysed for their quality, chemical composition and in vitro DM digestibility. The results showed that simple covering of the silo significantly affected the quality, chemical composition and in vitro DM digestibility of whole-plant corn silage from 0 to 40 cm under the surface of the silo, as indicated by the higher population of fungi and lactobacilli, the higher content of NDF, ADF and butyric acid, the higher pH value but the lower content of total sugars in the silage. Furthermore, the quality, chemical composition and in vitro DM digestibility of whole-plant corn silage were also affected by the depths under the top surface of the silo, as sampling depths increased, the contents of DM, total sugars and WSC, pH values and DMD decreased gradually, but the contents of lactic acid, VFA, NDF and ADF increased, with the populations of fungi and lactobacilli, the contents of crude protein, EE and ash remaining unaffected.

Gene Cloning and Bioinformatics Analysis of Outer Capsid Protein VP7 genefor Giant Panda Rotavirus CH-1 Strain

YAN Qi-gui;LEI Yan;ZHANG Zhi-he;WANG Cheng-dong;YANG Ai-guo;WANG Xu;ZUO Lan;XIAO Yang

2010, 41(6): 705-710. doi:

Abstract ( 1302 )

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The VP7 gene coding outer capsid protein of giant panda rotavirus（GPRV）strain CH-1 was cloned by RT-PCR from GPRV infected MA-104 cells，and sequencing and bioinformatic analysis were then conducted to investigate the structure and function of GPRV VP7 protein. The function of this protein was predicted by bioinformatics software and the phylogenetic tree of VP7 gene was constructed by CLUSTALW software. The results indicated that a length of 1 042 bp GPRV VP7 protein encoding gene was obtained（GenBank accession number：GU188284）. Bioinformatics analysis showed that the sequence contains the complete coding sequence of VP7 protein, which is 981 bp, encoding 326 aa. The molecular weight, isoelectric point, estimated halflife, instability index of GPRV VP7 were 37 354.8 Da, 4.76, about 30 hours and 26.71, respectively. Grand average of hydropathicity of VP7 protein was -0.015, while the hydrophobicity was between -2.344 and 3.567. The protein has one signal peptide between 1 and 24 site, two transmembrane regions 4—23 site and 33—53 site and 12 antigenic determinants predicted by online analysis. Motif searching showed that VP7 protein may have 2 Nglycosylation sites, 5 protein kinase C phosphorylation sites, 4 casein kinase Ⅱ phosphorylation sites, 1 tyrosine kinase phosphorylation site, 4 Nmyristoylation sites, 1 prokaryotic membrane lipoprotein lipid attachment site and 1 grampositive cocci surface proteins “anchoring” hexapeptide. The phylogenetic tree showed that the evolution distance of giant panda rotavirus VP7 gene is homogeneous to human rotavirus A. The research obtained the clone of GPRV VP7 gene, and displayed some new clues for further studying the biological functions of the gene and establishing diagnosis method of the virus.

Prokaryotic Expression of NSP2 Major Antigen Region of Highly PathogenicPRRSV and Development of an Indirect ELISA Based on the Expressed Protein

LIN Hua;GUO Wan-zhu;ZHANG Bo;CHEN Di-shi;CHEN Yang;WANG Xiao-yu;XU Zhi-wen;WANG Yin;ZHU Ling

2010, 41(6): 711-716. doi:

Abstract ( 718 )

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The present experiment was performed with the objective of establishing an indirect ELISA for detection of PRRSV with the purified NSP2 fusion protein as coating antigen. The major antigen region of NSP2 was successfully amplified by RT-PCR from a highly pathogenic PRRSV (SCMS08 strain), and inserted into pET-32a(+) vector. The recombinant plasmid was transformed into E. coli Rosetta 2 (DE3) and induced with IPTG. The fusion protein was treated with Ni-chelated chromatography under denaturing conditions. With the purified fusion protein as coating antigen, an ELISA for detection of PRRSV was established. SDS-PAGE showed that the fusion protein was expressed at high level in Rosetta 2. Western blot analyses showed that the expressed protein was recognized specifically by anti-His monoclonal antibody as well as PRRSV positive serum. In the optimized NSP2-ELISA, the fusion protein was coated at 1.7 μg·mL^-1 and swine serum samples were diluted at 1∶40. About 185 serum samples were detected by the method and IDEXX-ELISA kit, respectively. The agreement ratio between the two methods reached at 90.8%. The results indicated that NSP2-ELISA was specific, sensitive and suitable for routine diagnosis of PRRS and also for epidemiological surveys.

