Acta Veterinaria et Zootechnica Sinica

Research Advances in Invasion Mechanism of Bovine Viral Diarrhea Virus

GONG Xiaowei;ZHENG Fuying;LIN Guozhen;CAO Xiaoan;WANG Guanghua;YIN Hong;ZHOU Jizhang;CAI Xuepeng

2012, 43(4): 503-508. doi:

Abstract ( 349 )

PDF (742KB) ( 680 )

Related Articles | Metrics

Bovine viral diarrhea virus (BVDV) is recognized as one of the important pathogens of ruminant and swine. BVDV causes a series of clinical symptoms of persistent infection, immunosuppression, reproductive disorders and acute or chronic mucosal disease (MD). The disease impacts the economic development of cattle industry. It is difficult to treat and eradicate the disease from cattle because of the complex pathogenesis. With the development of molecular virology of classical swine fever virus and other members of the Flaviviridae, some achievements have been obtained about the research on BVDV, particularly at the cellular and molecular levels. In this paper, the pathogenesis and persistent infection of BVDV are explained, through review the mechanism of BVDV invasion cell, replication and interaction with host cell. It gives some new ideas for vaccine research and control strategy of bovine viral diarrhea (BVD).

Polymorphism of Exon1 of FSHR Gene and Its Relationship with Litter Size in Xiaomeishan Pigs

WU Jingsheng;;;ZHANG Genxi;DAI Guojun;YE Lan;WANG Jinyu;

2012, 43(4): 509-515. doi:

Abstract ( 351 )

PDF (701KB) ( 458 )

Related Articles | Metrics

The present study was to detect the polymorphism of exon1 of folliclestimulating hormone receptor (FSHR) gene and investigate the relationship between FSHR gene and litter size in pigs. The polymorphism of exon1 of FSHR gene was investigated by PCRSSCP and direct sequencing methods and the effect of exon1 of FSHR gene on litter size in Xiaomeishan pigs was analyzed using the least square analysis, simultaneously the variance components were analyzed and the selective reaction was predicted. The results showed that three SNPs (C70T, C74G and C81T) were detected in exon1 of FSHR gene, of which C74G caused amino acid change. Simultaneously three haplotypes and five genotypes were screened out. The difference of FSHR gene haplotypes and genotypes distribution among Xiaomeishan pigs, Fengjing pigs and Large White pigs reached extremely significant level (P＜0.01). According to the result of correlation analysis, in Xiaomeishan pigs, the TNB of the sows with genotype AA and AC after the second parity were 0.64 (P>0.05) and 0.36 (P>0.05) piglets less than those with genotype BC, and the TNB and NBA of the sows with genotype AA in all parities were 0.80 (P＜0.01) and 0.67 (P＜0.01) piglets less than those with genotype AC. The heredity of TNB and NBA in Xiaomeishan pigs was mostly influenced by the genetic dominant effect in exon1 of FSHR gene.

The Differential Expression of SLADQA Gene between Weaning Piglets Resistant or Sensitive to Escherichia coli F18

ZI Chen;LIU Lu;SU Xianmin;DU Zidong;HUAN Xuegen;YANG Jiansheng;SUN Shouyong;WU Shenglong;;BAO Wenbin;

2012, 43(4): 516-520. doi:

Abstract ( 410 )

PDF (731KB) ( 350 )

Related Articles | Metrics

The mRNA expression of SLADQA was assayed by realtime PCR in 11 different tissues in order to analyze the differential expression between resistant and sensitive postweaning Sutai piglets invaded with ETEC F18 aiming to explore the role of SLADQA gene in the resistance against ETEC F18. The results showed that the SLADQA gene was broadly expressed in 11 tissues with a similar expression rule. The expression level was the highest in thymus, lung and lymph nodes. The expression level was moderate in the jejunum, duodenum and spleen. The mRNA expression of SLADQA gene in resistant piglets was higher than that in sensitive piglets generally. In lymph node, lung, spleen, jejunum and duodenum, the mRNA expression level of SLADQA gene in resistant piglets were significant higher than that in sensitive piglets (P<0.05). The analysis suggest that higher expression level of SLADQA could provide optimal conditions for the formation of the SLA Class II molecule when the weaning piglets were affected by E. coli F18related toxins. SLADQA gene might not be the direct immune factor resisting the F18 of E. coli, but perhaps enhance humoral immunity and cell immunity to reduce the transmembrane signal transduction of ETEC F18 bacterial LPS and then led to the resistance to ETEC F18 in piglets.

