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Proviral Genomic Variation of EIAV Envelop gp 90 Gene Observed in Experimentally Infected Horse

Quankai WANG ; Hai PANG ; Xiangang KONG ; Hua XUAN ; Jingang ZHANG ; Jingliang LU ; Enge FEI

Chinese Journal of Veterinary Science.1999;19(3):207-211.

The present experiment was designed to observe the genetic variation of equine infectious anemia virus (EIAV) envelop gp 90 gene in infected horse. One horse was infected experimentally with P337-V70 strain and showed no clinical signs after being infected at twice with the same virus strain. Seventeen proviral sequences covering principal neutralizing domain (PND) of EIAV gp 90 gene were obtained from the buffy coat and liver of the horse through PCR amplification and cloning. Comparative analysis of the sequences revealed that some sequences contained the nucleotide insertions in the PND region. The insertions might be generated by direct repeat and strand displacement of sequence segment in their PND gene, showing different lenghts.

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Antibiotic Sensitivity Detection and Analysis of Escherichia coli Isolated Clinically in China

Liancheng LEI ; Dan ZHENG ; Wenyu HAN ; Fengtao JI ; Weidong XU

Chinese Journal of Veterinary Science.2005;25(5):470-473.

The susceptibility to 9 kinds of antibiotics of 393 animal pathogenic E. coli isolated from clinical samples was determined from 2000 to 2003. The resistance to TC, GM, CMP, AMP, RA, KM, FT, SM and CIP were 93.89%, 57.76%,78.63%, 77.86%, 92.11% ,47.33%, 46.82%, 76.84% and 74.81%, respectively. The isolates could be classified into eight classes according to the number of drugs to which srains were resistant. The resistance spectrum of the isolates varied from 2to 9 kinds of the above drugs. The strains that were resistant to seven kinds of drugs were more than 80 percent. The resistance rates of swine origin strains to GM,AMP and KM were higher than those of poultry origin strains, while the resistance rates of poultry origin strains to TC, CMP, SM and CIP were higher remarkably than those of swine origins. The frequency of resistance increased from 2000 to 2003,which was from 67.33% to 90.58% for CMP, 71.29% to 84.81% for AMP, 73.76%to 80.10% for SM and 61.88% to 88.48% for CIP. At the same time, the resistance rate of GM decreased from 61.39% to 53.92%. Thirty-seven strains (95 %) could make all the Kunming mice die within 72 h injected intraperitoneally with the culmice three times in vivo. The pathogenicity of wild strains with drug resistance acquired naturaly to mice did not decline.

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Expression of Bovine Interleukin-2 Gene in Pichia pastoris

Fang CHEN ; Hongli SUN ; Xiangrong CAO ; Zhen LI ; Ruisong YU

Chinese Journal of Veterinary Science.2005;25(2):178-182.

The Interleukin-2 gene cDNA was cloned into the Pichia pastoris expression vector pPICZB,which is under the control of the alcohol oxidase promoter AOX1. The linearized recombinant plasmid of BoIL2-pPICZB,digested by Sac I ,was transformed into X-33 strains by electroporation. The multi-copy insert transformants were screened by Zeocin-resistance and induced by 1% methanol. The intracellular expression products were tested by SDS-PAGE analysis and Western blotting. Purified recombinant BoIL2 was gained by metal-chelating affinity chromatographic (MCAC). Assay with murine CTLL-2 cells showed that the recombinant BoIL2 exhibited the biological activity.

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The Influence of Sera,Gonadotropins,EGF and Water on In vitro Maturation of Goat Oocytes

Chao WANG ; Zhiqing WEI ; Xiangchen LI ; Zhixin AN ; Zhiping ZHANG ; Jianhong SHU ; Yong ZHANG

Chinese Journal of Veterinary Science.2004;24(6):599-602.

