Acta Veterinaria et Zootechnica Sinica

Genetic Polymorphic Analysis and Forecast of Heterosis of Sheep in Ningxia Autonomous Region

QU Dong-yan;CHEN Ren-jin;MAO Yong-jiang;SUN Chen-chen;GEN Yan;TIAN Dai-jun;CHEN Liang;ZHAO Xiao-yong;YANG Zhang-ping

2009, 40(1): 1-6. doi:

Abstract ( 1463 )

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The genomes of six populations (Dorset, Texel, Black-Suffolk, Tan sheep,Small-tailed Han sheep, hybrid of Tan and Small-tailed Han sheep) were screened with 7 microsatellite loci, and genetic diversities and genetic distance were estimated in this study. The results showed that, 108 alleles were detected in six populations, all the loci showed polymorphism in each population. The expected heterozygosity of all populations were much higher than the observed heterozygosity. The coefficient of phenotype differentiation (F_st) between populations was very low, the value was 5.8%, which was consistent with the coefficient of gene differentiation (G_st). The inbreeding coefficients for all populations (F_it) amounted to 56.90%. The inbreeding coefficients within breeds (F_is) was 54.36%. The heterosis between populations was also forecast according to DC distance and Nei’s genetic distance (DA), hybrid with Tan and small-tailed Han sheep as female parent was the best, then was Tan sheep, and Smalltailed Han sheep was the worst. The selective order for male parent should be Texel, Black-Suffolk, and then Dorset concerned with hybrid effects.

Relationship between Microsatellite Markers and Producing Ability on Litter Trait of Hu Sheep

SUN Wei;CHANG Hong;JIN Yin;WANG Peng;QIAN Jian-gong;WU Wen-zhong;CHEN Ling;WANG Wei

2009, 40(1): 7-14. doi:

Abstract ( 1468 )

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Four microsatellite loci (OarAE101，BM1329，BM143，OarHH55) linked to Fec^B gene mapped to chromosome 6 and one microsatellite locus (OarHH35) on chromosome 4 were selected to study the litter trait in Hu sheep. Results showed that all 5 microsatellite loci reached high polymorphism (PIC＞0.5), and the five microsatellite loci can be used for genetic diversity evaluation in Hu sheep breed. The result of variance analysis showed that there were extremely significant difference among the different genotypes for the Producing ability of litter size in OarAE101 locus and BM143 locus（P＜0.01）, there were significant difference among the different genotypes for the Producing ability of litter size in OarHH35 locus（0.01＜P＜0.05）,and there were no significant difference among the different genotypes for the Producing ability of litter size in BM1329 and OarHH55 loci（P＞0.05）.Least squares mean for the Producing ability of litter size for OarAE101 107/113 was the most， and it is not significantly higher than those for OarAE101 103/113、107/107，101/111，113/113，107/111 and 103/103 (P＞0.05), however it is significantly higher than those for OarAE101 101/101 and 103/107（0.01＜P＜0.05）, and it is extremely significant higher than those for OarAE101 115/115 and 111/111 （P＜0.01）in Hu sheep. Least squares mean for the Producing ability of litter size for OarHH35 139/139 was the most， however it is not significantly higher than those for OarHH35 139/141，139/127 and 137/125 (P＞0.05), and it is extremely significant higher than those for OarHH35 139/125，127/127，125/125（P＜0.01）in Hu sheep. Least squares mean for the Producing ability of litter size for BM143 118/118 was the most，however it is not significantly higher than those for BM143 118/106，118/110，106/104 and 112/114 (P＞0.05), it is significantly higher than those for BM143 118/116 and 102/106（0.01＜P＜0.05）, and it is extremely significant higher than those for BM143 102/104，112/102 and116/112（P＜0.01）in Hu sheep. And this study will have an important role in MAS and molecular breeding in Hu sheep in future.

The cDNA Cloning, Sequencing Analysis and Expression of Xinjiang Fine-wool Sheep Inhibin α Subunit

ZHANG Cheng-yun;WU Zhi-huan;ZENG Xian-yin

2009, 40(1): 15-19. doi:

Abstract ( 1367 )

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The inhibin α subunit was amplified from ovary of Xinjiang Fine-wool sheep by RT-PCR using two pairs of primers which were designed and synthesized according to the sheep inhibin α subunit gene sequence in GenBank.The gene about 812 bp by RT-PCR was inserted into pMD18-T vector and sequencing, the result of sequence analysis showed that the gene is Xinjiang Fine-wool sheep inhibin α subunit precursor. The gene of mature inhibin α subunit was cloned by PCR, which was double-digested by NcoⅠand HindⅢ and directionally cloned into the pET32a vector. The recombinant expression plasmid in the E.coli BL21 (DE3) was induced with 1 mmol·L-1 IPTG. SDSPAGE analysis showed that the induced expressed protein was about 32 ku. Western blot revealed that the expressed protein was the target fusion protein, which indicated that the recombinant prokaryotic expression vector expressed the fusion protein successfully. The cloning and expression of Xinjiang Fine-wool sheep inhibin α subunit made a foundation for the study of function and application.