Genetic Variation Analysis and Animal Pathogenicity Test for PRRSV TP Strain P4, P60, P90 Genome

ZHAO Li-chan;SU Run-huan;DENG Yu-xiu;WANG Dong-dong;YANG Ming-liu; LI Chun-mei;LUO Xiao-fei;SONG Yan-hua

2010, 41(6): 717-725. doi:

Abstract ( 831 )

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The aim of this experiment was to study the genetic variability of the three different generations of porcine reproductive and respiratory syndrome virus (PRRSV) TP strain, P4, P60, P90. RT-PCR, 5′ RACE and 3′ RACE were employed to amplify the virus genome of the three generations of PRRSV TP strain, and the genomic analyses were conducted using some bioinformatics software including the clustalx and DNAStar, and the animal pathogenicity test. The sequences of the TP strain P4, P60, P90 were 15 346, 15 394 and 15 393 bp, respectively. The variations of the ORF1a gene and 3′UTR were obvious in these three generations genome sequences, especially the poly(A) tails in the 3′UTR region，they were 26, 74 and 73 bp, respectively. Comparing the genome sequences of TP P90 strain with TP P4 strain, there found 58 nucleotide mutations that resulted in 27 amino acid changes. The antigenic index and surface probability of GP5 protein of TP P90 strain were increasing than P4 and P60. Moreover, the virulence test indicated that the pathogenicity of TP P4 is stronger than the TP P60, P90 strain. The results showed that the genome sequences of the three different generations has some variations, especially between passage 4 and 60. The PRRSV TP P4, P60, P90 all belong to the PRRSV America strain branch, and display the highest nucleotide identity to the high pathogenic PRRSV JXA1 strain, 99.5%, 99.2% and 99.3%, respectively.

Antigenic Distribution and Expression Regularity in the Different Doses ofpcDNA-DPV-gC DNA Vaccine in Tianfu Ducklings

SHEN Fu-xiao;CHENG An-chun;WANG Ming-shu;JIANG Jin-feng;JIA Ren-yong;ZHU De-kang;CHEN Xiao-yue;SUN Tao;YANG Jin-long;

2010, 41(6): 726-734. doi:

Abstract ( 1055 )

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The objective of the present study was to develop and apply an indirect immunohistochemistry method for detecting the antigenic distribution and expression regularity of pcDNA-DPV-gC in the experimentally vaccinated duckling tissues. Three-week-old Tianfu ducking were inoculated with different doses (50, 100 and 200 μg) of DPV gC DNA vaccine by intramuscular injection. At intervals of 4 h, 12 h, 1 day, 3 days, 5 days, 7 days, 2 weeks, 4 weeks, 6 weeks, and 10 weeks postvaccination (p.v.), two ducklings were randomly euthanatized and their organs (liver, spleen, lung, kidney, pancreas, brain, thymus, Harderian gland, bursa of Fabricius, duodenum, caecum, rectum and muscle of injected spot) were collected. Meanwhile, the indirect immunohistochemistry method was established to detect the antigenic distribution and expression regularity of pcDNA-DPV-gC in the experimentally vaccinated ducking tissues. The results showed that:①The positive staining were found in the liver, duodenum, caecum, rectum and muscle of injected spot at 1 day p.v., and the immunoreactivity in the muscle of injected spot was stronger than other organs. In addition, a drastic reduction of DPV gC antigen level were observed after the highest immunoreactivity at 2 weeks p.v., but still had a detectable vaccine antigen level in the liver, brain, duodenum, caecum and rectum at 10 weeks p.v.. Interestingly, there was scarcely detected positive signal in the pancreas throughout the immunologic process; ②The positive staining reaction for pcDNA-DPV-gC had a conspicuous discrepancy among the different tissues. The positive immunogenicity was mainly found in the liver, spleen, bursa of Fabricius, brain, duodenum, caecum, and rectum, which served as the principal sites for antigen localization. The DPV gC encoding glycoprotein could be expressed in the various types of cells, especially in the lamina propria mucosae cells of intestinal tract, the lymphocyte of splenic white pulp and red pulp and the nerve cement cells of pallium. ③According to the immunogenicity intensity and duration time, the positive staining in all tissues were in below order: duodenum, caecum, rectum > liver > bursa of Fabricius > spleen > brain > muscle of injected spot > thymus > lung > Harderian gland > kidney > pancreas. ④The signal intensity and duration time of different doses were in below order: 200 μg group > 100 μg group > 50 μg group. Different doses of pcDNA-DPV-gC could continuously express DPV gC encoding glycoprotein as early as 1 day p.v., and exist in those at least 10 weeks p.v. The results demonstrated that the immunization time of pcDNADPV-gC was a long period located in the experimentally vaccinated ducking tissues.

