Acta Veterinaria et Zootechnica Sinica

Progress on the Molecular Patterns and Pathways of Host Recognitionto Viral Nucleic Acids

JING Zhi-zhong;HE Xiao-bing;FANG Yong-xiang;CHEN Guo-hua;ZENG Shuang;JIA Huai-jie

2011, 42(3): 311-322. doi:

Abstract ( 604 )

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Molecular mechanism of pathogen recognition by the host innate immune system has become an exciting and growing area of research focus in recent years. It also promotes the development of innate molecular immunology This review mainly introduces the cellular localization, molecular construction and interaction pathway of the multiple pattern recognition receptor (PRR) families, including Toll-like receptors (TLRs), RIG-Ilike receptors (RLRs), NOD-like receptors (NLRs) and DNA-dependent activator of interferon-regulatory factors (DAI), contributing significantly to virus detection by sensing viral nucleic acids. And systemically discusses the host defense pathway of innate molecular immunity how to discriminate and eliminate virus infection.

Advances in Antimicrobial Molecular Mechanism of Organic Acids

ZHANG Jun;TIAN Zi-gang;;WANG Jian-hua;WANG An-ru

2011, 42(3): 323-328. doi:

Abstract ( 726 )

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Organic acids, as growth promoter, have antimicrobial effects on animals, through different mechanisms. In this review, five possible mechanisms of organic acids are discussed including: energy competition permeabilizing through outer membrane, increase of cellular osmolarity, inhibition of macromolecule synthesis and induction of antimicrobial peptide in host cell, at levels of molecular mechanism, respectively. This review concludes that, the intracellular accumulation of anions of organic acid is a primary contributor to antimicrobial effects, which can be influenced by differences or shifts between intracellular and extracellular concentration of pH and anion concentration of extracellular acids.

Identification of Transcription Regulation Activity in 5′Flanking Regionof Pig THRSP Gene

CHEN Hua;CHEN Hong-quan;ZHOU Qian-qian;ZHANG Yi-peng;WEI Han-qing;LI Chun-miao;;ZHANG Xiao-rong;PENG Ying-lin

2011, 42(3): 329-334. doi:

Abstract ( 652 )

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The aims of this study were to identify TP526promoter activity of pigs THRSP gene and to understand functions of 5′ regulation region of THRSP gene. The DNA samples were extracted from pig ear tissue. The TP526promoter of thyroid hormon einducible nuclear protein Spot14 (THRSP) gene was amplified using PCR technology, inserted into the luciferase reporter vector (pGL3-basic) to construct pGL3-TP526/promoter. 293T cells were cotransfected with a plasmid containing the pGL3-TP526/promoter and pGL3-basic, respectively. The expression of luciferase gene in 293T cells was tested using the Dual-Luciferase Reporter Assay kit. The results showed that the fluorescence intensity ratio of firefly to sea pansy luciferase with （1.816 2±0.253 3） of pGL3-TP526/promoter was significant higher than that with （0.126 7±0.020 3） of pGL3-basic (P<0.01). It implied a significant promoter activity of TP526.

Studies on Polymorphisms of Interferon-γ Gene and Its Associationwith the Early Growth Performances and Reproduction in Porcine

WANG Da-li;ZHAO Wei;SUN Hai-tao;SUN Bo-xing;HUANG Da-peng;ZHAO Zhi-hui

2011, 42(3): 335-342. doi:

Abstract ( 629 )

