Acta Veterinaria et Zootechnica Sinica

Polymorphism of Neuropeptide Y Gene and Its Relationship with Litter Size of Small Tail Han Sheep

ZHANG Baoyun;CHU Mingxing;WANG Pingqing;FANG Li;DI Ran;FENG Tao

2009, 40(12): 1705-1711. doi:

Abstract ( 676 )

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The objectives of the present study were to elucidate the polymorphism of ovine neuropeptide Y (NPY) gene and its relationship with litter size of Small Tail Han sheep, and to provide a scientific basis for markerassisted selection for prolificacy in sheep. The NPY gene was studied as a candidate gene for prolificacy in some sheep breeds. Single nucleotide polymorphisms (SNPs) of all three exons (exon 1, 2 and 3) of NPY gene were detected in Small Tail Han, Hu, Texel and Dorset sheep by polymerase chain reactionsingle strand conformation polymorphism. Only the product amplified by primer P1 displayed polymorphism. Five genotypes were detected in Small Tail Han sheep, three genotypes in Hu and Dorset sheep, but only one genotype in Texel sheep. Sequencing results revealed that there were three alleles (CR, TR and TW) in Small Tail Han sheep, two alleles (CR and TR) in Hu and Dorset sheep, only one allele (CR) in Texel sheep. Compared with CR allele, there was one single nucleotide mutation (C→T) at 93 bp position of coding region of NPY gene in the TR allele. Compared with CR allele, besides C93T mutation, there was one double nucleotide mutation (GA→AT) at 130 and 131 bp position of coding region of NPY gene in the TW allele. This mutation caused an Asp→Ile replacement at position 16 of amino acid sequence of sheep NPY mature peptide. For C/T polymorphic site, the differences of the litter size between three genotypes were nonsignificant in Small Tail Han sheep (P>0.05). For GA/AT polymorphic site, the ewes with genotype RW had 0.56 (P<0.05) lambs more than those with genotype RR in Small Tail Han sheep. In the five combined genotypes of Small Tail Han sheep, the difference of the litter size between TTRW and CTRW genotypes was nonsignificant (P>0.05), the differences of the litter size between TTRR, CTRR and CCRR genotypes were nonsignificant (P>0.05), least squares means of litter size for TTRW and CTRW genotypes were significantly higher than those for the other genotypes (P<0.05). These results preliminarily indicated that allele W of GA/AT polymorphic site of NPY gene is a potential DNA marker for improving litter size in sheep.

CIB1 cDNA Cloning from Small Tail Han Sheep and Its Expression in E.coli

YU Yan;;WANG Chuduan;LI Hongbin;WEI Caihong;SUN Dan;LU Jian;LIU Kaidong;LV Yanfei;DU Lixin

2009, 40(12): 1712-1717. doi:

Abstract ( 759 )

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In order to clone cDNA of Small Tail Han sheep CIB1 gene, and express the fusion protein in E.coli, a pair of specific primers were designed according to the nucleotide sequence of the published CIB1 gene. Fulllength cDNA of CIB1 gene was cloned from Small Tail Han sheep by RTPCR. CIB1 was cloned into pMD19T vector and identified by sequencing. CIB1 gene was cloned into pET32a vector to construct a prokaryotic expression vector pET32aCIB1 and transformed into BL21（DE3）pLysS cells. The recombinant plasmid pET32aCIB1 was induced in Escherichia coli using IPTG. The results showed that the identity at nucleotide level were more than 90% with CIB1 gene from cattle, pig, rhesus monkey, human, mouse, rat, chimpanzee and so on, and the amino acid similarity were higher than 93% correspondingly. The CIB1 cDNA containing a 576 bp open reading frame (ORF) encoding 191 amino acids. The expressed recombinant fusion protein is 38 ku in size. These results provided basis for the future research on ovine CIB1 protein function.