Morphometry Study on the Structure of Thyroid Gland at Different Growth Stages in Pigs

WANG Yanli;LI Yanling;HAO Zhu;WANG Ying;XU Ningying

2012, 43(4): 521-526. doi:

Abstract ( 375 )

PDF (2301KB) ( 541 )

Related Articles | Metrics

To study the morphological changes of the thyroid gland at different growth stages of pigs, the Large Yorkshire pigs at birth, the body weight of 20, 40, 80 and 100 kg were slaughtered, the weight, length, width of thyroid glands were measured, the number of proliferated thyroid cells were counted and the expression of thyroid hormone in serum were analyzed. Meanwhile, the thyroid tissue sections were observed, the morphometric indicators of follicular lumen and follicular epithelial cells of thyroid were measured and analyzed by Nikon NISElements Documentation analysis system. The results showed that the number of proliferating thyroid cells, the expression of thyroid hormone, number density of follicle lumen, height of follicular epithelial cells gradually decreased with the growth of pigs; while the sectional area of follicular lumen, equivalent diameter of sectional area of follicular lumen and density of sectional area of follicular lumen gradually increased; and the variation of sectional area of epithelial cell nuclear, equivalent diameter of epithelial cell nuclear and number density of epithelial cell nuclear were not significant through all the growth stages of pigs. Correlation analysis showed that the expression of FT3 in serum was significantly negatively correlated with the thyroid width, and positively correlated with the height of follicular epithelial cells; while the expression of FT4 and TT4 were significantly negatively correlated with the body weight, thyroid weight, length of thyroid, width of thyroid, sectional area of follicular lumen, equivalent diameter of sectional area of follicular lumen, and positively correlated with the height of follicular epithelial cells and number density of epithelial cell nuclear. The morphological measurements provide accurate quantitative data for the indepth study of structural changes at different growth stages in porcine thyroid.

The Expression Analysis of DLK1 and MSTN Genes in Fetus during Mid and Latepregnancy of Sheep

WEI Caihong;LIU Gang;DU Lixin;LI Fadi

2012, 43(4): 527-533. doi:

Abstract ( 429 )

PDF (1698KB) ( 409 )

Related Articles | Metrics

In order to investigate the expression of DLK1 and MSTN genes in different skeletal muscles of fetus at different pregnant stages in Texel and Ujumqin sheep, and observe the muscle development regularity during the early period. The weight of muscles including semitendinosus，semimembranosus，longissimusdorsi，quadriceps femoris and biceps femoris muscle were compared by variance analysis methods, and the expression of the two gene were analysed in the five kinds of muscles of fetus on d85, d100, d120 and d135 during pregnant . The results indicated that the weight of semimembranosus，longissimusdorsi，quadriceps femoris and biceps femoris muscles in Texel sheep were significantly higher than that in Ujumqin sheep (P<0.05 or P<0.01) on d85 and d100. The results of quantitative realtime PCR indicated that the expression of DLK1 gene increased with the fetus development, and the expression of DLK1 gene in Texel sheep was higher than that in Ujumqin sheep in semitendinosus(d120)，semimembranosus（d120），longissimusdorsi（d100 and d135），quadriceps femoris（d85）and biceps femoris (d135，P<0.05). The expression of MSTN gene in Texel sheep was higher than that in Ujumqin sheep. The expression of MSTN gene in longissimusdorsi and quadriceps femoris were significant different between Texel and Ujumqin sheep. The result indicate that the expression of DLK1 and MSTN genes in muscles at different pregnant stages may be related to muscle development at early stage.

Study on the Antioxidant Ability in Longissimus doris of Different Breeds of Beef Cattle

XIE Xiangxue;MENG Qingxiang;REN Liping;ZHANG Xinzhuang;LIU Ping;

2012, 43(4): 534-552. doi:

Abstract ( 460 )

PDF (1013KB) ( 470 )

Related Articles | Metrics

The objective of this study was to investigate the lipid oxidation characteristics of different cattle breeds and provide data for the high quality beef production. The longissimus dorsi of five cattle breeds including Limousin, Simmental, Luxi, Jinnan and Qinchuan cattle were selected for comparison. Intramuscular fat content, drip loss(24 hour), lipid peroxides（PV）, meat colour and thiobarbituric acid reactive substances（TBARS）(072 h)at 04 ℃ were measured. The results showed that the intramuscular fat content and drip loss(24 hour) of longissimus dorsi had no significant differences among different cattle breeds ; The a* and b* values in all cattle breeds decreased gradually during 072 h; After 4 h, the a* value of Limousin was the highest among all breeds. The a* value of Limousin at 8 h and 12 h was significantly higher (P<0.05) than that of Luxi and Qinchuan. After 24 h, a* value of Limousin was significantly higher (P<0.05) than that of Simmental; The values of PV and TBARS gradually increased with the increase of storage time for all cattle breeds. Limousin had the slowest increase for PV and TBARS, and Qinchuan and Luxi cattle showed the faster increase for PV and TBARS. In summary, there are no significant differences in intramuscular fat and drip loss among all breeds cattle, however Limousin cattle can keep meat colour and antioxidant ability for longer time during the storage.

Polymorphisms of MTNR1B Gene and Its Association with Laying Traits in Wenchang Chickens

LI Xiaoning;WANG Hui;YU Xijiang;JIANG Yunliang;SONG Qian;YAN Chao;TANG Hui

2012, 43(4): 540-545. doi:

Abstract ( 365 )

PDF (607KB) ( 465 )

Related Articles | Metrics

The objective of this study was to investigate the polymorphism of the melatonin receptor gene(MTNR1B) and its association with laying traits in Wenchang chickens. Single nucleotide polymorphisms (SNPs) were detected in 5′ regulatory region, exon 1 and exon 2 of MTNR1B by direct sequencing, and the association of SNPs with laying traits was also analyzed. The results showed that no base mutations or deletions were detected in exon 1 and exon 2, while 15 SNP loci were found in 5′ regulatory region. Dominance effects at -778 locus were 2.3 and 6.2 eggs for egg number from 22 to 31weekold and total egg number until 46weekold, respectively. At -216 locus For egg number from 32 to 46weekold and total egg number until 46weekold, additive effects were 2.5 and 3.2 eggs, respectively, and dominance effects were 3.5 and 6.8 eggs, respectively. Hens with the haplotype AA had the least egg number from 32 to 46weekold and egg number until 46weekold and hens with the haplotype GC had the earliest age at first laying. Hens with combined genotype AC/GA had the most egg number during the periods from 22 to 31weekold, 32 to 46weekold. In conclusion, exons of MTNR1B are highly conservative, while SNPs in 5′ regulatory region are maybe as candidate markers for laying traits.