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To study the possibility of Interferon-γ (IFN-γ) gene as a genetic marker in porcine resistance breeding, the single nucleotide polymorphisms (SNPs) of the IFN-γ gene were detected in 481 pigs including three breeds of Junmu1 White, Yorkshire and Duroc pigs. The correlations between SNPs and the early growth performances of piglet and reproduction of sow were analyzed. The results showed that three mutation sites were determined in intron 1, intron 3 and exon 4 of IFN-γ gene. The mutation sites were T825C, T2370C and G5301A. The results showed that 70 d weight of piglets with genotype AB(T825C site) was higher than that of piglets with genotype AA and BB (0.01<P<0.05), but the difference between individuals with AA and BB genotypes was not significant (P>0.05). The average daily gain(ADG) of weaning-to-70 d piglet with AB genotype was higher than that of piglets with AA and BB genotypes (P<0.01), but difference between individuals with genotype AA and BB was not significant (P>0.05). Difference of birth weight, weaning weight and birth-to-weaning weight among pigs in all genotypes were not significant (P>0.05).The effect of polymorphism at the sites of T2370C and G5301A on the early growth performance of pigs were not significant (P>0.05). The litter size, litter size born alive of pigs with genotype EF (G5301A) was higher than that of pigs with genotype EE in the Junmu-1 White pigs （0.01<P<0.05）. The difference of the litter size, litter size born alive, litter size at weaning, litter weight, litter size at 21 days among pigs with all genotypes at the sites of T825C and T2370C were not significant(P>0.05) In the conclusion, we can found some effects of SNPs (T825C) on 70 d weight and ADG of weaning-to-70 d, and some effects of SNPs (G5301A) on litter size and litter size born alive of sows.

Cloning and Prokaryotic Expression of PRL Gene in Shitou Goose

JIA Ru-min;WU Hui-ying;LIU You;ZHANG Li

2011, 42(3): 343-348. doi:

Abstract ( 688 )

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To further study the physiological regulatory mechanisms of PRL on the reproductive performance of Shitou goose. The cDNA of goose prolactin (PRL) gene was amplified from anterior pituitary gland of Shitou goose by RT-PCR. Then the PCR product was cloned into the pMD18.T vector to construct the pMD18-PRL for sequencing. And then the cDNA was subcloned into the prokaryotic expressing vector pET32a (+). Subsequently, the recombinant plasmid pET32-PRL was transformed into the E-coli BL21 (DE3) expression bacteria, then the recombinant PRL was induced by IPTG. The purified recombinant PRL was detected by SDS-PAGE and Western Blot. The results showed that coding region of PRL gene was comprised of 600 nucleotides（GenBank No. GQ856665） and encoded 199 amino acids putative protein which shared highly conservation with that of other birds. It was found that PRL protein was comprised of several α-Helixes, β-Sheets and Coils. It was inferred that the superiority region of antigenic epitope were in the N′ 70-76，95-102，150-155 and 207-213 sections. A high proportion of soluble PRL fusion protein about 53.6% of total bacterial protein was obtained in E.coli BL21 (DE3). The molecular weight of the fusion protein was about 41 ku. The Western Blot result showed that the recombinant protein was recognized by antiserum specifically. The result of this study will provide basis for further study of the biological function of prolactin protein and the effects of PRL on the broodiness and production traits of Shitou goose.

Preparation of Antiserums against Chicken Perilipin1 and TissueExpression Analyses of Perilipin1

WANG Yan-bo;WANG Ning;WANG Li;WANG Yu-xiang;LI Yu-mao;LI Hui

2011, 42(3): 349-355. doi:

Abstract ( 617 )

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The aim of this study was to analyze the expression characteristics of chicken Perilipin1 in different tissues and the function of Perilipin1 in lipid metabolism by preparing the antiserum against chicken Perilipin1. Chicken Perilipin1 gene was amplified by RT-PCR and inserted into pET-28a vector. The recombinant His/Perilipin1 was expressed in E-coli with IPTG induction and renatured by Protein Refolding Kit. The renatured His/Perilipin1 was used as immunogen to immunize the rabbit for preparing polyclonal antibody. The titer and specificity of this antibody were detected by ELISA and Western blot, and the expression characteristics of chicken Perilipin1 in different tissues was analyzed by Western blot. ELISA and Western blot results showed that the high titer and specificity of the antiserum against Perilipin1 was obtained The tissue expression characterization analysis by Western blot showed that Perilipin1 only expressed in chicken adipose tissue, but no signal was detected in liver, duodenum, breast muscle, leg muscle, gizzard, heart, spleen and kidney. Furthermore, the Perilipin1 expression characteristics between fat and lean broiler lines were also detected, and the results showed that Perilipin1 expressed significantly higher in fat line broilers adipose tissue than that in lean ones (P<0.05). These results of this study suggest that there is a close relationship between the expression of Perilipin1 and the deposition of abdomen adipose in fat and lean broiler lines.