Effects of Serum on Expression Models of the Genes Involved in Adipogenesis during Porcine Preadipocyte Differentiation

LIU Haifeng;LI Xuewei;WANG Tao;ZHANG Xu;SHUAI Surong;LI Mingzhou

2009, 40(12): 1718-1726. doi:

Abstract ( 765 )

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To investigate the effects of serum on the preadipocyte during the process of induced differentiation and describe the expression patterns of genes related to adipogenesis in porcine, preadipocytes were isolated from subcutaneous adipose tissue of two days old piglets (Large White × Rongchang pigs) by using collagenasedigestion approach. The preadipocytes were cultured in different media supplemented with 50 nmol·L1 insulin, 100 nmol·L1 dexamethasone, 0.25 mmol·L1 1methyl3isobutylxanthine and 100 nmol·L1 rosiglitazone. In addition, the different effects between the two methods were also been compared, which included the effects of serum (control group) and serumfree (experimental group) on differentiation of preadipocytes, respectively. Total RNA were extracted from the cells and the mRNA expression level of genes involved in adipogenesis were measured by realtime quantitative RTPCR method. The results indicated that the mRNA expression levels of PPARα, C/EBPα, FASN, ACOX1, GPAT and ENPP2 were significantly downregulated (P<0.01) under the condition of serum added, in contrast, the mRNA expression level of PPARγ and FABP4 were significantly upregulated (P<001). The mRNA expression level of FASN, GPAT and ACOX1 in the experimental group were higher than that in the contrast group at P<0.01 level. Strikingly, the significant positive correlations among the mRNA expression levels of PPARα, PPARγ, C/EBPα and ACOX1 were observed in the experimental group. The results indicated that the serum has a fairly strong inhibitory discoloring effect on porcine preadipocyte during the process of induced differentiation. PPARα, PPARγ, C/EBPα and ACOX1 have the similar expression patterns. The serum is essential to accelerate the deposition of intracellular fat.

Cloning, Sequence Characterization and Expression Analysis of Porcine TIMP2 Gene

HUANG Honggang;XIAO Bang;REN Hongyan;TANG Zhonglin;YANG Shulin;CUI Wentao;MU Yulian;CHU Mingxing;LI Kui

2009, 40(12): 1727-1734. doi:

Abstract ( 733 )

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In this study, the molecular properties and potential functions of porcine TIMP2 gene were analyzed and characterized in Wuzhishan laboratory miniature pig. Porcine TIMP2 cDNA was obtained by reverse transcription polymerase chain reaction (RTPCR), using in silico cloning strategy based on porcine ESTs database, the nucleotide and amino acid sequences were characterized with bioinformatics methods, and its expression patterns in different tissues and development stages were also studied by RTPCR. The length of the cloned porcine TIMP2 cDNA was 909 bp including a 663 bp entire open reading frame (ORF), which encodes a 221 amino acids protein. Multiple sequences alignment and phylogeny analysis indicated that porcine TIMP2 protein shared high similarity with that of other species, such as Homo sapiens (97%), Mus musculus (97%) and Rattus norvegicus(97%). Protein sequence analysis showed that the deduced porcine TIMP2 protein sequence (pI=7.65， MW= 24.5 ku) contained 27AA leader sequence and 194 AA mature peptide, the leader sequence in pig has one more amino acid (Leu residue) than that in other 6 species. A NTRTIMP function domain and a conserved VIRAK motif at Nterminus were predicted by bioinformatic analysis and homologous comparison, and the 12 Cys residues in the sequence were predicted to constitute 6 disulphide bonds. The expression of porcine TIMP2 mRNA varied greatly in different tissues as well as different developmental stages, it was highly expressed in testis, pituitary body, stomach, large intestine, ovary, uterus and 90 days embryonic longissimus muscle, but lowly in heart, small intestine, brain, liver and adult longissimus muscle, and a moderate expression level was detected in kidney, thymus, spleen, thyroid and 33 days embryonic longissimus muscle. The result indicated that the porcine TIMP2 mRNA was highly expressed in organs and phases with relatively high tissue growth and/or remodelling activities, and it was in accordance with the innate biological function of TIMP-2.