In present study,comparisons were carried out to develop and improve in vitro mauration(IVM) systems of goat oocytes. Oocytes were cultured in TCM-199 supplemented with(1)10% sera (either estrous goat serum (EGS) or fetal bovine serum(FBS)) + 20 mg/L luteinizing hormone (LH) + 10 mg/L follicle stimulating hormone (FSH) + 1 mg/L estradiol (E2); (2)10% EGS with different gonadotropins(LH: FSH) at a concentration of 5 mg/L: 0.5 mg/L or at 20 mg/L: 10 mg/L with0. 075 IU/mL human menopausal gonadotropin(HMG),1 mg/L estradiol 17β; (3)10% EGS+ 0. 075 mg/L HMG+10-20 μg/EGF. The culture was also performed by M199 supplemented with 10% EGS+0. 075 mg/L HMG+ 10-20 μg/L EGF into ultra-filtrated water which was derived either from self-producing or from purchased for use. The oocytes were cultured at 38 ℃,5% CO2 in air for 24 h,and the meterphase Ⅱ stage oocytes were examined under dissecting microscope. The results showed that EGS was better than FBS in supporting goat oocyte IVM. An addition of HMG in M199 could improve oocyte maturation and induce a higher percentage of metaphase Ⅱ oocytes compared to gonadotropins. 10-20 μg/L EGF improved goat oocyte maturation but the influence was not significant. Fresh,high quality water was vital for oocyte IVM. In conclusion,under our conditions with IVM ,the best result in maturation of goat oocyte has been M199 supplemented with 10% EGS+0.075 IU/mL HMG+10-20 μg/L EGF and prepared in fresh purified water.

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Effect of Zinc on Peripheral Blood T-lymphocyte in the Duckling

Hengmin CUI ; Jing FANG ; Xi PENG ; Junliang DENG

Chinese Journal of Veterinary Science.2004;24(1):66-68.

150-day-old Tianfu ducklings were divided into three groups,and fed with diets as follows :Zn deficient (22. 9 mgZn per kg diet),controls(100 mg Zn per kg diet) and Zn toxic (1 300 mg Zn per kg diet) for seven weeks (Zn deficiency,ZD)or four weeks (Zn toxieity,ZT). The ANAE+ positive ratios of the peripheral blood T-lymphocytes were much lower (P＜0.01 ) in Zn deficient and toxic groups than in the control group. The results showed that Zn deficiency and toxicity would suppress the development of T-lymphocytes and reduce the peripheral blood T-lymphocyte populations. Potential mechanisms underlying these observations are also discussed.

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Polymerase Chain Reaction Detection for Porcine Endogenous Retrovirus in 4 Pig Cell Lines

Maomin LU ; Hong JIN ; Ruichun DENG ; Jianguo WANG ; Shu YANG ; Jingfeng XIONG ; Zhuang DING ; Jingang ZHANG

Chinese Journal of Veterinary Science.2003;23(1):1-3.

Porcine endogenous retroviruses (PERV) can infect human cell in vitro, which raised widely concerns re-garding the transmission of PERV to xenograft recipients. It's essential to establish a method for detection of PERV.3 pairs of primers were synthesized according to the sequence of gag, pol and env gene of PERV. Polymerase chainreaction (PCR) and reverse transcription PCR (RT-PCR) assays were performed for detection of PERV provirusDNA and PERV specific mRNA. The results showed that provirus DNA and mRNA of PERV existed and expressedin all 4 tested cell lines. The sizes of amplified fragments are identical with the predicted. These methods may be suit-able for monitoring PERV in other cells or tissue.

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Influence of the growth traits and virulence genes of Enterococcus faecalis isolate N41 by cocultivation with Salmonella

Erli WU ; Chunhao HE ; Yue JIN ; Lingling LIU ; Hui HU ; Han CAO ; Yabin WANG ; Liying CHEN

Chinese Journal of Veterinary Science.2017;37(8):1528-1533. doi:10.16303/j.cnki.1005-4545.2017.08.17

To explore the impact of Salmonella on Enterococcusfaecalis isolate N41 under the coexisting,the growth traits and the transcription of 26 virulence genes of N41 at various growth phases were detected.Salmonella and/or N41 were inoculated and done plate counting,then growth curves were drawn and bacterial total RNA were extracted at given time points,quantitive realtime PCR was used to analyze the RNA transcription levels of 26 virulence genes of N41.The result showed that comparison with single incubation,the bacteria concentration of N41 dropped about 3.9 times,and Salmonella dropped about 110-times.Among 26 virulence genes of N41,the RNA transcription levels of 12 virulence genes such as ebpA,ebpC,rnjB,ace,ebpR,psr and so on were promoted at the four growth phases,but the RNA transcription levels of SlyA and sprE were dropped.Except that the RNA transcription levels of CylL-S,CylL-L,efaA and AS had no significant changes at the four growth phases,the mostly rest genes increased dramatically at post-log phase.This indicated that when incubated together,N41 inhibited the growth of Salmonella significantly,and Salmonella promoted the transcription levels of virulence genes of N41 as a whole.The results lay a foundation for further research on the interaction between bacteria species and the pathogenicity mechanism of Enterococci.