Association of Toll-like Receptor 2 Polymorphisms with Somatic Cell Score in Bovine

BAI Jie;;LIN Jia-peng;YUAN Fang;HOU Min;LI Wen-rong;LIU Ming-jun;

2011, 42(3): 356-362. doi:

Abstract ( 1417 )

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This present study was aimed to investigate the effect of TLR2 on submastitis in bovine. 15 Holstein and Xinjiang Brown cattle with the highest and lowest SCS were chosen to sequence the TLR2 gene, respectively, and then the SNP sites were detected with PCR-RFLP. Finally, the associations between genotypes and SCS were analyzed. The results showed that: (1) 3 SNP sites (E+189, E+631 and E+2260) were discovered in TLR2. (2) At E+189 site of TLR2, the SCS of individuals with BB genotype was lower than that of individuals with AA and AB genotypes (P<0.05). It suggested that BB genotype played an important role in mastitis resistance of cattle. (3) At E+631 site of TLR2, SCS of individuals with BB genotype was lower than that of individuals with AA genotype, but not significant (P>0.05). (4) When E+2260 site was mutated and translated into termination codon, the SCS of individuals with AB genotype increased. It indicated that this mutation was harmful to mastitis resistance of cattle. (5) The SCS in Xinjiang Brown cattle was lower than that in Holstein. It indicated that mastitis resistance of Xinjiang Brown cattle was stronger than that of Holstein. (6) Mutation rate of three sites (E+189, E+631 and E+2260) in Xinjiang Brown cattle were higher than that in Holstein（P<0.01）. Conclusion: E+189 and E+631 SNP sites affect the mastitis resistance of cattle. The possible reason of the stronger resistance in Xinjiang Brown cattle is that there are some difference in distribution of two breeds between E+189 and E+631 sites.

Genetic Variation at C878T in Bovine SCD1 Gene and Its Associationwith Some Fat Traits in Chinese Simmetal Cattle

WU Xiu-xiang;SHI Xue-kui;CHANG Ling-ling;YANG Zhang-ping;LI Jun-ya;MAO Yong-jiang;GAO Hui-jiang

2011, 42(3): 363-368. doi:

Abstract ( 687 )

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This experiment was conducted to study the effect of SCD1 gene.To explore the effect of genetic variance in SCD1 gene on carcass and meat traits,682 cattle were selected randomly,including Chinese Simmetal cattle (Simme),Leiqiong cattle (LQ),Yunnan Gaofeng cattle (GF),BMY cattle (BMY),Minnan cattle (MN),Luxi cattle (LX),Bohai Black cattle (BH) and Holstein of South Chinese (Hols.),the polymorphism of specific fragment of SCD1 gene was analyzed by PCR-SSCP technique.A nucleotide mutation had been found at site 878,leading to an amino acid change （Glycine to Valine） at site 293,it was C878T.Three genotypes (CC,CT and TT) had been found at this site,and the frequency of genotype TT was highest in Chinese Simmetal population with the value of 0.114,and lower in Luxi cattle,Bohai Black cattle and Holstein with the value of 0.050,0.063 and 0.040,respectively.But genotype TT had not been found in the four tropic populations including Leiqiong cattle,Yunnan Gaofeng cattle, BMY cattle and Minnan cattle.The individuals with CC genotype of SCD1 showed a higher content of IMF and Mesentery fat weight traits (P<0.05),but lower value of backfat thickness trait (P<0.01),compared to the individuals with TT genotype.There was no significant difference among different genotypes with other traits.The results indicate that the C878T mutation of SCD1 gene has a great genetic effect on carcass and meat traits and could be a useful genetic marker for Chinese beef breeding.