Comparison of Somatic Cell Nuclear Transfer of Buffalo and the Effects of Time Interval before Activation on Nuclear Transfer by WholeCell Intracytoplasmic Microinjection

YANG Sufang;CHEN Zihong;WEI Jingwei;LU Fenghua;SHI Deshun

2009, 40(12): 1735-1740. doi:

Abstract ( 1036 )

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The aim of the study was to compare two methods of somatic cell nuclear transfer(SCNT), and explore the effects of time interval before activation on nuclear transfer(NT) by WholeCell Intracytoplasmic Microinjection (WCICSI). NT effect of Perivitelline Microinjection (PM) and WCICSI are compared, when the fetus fibroblasts were used for donor cells. And effect of activation time interval after donor cell by WCICSI on buffalo SCNT was also examined. The results showed that the embryo reconstructing efficiency by WCICSI was higher than that by PM (87.1% vs 81.1%, P<0.05), but the cleavage rate of reconstructed embryos was lower in former than that in latter (49.5% vs 71.8%, P<0.01). The blastocyst development rate of reconstructed embryos and the total efficiency of nuclear transfer were not significantly different each other (21.6% vs 19.3%, P>0.05;18.8% vs 15.7%, P>0.05). The blastocyst development rate of reconstructed embryos derived from cumulus cells and fibroblasts were the highest when the reconstructed embryos were activated at 3 h after injection. There was no significant difference in the development of blastocyst derived from granular cells between the time interval from 1.5 to 3 h, but the best time interval before activation was 3 h. The results indicated the WCICSI method can be employed in buffalo SCNT. Development rate of reconstructed embryos is best when activation at 3 h after buffalo cytoplasmic injection of donor cells.

Cloning and Expression of Bclaf1 cDNA Partial Sequence in Alpaca Skin

LI Yan;DONG Changsheng;HE Junping;LI Peng

2009, 40(12): 1741-1746. doi:

Abstract ( 706 )

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To identify the fragment cDNA encoding alpaca Bclaf1, and study the expression characteristic of the Bclaf1 in alpaca skin, a cDNA library was screened from alpaca skin and ESTs were analyzed, the cellular localization of Bclaf1 in alpaca skin was studied by immunohistochemistry. The multiple sequence alignments revealed that the similarity of Bclaf1 of alpacas was 100%, 92.9%, 86.0%, 81.1% and 77.9% to Bclaf1 of rattus norvegicu, mus, canis families, homo sapenis, pan troglodytes, respectively. The result showed that no positive cells were found in hair follicles of juvenile alpaca, the positive expression was focused in outer rootsheath of hair follicles of adult alpaca, white alpaca and brown alpaca; the expression in hair follicles was significantly different between the white and brown alpacas based on the average optical density. The expression and localization of Bclaf1 in alpaca skin were changed with age and coat color pattern.

Effects of Slowrelease Compound Acidifiers on Gastrointestinal pH and Intestinal Function in Weaning Piglets

YAN Jiayou;JIA Gang;WANG Kangning;WANG Lizhi

2009, 40(12): 1747-1754. doi:

Abstract ( 1210 )

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The experiment was conducted to study the effects of the addition of encapsulated and nonencapsulated compound acidifiers in a diet based on cornsoybean mealextruded soybean on gastrointestinal pH, growth performance, villus height and crypt depth of jejunum, intestinal digestive enzymes activities, microbial population and intestinal mucosal secretory immunoglobulin A (SIgA) in weaning piglets. Sixtyfour 28dayold, crossbred piglets (Landrace×Large White), weighing an average of (7.00±0.10) kg, were randomly assigned into four treatments with four replicates and four piglets (2 male and 2 female) per pen, according to singlefactor design principle. The feeding trial lasted for 35 days. The results showed that microcapsule slowrelease compound acidifiers significantly reduced the pH in stomach and small intestine（P<001）. In addition, microcapsule slowrelease compound acidifiers significantly increased the ratio of the villus height to crypt depth of jejunum（P<001）, and stimulated the sucrase activity and lactase activity in the early weaning period（P<005）as well. During the later weaning period, microcapsule compound acidifiers significantly increased the counts of lactobacillus and decreased the counts of Escherichia coli in the cecum and the colon（P<001）. It was also noted that there was an tendency of lower secretion of intestinal mucosal secretory IgA（P>005）. These results indicated that the microcapsule slowrelease compound acidifiers could optimize the intestinal microflora and improve the intestinal morphology and function by reducing the gastrointestinal pH, so as to enhance the intestinal digestibility and adaptation, and consequently improve the growth performance in weaning piglets.