Distribution and Quantitative Assay of Sheep β-defensin-1 mRNA in Differently Developmental Stages

SHENG Jin-liang;CHEN Chuang-fu;YANG Xia;WANG Yuan-zhi;ZHANG Hui;CAO Xu-dong;SHI Qian-wei;ZHAO Min

2009, 40(1): 20-25. doi:

Abstract ( 787 )

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Detecting distribution and expression level of sheep beta-defensin1 in differently developmental stages. Sheep beta-defensin1 mRNA was detected by traditional RT-PCR and fluorescence quantitative RT-PCR. sheep betadefensin1 was mainly expressed in digestive and respiratory tracts，included tongue，esophagus，rumen，abomasum，duodenum，jejunum，ileum，colon，appendix，trachea and lung，but not in heart，liver，kidney，lymph node and alveolar macrophage. The expression level of beta-defensin1 in abomasum，duodenum and jejunum was sBD1 mRNA expression level (neonatal)＞sBD1 mRNA expression level(adult)＞sBD1 mRNA expression level(fetal sheep). Sheep beta-defensin1 was consistently expressed in development process and its expression level in gastrointestinal tract was sBD1 mRNA expression level (neonatal)＞sBD1 mRNA expression level(adult)＞sBD1 mRNA expression level(fetal sheep).

The Effects of Cytoskeleton Inhibitor and Electro-activation Solution on the Parthenogenetic Activation and Preimplantation Development in Rabbit Oocytes

XU Ying;;LEI Lei;YANG Li-guo;CHEN Da-yuan

2009, 40(1): 26-31. doi:

Abstract ( 872 )

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The present study aimed to investigate effects of cytoskeleton inhibitors and electro-activation solutions on the parthenogenetic activation and preimplantation development in rabbit. MII oocytes were treated with cytochalasin B or cochicines before electro-activation. The results showed that there was no significant different among electro-activation solutions considering oocytes parthenogenetic activation (P>0.05). Furthermore there was no significant difference of 2-cell or blastocyst development when rabbit oocytes were treated with or without cytochalasin B(7.5 μg·mL^-1) before electro-activation (82.2% vs 76.4%, and 43.4% vs 51.5%, respectively). (P>0.05). The developmental rate of 2-cell and blastocyst were significantly decreased (64.9% and 23.8% respectively) when oocytes were treated with colchicines(1.0 μg·mL^-1) before electro-activation comparing with that of no treatment (P<0.05). The results from cytochalasin B plus cochicine treatment were similar to that of cochicines treated alone (62.5% 2-cell rate and 30.9% blastocyst rate). Immunofluorescent staining and laser scanning confocal microscopy results indicated that there was no abnormality of microtubule or microfilament in blastocysts which derived from cytoskeleton inhibitor treated and following electronic activated oocytes.

Histological Observation on Ovary of Early Fetal Pig

CHENG Zhi-jun;TAO Yong;XU Wei;WANG Yong;ZHANG Xiao-rong

2009, 40(1): 32-37. doi:

Abstract ( 854 )

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From 33 to 61 day during gestation is critical stage of oogenesis for fetal pigs. To elucidate the histological change of ovaries in fetal pig, ovaries of 33, 40, 46, 54, 61 day-old fetal pigs were collected and H-E stained. The results showed that oogonia and medullary cords could be seen in 33 day-old fetal ovaries, which were coated by one or two layers of coelomic epithelial. Synpliasma-like cells and oocytes at preleptotene or leptotene occurred in 40 d fetal ovaries. Synpliasma-like cells increased and oocytes entered preleptotene or leptotene in 46 d fetal ovaries, when medullary cords separate into fragments. Superficial epithelium in 54 d fetal ovaries was made of one layer cells and boundary between cortex and medulla was differentiable. Syncytium-like cell groups appeared in middle cortex and primordial follicles appeared in deep cortex. The cortex and medulla in 61 d fetal ovaries was distinguished. The syncytium-like cell groups occurred in middle cortex and primordial follicles occurred in deep cortex. In medulla, however, follicles degenerated and oocytes had nucleus pyknosis.