Molecular Cloning and Construction of Luciferase Reporter Recombinant of CD8α Gene Promoter in Duck

XU Qi;CHEN Yang;HUANG Zhengyang;ZHAO Wenming;ZHANG Yang;LI Xinyu;ZHAO Rongxue;LI Xiu;DUAN Xiujun;CHEN Guohong

2012, 43(4): 546-552. doi:

Abstract ( 336 )

PDF (2296KB) ( 564 )

Related Articles | Metrics

This experiment was conducted to explore the potential regulatory sequences of duck CD8α gene and the mechanism of transcription regulation. The CD8α gene promoter was cloned by genome walking, the sequence of the promoter was analyzed by online software of bioinformatics, and then the promoters from DHVresistant and DHVsusceptible duck were subcloned into the luciferase expression vector pGL3Basic directly, and their luciferase activity was detected. The DNA sequence of 2 480 bp was amplified including 56 bp in exon1 and 2 424 bp of promoter region. There were TATAbox, GCbox, CAATbox and transcriptional start site in the sequence. 41 transcriptional factor binding sites including CdxA, Nkx2, GATA1, SRY, and so on, were detected. The luciferase reporter gene recombinant pGL3 CD8α including correct target gene was constructed and identified by restrictive endonuclease enzyme cutting and sequencing, the pGL3Gy and pGL3Gk plamids had strong luciferase activity. The result lay a foundation for exploring the promoter activity and transcriptional regulation mechanism of duck CD8α gene.

Proliferative Effects of Growth Factors on Bone Marrow Mesenchymal Stem Cells of Rabbits in vitro

HE Xiaoe;REN Weiqing;DENG Lixin;HE Jundan;QIN Jiachen;LV Jingyu;CAO Guibin;WANG Xinzhuang

2012, 43(4): 553-557. doi:

Abstract ( 329 )

PDF (1403KB) ( 373 )

Related Articles | Metrics

To study the effects of basic fibroblast growth factor (bFGF) on growth and proliferation of rabbit bone marrow mesenchymal stem cells (BMSCs) in vitro, rabbit BMSCs were cultured and identified in vivo, treated with bFGF at final concentration of 5, 10, 20, 40, 80, 100 μg·L-1. Cell growth and proliferation were detected by MTT assay and flow cytometer. MTT assay showed that proliferation promotion effect of bFGF nearly reached the peak at 80 μg·L-1(P＜0.01). Compared with the negative control group, significant proliferation promotion effect was detected in the bFGF groups from 72 h(P＜0.01); Flow cytometer result showed that proliferation index of bFGF groups BMSCs was higher than that of control group(P＜0.01). Basic fibroblast growth factor can promote proliferation of rabbit BMSCs in vitro, 80 μg·L-1 is the optimum concentration for in vitro culture which can be used as the best culture condition of rabbit BMSCs proliferation in vitro.

Effects of Active Immunization against Inhibin on Hormonal Level and Ovarian Development in Rats:Comparison of Montanide and Freund’s Adjuvant

CAO Xiaohan;WANG Zhenhua;HAN Xingfa;ZHU Shiya;ZHANG Fugang;ZENG Xianyin

2012, 43(4): 558-563. doi:

Abstract ( 357 )

PDF (1294KB) ( 379 )

Related Articles | Metrics

The aim of this study was to investigate the effect of active immunization using inhibin α subunit mature peptide(roINH) with Montanide(MON) and Freunds (FA) adjuvants on ovarian development in Xinjiang Finewool sheep. 72 Sprague Dawley female rats of (910)weekold were divided into 3 groups and actively immunized against the recombination fusion protein of Xinjiang Finewool sheep roINH with roINH+MON adjuvant (MON group) or roINH+FA adjuvant (FA group), or injected with physiological saline (control group), respectively. Each rat received a primary immunization and 2 booster immunization at 20day interval. The results showed that, the antibody titer of MON group was significantly higher than that of FA group (P<0.05), compared to controls, mean concentration of FSH was higher (P<0.01) in the MON and FA groups, and there was a significant increase in the peak of P in MON and FA groups (P<0.05), respectively. For the entire period of the study, mean concentration and peak of FSH and P were of no significant difference between the two immune groups (P>0.05); mean concentration and peak of LH between the two immune groups had no significant difference(P>0.05). There was no significant increase between MON and FA groups in the number of mature follicle (P>0.05), which were both significant higher than that in control group (P<0.05). Compared to FA group, the inflammatorial size was smaller and the inflammatory reaction was weaker in MON group. These results clearly demonstrate that active immunization against inhibin α subunit mature peptide in MON or FA adjuvants both can enhance immune effect, furthermore, the inflammatory reaction of MON adjuvant is less than that of FA adjuvant.