Effects of Linseed Oil on Meat Quality and Fatty Acid Compositon of Adipose Tissue in Finishing Pigs

CAI Chuanjiang;CHE Xiangrong;ZHAO Kebin;WANG Lixian ;YAN Hua;CHENG Duxue;WU Jing;

2009, 40(12): 1755-1760. doi:

Abstract ( 723 )

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This experiment was conducted to study the effect of Linseed oil (LO) on meat quality and fatty acids composition of adipose tissue in finishing pigs. Eightyfour Large White pigs（21 barrows）, average weight of 63 kg, were randomly assigned into three dietary treatments with four replicates and seven pigs（1 or 2 barrows）with each replicate. The groups received one of the three dietary treatments: 0%, 15% and 3.0% LO, respectively. The feeding trial lasted for 50 days. There was no significant effect of LO added into diets on meat quality and carcass traits（P>0.05）；The inclusion of LO in finishing pig diets significantly increased αlinolenic acid (ALA) (P<0.01), EPA(P<0.01) and DPA（P<0.05）levels，and total n3 polyunsaturated fatty acid (PUFA) (P<0.01) concentration, and significantly reduced the levels of C20:4 n6（P<0.01）and C22:4 n6（P<0.01）in adipose tissue, but the total n6 PUFA content was not significantly affected by the diet（P>0.05）. These results indicated that pigs could not only deposit αlinolenic acid into adipose tissue, but also elongate and desaturate it; the 1.5% LO diet produced an n6n3PUFA ratio in the subcutaneous fat of 4.11,close to the recommended ratio for the human diet, and the ratio of 2.51 for the 3.0% LO diet, which can significantly improve the health value of adipose tissue.

Effect of Duodenal Soybean Small Peptides Infusion on Mammary Uptake of Amino Acid and Expression of APN in Lactating Goat

LIU Hui;;WANG Ling;;LI Shengli;WANG Libin;

2009, 40(12): 1761-1768. doi:

Abstract ( 1023 )

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The aim of this study was to investigate effect of duodenal soybean small peptides (SSP) infusion on mammary uptake of amino acids (AA). Eight Chinese Saanen goats with duodenal fistula and catheter were used in a crossover design trial. Goats were infused 0, 60, 120, 180 g·d1SSP. The experiment period lasted for 14 d. The results showed: 1) milk protein yield and content were increased, increment of milk protein yield were significant (P<0.05). Milk fat yield and content were decreased with the increased of the amount of SSP infusion (P<0.05). 2) Mammary plasma flow were no changed by SSP infusion though the average was slightly higher (P>0.05). Mammary plasma flow/milk yield was decreased by SSP infusion and there was significant difference between the 120 g·d1treatment and the control (P<0.05). 3) Compared with the control treatment, uptakes of most free amino acids were increased in the 60 and 120 g·d1 treatments, and decreased in 180 g·d1treatment. The uptakes of all peptidebound essential amino acids (PBEAA) were increased except for PBIle. Uptake of PBVal, PBLeu, PBPhe, PBThr, PBMet and PBLys was the highest in the 120 g·d1 treatment. Among peptidebound nonessential amino acids (PBNEAA), uptake of PBSer, PBTyr, PBPro was increased (P<0.05). However, that of PBGly was drop down (P>005). 4) Secretion of all essential AA (EAA) was increased by SSP infusion apart from Lys, that of those EAA was increased with increment of the amount of SSP infusion from 0102 g·d1 treatment (P<0.05), however increment of their secretion in milk were not significant in the 180 g·d1 treatment and were lower than in the 120 g·d1 treatment. 5) The expression of APN was improved by SSP, that in 60, 120, 180 g·d1 treatment was 13.55, 18.69, 10.01fold in control treatment. Conclusion: SSP might be used as substrate of milk protein synthesis by mammary gland, and improved milk protein synthesis and secretion. However, when the concentration of AA was stature, increment of AA resulted in absorption inhibition. Increment of APN expression accorded with the variety of PBAA uptakes by mammary gland, which suggested APN was one of peptidase regulated small peptides absorption in mammary tissue.