Polymorphisms of the Gene ELA-DRA*exon 2 in Przewalski’s Horse

YU Li-juan;Entemake;LING Ying-hui;MA Yue-hui;Mahmut·HALIK

2011, 42(3): 369-374. doi:

Abstract ( 591 )

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To understand the polymorphism and allele frequency distribution of ELADRA*exon2 in Przewalski’s horse(Equus Przewalskii), Kazakh and Yanqi horse, ELA-DRA*exon2 in three types of horses was genotyped by PCRSSCP methods．Both the nucleotide and the amino acid sequences of the different alleles were analyzed in these horses． The results showed that seven genotypes were detected in 127 horses: three homozygotes named AA, BB and CC, four heterozygotes named AB, BC, AC and AD. AA genotype was dominant in Kazakh and Yanqi horses. CC was the dominant genotype in Przewalski’s horse(Equus Przewalskii). The chi-square analysis suggested that the allele and genotype frequencies were in Hardy-Weinberg equilibrium in the three horse types. The analysis of PIC and He showed that the three horse types belonged to intermediate polymorphism. The values of PIC and He in Przewalski’s horse(Equus Przewalskii) were lower than that in Kazakh and Yanqi horses.

Effects of Freezing-thawing on Muscle Quality and Microstructure ofZaosheng Cattle Meat

Ayimuguli;CAI Yong;CHEN Shi-en;SHEN Xiao-rong

2011, 42(3): 375-380. doi:

Abstract ( 592 )

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In order to study the effects of repetitive freezing-thawing on beef quality, 2.0-2.5 years old bullock Zaosheng cattle were selected, longissimus dorsi (LD) were collected and cut into 6 samples randomly, weighted and vacuum packed and repetitively freezing-thawing different times, then thawing loss (TL), cooking loss (CL), water loss(WL),shear force(SF), pH and microstructure were studied. The results showed that compared with fresh beef, repetitive freezing-thawing significantly increased the TL and CL; WL were increased significantly after freezing-thawing once; SF were significantly increased when freezing-thawing twice and then decreased; Freezing-thawing increased the space between muscle fiber, confused fiber bundles, and shortened sarcomere, vacuolated structure formation in the mitochondria, blurred even disappeared Z disc. The results demonstrated that repetitive freezing-thawing seriously damaged the structure of beef and declined the muscle quality of Zaosheng cattle.

Comparisons of the Quality and Nutritional Value of Whole-plant Corn Silageat Various Depths under the Top Surface of Silo in Summer

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

2011, 42(3): 381-388. doi:

Abstract ( 631 )

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This study evaluated the effect of simple covering of silo on the quality and nutritional value of whole-plant corn silage in summer. The silage samples were taken from 10, 20, 40, 80, 160 and 320 cm under the top surface of aboveground silo and analysed for their quality, nutritional composition and in vitro DM digestibility. The results showed that the whole-plant corn silage from 0 to 40 cm under the surface of the silo contained 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. Moreover, as sampling depths increased, the contents of DM, total sugars and WSC, pH values and DMD decreased gradually, but the contents of lactic acid, acetic acid, NDF and ADF increased, with the contents of crude protein and EE remaining unaffected. Therefore, both the simple covering of the silo and the depths under the top surface of the silo significantly affected the quality, nutritional composition and in vitro DM digestibility (DMD) of whole-plant corn silage in summer

Pathogenicity of Porcine Reproductive and Respiratory Syndrome VirusIsolates with Different NSP2 Variation in Piglets

WANG Xing-long;ZHOU Ye-fei;LI Yu-feng;LI Zhi-jun;WANG Xian-wei;JIANG Ping

2011, 42(3): 389-395. doi:

Abstract ( 661 )