Effects of Nacetylcysteine on Immunological Stress and Energy Status of Piglets Chronically Challenged with Lipopolysaccharide

YANG Zhenguo;ZHANG Wei;HOU Yongqing;DING Binying;WANG Lei;LIU Yulan;ZHU Huiling

2012, 43(4): 564-571. doi:

Abstract ( 334 )

PDF (561KB) ( 587 )

Related Articles | Metrics

: The aim of the current study was to evaluate the effects of dietary supplementation with Nacetylcysteine（NAC）on liver immunological stress and energy status in piglets after lipopolysaccharide (LPS) challenge. Eighteen healthy piglets with the same genetic origin and similar body weight (Duroc ×Landrace × Large white, (11.58 ± 0.26) kg BW) were randomly allocated into 3 treatments (control group, LPS group and NAC group). Each treatment included 6 replicates, one piglet per replicate. The piglets in the control group and LPS group were fed with the basal diet, while the NAC group was fed with the basal diet + 500 mg·kg-1 NAC. On d10, d13 and d20, the piglets in the LPS and NAC groups were injected intraperitoneally with 100 μg·kg-1 BW LPS, and the piglets in the control group were injected intraperitoneally with the same dose of physiological saline. On d10 and d20, blood samples were collected from precava vein after 3 h postinjection of LPS. On d21, all the piglets were sacrificed to collect liver for analysis. The results showed that: (1) Dietary supplementation of NAC attenuated the decrease of F/G （P<0.05）and the increase of ADG（P<0.05）.(2) Dietary supplementation of NAC significantly attenuated the increase of TNFα, IL6 and PGE2 in plasma and liver(P<0.05). (3) Dietary supplementation of NAC attenuated the increase of the expression of NFκB and HSP70 induced by LPS challenge (P<0.05). (4) Dietary supplementation of NAC attenuated the decrease of ATP, ADP and energy charge (EC), while attenuated the increase of AMP level and AMP/ATP induced by LPS challenge significantly (P<0.05). These results indicated, under the present experimental conditions, dietary supplementation with 500 mg·kg-1 NAC could significantly improve growth performance via ameliorating the immunological stress and energy depletion in piglets challenged with LPS.

comEffects of Different Rumen Degradable Protein Balance (RDPB) on Rumen Fermentation, Total Gas and Methane Production in vitro

LI Qiufeng;CAO Yufeng;LI Jianguo

2012, 43(4): 572-579. doi:

Abstract ( 725 )

PDF (410KB) ( 443 )

Related Articles | Metrics

This experiment was conducted to study the effects of RDPB（RDPB，g·kg-1 DM） on rumen fermentation, total gas and methane production in vitro. AnkomRFgas production system was used by random block design, with 6 different rumen degradable protein balance (RDPB) diets, namely diet 1（-20 g·kg-1 DM）, diet 2(-10 g·kg-1 DM), diet 3(0 g·kg-1 DM), diet 4 (10 g·kg-1 DM), diet 5 (20 g·kg-1 DM) and diet 6 (30 g·kg-1 DM), with 2 replicates, respectively, culturing for 24 h. The results showed that: (1) The fermentation broth pH, acetic acid / propionic acid, amount of gas production and CH4 production of and diet 3（P<0.01） or diet 4（P<0.01） showed the lowest level. (2) Volatile fatty acids, acetic acid, propionic acid, butyric acid and rumen ammonia concentration increased with the increase of the balance value of the degradation, therefore, changing the balance of dietary rumen degradable protein could affect the rumen fermentation. The balance of rumen degradable protein had no significant effect on the total amount of volatile fatty acids and protozoa. (3) Concentration of methane at different time points and 24 h methane production were affected by dietary rumen degradable protein balance. Methane production of diet 4 （P<0.05） was lower than that of other diets. Thus, in vitro culture conditions, dietary RDPB affected rumen pH, ammonia nitrogen (NH3N), volatile fatty acids (VFA) concentration, gas and CH4 production. The results indicated that appropriate RDPB was 010 g·kg-1 DM, considering its effect on rumen fermentation and methane production.

Influence of Porcine Circovirus Type 2 Infection on Immune Response of Pigs to Classical Swine Fever Vaccine

ZHAO Danping#;TAO Bin#;CHEN Ligong;LI Yanqin;SONG Qinye;LI Tanqing;YANG Fang;LI Hongyuan;YUAN Hongxing

2012, 43(4): 580-587. doi:

Abstract ( 375 )

PDF (942KB) ( 522 )