The Variation of Mast Cells and Histamine Concentration in Uterus and Oviduct of Goat in Estrous Cycle

WANG Li-qin;SHEN Ying;WANG Shu-ying;HUANG Li-bo;HOU Yan-meng;WANG Hui;WANG Jian-min

2009, 40(1): 38-43. doi:

Abstract ( 893 )

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The distribution, morphology and number changes of mast cells in goat uterus and oviduct during estrous cycle were observed using modified toluidine blue (MTB) staining and transmission electron microscopy. The histamine concentration in tissues of uterus and oviduct at different stages was also measured by modified fluorospectrophotometry. The results showed that the mast cells in uterus were mainly distributed in the myometrium, followed by the lamina propria, and sporadically scattered in the perimetrium. However, mast cells were not seen in the uterine caruncle and endometrium. In oviduct, mast cells were distributed in the mucosal plica, muscular layer and adventitia. Morphologically, round, oval or fusiform mast cells were present in all stages of estrous cycle. Significant degranulation in the mast cells was observed mainly during metaestrus. The amount of mast cells in the uterus and oviduct was in the order of MC（metestrus）>MC（proestrus）>MC（diestrus）>MC（estrus）. They were different significantly only in the myometrium（P<0.01）, while there was no difference in lamina propria and oviduct. Histamine concentration in uterine horn had a significant changes during metestrus with other stages (P<0.05), while there was no changes in the body of uterus, cervix and oviduct (P>0.05).

Research on Coculture of Mouse Sertoli Cells and Islets in vitro

YANG Run-jun;XU Shang-zhong;LI Jun-ya;GAO Xue

2009, 40(1): 44-51. doi:

Abstract ( 849 )

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On the basis of success in separating sertoli cells and islets, mouse sertoli cells as feeder layer were cocultured with islets, the islets growth characteristics and insulin secretion under the condition of single culture and coculture was observed respectively by morphology method and insulin fluoroimmunoassay method. The result indicated that the sertoli cells could provide a better microenvironment for the growth of islets, which could promote the proliferation of islets and significantly extend its survival time in vitro, and the survival rate of islets was above 90% after 19 days of coculture. The insulin secretion in coculture group was significantly higher than that in single cultured group from 7th days（P<0.01）, 15 days after coculture, the average insulin stimulation index in the two coculture groups was still 2.13±0.13 and 2.58±0.11 respectively.The results showed that sertoli cell as cytotrophoblast could maintain a higher level of insulin secretion and response to glucose, which was an ideal method for longterm culture of islets in vitro.

Ontogenetic Expression of rBAT and y+LAT2 mRNA in Intestine of Broiler

FENG Ding-yuan;TAN Hui-ze;ZOU Shi-geng;LIU Qing-shen;ZUO Jian-jun;DONG Ze-min;YE Hui;ZHANG Chang-ming

2009, 40(1): 52-58. doi:

Abstract ( 1491 )

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160 1-day-old parental male Arbor Acre (AA) broilers and 160 1-day-old parental male Lingnan Yellow broilers were randomly divided into 4 replicates, respectively. Ontogenetic expression of rBAT and y+LAT2 mRNA in duodenum, jejunum and ileum of AA broilers and Lingnan Yellow broilers were determined by relative quantitative RT-PCR. It was found that: (1) Ontogenetic expression of rBAT and y+LAT2 mRNA in duodenum and jejunum was different from that in ileum. (2) Ontogeny patterns of rBAT and y+LAT2 mRNA in duodenum and jejunum were consistent between AA broilers and Lingnan Yellow broilers. The abundance of rBAT and y⁺LAT2 mRNA in two breeds was increased from day 2 to day 30, then decreased on day 44 and increased again on day 58. While compared two breeds directly, the change range of rBAT and y⁺LAT2 mRNA in AA broilers was larger than that in Lingnan Yellow broilers. The abundance of rBAT and y⁺LAT2 mRNA in AA broilers was higher than that in Lingnan Yellow broilers from day 2 to day 30. (3) Ontogeny patterns of rBAT and y⁺LAT2 in ileum was different between two breeds. These results indicate that: (1) Ontogeny patterns of rBAT and y⁺LAT2 mRNA in duodenum and jejunum was different from that in ileum, indicating that the function of cationic amino acid absorption in proximal intestine may be different from that in distal intestine. (2) Ontogeny of rBAT and y⁺LAT2 mRNA in two breeds had same pattern in duodenum and jejunum, and there was difference at same development stage between AA broilers and Lingnan Yellow broilers and ontogeny pattern between duodenum (jejunum) and ileum indicating that rBAT and y⁺LAT2 mRNA expression could be regulated by developmental stage, breed and segment of intestine. (3) Ontogeny of rBAT and y+LAT2 mRNA in ileum were different between Lingnan Yellow broilers and AA broilers.

Effects of Trivalent Chromium from Different Source on Carcass Composition and Lipid Metabolism in Finishing Pigs

WANG Min-qi;XU Xiao-ling;LEI Jian;HE Yu-dan;XU Zi-rong

2009, 40(1): 59-65. doi:

Abstract ( 893 )