Studies on Histopathology and Ultrastructure of Subgroup J-avianLeukosis of Egg-type Chicken Breeder

CHAI Jia-qian;WU Zhao-na;YIN Qing;SUN Shu-hong;CUI Zhi-zhong

2010, 41(6): 735-740. doi:

Abstract ( 672 )

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This experiment was conducted to study the mechanism of Subgroup J-avian leukosis of Egg-type Chicken Breeder. The disease samples from an eggtype chicken (200-day old) flock in Jining, Shandong Province were studied by histopathological and ultrastructural pathologic methods. The antigen and antibody of avian leukemia were tested by ELISA test kit. The virus was isolated and detected by Indirect immunofluorescence monoclonal antibody test. The sick flocks showed clinical signs of depression, loss of appetite, and extreme emaciation and they finally got paralysed and died. The mortality rate was about 10%. In the necropsy of five dead chickens, almost all livers and spleens showed moderate to severe enlargement with diffuse graywhite nodules; Histopathological examination showed that focal tumor cells, with spherical eosinophilic granules in the cytoplasm, were found in the kidney, spleen, lung, liver. Diffused lymphocytes and lymphoblasts were found in those tissues. At the same time, the virus particles were appeared in the spleen tissues and bursa of Fabricius, about 100 nm in diameter, which are enveloped and spherical. Results showed that p27 antigen and serum antibody were positive in the disease samples, and ALV-J was isolated from them. On this basis, we concluded that the disease in the eggtype chickens was caused by subgroup J Avian Leucosis virus. As there are more reports of ALV affection in commercial layer chickens, special attention should be called.

Expression and Authenticate of Lymphocyte Proliferation Activity ofPorcine Interleukin-2/Porcine Interleukin-6 Fusion Protein

ZHEN Hong-hua;WANG Jin-liang;SHAN Hu;SUN Zhi-yuan;ZHAO Jin-hua;SHEN Zhi-qiang;

2010, 41(6): 741-745. doi:

Abstract ( 651 )

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In order to get the fusion protein which has the bioactivities of both porcine interleukin-2 (pIL-2) and porcine interleukin-6 (pIL-6) and explore the feasibility of the fusion protein as immunoadjuvant, the mature peptide genes of pIL-2 and pIL-6 were linked by a fragment which consisted of hydrophilic and low charge base pairs. Then they were subcloned to pBV220 for prokaryotic expression. The recombinant plasmid was transformed into E. coli DH5α, E. coli BL21(DE3) and E. coli Rosetta(DE3) and then induced at 42 ℃. SDS-PAGE analysis showed that the target protein pIL-6-2 was about 36.7 kDa in molecular weight which existed in inclusion bodies. The biological activity of the expressed protein was authenticated by MTT after being purified and refolded. The results showed that the effects of different concentrations of pIL-6-2 are significantly diverse in inducing the proliferation of lymphoblast cells from the spleen of mice. The concentration of 0.1 μg·mL^-1 pIL-6-2 exhibited the best effect. This study laid foundation for the application of this protein as a novel efficient immune reagent.