Related Articles | Metrics

To study the influence of porcine circovirus type 2 (PCV2) infection on immune response of pig to classical swine fever (CSF) vaccine, twenty 28dayold healthy weaning piglets were randomly divided into four groups of five (Group VI, IV, V and C). The piglets in Group VI were intramuscularly injected with CSF vaccine two days before being inoculated with PCV2, piglets in Group IV were infected by PCV2 two days before being injected with CSF vaccine, piglets in Group V were only vaccinated by CSF vaccine and Group C as the blank control without inoculating anything. Piglets were vaccinated twice at an interval of twentyone days and blood samples were collected periodically so as to detect CSFVspecific serum antibody, lymphocyte proliferation activity and mRNA expression levels of IFNγ, IL2 and IL4 in peripheral blood lymphocyte (PBLC). PCV2 infection, either before or after vaccination, reduced CSFVspecific serum antibody production, seroconversion and lymphocyte proliferation activity. After the primary vaccination, expression of IFNγ, IL2, IL4 and IL10 mRNA decreased seriously in PBLC from piglets in Group VI and Group IV, especially in Group VI. Following the secondary vaccination, expressions of IFNγ and IL10 mRNA in PBLC from Group VI and Group IV appeared seriously unbalanced and those of IL2 as well as IL4 mRNA were deficient, especially in Group IV. In conclusion, PCV2 infection before or after vaccination with CSF vaccine affected porcine humoral and cellular immunity and led to expression suppression and unbalance of cytokines in PBLC.

Preparation and Preliminary Application of Monoclonal Antibodies against Nucleoprotein of Peste des Petits Ruminants Virus

ONG Yunfeng;ZHOU Xiaoli;YANG Junxing;WANG Qiong;ZHU He;YE Lingling;DONG Jun;LV Jianqiang;WANG Jinping;CHEN Chaoyin;HUA Qunyi;YANG Shibiao;YIN Shanglian;XU Weijia;AI Jun

2012, 43(4): 588-595. doi:

Abstract ( 376 )

PDF (893KB) ( 523 )

Related Articles | Metrics

The objective of this study was to prepare specific monoclonal antibodies (mAbs) against nucleoprotein of peste des petits ruminants virus (PPRV) , characterize its properties, and use it in ELISA. After immunization of BALB/c mice with purified recombinant protein BacmidPPRVN, two monoclonal antibodies against PPRV nucleoprotein were successfully prepared by polyethyleneglycol (PEG)mediated fusion of sensitized lymphocytes and myeloma cells and named 5B11 and 3H103B8, respectively. The specificity and biological characterization of the mAbs were identified. A competitive ELISA using PPRV recombinant nucleoprotein as coating antigen and mAb 5B11 as competitive antibody was established. Our results about isotypes and subclasses indicate that 5B11 and 3H103B8 belong to IgG2b, the titres of ascitic fluids of 5B11 was up to 1:819 200 and 3H103B8 was 1:12 800. These mAbs could specifically recognize recombinant protein BacmidPPRVN antigens in a serologic test. Antibodies additivity assay demonstrated that 5B11 and 3H103B8 recognized the different epitopes of PPRV nucleoprotein. The number of chromosome of the hybridoma cell lines was 99 to 104. A total of 222 serum samples were detected in parallel by cELISA and reference cELISA kit.The coincidence rate of the two assaya was 98.15%. We successfully prepared two specific monoclonal antibodies against PPRV nucleoprotein and established the competitive immunofluorescent method for detecting PPRV.

Antibody Dynamics and Protective Efficacy of Recombinant Fowlpox Virus Coexpressing Cytokine IL6 Gene and HA Gene of H5 Subtype Avian Influenza Virus

CHEN Sujuan#;DING Yanhong#;QIAN Cheng;CHAI Mao;PENG Daxin;LIU Xiufan

2012, 43(4): 596-601. doi:

Abstract ( 359 )

PDF (350KB) ( 367 )

Related Articles | Metrics

In order to evaluate the antibody dynamic in different immune routes and immune protection of the recombinant fowlpox virus coexpressing cytokine IL6 gene and HA gene of H5 subtype avian influenza virus (rFPVAIH5AIL6). Groups of 10dayold SPF chickens were inoculated by subcutaneous route in the back of neck and wings route with inoculums containing rFPVAIH5AIL6. There was no significant difference in body weight between the group of rFPVAIH5AIL6 and the normal control group, but the body weight in group of wild type strain was lower than that of the normal control group. The result of HI antibodies showed that the chickens inoculated subcutaneously by wing route produced higher level HI antibody. HI antibody induced by rFPVAIH5AIL6 was higher than that induced by rFPVAIH5A. HI antibody reached a peak at 21 days post vaccination，decreased at 28 days post vaccination，and remained at a certain level at 49 days post vaccination. Another Group of 10dayold SPF chickens was inoculated by subcutaneous route in wings with inoculums containing rFPVAIH5AIL6. The chickens were challenged with H5 subtype avian influenza virus at 21 days post vaccination. The protection indexes of groups immunized with rFPVAIH5AIL6 and killed vaccine were 95%, significantly higher than that of other groups. The result of virus shedding showed that the shedding rate of chickens immunized with rFPVAIH5AIL6 was the lowest. In summary, chicken IL6 could improve the immune efficacy against AI when it was coexpressed with HA gene in recombinant fowlpox virus. This study paves the way for further development of a new AIV recombinant vaccine.