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This study was conducted to evaluate the pathogenicity of different porcine reproductive and respiratory syndrome virus (PRRSV) isolates. Based on the NSP2 sequence of PRRSV isolates, four different isolates named as YX0907, BB0907, SY0909 and NT0801 were selected to study pathogenicity in piglets Among them, YX0907, BB0907 and SY0909 isolates have 30 aa deletion in NSP2 and NT0801 isolate has no amino acids deletion. Twenty-three 45-day old piglets, free of PRRSV, porcine circovirus 2 and Haemophilus parasuis infection, were selected and randomly separated into 5 groups. The 4 experimental groups were inoculated individually with 4 PRRSV isolates by injection of 2 mL of 2×10^4.32TCID₅₀·mL^-1 per pig. Three pigs in the control group were mock inoculated. After challenging, the rectal temperature of every pig was daily recorded. The sera were collected at 7, 14 and 21 day post inoculation (dpi) for viremia detection. The pigs dead and sacrificed at 21 dpi were autopsied and the gross lesions were valued. The results showed that BB0907 isolate had the highest virulence, which caused obvious fever and clinical signs with great pathological changes and 3 of 5 pigs were dead after infection. Compared with BB0907, YX0907 isolate was less virulent. The pigs in BB0907 inoculated group also showed apparent clinical syndrome and gross lesions and 2 pigs were dead after inoculation. But NT0801 isolate has relatively lower pathogenic ability and causes pigs had 1.2 days fever with slight clinical signs. SY0909 isolates has the lowest virulence, some pigs in this group only experienced 1.2 days fever without clinical signs. Meanwhile, the pigs in the control group performed well and without any clinical signs. These results revealed that the order of virulence of these four isolates is BB0907> YX0907> NT0801> SY0909, which indicating that the isolates with the same 30 aa deletion have different pathogenic ability and the 30aa deletion has no relationship with the virulence of isolate.

The Separation of Subgroup A and J ALV in Soft Tissue Sarcomasof “817” Broiler Hybrids

LIU Shao-qiong;WANG Bo;ZHANG Zhen-jie;WANG Jian;SUN Shu-hong;CUI Zhi-zhong

2011, 42(3): 396-401. doi:

Abstract ( 657 )

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The jugular soft tissue sarcomas were obtained from “817” broiler hybrids The tissue and cell lysis solution was prepared and inoculated in CEF cells for 12 days′ culture, as well as “817” Broiler Hybrids and SPF chickens, ALV-A and ALV-J viruses were identified by IFA from the culture and then to be named as SD1005. The duplicate soft tissue sarcomas of “817” Broiler Hybrids were prepared to culture and touch the piece, and then ALV-A and ALV-J were identified by IFA. In the meantime, ALV-A and ALV-J were discovered co-infection in the same cell. ALV-A and ALV-J were identificated with two pairs of primers toward ALV-A and ALV-J by PCR respectively. The result of autogenous variation analysis of env gene (1 710 bp) to ALV-J indicated that the homology between SD1005 and 14 reference strains at home and abroad was 88.5%-94.0%, the highest homology of which with U.S.A strains 0661 and ADOL-Hc1 were 94.0%, 93.8%. The present results indicated that there were mixed viruses in the soft tissue sarcomas from “817” broiler hybrids, with acute and high tumorigenicity. However，whether it was caused by ALV-A and ALV-J and the underlying mechanism of its tumorigenicity needs to be investigated further.

The Adjuvant Effects of Chicken β-defensin-1 on Infectious Bursal Disease Virus VP2 Gene DNA Vaccine

ZHANG Hui-hua;YANG Xiao-mei;XIE Qing-mei;MA Jing-yun;LUO Yan-na;CAO Yong-chang;BI Ying-zuo;

2011, 42(3): 402-408. doi:

Abstract ( 604 )