Genetic Relationship and Sequence Variation of Haemaphysalis longicornis and H. conicinna based on ITS-2, COⅠ and COⅡ Gene Sequences

GU Xiao-bin#;YU Zeng-ying#;YANG Guang-you;SUN Jia-gang;WEI-Hong;LI Kai-jun;WANG Shu-xian

2010, 41(6): 746-754. doi:

Abstract ( 639 )

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This experiment was conducted to clarify the genetic relationship between Haemaphysalis longicornis and H. conicinna. The second internal transcribed spacer (ITS-2) of the rRNA gene, cytochrome c oxidase subunit Ⅰ (COⅠ) and cytochrome c oxidase subunit Ⅱ (COⅡ) of the mitochondrial gene were sequenced. Then, the nucleotide variability of the genes between the two tick species was compared and phylogenetic relationship was analyzed. The lengths of ITS-2, COⅠ and COⅡ in H. longicornis were 361, 479 and 647 bp, and those in H. conicinna were 354, 474 and 661 bp, respectively. The gene homology of ITS-2, COⅠ and COⅡ between H. longicornis and H. conicinna were 84.2%, 89.0% and 88.4%, respectively. Phylogenetic trees reconstructed using the NJ method based on genes ITS-2, COⅠ and COⅡ indicate that H. longicornis and H. conicinna are clustered in the same clade. The results suggest that the length of ITS-2, COⅠ and COⅡ of H. longicornis and H. conicinna are different, and H. longicornis and H. conicinna should be two different valid species with close relationship in genetics, which supports the morphologic taxonomic status of H. longicornis and H. conicinna.

Changes of Cytokines in Immune Organs of Chickens Infected with H5N1 Avian Influenza Virus

ZHENG Shi-min;GAO Xue-li;LIU Chao-nan;LIU Ming

2010, 41(6): 755-760. doi:

Abstract ( 687 )

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The aim of the present study was to investigate role of cytokines in avian influence immune pathogenesis, technology of cell culture and cytokines activity detection was used to research the dynamic changes of inductive activity of interleukin-2 (IL-2), interferon (IFN) and tumor necrosis factor (TNF) in thymus and spleen of SPF chickens infected with H5N1 avian influenza virus（AIV）at 7 and 21 days old. The results showed that: IL-2 and IFN inductive activity of T-lymphocytes in thymus and spleen of SPF chickens infected with AIV were decreased compared with that of control, but TNF inductive activity from spleen of SPF chickens infected with AIV was increased compared with that of control. The results indicated that above-mentioned cytokines played a very important regulation role in immune pathogenesis of avian influenza.

Effect of Dietary High Molybdenum on the Cell Cycle and Apoptosis of Thymus in Broilers

YANG Fan;CUI Heng-min;XIAO Jie;PENG Xi;CUI Wei;CHENG An-chun;CHEN Tao;BAI Cai-min

2010, 41(6): 761-768. doi:

Abstract ( 754 )

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The experiment was conducted with the objective of examining the effects of high molybdenum on the thymus in the broilers by the methods of experimental pathology and flow cytometry(FCM)．360 one-day-old Avian broilers were randomly divided into four groups，and fed on diets as follows：Control diet (Mo 13 mg·kg^-1) and High molybdenum diets(Mo 500 mg·kg-1, High molybdenum groupⅠ; Mo 1 000 mg·kg-1, High molybdenum groupⅡ; Mo 1 500 mg·kg^-1, High molybdenum group Ⅲ) for 6 weeks. The results showed that decreased lymphocytes, enlarged Hassall′s corpuscles and increased myoid cells were observed in high molybdenum groupsⅠ,Ⅱand Ⅲ in comparison with those of control group histopathologically. The weight and the relative weight of thymus were decreased. The statistical analyses by FCM indicated that the G0/G1 phase of thymocyte was increased (P<0.05 or P<0.01) and the G2+M phase, S phase of thymocyte and the PI (Proliferating index) were decreased (P<0.05 or P<0.01) in high molybdenum groupsⅠ,Ⅱ and Ⅲ. Meanwhile, the percentage of cellular apoptosis was higher in high molybdenum groupsⅠ,Ⅱ and Ⅲ than that in control group. Also, the TUNEL staining was consistent with the result of FCM. It was concluded that dietary molybdenum in excess of 500 mg·kg^-1 inhibited the development of thymus, caused pathological changes in the thymus and impaired the function of cellular immunity in broilers.