New Features of Tumors Induced by Avian Leukosis Virus Subgroup J in Layer Flock

YU Linlin;JIANG Yanping;WANG Yue;CHEN Hongbo;WANG Feng;WANG Xiaowei;WANG Guihua;CHENG Ziqiang

2012, 43(4): 602-608. doi:

Abstract ( 278 )

PDF (3269KB) ( 698 )

Related Articles | Metrics

In August 2009, A Hyline layer chicken flock was infected at Zoucheng in Shandong Province, and mortality was up to 7% at age of 160day. Three of illness chickens were presented and diagnosed as avian leukosis virus subgroup J (ALVJ) infection by gross and histopathological observation, PCR and immunohistochemistry, etc. Histopathological observation showed that hemagioendotheliosarcoma existed single in No.1 chicken; fibrosarcoma was concurrent with myelocytomas in No.2 chicken. No cytopathic effect was found on the cultured DF1 cells inoculated with sick bird liver samples and of which supernatants fluid was positive for ALV p27 antigen by ELISA test. Phylogenetic analysis showed that gp85 nucleotide sequences of the isolated strain had the highest homology with that of the prototype HPRS103 (94.1%). Our study demonstrated that ALVJ is a multipotential oncogenic virus. The mechanism of tumor spectrum expansion induced by ALVJ in layer flock needs to be further studied.

Construction of a cDNA Library from Male Adult Toxocara canis and Analysis of Expressed Sequence Tag (EST)

MA Guangxu;ZHOU Qi;WEI Zhipeng;WANG Deheng;CHEN Yuming;ZHOU Rongqiong

2012, 43(4): 609-614. doi:

Abstract ( 331 )

PDF (765KB) ( 332 )

Related Articles | Metrics

To construction of a cDNA library for male adult T. canis, total RNA were extracted using Trizol reagent from Invitrogen (GIBCO BRL) according to the manufacturers instruction. The cDNA was synthesized and the ds cDNA was ligated into the λTripEx2 Vector. The ligation products were packaged with Gigapack III Gold Packaging Extract (Stratagene). Then the mixtures were transformed into Escherichia coli XL1Blue host cell culture to determine the titers of the unamplified and amplified libraries. The library qualification evalution showed: the titer of the primary cDNA library was 5.25×106 pfu·mL－1 with a recombination rate of 99.47%, and the titer of the amplified cDNA library was 6.90×109 pfu·mL－1. PCR amplification of randomly picked clones revealed that the inserted cDNA fragments ranged from 500 to 2 000 bp with an average length of 1 000 bp. The cDNA library was constructed successfully, and 189 efficient ESTs (expressed sequence tags) of 5′ end were obtained from this cDNA library. Cluster analysis of these ESTs identified 101 unique sequences containing 27 Contigs and 74 Singletons. All the unique sequences were deposited under dbEST in GenBank (GenBank: HO348195HO348295). BLASTX searches revealed that 45 Unigenes (44.55% of the total) or 88 ESTs (46.56% of the total) were novel genes that had no significant matches to any protein sequences in the public databases. The rest 56 Unigenes (55.44% of the total) or 101 ESTs (53.44% of the total) were closely matched to the known genes or sequences deposited in the public databases. This work will provide a valuable resource for further research on gene function and molecule mechanism of T. canis.

Analysis of Benzalkonium Bromide and Chlorhexidine Induced Escherichia coli Resistance

ZHANG Jing;LIU Qingqing;MA Baorui;WEI Shuyong;WU Junwei

2012, 43(4): 615-619. doi:

Abstract ( 360 )

PDF (1513KB) ( 683 )

Related Articles | Metrics

To explore the correlation of resistance between disinfectants and antimicrobial drugs, Escherichia coli control strain ATCC25922 was induced in vitro by subinhibitory concentration of benzalkonium bromide and chlorhexidine, the MIC of various antimicrobial drugs were detected and the expression of acrA which is the regulator gene of fusion protein AcrA of Escherichia coli efflux pumps were tested by realtime fluorescent quantitative PCR before and after induction. The results showed that after induction by benzalkonium bromide and Chlorhexidine, Escherichia coli was resistant to Enrofloxacin, Sulfadimidine sodium, Oxytetracycline and Amoxicillin, and acrA gene was highly expressed. The results indicated that there was correlation between disinfectants and antimicrobial drugs, and the mechanism is probably efflux pump.

Tandom Expression and Bacteriostatic Activity in vivo of Plectasin

GUAN Tao;YU Bing;CHEN Daiwen;HAN Guoquan;HUANG Zhiqing;MAO Xiangbing;ZHENG Ping;MAO Qian

2012, 43(4): 620-626. doi:

Abstract ( 329 )

PDF (661KB) ( 440 )

Related Articles | Metrics

: The study was conducted to construct the tandom expression strains of the plectasin，and to analyze the bacteriostatic activity of the tandom expression products. According to the codon preference in E. coli expression system, the gene of plectasin was synthesized and inserted into plasmid pGEX4T1 to construct the recombinant plasmid pGEXPT/pe. The coadhesive end restriction and ligation strategy was used to add the repeat unit one by one. The recombinant vectors pGEXPT/pes2 and pGEXPT/pes3 with two and three repeat units of plectasin were constructed respectively. The expression vectors of tandom repeat plectasin gene were respectively transformed into E. coli DH5α and induced with IPTG. The expressed protein was detected by SDSPAGE and Western Blot analyses. Then the bacteriostatic activity was determined. The results showed that the fusion proteins were correctly expressed. DNA sequencing indicated that 135, 261 and 387 bp of gene fragments were obtained after amplifying by PCR. SDSPAGE results showed that the expressed fusion proteins had molecular weight of 30, 35 and 40 kDa bands and accounted for 64.5%, 26.4% and 25.3% of the total bacterium proteins, respectively. The Western Blot analysis demonstrated that all of the target proteins had immunological activity. The results of bacteriostatic activity in vivo indicated that all of the expressed fusion proteins were definite inhibitors of E. coli DH5α. However, bacteriostatic efficacy of the fusion expression products significantly increased and antibacterial time extended with the increasingly number of plectasin gene. The successful construction of the tandom expression strains and the inhibition of the expression products to host bacteria laid a foundation for product engineering of efficient bacteriostatic effect of plectasin.