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The aim of present study was to investigate the adjuvant effects of chicken β-defensin-1 (also named avian β-defensin-1, AvBD1) on DNA vaccine encoding chicken infectious bursal disease virus (IBDV) VP2 gene. The eukaryotic expression vectors pcDNA3.1(+)-AvBD1 and pcDNA3.1(+)-VP2 were constructed. Five groups of 14-day-old chickens were intramuscularly injected with PBS, empty vector pcDNA3.1(+), pcDNA3.1(+)-VP2, a mixtures of pcDNA3.1(+)-AvBD1 and pcDNA3.1(+)-VP2, and IBD attenuated vaccine, respectively. Serum of chicken in each group was collected from the peripheral blood at different time post inoculation, and VP2specific antibody was measured by ELISA. Content of CD3⁺, CD4⁺ and CD8⁺ T-lymphocytes in each group of chickens were tested by cytospectrometry. Results showed that VP2-specific antibody level of group immunized with a mixtures of pcDNA3.1(+)-AvBD1 and pcDNA3.1(+)-VP2 was the highest among all the groups. The VP2-specific antibody level of group immunized with pcDNA3.1(+)-VP2, as well as group immunized with IBD attenuated vaccine, was significantly higher than that of the group immunized with PBS and empty vector pcDNA3.1(+). The percentages of CD3⁺, CD4⁺ and CD8⁺ T-cell subtypes between groups of pcDNA3.1(+)-VP2 and pcDNA3.1(+)-VP2 combination with pcDNA3.1(+)-AvBD1 was significantly different (P<0.05), the latter was higher, at 7 days post boost. The protection from IBD challenged by immunized chickens with DNA vaccines encoding IBDV VP2 gene alone was lower than that by immunized with a mixtures of pcDNA3.1(+)-AvBD1 and pcDNA3.1(+)-VP2. The results indicated that AvBD1 can be used as an adjuvant to enhance the IBDV VP2 DNA vaccine immunity.

Subculture of Chicken Embryo Cecum Epithelial Cell

GU Shao-peng;ZHAO Su-fen;WANG Yun-sheng;ZHENG Ming-xue;HOU Jin-huan

2011, 42(3): 409-415. doi:

Abstract ( 621 )

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Conditions and characteristics of chicken embryonic cecum epithelial cells (CEC) in subculture were studied in order to provide an in vitro model for the research of E-tenella’s injury mechanism and coccidiostats. Primary chicken embryonic CEC were isolated by digestion combined or step by step with trypsin and EDTA. The digestion method was selected by evaluating the coverage rate of adherent cells. The optimal concentrations of L-GLN and insulin in subculture medium were identified. The CEC were further purified by differential attachment. Characteristics of passage cells were investigated by morphological and SEM observation, immunohistochemistry, histochemistry, PAS, and flow cytometry. Under the conditions of digestion combined with trypsin and EDTA and removal of fibroblasts using differential attachment for 15 min, the CEC grew better in subculture medium with 72.5 mg·L^-1 L-GLN, 0.05 mg·L^-1 insulin, and 5% FBS. The cultured cells had the typical morphological features of epithelium. 24-72 h after passage, the cell purity was more than 95%, and the coverage rate of adherent cells was more than 85%. Results of apoptosis detection showed that high viability was obtained on 5 passages. While the apoptosis rate was significantly increased and coverage rate of adherent cells was decreased after the 6th passage. The subculture conditions were optimized for harvest stable chicken embryonic CEC with large quantity, high viability and purity.

Immunohistochemical Analyse of Apoptosis-related Proteins in the Embryonicand Post-embryonic Development of the Duck Bursa of Fabricius

FANG Jing;CUI Heng-min;CUI Xue;CHEN Yue

2011, 42(3): 416-422. doi:

Abstract ( 558 )