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96 pigs with an initial average body weight 65 kg were used to study the effects of trivalent chromium from different source on carcass composition and lipid metabolism. The pigs were assigned into four treatments with three replicates each, and were feed with four kinds of diets including a control diet and the control diet supplemented with 200 μg·kg^-1 chromium from either chromium chloride（CrCl3）, chromium picolinate（CrPic）and chromium nanocomposite （CrNano）for 40 days. The results showed that, compared with control group, supplemental CrNano improved average daily gain and feed conversion rate（P<0.05）, increased carcass lean percentage and loin eye area （P<0.05）, and decreased carcass fat percentage and backfat thickness （P<0.05）. CrPic supplementation also had significant effect on carcass fat percentage and loin eye area （P<0.05）. Hormone sensitive lipase in subcutaneous fat, total protein and free fat acid in serum was increased with the supplementation of Cr from either CrNano or CrPic（P<0.05）. The supplementation of Cr from CrNano or CrPic decreased serum glucose and insulin level（P<0.05）. Urea nitrogen, serum triglyceride and cholesterol were decreased（P<0.05）, and serum high density lipoprotein, insulin like growth factor I and lipase activity were increased with the supplementation of CrNano(P＜0.05). These results indicated that chromium nanocomposite exhibited more effects on growth and carcass composition of pigs than traditional chromium sources, and it could stimulate lipid catabolism and modulate lipid metabolism.

Effects of Ratios of Structural to Nonstructural Carbohydrate in Diets on Digestion and Metabolism for Sheep

WU Qiu-jue;HAO Zheng-li;LI Fa-di;ZHANG Xiao-qing;LI Yong;YE De-he

2009, 40(1): 66-71. doi:

Abstract ( 822 )

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This study was conducted to investigate the suitable ratios of structural to nonstructural carbohydrates(SC/NSC) in diets for sheep. Six Gansu high mountain fine wethers fitted with permanent ruminal and proximal duodenal cannulae, about one and half years old, and weighing 25-30 kg， were used according to a double 3×3 Latin square design with the three feeding cycles of 19 days each ( 10 days for adapting and 9 days for sampling ) to study the effects of ratios of SC/NSC in the three diets of Ⅰ，Ⅱand Ⅲ(1.57, 1.95 and 2.29, respectively) on apparent digestion in stomach, mid-and hind-gut and in the total tract, and also on the retention of nitrogen. The results showed that DMD and OMD in total tract for dietⅠ were higher than those for diet Ⅲ(P<0.01). No significant difference on NDFD and ADFD in total tract among the three diets was observed (P>0.05). Differences of DMD and NDFD in stomach， in midand hindgut among the three diets were not significant (P>0.05). Digested N were higher for diet Ⅰthan that for diet Ⅱand Ⅲ( P<0.01), and for diet Ⅱthan those for diet Ⅲ also( P<0.01). There were the tendencies that ND in stomach of dietⅡ(P=0.278), and ND in total tract(P=0.244) and retention of N（g·d^-1， P=0.091) of diet Ⅰ,Ⅱ were superior to other diets. These results indicated that the effects of the ratio of 1.57 for SC/NSC (dietⅠ)on digestion and metabolism was the best.

Development of a Multiplex Real-time RT-PCR for Simultaneous Detection of Classical Swine Fever Virus and North American Genotype Porcine Reproductive and Respiratory Syndrome Virus

CHENG Dan;;ZHAO Jian-jun;LI Na;SUN Yuan;ZHOU Yan-jun;ZHU Yan;TIAN Zhi-jun;TU Chang-chun;TONG Guang-zhi;QIU Hua-ji

2009, 40(1): 72-77. doi:

Abstract ( 932 )

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A multiplex real-time RT-PCR for simultaneous detection of classical swine fever virus (CSFV) and porcine reproductive and respiratory syndrome virus (PRRSV) was developed and evaluated. The detection limit of the assay was 3.2 TCID50 for CSFV and 1.8 TCID50 for PRRSV. Out of the 155 field samples tested, 73 were detected positive for PRRSV, and 16 were positive for CSFV, and 13 were shown to be co-infections with the two viruses. The agreement between the multiplex real-time RT-PCR and conventional RT-PCR was 99.4%. The method was shown to be sensitive, specific, and reproducible.

The Effects of PCV2 Experimental Infection on Transcriptional Levels of IL-2, IL-4, IL-10, IL-12p40, IFN-γ and TNF-α mRNA of Peripheral Blood Lymphocytes

SI Xing-kui;GUO Xin;YANG Han-chun

2009, 40(1): 78-82. doi:

Abstract ( 1624 )

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The transcriptional levels of IL-2, IL-4, IL-10, IL-12p40, IFN-γ and TNF-α of peripheral blood lymphocytes derived from piglets experimentally inoculated with PCV2 at 40 days old were detected by quantitative competitive RT-PCR (qcRT-PCR). The results showed that IL-2 and IL-12p40 mRNA levels were notably increased in the affected pigs at 14 (P<0.01, P<0.05) and 21 days post-inoculation (DPI) (P<0.01, P<0.05), respectively. IFN-γ mRNA levels were notably decreased at 7 DPI (P<0.05)，then upregulated statistically at 28 (P<0.01) and 42 DPI (P<0.05). IL-4 mRNA levels of the inoculated pigs were statistically higher than the control pigs at 28 DPI (P<0.01). Whereas IL-10 mRNA levels were significantly increased at 7 (P<0.01) and 14 DPI (P<0.05), then clear decrease were detected at 35 DPI (P<0.05). TNF-α mRNA expressions appeared not obviously be affected by PCV2 infection. In conclusion, the results indicated that PCV2 infection could cause imbalance of Th1/Th2 driven immune response, resulting in the impairment of humoral immunity along with the enhancement of cellular immunity.