Effects of SteamFlaked Corn on the Production Performance and Excretion of Nitrogen and Phosphorus in Dairy Cows

CHEN Tao;GAO Yanxia;CAO Yufeng;LI Jianguo;LI Weijun

2009, 40(12): 1769-1775. doi:

Abstract ( 1007 )

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The experiment was conducted to study the effects of steamflaked corn (SFC) in the diet on the performance and the excretion of nitrogen and phosphorus in dairy cows, using a 4×4 replicated Latin square design. Steamflaked corn contents in the experimental ration were dividedly 0%, 10%, 20% or 30% of the concentrate. There were four 28d experimental periods. The results indicated that dry matter intake was unaffected by diet (P>0.05), but steamflaked rather than dryground corn (DGC) in diets could significantly increase the feed efficiency (4% FCM/DMI) (P<0.05). Milk yield was increased (P<0.05) for cows fed SFC diet compared with cows fed DGC diet. FCM yield for cows fed concentrate diets containing 20% or 30% SFC was separately significantly increased 2.78(P<0.01) and 2.87 kg·d1 (P<0.01) than the control diet. The percentages of milk fat, protein, lactose and nonfat solid (SNF) were unaffected by diet（P>0.05）, but tended to increase except milk fat; The apparent digestibility of nutrients were unaffected by diet（P>0.05）. The MUN were decreased (P<0.05) for cows fed SFC diets. Comparing with the control group (0%SFC), the excretion of nitrogen of the cows fed with the SPC diets w were decreased by 4.23%（P>0.05）,9.37%（P<0.05）and 10.29%（P<0.05）,and the excretion of phosphorus were decreased by 1.93%（P>0.05）,2.59%（P>0.05）and 5.19%（P>0.05）, respectively. In conclusion, compared with DGC diet, SFC diet had decreased the excretion of nitrogen and phosphorus while the performance was significantly increased.

Changes of Inflammatory Reactions of Porcine Skin Dendritic Cells Infected with Porcine Circovirus Type 2 in vivo

WANG Zhensheng;NIE Xiaohua;LI Huanrong;SUN Yingjian;SUI Lihua;CUI Defeng

2009, 40(12): 1776-1781. doi:

Abstract ( 655 )

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Porcine dendritic cells(DCs) from fortydayold SPF piglets inoculated oralnasally with porcine circovirus type 2 （PCV2） B1 strain were collected at different days post inoculation (DPI). Realtime FQPCR was used to detect the kinetics of mRNAs of IL10, TNFα, IFNα, IL8, chemokine CC motif receptor1 (CCR1) and CCR5 from skin DCs. The results showed that the transcriptions of proinflammatory cytokines mRNA decreased with significant levels of IFNα at 3DPI, TNFα and IL10 mRNA were increased significantly at 7DPI(P<0.05). The transcriptions of chemokine IL8 mRNA decreased with almost significant levels. MCP1 mRNAs were down at 3DPI and 14DPI, up at 7DPI, while the transcriptions of MIP1β mRNA were up at 3DPI and 7DPI, and back to the normal levels at 14DPI. The transcriptions of CCR1 and CCR5 were up at 3DPI, 7DPI and 14DPI, with significant increase at 7DPI (P<0.05). Taken together, PCV2 could significantly weaken the inflammatory reactions of porcine skin DC, and make its immune response out of control. These will affect the humoral immunity and cellular immunity of the animals.