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This experiment was conducted to study the expression of Bcl-2，Bax，Fas and FasL proteins during the embryonic and post embryonic development of the Tianfu duck bursa of Fabricius. 20 Tianfu ducks were divided into 4 groups as follows: on embryonic day 24 (E24), and post-embryonic week 3, 8 and 29 (P3, P8, P29). Immunohistochemisty technique was used in this study. The results showed that Bcl-2, Bax, Fas and FasL were expressed in the follicle lymphocytes, and FasL was also displayed in the interfollicular epithelia.The Bcl-2 positive ratios of lymphocytes in the follicle cortex and medulla progressively decreased (except P3-8 in the medulla). The Bax positive ratios of lymphocytes remained stable from E24P8, but dramatically rose on P29The values of Bcl-2/Bax of lymphocytes showed a decreased tendency; however, they remained unchanged in the cortex and rose in the medulla from P3-8, respectively. The Fas positive ratios of lymphocytes increased from E24-P3 and P8-29, but decreased from P3-8. The changing pattern of FasL positive ratios of lymphocytes was similar to that of Fas, however, it remained stable from P3-8. Furthermore, the Bcl-2 positive ratios and the values of Bcl-2/Bax of lymphocytes in the follicle cortex were significantly higher from E24-P8 and lower on P29 than those in the follicle medulla, respectively. The Bax positive ratios of lymphocytes in the follicle cortex were not significant different from those in the medulla from E24-P8, but higher than those in the medulla on P29. The Fas and FasL positive ratios of lymphocytes in the follicle cortex were significantly lower from E24-P8 and higher on P29 than those in the medulla. This study indicated that Bcl-2, Bax, Fas and FasL proteins were important modulators for the lymphocyte apoptosis of the duck bursa of Fabricius during the embryonic and post embryonic development.

Effect of High-level Copper Diet on Expression of TrxR2 mRNA and Reduction Activity of TrxR2 of Liver in Broiler

LIU Hao-peng;TANG Zhao-xin;SU Rong-sheng;HU Jing-jing;HAN Qian-biao;HU Kai;LIU Chuan-dun;WAN Ting

2011, 42(3): 423-428. doi:

Abstract ( 621 )

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In order to examine the effect of high dietary copper on expression of TrxR2 mRNA in liver，and the reduction activity of TrxR2, the experiment was conducted with two hundred healthy 1 day-old Cobb broilers which were randomly divided into four groups based on diets as follows: control group (Cu 11 mg·kg^-1 as the group Ⅰ) and high copper groups (Cu content 110, 330, 550 mg·kg^-1, named group Ⅱ、Ⅲ and Ⅳ). The liver was taken from broiler in each group at the 10th, 30th and 50th days, respectively, reduction activity of the liver TrxR2 was determined by the method of DTNB, and the testing material was liver mitochondria which has been extraction from the liver, and the expression of the liver TrxR2 was determined by the method of semi-quantification and reverse transcription polymerase chain reaction (RT-PCR). The results showed that the reduction activity of liver TrxR2 decreased(P<0.05) and the expression of TrxR2 mRNA was reduced (P<0.05)at 50 d of Group Ⅳ, the reduction activity of liver TrxR2 was increased(P<0.05) at 30 d and reduced (P＞0.05)at 50 d of Group Ⅲ, The experiental results indicated that fed high copper diets (330-550 mg·kg^-1) could decrease TrxR2 mRNA expression in the liver and increase the reduction activity of TrxR2 first, and decrease it on 50 days.

Screening and Identification of Swainsonsine-producing Endophytic Fungi from Oxytropis.flabra

LU Wei;LU Hao;ZHAO Bao-yu;RONG Jie;CHEN Ji-ping;WANG Rui;GAO Wen-chao;CHEN Zhan-li

2011, 42(3): 429-436. doi:

Abstract ( 644 )

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To screen and obtain swainsonsine-producing endophytic fungi in Oxytropis.flabra, explore the relationship between endophytic fungi and swainsonine toxin in Oxytropis.flabra． In this research, we isolated and purified endophytic fungi of 6 Oxytropis.flabra samples from the west Inner Mongolia. Swainsonine was detected by thin layer chromatography and gas chromatography in fermentation liquor of the isolated fungus, then screened the Swainsonsine-producing endophytic fungi. Combined colonial morphology and micro-morphology to identify the species of isolated fungus. Finally got 6 species of endophytic fungi through identification. 2 of the 6 species endophytic fungi can produce swainsonine, named them XJ-J-1 and XJ-H-1 respectively. The Swainsonine yield was determined by gas chromatography, 4.209 2 mg·L^-1 in the fermentation liquor of XJ-J-1 and 16.733 8 mg·L^-1 in the fermentation liquor of XJ-H-1. The 2 strains endophytic fungi have the same genotype but different phenotype, both of them are Fusarium proliferatum.The results show that the swainsonine-producing endophytic fungi can be isolated from the Oxytropis.flabra .