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Establishment and preliminary application of multiplex PCR for detecting toxin-typing of goat Clostridium perfringens strains

Changlei BAO ; Mingzhe FU ; Yapeng HE ; Tao BAI ; Jianxuan WEI ; Yanming ZHANG ; Xingang XU

Chinese Journal of Veterinary Science.2017;37(8):1523-1527. doi:10.16303/j.cnki.1005-4545.2017.08.16

According 1o the genome sequences of α.β,e,ι toxins of Clostridium perfringens in GenBank,four pairs of primers targeting α,β,ε,ι toxin genes were designed.After the multiplex PCR reaction condition was optimized,the multiplex PCR for identification and toxintyping of C.perfringens strains was developed.The specificity test showed that the expected fragments of C.perfringens reference strains including A.B,C,D,E five toxin types were amplified successfully from genomic DNA of C.perfringens,respectively.However,a band could not be amplified from Clostidrium novyi and Clostridium septicum as negative control groups.The sensitivity test showed that the limit detection of multiplex PCR was 9.0,17.8,12.2,13.8,18.5 pg DNA of A,B,C,D,E five toxin types C.perfringens,respectively.Repetitive testing showed that the established method had a good repeatability.Nine type A strains of and 1 type C strains of C.Perfringens from 21 clinical samples of dead goat were detected by the multiplex PCR developed in this study.This study establishes the multiple PCR method which not only can detect C.perfringens rapidly but also can identify five toxin types of C.perfringens.

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Comparative analysis of the intestinal microbial flora diversity of adult and young African lions

Xiaoying JIA ; Wei WANG ; Wenlong DONG ; Jiazhen WANG ; Shixin ZHU ; Yujie LOU ; Yunhang GAO

Chinese Journal of Veterinary Science.2017;37(8):1507-1511. doi:10.16303/j.cnki.1005-4545.2017.08.13

In this study,the intestinal microbial flora diversity of adult and young African lions in the same breeding environment was detected by PCR-DGGE technique.Total bacterial DNA was extracted and 16S rDNA V3 region was amplified,then conducting PCR-DGGE.Subsequently,the specific bands of DGGE were cloned and sequenced.The bacterial species were identified by comparing the sequence through BLAST.The results indicated that the intestinal microbial flora of adult African lions includes Clostridium,Lachnospiraceae bacterium,Anaerovorax,Lactococcus,Peptostreptococcus and Blautia.While the intestinal microbial flora of young African lions is lesser,most bacteria are common to adult and young lions,such as Bacteroidetes bacterium and rumen bacterium.The UPGMA clustering analysis of the DGGE fingerprint showed the similarities of the bacteria structures between adult and young African lions were only 34％.These results revealed that the intestinal microbial flora has significant difference in different stages of African lions.This study lays a foundation for the development of microecological agents in different growth stages of wild animals.

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Investigation of avian diseases eradication in parent breeding flocks in China in 2015

Xue HAN ; Qian ZHANG ; Xiaoxue GU ; Yuliang LIU ; Yang LIU ; Yiming BI ; Xinyan ZHAI ; Chuanbin WANG

Chinese Journal of Veterinary Science.2017;37(8):1490-1494. doi:10.16303/j.cnki.1005-4545.2017.08.10

In order to understand the management measures,technique abilities and difficulties in controlling and eradicating avian diseases in parent breeding flocks,the current situation of avian disease eradication in breeding flocks was investigated by China Animal Disease Control Centre in 19 provinces in July 2015.Questionnaires investigation was conducted and the feedbacks were received from 214 parent breeding flocks.This study summarized and analyzed the information of farm management,breeding resources,avian disease surveillance and disease eradication in these flocks,which will provide the basic data to promote the avian diseases control and eradication in China.

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