Expression of the Whole Capsid Protein Gene P12A of Asia Ⅰ Foot-and-mouth Disease Virus in Insect Cells and Analysis of Its Immune Activity

CAO Yi-mei;SUN Jia-chuan;LU Zeng-jun;SUN Pu;GUO Jian-hong;LIU Zai-xin

2009, 40(1): 83-88. doi:

Abstract ( 795 )

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Structure protein gene P12A of FMDV from serotype Asia Ⅰ was inserted into the baculovirus transfer vector pMelBac-B with a melittin secretion signal sequence to gain recombinant plasmid of pMel-P12A. The recombinant transfer plasmids and linearized baculovirus DNA were co-transfected into sf9 insect cell to produce recombinant baculoviruses. After amplification of recombinant baculoviruses on cell passages, the recombinant viruses were seeded on sf9 cell with 10 MOI (multiplicity of infection), and cells were harvested 72 hours post infection. The expressed products of P12A gene in insect cells were detected by Western blotting, Immunofluorescence and sandwich ELISA assay. The results demonstrated that the P12A gene was successfully expressed in insect cells. The products were 82 kDa protein and could be recognized by anti-FMDV serum. Some expressed proteins were secreted into cell culture medium, and some were still in the cell lysate, as shown in sandwich ELISA. Furthermore, the expressed proteins were injected to guinea pigs intramuscularly and anti-FMDV antibodies were detected by indirect ELISA at 15 days post-infection. The result showed that the baculovirus-expressed P12A protein was able to induce anti-FMDV antibodies in the second week after vaccination. This study laid a good foundation for the research of the assembly of FMDV empty capsids in vitro and the development of subunit vaccine.

Biological Characteristics and Genetic Diversities Analysis of Infectious Bronchitis Virus

WU Pei-pei;TANG Ying-hua;LIU Wen-bo;CHEN Yi-ping;ZHANG Xiao-rong;WU Yan-tao;LIU Xiu-fan

2009, 40(1): 89-92. doi:

Abstract ( 1371 )

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Seven infectious bronchitis viruses(IBV) isolated from chicken in Jiangsu province between 2006 and 2007 were identified as nephropathogenic strains by dwarfing embryonated chicken eggs, chicken embryonated tracheal organ culture (TOC) assay, inhibition with NDV test and chicken recurrent infection. S1 gene of them were amplified by reverse transcriptionpolymerase chain reaction (RT-PCR), subsequently, the amplified fragments were cloned into TA vector and sequenced. Analysis of the phylogenetic tree based on S1 gene showed that Jiangsu IBV isolates were clustered into one group, and attributed to three subgroups. Homology of S1 nucleotide sequence among seven viruses varied from 94.6% (between CK/CH/JS/06I and CK/CH/JS/06Ⅲ) up to 99.4% (between CK/CH/JS/07Ⅲ and CK/CH/JS/07Ⅳ). The viruses in this study shared high similarity with the viruses isolates in China recent years, but shared low similarity with vaccine strains (H120 and H52) and other serotype index strains (T, Mass, 793B, 4/91).The genetic distance differences between IBV epidemic and vaccine strains lead to outbreak of the IB and circulate in immunized chicken.

Construction of Eukaryotic Expression Plasmids Encoding Chicken Interleukin-6 and Study on Its Immunoenhancement on Newcastle Disease LaSota Vaccine

LIU Rui-na;ZOU Nian-li;WANG Hong-ning;LIU Ping;HUANG Yong

2009, 40(1): 93-97. doi:

Abstract ( 795 )

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The chicken interleukin-6 (IL-6) was amplified from RNA of ConA-activated chicken splenic lymphocyte by RT-PCR, the gene was inserted into pCI-neo vector, and the recombinant plasmid pCI-IL-6 was acquired. The expression of pCI-IL-6 in CEF and its immunoenhancement effects on Newcastle disease LaSota vaccine were studied. The results showed that pCI-IL-6 can be expressed in CEF, the HI antibodies and percentage of CD4⁺,CD8⁺ and CD3⁺T lymphocyte of co-immunization group of pCI-IL-6 and LaSota vaccine were higher than those of LaSota vaccine group at 14 days post immunization, the difference of HI antibodies was significant （P<0.05）at 14，21 and 35 days post immunization and very significant （P<0.01）at 28 days post immunization respectively. The difference of percentage of CD4⁺,CD8⁺ and CD3⁺T lymphocyte was significant （P<0.05）or very significant （P<0.01）at 14 days post immunization.Challenge test at 35 days post immunization showed that the protection ratio of co-immunization group of pCI-IL-6 and LaSota was 89.5%, while those of LaSota vaccine group was 76.5%. The results showed that chicken IL-6 was expressed in vivo and can improve the immunization level of NDV LaSota vaccine obviously, showing immunoenhancement effect. This paper provided data for the further research and utilization of chicken IL-6.