The Molecular Genetic Study of Cytochrome b (Cyt b) Gene of Tibet Mini-pigs

2010, 41(6): 769-773. doi:

Abstract ( 1009 )

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To analyze the genetic differentiation of Tibet mini-pigs and its relationships with other pig breeds including European and domestic pigs, the sequences of nucleotides and amino acids of Cyt b were sequenced. The results showed that there were 9 mutation sites in Cyt b gene of Tibet mini-pigs and the nucleotide diversity was 0.078 9. Compared with pig breeds from Europe, there were 16 mutations in domestic pigs in China. There was significant difference between T-C switch at site 420 and G-A switch at site 883. The amino acid sequence analysis indicated that Tibet mini-pigs, compared with pig breeds from Europe, were different at three amino acid sites, especially at site 295, the partial Tibet mini-pigs together with other pig breeds were Valine, while another pig populations were Isoleucine at this site. The results indicated that the partial Tibet mini-pigs had very closely blood relationships with Bama Miniature pigs, Guizhou Xiang pigs and WZS pigs, and there was genetic differentiation in Tibet mini-pig group.

Comparision on Pathogenicity of Reticuloendotheliosis Virus to Different Ages′ SPF Chickens

SUN Shu-hong;ZHAO Peng;LIU Shao-qiong;CUI Zhi-zhong

2010, 41(6): 774-777. doi:

Abstract ( 657 )

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This experiment was conducted to investigate the pathogenicity of reticuloendotheliosis virus (REV) on 1 and 8-day-old SPF chickens. SPF chickens were inoculated with 1 000 TCID₅₀·bird ^-1 of REV-C99, a cloned strain. The pathogenicity of REV was evaluated by the measurement of growing rate of chicks and the antibody titers to the Newcastle disease virus(NDV). The results showed that REV challenge at a dose of 1 000 TCID₅₀·bird^-1 resulted in significantly (P<0.05) suppressed HI titers of NDV untill 13-week after vaccination for 3 times. For the chicks challenge with REV at 8-d of age, however, there was no significant difference in NDV titers. The results indicated that REV challenge at 1-d of age would retard the growth of chicks and suppress the humoral immunity. The result suggests that harmful effect of REV infection is agerelated and the control of REV at early age is important.

Study on Structure of Young Male Alpaca Pituitary

HE Xiao-yan;HAO Huan-qing;CAO Jing;LIU Dan-dan;ZHU Zhi-wei;DONG Yan-jun;BAI Rui;YU Xiu-ju

2010, 41(6): 778-784. doi:

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To research the morphology of alpaca pituitary and provide morphological basis for the endocrine function of alpaca pituitary. In this study, 7-month-old male alpaca pituitary was observed by the light microscope and transmission electron microscope. The results showed that alpaca pituitary located in pituitary fossa of the skull sphenoid were similar to the buffalo pituitary. It was ellipse, dorsal calm, gastro convex, and was composed of adenohypophysis(pars distalis, pars tuberalis, pars intermedia)and neurohypophysis, and existed pituitary cleft and undeveloped pituitary coelom. Pars distalis was 40%-45% of sagittal aera, and pars intermediate and pars nervosa rather developed, the ratio of oxyphil cell, basophilic cell and chromophobe cell was 8∶1∶11 in the pars distalis of alpaca pituitary. Somatotroph, thyrotroph, corticotroph, gonadotroph and follicular cell of the pars distalis of alpaca pituitary were found by transmission electron microscope, and somatotrophs was most. The results showed that pituitary structure of the 7-month-old alpaca was similar to that of most mammals, but had its unique characteristics.

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