Genetic Variation Analysis of Subgenotype Ⅶd Newcastle Disease VirusesIsolated from Jiangsu Province in 2008

WU Shuang;HUANG Weiping;WANG Weiwei;HU Shunlin;WANG Xiaoquan;LIU Xiufan

2009, 40(12): 1782-1787. doi:

Abstract ( 686 )

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In order to explore the molecular epidemiology of subgenotype Ⅶd Newcastle disease virus (NDV) in Jiangsu Province, 16 NDVs isolated from different regions in 2008 were characterized phylogenetically. Fusion (F) and hemagglutininneuraminidase (HN) genes were amplified by RTPCR and sequenced. The phylogenetic trees based on the sequences of the F and HN genes, respectively, were almost identical. These results suggesting that no recombination was present among these NDVs. In addition, the isolation frequency of the variant strain with E347K mutation on HN increased significantly in more recent years. More attention should be paid to the emergence of the variant genotype Ⅶd NDV strains.

Development of an Indirect ELISA for Detecting Antibodies against Cysticercus cellulosae Based on Its Recombinant 18 ku Protein Expressed in E. coli

WU Guohua;ZHENG Yadong;JIA Wanzhong;ZHANG Shaohua;JING Zhizhong;LUO Xuenong;LIU Shiquan;CAI Xuepeng

2009, 40(12): 1788-1793. doi:

Abstract ( 717 )

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A 18kDacoding gene of Taenia solium metacestodes was amplified by RTPCR and subcloned into pGEMT vector for sequencing. A recombinant plasmid pGEXCE18 was constructed and transformed into E. coli BL21 for in vitro expression. SDSPAGE and Westernblot were employed for analyzing the recombinant protein, which was used for development of an indirect ELISA for detection of anticysticercosis antibodies. The results showed that the interest protein was 35 kDa in size, accounting for 28% of total bacteria proteins, and could be recognized by positive sera against cysticercosis. Using the constructed indirect ELISA and a commercialized ELISA kit, paralleled analysis of 178 serum samples indicated that the concordant rate was 9883% and the ELISA showed good performance in specificity and sensitivity, supporting its application for cysticercosis diagnosis.

Pathogenicity and Development of Transgenic Eimeria tenella Expressing both Yellow Fluorescent Protein and TgDHFRTS Genes

YAN Wenchao;WANG Tianqi;SUO Xun;HAN Lifang;DING Ke;DONG Faming;ZHANG Longxian

2009, 40(12): 1794-1798. doi:

Abstract ( 698 )

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The pathogenicity and development of transgenic E. tenella strain TE1, which could express yellow fluorescent protein (YFP) and dihydrofolate reductase thymidylate synthase derived from Toxoplasma gondii (TgDHFRTS), in chickens were compared with that of the parental strain BJ to determine the effect of foreign gene on transgenic parasite. Results indicated that the fecundity of transgenic parasite (TE1) reduced at least 4 times than that of BJ strain. Low dosage of TE1 strain induced less pathogenic to chickens than BJ strain, but chickens inoculated with high dosage of TE1 oocysts displayed severe pathogenicity and certain mortality as well as BJ strain. In addition, trophozoites, the first generation and the second generation meronts and merozoites, microgamonts and macrogamonts of transgenic parasite TE1 were seen under flurescent microscope as that of BJ strain. More interestingly, not all four sporonts in the sporulating transgenic oocysts express YFP. These findings suggest that expressions of YFP and TgDHFRTS genes reduced pathogenicity and reproductive potential of transgenic E. tenella to some extent. Recombination between homologous or nonhomologous chromosomes appears during zygotic meiosis in E. tenella.

Apoptosis and Localization of Apoptotic Related Protein in Testis of Alpaca (lama pacos)

HE Junping;JIANG Junbing;DONG Changsheng

2009, 40(12): 1799-1804. doi:

Abstract ( 693 )