Indirect ELISA for Detection of Antibody against Swine Bordetella bronchiseptica Using Recombinant fimD Expressed in E. coli

HE Hua;PEI Jie;WU Bin;ZHAO Zhanqin;ZHANG Jianmin;LUO Yong;XIANG Min;LU Shun

2009, 40(1): 98-102. doi:

Abstract ( 824 )

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According to the Bordetella bronchiseptica fimD sequence（X75811） of GenBank，we designed a pair of primer, and 1 098 bp DNA fragment of fimD was amplified from swine Bordetella bronchiseptica identificated in our laboratory. The acquired PCR products were inserted into pET-28a and expressed in BL21(DE3). Results of Sodium docecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot assay showed that the fimD gene was expressed in the form of inclusion body,and the recombinant protein was about 42 kD,and can react with the swine antisera to Bordetella bronchiseptica. The fusion protein was purified and used as coating antigen to develop the indirect Enzyme-Linked Immunosorbent Assay(ELISA). When using this ELISA to detect 668 clinical sera, 30.7% positive sera were detected. Parallel examination of the same 102 serum samples was conducted by using the ELISA and micro-agglutination test(MAT), the result showed that the ELISA is more sensitivity than MAT.

The Virulence Change of Streptococcus suis Resistant to Antimicrobials

WANG Li-ping;LU Cheng-ping

2009, 40(1): 103-108. doi:

Abstract ( 857 )

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The study was designed to investigate the relationship between virulence and antimicrobial resistance of Streptococcus suis. The difference of the growth rate, LD₅₀, haemolytic ring, histological damage and PCR of virulent factors such as MRP, CPS and EF between parental and resistant strains were determinated. The results showed that Streptococcus suis HA9801 strain can be induced into resistant strain when exposing to penicillin and erythromycin at sub-MIC concentration. Resistant strains is still virulent, as demonstrated experimently in mice by quantitative evidence of tissue damage，LD50 and in vitro by growth rate, haemolytic ring and virulent factors( MRP，CPS and EF) by PCR method between parental strain and induced-resistant strains. The results draw attention to the risk that the pathogenicity of the strains can not be reduced in Streptococcus suis type 2, the HA9801 resistant strain, to some extent, the strain can still cause infection.

Effects of the Compound Traditional Chinese Medicine Weeping Forsythia Capsule on Immunity Function of Chickens Infected by IBDV

GAO Hai;LIU Kai-yong;LI Xiu-lan;ZENG Zhen-ling;CHEN Jian-xin;CHEN Zhang-liu

2009, 40(1): 109-116. doi:

Abstract ( 919 )

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One hundred and sixty 7-day-old chickens were randomly divided into four groups with 40 chickens for each.GroupⅠserved as control; Chickens in groupⅡand Ⅲ were infected with 0.4 mL of IBDV by nasal drip and eye droppings when they were 8 day-old; Chicken in group Ⅲ and Ⅳ were treated with the compound Chinese traditional medicine Weeping Forsythia by oral administration of a single dose (20 g·kg^-1). And on the following 7th, 14th, 21st, 28th day post inoculation, the phagocytosis ability of blood corpuscle, the percentage of lymphocyte and lymphocyte proliferation rate were determined, the weight of whole body，spleen and bursa were measured for calculation of immune organ index. After 7 days post inoculation，the bursa and spleen in chickens were collected for observing histological structure and the ultra-structure. The content of CD3⁺, CD4⁺ and CD8⁺T lymphocyte subpopulation in the peripheral blood and splenic cell suspension were detected by flow cytometry with double-colorstaining method. The results showed that the compound traditional Chinese medicine Weeping Forsythia Capsule increased the phagocytosis ratio and index of blood corpuscle, the percentage of lymphocyte transformations and lymphocyte proliferation rate, weight and growth speed of bursa and spleen. The compound traditional Chinese medicine Weeping Forsythia Capsule could increase immunological function and the content of CD3⁺, CD4⁺，CD8⁺, CD4⁺/ CD8⁺T lymphocyte subpopulation in the peripheral blood and splenic cell suspension and prevent damages of immune organ of chicken infected by IBDV. However, all the values of IBDV infected group were lower than that of control group. The values of group treated with the compound traditional Chinese medicine Weeping Forsythia Capsule were higher than that of IBDV infected group.