Modulation of Retinoic Acid in LPSinduced Inflammation in Primary Culture of Rat Mammary Epithelial Cells

GU Beibei;;MIAO Jinfeng;LU Jinye;ZOU Sixiang

2012, 43(4): 627-633. doi:

Abstract ( 337 )

PDF (2032KB) ( 412 )

Related Articles | Metrics

Lipopolysaccharide (LPS) induced primary culture of rat mammary epithelial cells (rMEC) inflammatory model was established and the protective mechanism of retinoic acid was studied. rMEC s were separated through digestion by collagenase and hyaluronidase. When cells were grown to 90% confluence in 6well plates, (1) cells were treated with different concentrations of LPS; (2) cells were treated with 1 μmol·L-1 retinoic acid before stimulation with 10 μg· mL-1 LPS. Cells and supernatants were collected. In this study, inflammatory models of primary culture of rMEC were successfully established. Stimulation of rMEC with LPS elicited a marked increase in mRNA expression for inflammatory mediators. Treatment of retinoic acid significantly decreased the mRNA expression of those mediators. Retinoic acid could reduce LPSinduced inflammatory reaction. These results further indicate that retinoic acid may protect mastitis from proinflammatory cytokinemediated damage of mammary epithelial cells.

Effect of Lactobacillus rhamnosus LGA on βdefensin 9 Expression in Cultured Chicken Small Intestinal Epithelial Cells

LI Guanhong;HONG Zhimin;JIA Yongjie;YI Zhonghua;QU Mingren;LIU Siguo

2012, 43(4): 634-641. doi:

Abstract ( 372 )

PDF (801KB) ( 750 )

Related Articles | Metrics

This study aimed to investigate the effect of probiotic Lactobacillus rhamnosus LGA (L. rhamnosus LGA) on the βdefensin 9 (AvBD9) expression in cultured chicken small intestinal epithelial cells. L. rhamnosus LGA was selected to examine the timeand doseresponse of AvBD9 gene expression upon stimulation of chicken intestinal small epithelial cells with L. rhamnosus LGA. The AvBD9 mRNA expression was determined by fluorescence quantitative PCR (FQPCR). The results showed that AvBD9 expression was upregulated by stimulation of L. rhamnosus LGA at different concentrations (2×105, 2×106, 2×107 cfu·mL-1), and expression difference was observed between treatments at three bacterial concentrations. Heatinactivated L. rhamnosus LGA also stimulated the expression of AvBD9, and showed stronger induction response than live bacteria (P<0.05). The L. rhamnosus LGA promoted AvBD9 mRNA transcription in timedependent manner. The AvBD9 mRNA expression peaked at 12 h of incubation upon treatment of epithelial cells with lactobacillus. AvBD9 peptide was detected by Western blot in the supernatants of cultured epithelial cells treated with L. rhamnosus LGA, indicating that AvBD9 peptide releases into extracellular medium and exerts its biological actions. Probiotic L. rhamnosus LGA stimulates AvBD9 expression in the epithelial cells of chicken small intestine during lactobacillusepithelial cell interaction. This study reveals the new possible functional mechanism by which the probiotic Lactobacilli exert their beneficial effects to the host through the antimicrobial peptide expression in the small intestinal epithelium.

The Differential Expression of Cadherin 1 (CDH1) in the Growth Plate of Tibial Dyschondroplasia in Broiler Chickens at the Early Stage

TIAN Wenxia;;;LI Jiakui;WANG Rui;QIN Ping;NING Guanbao;QIAO Jiangang;LI Hongquan;BI Dingren;PAN Siyi;GUO Dingzong

2012, 43(4): 642-646. doi:

Abstract ( 368 )

PDF (1054KB) ( 378 )

Related Articles | Metrics

The aim of this study was to explore the differential expression of cadherin 1 (CDH1) in the growth plate of broiler chickens with tibial dyschondroplasia (TD) induced by thiram at the early stage. Chickens were dissected and growth plates were fixed in 4% paraform respectively in 4 ℃ at days 1, 2 and 6. RNA was extracted from the growth plates of control and thiramfed chickens. Then differential expression of CDH1 was identified by realtime PCR and immunohistology. The results showed that expression of CDH1 was upregulated on the growth plate of thiramdiet fed group. CDH1 protein synthesis mainly lied in cytoplasm of the prehypertrophic and hypertrophic chondrocyte in control or TD growth plates. No expression was discovered at proliferative zone and low expression was detected at calcification zone. Remarkably increased CDH1 was detected in TD growth plates at days 1, 2 and 6 after feeding thiram diet, which mirrored with their mRNA differential expression, and highly increased CDH1 staining was clearly seen at days 2 and 6 respectively. It was suggested that CDH1 was associated with cell adhesion, blood vessel invasion and it also regulated Wnt/βcat signal transmission with other regulator among endochondral bone formation.