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To investigate apoptosis and localization of apoptotic related proteins in alpaca testis during the process of postnatal development and spermatogenesis, testicular tissues were collected from male alpacas at neonatal, 12 and 24 months of age. Apoptotic activity was studied by TUNEL (terminal deoxynucleotidyl transferasemediated dUTP nick end labeling) method, the distribution of apoptotic related proteins Caspase3 and Bcl2 in testes were examined by immunohistochemical method. Results showed that TUNELpositive cell wasn′t observed in neonatal testis. While Caspase3positive and Bcl2positve cells were observed in the interstitial compartment of neonatal testes. These indicated that apoptotic proteins involved in the apoptotic process of interstitial cells, and gave a space for seminiferous development. In the testes of 12monthold alpacas, TUNELpositive cells were observed in the central part of seminiferous, Caspase3positive and Bcl2positive cells were observed in the interstitial compartment and central part of seminiferous. These founding gave a clue that apoptosis and apoptotic proteins involved in the process of lumen′s formation of seminiferous of puberty (12monthold) alpaca testis. At 24 months of age, TUNELpositive cells were observed in the spermatogonia, spermatocytes and spermatozoa, Caspase3positive and Bcl2positive cells were observed in the interstitial compartment and all stages of germ cells. Caspase3 localized mainly in the spermatogonia and early spermatocytes, while Bcl2 took a preferential adluminal compartment staining pattern in the seminiferous tubules. These results indicated that apoptosis mainly occurred in spermatogonia and early stage of germ cells development, Bcl2 might inhibit the germ cell death of postspermatocyte during the process of spermatogenesis. Apoptosis involved in the process of postnatal testes development and spermatogenesis in alpaca. Apoptotic related proteins Caspase3 and Bcl2 involved in the regulation of apoptosis during the process of postnatal development and spermatogenesis in alpaca.

Effects of Dietary Copper of Different Sources and Dosages on Hydrogen Peroxide Generation of Mitochondria from Broiler Hepatocyte

CAO Huabin;SU Rongsheng;GUO Jianying;PAN Jiaqiang;LI Ying;TANG Zhaoxin

2009, 40(12): 1805-1811. doi:

Abstract ( 1013 )

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The experiment was conducted with the objective of examining the effects of dietary copper of different forms and dosages on hydrogen peroxide (H2O2) generation of mitochondria from broiler hepatocyte. Copper sulfate and copper methionine have been used as the copper source, and four concentration copper groups were set for each copper source. 288 broilers (Cobb 500, Gallus domesticus) aged 1 day were randomly divided into 8 groups with 36 each and fed with diets as follows: Controls (Cu 11 mg·kg1, group Ⅰ), and high copper (Cu 110 mg·kg1, group Ⅱ; Cu 220 mg·kg1, high copper group Ⅲ; Cu 330 mg·kg1, high copper group Ⅳ) for 60 days under normal condition, three samplecollections were processed at 12dayold, 36dayold and 60dayold respectively to investigate the changes of H2O2 generation of mitochondria from broiler hepatocyte. In result, compared with those of control group, H2O2 generation of mitochondria in high copper group Ⅱ, Ⅲ and Ⅳ of two copper sources were increased (P<0.05 or P<0.01); At the experimental time of the 36th and 60th day, H2O2 generation of mitochondria from broiler hepatocyte in each group of copper methionine were increased compared to those of each group of copper sulfate (P<0.05 or P<0.01). The location of H2O2 generation of mitochondria from broiler fed with high dietary copper were mainly concentrated in mitochondrial complex Ⅳ. The results obtained above indicated that high dietary copper could induce oxidized stress damage in liver; at the same level of dietary copper supplement, the organic copper will lead to much faster H2O2 generation velocity of mitochondria from broiler hepatocyte than inorganic copper. The results can also suggest that mitochondrial complex Ⅳ might be one of attack target spots under high dietary copper stimulation of organism.

Effect of 0.2 mg·kg1 Selenium in Broiler Diets on the Activity of GPx in Hepatic Tissue: a Meta-analysis

WANG Xiaolong;ZHU Dongze;XU Kai;WEI Ying;YANG Chuanping

2009, 40(12): 1812-1816. doi:

Abstract ( 653 )