Effect of Crude Polysaccharides from Mycelium of Cordyceps sinensis on Expression of p53 Gene and Induction of Apoptosis in SP2/0 Cells

LIU Yan-wei;LIU Na;HAN Bo;TIAN Xiang-rong;SU Jing-liang;ZHAO De-ming

2009, 40(1): 117-121. doi:

Abstract ( 1300 )

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The SP2/0 cells were treated with crude polysaccharides from mycelium of Cordyceps sinensis in vitro to explore the effect of crude polysaccharides on induction of apoptosis and expression of p53 gene in SP2/0 cells. MTT assay was used to measure the anti-proliferative effect of crude polysaccharides. Apoptotic morphologic changes of the treated cell were observed by Hochest 33342 staining. Induction of appoptosis was also proved by agarose gele electrophoresis. Cell cycle were analyzed by flow cytometry. Expression of p53 was measured with real-time quantitative PCR. The results were as follows: Crude polysaccharides could inhibit the growth of SP2/0 cell in a dose dependent manner; Typical apoptosis was observed under fluorescence microscope and a fragmented DNA ladder was visualized by agarose gel electrophoresis. In the flow cytometry analysis, arrest of cell cycle was observed in polysaccharides treated cells. Real time quantitative PCR analyses revealed the variation of p53 gene expression in the crude polysaccharides treated Sp2/0 cells.These results suggested that the inhibition of SP2/0 cell by crude polysaccharides may be related to induction of apoptosis and expression of p53 gene in SP2/0 cells.

PON1 Gene SNPs and Association with Growth and Carcass Traits in Beef Cattle

JI Ai-guo;ZHOU Zheng-kui;ZHANG Lu-pei;YANG Run-jun;XU Shang-zhong;WANG Shu-hui;GAO Xue;LI Jun-ya

2009, 40(1): 122-128. doi:

Abstract ( 887 )

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In order to estimate the allelic and genotypic frequencies at the PON1/EcoRⅤand PON1/AluI loci and association analysis between these SNPs and growth and carcass traits. The cDNA of bovine PON1 gene was cloned, and the SNPs in its exons were detected. Genotyping was performed on 30 Angus, 30 Hereford and 28 Simmental cattles using the GLM procedure of SAS and the least square means of the genotypes were compared by the Tukey′s test. 1 205 bp cDNA sequence were obtained, including 1 068 bp ORF region and which encodes 355 amino acids residues, and was accessed by GenBank, the accession number is EU289337. Two SNPs were detected in exon6 only. Individuals with AG genotype at the PON1/EcoRⅤ locus have higher weight at the time of entry into the fattening corrals (329.97±6.08)kg and close to the time of slaughter (577.56±8.32)kg and net meat weight (275.89±4.05)kg, fitted tenderness (3.10±0.19)kg (P<0.05); Individuals with AA genotype at PON1/Alu I locus has higher weight at the time of entry (333.37±8.93)kg and slaughter (576.82±13.18)kg and net meat weight (275.49±6.43)kg, and average daily gain (0.68±0.02)kg·d^-1 (P<0.05). The meat color score is also significant higher (P<0.05). Between genotypes and breeds, there were significant difference observed except TBW, REMG, MBS, REA and MCS (P<0.05). As a metabolism gene, genotypes of the SNPs of PON1 gene might be reflecting BFT directly, such as AeAeGaGa genotype.

Measures of Genetic Connectedness among Herds in Chinese Holstein

CHEN Jun;SUN Dong-xiao;WANG Ya-chun;ZHANG Yi;ZHANG Yuan

2009, 40(1): 129-132. doi:

Abstract ( 1517 )

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Genetic connectedness between herds is one of the most important components for the accuracy of dairy cattle genetic evaluation. In this study, two methods for the measure of genetic connectedness with mixed linear model were used, CR (Genetic Connectedness Rating) and GLT (total number of direct genetic links between group) were used to analyze the connectedness among the Chinese Holstein herds from Beijing, Shanghai and Tianjin. Genetic connectedness between the herds from Beijing and Tianjin is 23.95%, Beijing and Shanghai is 17.10%, Shanghai and Tianjin is 14.28%. Genetic connectedness between herds from Beijing and Tianjin is the highest and that between Shanghai and Tianjin is the lowest. Correlation coefficient of the two methods is 0.808. Some suggestions on genetic evaluation of dairy cattle were also given in order to get greater genetic gain.

Effect of Cytokine on in vitro Maturation of Oocytes and Early Embryos Development in Goat

ZHAO Zhen-hua;ZHOU Min-min;QIAN Hong-juan;WANG Xing-long

2009, 40(1): 133-137. doi:

Abstract ( 821 )