Detecting Copy Number Variations in Porcine KIT by TaqMan-MGB Probe

WANG Tao;TANG Hui;ZENG Yong-qing;LI Xiao-ning;YAN Chao;DONG Lin-song;YU Xi-jiang

2011, 42(3): 437-441. doi:

Abstract ( 638 )

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The main objective of the present study was to establish a simple, rapid method for detecting copy number variations (CNVs) in porcine KIT gene TaqMan-MGB probes and primers were designed according to sequence of exon 2 of KIT gene, and a real-time fluorescence quantitative PCR procedure was established for the CNVs detection. The results showed that amplification curves of KIT obtained by TaqMan-MGB probe were a series of parallel curves corresponding to 2-fold serial dilutions of DNA samples, Ct values between groups were significantly different (P<0.001), and the coefficient-of-variations were low (from 0.12% to 0.26%). The amplification efficiencies of the KIT and ESR were approximately equal. The CNVs in KIT of 50 pigs were estimated by cluster analysis, assigned to 2, 3, 4, 5 or 6 copies, respectively. The real-time quantitative PCR using TaqMan-MGB probe is a simple, rapid method with high resolution and stability to measure CNVs in KIT, and it could be carried out in common laboratories.

Effect of Sequential Media G1/G2 on Development and Quality of Goat SCNT Embryos

LIU Jun;ZHENG Cong-ying;LI Chang-lei;DU Shan;LI Liang-liang;MA Bao-hua;ZHANG Yong

2011, 42(3): 442-447. doi:

Abstract ( 648 )

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The aims of this study were to investigate the effects of sequential media G1/G2 and modified synthetic oviductal fluid (mSOF) culture systems on developmental competence, cell apoptosis and the pregnant rate of goat somatic cell nuclear transfer (SCNT) embryos. Goat NT embryos were cultured as following: (1) 72 h in G1 then 120 h in G2 (group G1G2), both the culture media supplemented with 0.8% BSA (bovine serum albumin); (2) 72 h in mSOF supplemented with 0.8% BSA then in mSOF supplemented with 10% FBS (fetal bovine serum) for 120 h (group mSOF-FBS). The results indicated that there was no significant difference between the sequential media G1/G2 and mSOF-FBS in blastocyst rates ((27.7±3.1)% vs (25.3±1.0)%，P＞0.05). Both of total cell number of blastocysts and apoptotic nuclei in blastocyst from G1/G2 were significantly lower than that in mSOF-FBS ( (93.2±4.5) vs (109.1±6.2) and (4.9±0.2)% vs (11.3±0.1)%, respectively, P<0.05 ). However, the pregnant rates on Day 30 in group G1/G2 were higher than those in group mSOF-FBS（21.4% vs 8.0%, P<0.05 ). In conclusion, compared to mSOF-FBS, the sequential media G1/G2 could better support the development of goat somatic cell nuclear transfer embryos.

Cloning and Phylogenetic Analysis of the 18S rRNA of Histomonas meleagridisIsolates from Hunan Province

LIU Jin-hui;PENG Jun-yu;SONG Hai-yan;LIU Yi

2011, 42(3): 448-452. doi:

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Histomonas meleagridis were identified by PCR form Hunan Province, and their phylogenetic relationship was analysed by using 18S rRNA sequences The 18S rRNA sequences were amplified from each H. meleagridis sample and the amplicons were cloned into pGEM-T Easy vector, respectively. The inserts were successfully sequenced, and the length of 18S rRNA of the 10 H meleagridis isolates were 532 bp. Sequence comparison revealed that the similarity in18S rRNA sequences between Hunan isolates and the H meleagridis available in GenBankTM were more than 97.5%, respectively The 10 isolated strains grouped together with the reported H. meleagridisThe existence of H. meleagridis in Hunan Province was confirmed at the level of molecular biology. The results indicated that PCR was suitable for the diagnosis and population genetics of H. meleagridis.

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