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This study was intended to combine previous results of different independent researches and judge them integrally to renew our knowledge of the corelationship between selenium and the activity of GPx from the aspect of evidencebased medicine. A Metaanalysis was facilitated, by searching multidatabases and related resources, and performed by combining and quantitative previous multiresults of different independent researches. The different characteristics of each research were checked. The results of Metaanalysis showed that there was a significant difference of GPx activity in chicks′ hepatic tissue between experimental and control groups (P<0.01) and there was a positive corelationship between seleniumand the activity of GPx. But the result of Metaanalysis also indicated that there were the facts that the effect of selenium on GPx activity had been overestimated or underestimated in the past, and an extra 26.79 μg·mg1 of GPx activity in comparison with control group in chick′s hepatic tissue had statistic importance when 02 mg·kg1 selenium was supplied in diets. The results of the corelationship between selenium and the activity of GPx generated by classical methods were credible, while it was unprecious to predict the degree of the corelationship and contrary to each other sometime. This historical phenomena just right reflected the necessary of Metaanalysis and this is in keeping with the development of veterinary medicine.

Effects of PreTreatment with Taxol on Vitrification of in VitroMatured Porcine Oocytes

WANG Lei;;YANG Shanting;;DAI Jianjun;WU Caifeng;ZHANG Tingyu;ZHANG Defu;

2009, 40(12): 1817-1822. doi:

Abstract ( 680 )

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The aim of this study was to improve the effect of vitrification of porcine matured oocyte. This study was designed to examine the effect of Taxol pretreatment on vitrification of porcine oocytes matured in vitro by a cryoloop method. The results showed that the vitrified oocytes pretreated with 1.0 μmol·L1 Taxol attained higher normal morphology rate(89.93%) and FDA stain survival rate(83.33%) than the control(79.12% and 70.97%) (P<0.05). But when the concentration of Taxol was increased, it came to weaken the effect of vitrification. The proportions of oocytes with normal morphology and survive rates could reach to 90.21% and 84.13%, when they were treated for 30 minutes before vitrification. So the best parameter was 1.0 μmol·L1 Taxol for 30 minutes before vitrification. Pretreatment with CB or Taxol had a significant positive effect on vitrification of porcine matured oocytes(P<0.05), but there were no significant differences between the two chemical materials(P>0.05).

Isolation and EGFP Gene Transfection of Porcine Mammary Gland Cells

ZHENG Yuemao;HE Xiaoying

2009, 40(12): 1823-1825. doi:

Abstract ( 688 )

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This experiment was conducted to isolate porcine mammary gland epithelial and fibroblast cells from adult porcine mammary gland tissue, and transfect the enhanced green fluorescent protein (EGFP) gene into the cells. Porcine mammary gland epithelial and fibroblast cells were isolated from adult porcine mammary gland tissue using their culture system. EGFP gene was transfected into the cells by lipofection technology. We succeeded in isolating porcine mammary gland epithelial and fibroblast cells from adult porcine mammary gland tissue, and the EGFP gene transfected epithelial and fibroblast cells were obtained. The epithelial cells were short shuttlelike or guboidal, which were connected with each other closely and properly. The cell nucleus were circular or oval which had 24 cores. The fibroblast cells were long shuttlelike. This result shows that porcine epithelial and fibroblast cells could be isolated from adult porcine mammary gland tissue and EGFP could be expressed in the cells.

Development and Evaluation of an ELISA Based on Synthetic Peptide for the Detection of Footandmouth Disease Virus Antibody

YANG Suzhen;YANG Jifei;BAO Dengke;LIU Qingtang;LUO Jun;FAN Jianming;LI Xuewu;DENG Ruiguang;ZHANG Gaiping

2009, 40(12): 1826-1830. doi:

Abstract ( 708 )

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The specific synthetic peptide of footandmouth disease virus（FMDV） main protective antigen VP1 was synthesized by a solidphase method, and was conjugated with carrier protein BSA. An ELISA for detecting FMDV antibody was developed and evaluated by using the conjugated peptide as the coating antigen. The sensitivity and specificity of it were 95.12% and 100%. Comparing this assay with two commercial ELISA kits by detecting 199 serum samples,results showed that the agreement of it with UBI FMD VP1 ELISA kit and liquid phase blocking ELISA kit was 98.49% and 96.98%, respectively. The results indicated that the ELISA using synthetic peptide as coating antigen is specific, sensitive, stable and easy, and can be used to monitor antibody level of FMD.

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