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Effects of GDNF, LIF and SCF separation or combination on in vitro maturation of goat oocytes and early embryos development were studied. The results showed that there was significant difference between control group and experiment groups in maturation rate(62.0%，75.5%，63.7% vs 53.0%，P<0.05), but no significant difference between different solutions（P＞0.05）. LIF and GDNF groups were significantly different from control group(48.1%，43.3% vs 26.2%，P<0.05), no difference between control group and SCF group. Experiment groups significantly improved the blastocyst formation rate（21.6%,27.3%,26.2% vs6.3%，P<0.05）. The 10 IU·mL^-1 LIF, 15 IU·mL^-1 GDNF and 15 IU·mL^-1 SCF were the best solutions for goat IVP- Maturation rate of goat oocytes was improved significantly in the groups with GDN＋LIF＋SCF and GDNF＋LIF than that in the groups with GDNF＋SCF，LIF＋SCF，GDNF，LIF (83.3%,80.3% vs 72.8%,65.4%,63.3%,77.9%，P<0.05).GDN＋LIF＋SCF and GDNF＋LIF groups were better than LIF＋SCF and GDNF＋SCF groups at cleavage rate(50.2%,48.9%,47.9%,43.7% vs 21.4%，P<0.0) and blastocyst rate(31.3%,27.6%,25.5%,24.3% vs 13.6%，P<0.05). The results obtained in this study indicated that GDNF, LIF and SCF significantly improved oocytes maturation and the blastocyst formation rate in IVP of goat.LIF and GDNF have coeffects on in vitro maturation of goat oocytes.

Pathogenic Experiment of Haemophilus parasuis to Certain Experimental Animals

GAO Peng-cheng;CHU Yue-feng;ZHAO Ping;HE Ying;LU Zhong-xin

2009, 40(1): 138-141. doi:

Abstract ( 1196 )

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The pathogenicity of Haemophilus parasuis to experimental animals including specific pathogen-free(SPF) mice, SPF rats, SPF syrian hamsters, SPF guinea pigs, clean guinea pigs and rabbits was identified in this study. These experimental animals were inoculated intraperitoncally with 2×10⁸ to 2×10¹⁰CFU culture of strain Nagasaki. Guinea pigs (SPF and Clean) and KM mice (SPF) inoculated with 2×10⁹CFU organisms suspended were died and other animals survived. The dead guinea pigs showed pathological changes of Glass’s disease，but there were no lesions in all dead mice. H. parasuis were isolated from the organs of the dead guinea pigs, including the brain, liver, heart blood, lung and peritoneal fluid. These results suggested that the guinea pig was susceptive to H. parasuis, and can be the candidate for developing animal model for investigation on the pathogenesis of H. parasuis.

Research of Drugresistance of Escherichia coli Isolated from Bovine with Endometritis and the Relationship betweenAntibiotic Resistance and Integron

ZHAO Hong-xia;LI Pei-feng;WU Cong-ming;FAN Hong-liang;SHEN Jian-zhong

2009, 40(1): 142-144. doi:

Abstract ( 810 )

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The drug-resistance of Escherichia coli isolates from dairy cows with endometritis in Huhhot and the relationship between antibiotic resistance of these strains and integron which they carrying were researched. The drugresistance of the isolates was defined by detecting MIC of every strain; The integrons in these E.coli isolates were investigated by PCR technique. 56 isolates of 10 serogroups were pathogenic strains. 21 of 57 strains were identified as positive for class I integron, account for 36.8%. The integronbearing isolates were all multi-resistance strains, and showed tolerance to more than eight kinds of antibacterial drugs. The results showed that the resistance rates of strains positive for integron I were remarkably higher than that of isolates negative for integron I. The resistance rates of the isolates to ceftiofur, furazolidone and quinolones were all lower than 35%; They were all resistant to sulfonamides and trimethoprim, the resistant rates were above 97%. The results revealed that the drugresistance and multiresistance of pathogenic E.coli from bovines with endometritis in Huhhot were strongly correlated with the integron I.

Molecular Epidemiology Analysis of Newcastle Disease Outbreaks in Waterfowl during 1997 to 2005 in China

LIU Hua-lei;ZHENG Dong-xia;SUN Cheng-ying;XU Tian-gang;WANG Yong-kun;WU Yan-gong;WANG Zhi-liang

2009, 40(1): 145-148. doi:

Abstract ( 1487 )

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Ten representative isolates of Newcastle Disease Virus (NDV) obtained from outbreaks in waterfowl in China during 1997 to 2005 were characterized both pathotypically and genotypically. Pathogenicity tests (MDT and ICPI) showed that all ten isolates were velogenic strains. The main functional region of the F gene comprising 535 nucleotides was amplified by RTPCR and sequenced. The amino acid sequence of the fusion protein cleavage site in all ten isolates was 112RRQKRF117, which is a typical sequence of velogenic strains and is in agreement with the results of in vivo pathogenicity tests. For genotyping, a phylogenetic tree based on nucleotides 47-435 of the F gene was constructed. Phylogenetic analysis showed that, among the 10 isolates, 8 isolates were of the genotype Ⅶd virus, 1 belonged to genotype Ⅸ virus, 1 belonged to genotype Ⅶc virus. Results indicate that the strains of genotype Ⅶd NDV have been the major pathogen, responsible for most epizootic ND outbreaks in waterfowl in China since 1997.

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