Temperature and pH Decline of Cattle Carcass and Its Effect on Drip Oozing Rate during Conditioning

ZHAO Huiping;SUN Zhichang;LI Yongpeng;ZHANG Songshan;ZHENG Shixue;SUN Baozhong

2012, 43(4): 647-652. doi:

Abstract ( 357 )

PDF (795KB) ( 393 )

Related Articles | Metrics

In order to investigate the effect of temperature and pH decline on weep of beef, the temperature and pH decline of front(neck), middle(striploin) and rear(rump) of cattle carcass from one hundred Qingchuan cattle was determined, and weep of meat surface after conditioning was also analyzed. The results showed that the rate of temperature decline of middle carcass was the highest, and was 1.76 and 1.45 times of front and rear carcass, respectively. The pH of front carcass decreased slowest, while the rate of pH decline of middle and rear carcass was 6.56 and 4.88 times of front. The rank of ultimate pH was front>middle>rear carcass. There was significant positive relationship between Temperature at pH6 and weep of carcass, with R2 exceeded 0.8. The Temperature at pH6 had significant effect on weep of beef. These results suggest that the relative variation between temperature and pH have significant effect on water holding capacity of beef.

Ultrastructure Observation of Cysticercus pisiformis

PAN Yaoqian;LIU Xingyou;ZHAO Zhensheng;CHEN Jinshan;ZHANG Huirong;ZHU Guangrui;XIA Yinke;YIN Mei;TANG Hairong

2012, 43(4): 653-658. doi:

Abstract ( 356 )

PDF (2631KB) ( 431 )

Related Articles | Metrics

The aim of the study was to observe the ultrastructure of Cysticercus pisiformis under different state. The Cysticercus pisifomis were obtained in the procedure of slaughter inspection from meatpacking plant. The Cysticercus pisiformis was taken out by cutting cyst directly, or cultured with physiologic saline containing 10% bile until evagination. Then it was observed and recorded by scanning electron microscopy. Observed from top of scolex of Cysticercus pisiformis locate in cyst，the rostellum looked like a umbrella. The cutaneous muscular columns with hamula covered the front end of scolex. Viewed from the side, a row of antlersliking hamula adhered to the scolex. Four suckers were looked like cavity and distributed around the scolex behind the rostellum. Primary neck segments were more width and body segments less fine，in which there were lots of wrinkles in their surface respectively. The Cysticercus pisiformis cultured was divided obviously into scolex, neck, body segment and zone of cyst. When cutaneous muscular column of rostellum contracted, the antlerliking hamula extended to periphery, and the sucker also had ringlike and vertical shape contraction. It was also found evidently that the ringliking contraction appeared in neck and the lengthways contraction in body segments. The Cysticercus pisiformis under different state has distinctly dissimilar ultrastructure.

Establishment of Rapid Reverse Transcription Loop Mediated Isothermal Amplification for Detection of Avian Novel Flavivirus

LI Zhaolong;CHEN Shilong;LIN Fengqiang;WANG Shao;CHENG Xiaoxia;ZHU Xiaoli;CHEN Shaoying;

2012, 43(4): 659-663. doi:

Abstract ( 469 )

PDF (942KB) ( 390 )

Related Articles | Metrics

A reverse transcription loop mediated isothermal amplification (RTLAMP) assay was developed for detection of avian novel flavivirus virus. According to the sequences of avian novel flavivirus virus E protein gene, six primers specific to the eight sites of novel flavivirus virus gene were designed, and the reaction conditions were optimized. The results showed that the assay had no cross reaction with NDRV, GPV, MDRV, NDV, ILTV and FPV. The assay was performed in water bath within 45 minutes and the amplification result was visualized by adding SYBR Green Ⅰ or inspecting the white sediment. The detection limit of RTLAMP assay was 1 pg, which was 100 fold higher than that of one step RTPCR. The detection in clinical samples was 100% compliance with RTPCR. These results suggested that the newly developed RTLAMP assay is a simple and specific method for rapid detection of avian novel flavivirus virus in field conditions.

Transcription Profiles of Etron2 Gene during the Different Development Stages

QI Nanshan#;SUN Mingfei#;WU Caiyan;LIAO Shenquan;LV Minna;YUAN Jianfeng;YU Jingshu;LI Xiangrui;CAI Jianping

2012, 43(4): 664-668. doi:

Abstract ( 370 )

PDF (749KB) ( 393 )

Related Articles | Metrics

The objective of this study was to study the transcription profiles of Etron2 gene during the different development stages. The parasites at different developmental stages were harvested and their total RNA were substracted by realtime PCR using Etactin gene as a reference for establishing standard curves to evaluate the copy number of Etron2 genes. All of quantitative analyses were reliable because the melting curve showed a single peak. Etron2 gene transcription reached a peak during the unsporulated oocysts stage, then merozoites, sporulated oocysts and the lowest during the sporozoites stage. It is likely that the transcription messages were stored during the unsporulated oocysts stage, and then the Etron2 protein can be translated and secreted into the rhoptry soon when the oocysts were sporulated, and it may play an important role in the process of invasion.

Table of Content