• Title/Summary/Keyword: Newcastle disease virus (NDV)

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Production of recombinant nucleocapsid protein of Newcastle disease virus in Escherichia coli for a diagnostic ELISA

  • Kim, Hyun-Il;Park, Kyoung-Phil;Park, Chan-Hee;Cho, Hyun-Ah;Yang, Ho-Suk;Hahn, Tae-Wook
    • Korean Journal of Veterinary Research
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    • v.49 no.1
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    • pp.39-44
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    • 2009
  • Transmission of avian viruses both bird-to-bird and from birds to non-avian species is a major health concern. Newcastle disease virus (NDV) is an economically important avian virus that poses substantial risks to the poultry industry. Rapid and sensitive diagnostic methods, such as the enzymelinked immunosorbent assay (ELISA), are required to track such infections. To develop an ELISA for detecting anti-NDV antibody in avian sera, the nucleocapsid protein (NCP) gene of the NDV La Sota strain was cloned and expressed in Escherichia coli and the 513-amino acid recombinant NCP was purified by Ni-NTA affinity chromatography. To evaluate its ability to replace NDV whole virus antigen as a coating antigen, NCP-coated and whole NDV-coated ELISAs were tested and compared using a panel of NDV positive antisera from chickens. Results using purified NCP were highly correlated with those obtained using whole NDV (r= 0.927), demonstrating that recombinant NCP expressed in Escherichia coli is a suitable substitute antigen for whole NDV in a diagnostic ELISA.

Antiviral Activity of Methylelaiophylin, an ${\alpha}$-Glucosidase Inhibitor

  • Lee, Do-Seung;Woo, Jin-Kyu;Kim, Dong-Hern;Kim, Min-Young;Cho, So-Mi K.;Kim, Jae-Hoon;Park, Se-Pill;Lee, Hyo-Yeon;Riu, Key Zung;Lee, Dong-Sun
    • Journal of Microbiology and Biotechnology
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    • v.21 no.3
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    • pp.263-266
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    • 2011
  • Methylelaiophylin isolated from Streptomyces melanosporofaciens was selected as an ${\alpha}$-glucosidase inhibitor with an $IC_{50}$ value of 10 ${\mu}M$. It showed mixed-type inhibition of ${\alpha}$-glucosidase with a $K_i$ value of 5.94 ${\mu}M$. In addition, methylelaiophylin inhibited the intracellular trafficking of hemagglutinin-neuramidase (HN), a glycoprotein of Newcastle disease virus (NDV), in baby hamster kidney (BHK) cells. Methylelaiophylin inhibited the cell surface expression of NDV-HN glycoprotein without significantly affecting HN glycoprotein synthesis in NDV-infected BHK cells.

Production Newcastle Disease Virus Using Vero Cell Culture (Vero 세포배양을 이용한 뉴캐슬병 바이러스 생산)

  • 이광원;김익환김동일
    • KSBB Journal
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    • v.10 no.3
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    • pp.292-297
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    • 1995
  • Studies on the production of Newcastle disease virus(NDV) were carried out to optimize culture conditions such as initial pH, temperature, serum concentration, multiplicity of infection(M.O.I.) as well as the addition of polycation, antioxidant, and DMSO. Initial pH from 7.2 to 8.1 showed little difference on NDV production but the initial pH below 6.8 resulted in the negative effect. The highest NDV titer was obtained at 0.1 M.O.I. In addition, the maximum production of virus was achieved at 2% FBS and optimum temperature was found to be $34^{\circ}C$. Treatment of polycoation increased the virus production. When ascorbic acid was added as an antioxidant, NDV production was also enhanced. Utilization of DMSO, a well-known permeabilizing agent, showed an inhibitory effect on the propagation of NDV.

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Monoclonal Antibodies Against a Paramyxovirus Isolated from Japanese Sparrow-Hawks(Accipiter virugatus gularis) (일본 새매 (Accipiter virugatus gularis)로부터 분리된 Paramyxovirus에 대한 단 Clone성 항체)

  • Hoshi;Mikami, S.T.;Onuma, M.;Izawa, H.
    • Korean Journal of Poultry Science
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    • v.10 no.1
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    • pp.60-66
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    • 1983
  • Monoclonal antibodies against Taka virus, a variant of Newcastle disease virus (NDV), were produced to compare the antigenicites of several avian paramyxoviruses including NDV. It was also used to study the activesite(s) of haemagglutin (HA) and neuraminidase activities of NDV. Five independent hybrid cell lines, which produced monoclonal antibodies against haemagglutinin-neuraminidase (HN) molecule of Taka virus, were established. From the results of the cross haemagglutination-inhibition(HI) test the monoclonal antibodies, the HN molecule of Taka virus seemed to have at least three different antigenic determinats; one was specific for all NDV strain tested, the second was only for Taka virus and the third was for Take virus, Banger and Yucaipa Furthermore the differences in the ratio of HI to neuraminidase-inhibition titers suggested that the active sites involved in HA and neuraminidase activities might be different from each other. However, since each of five monoclonal anitbodies was not especially specific for either HA or neuraminidase, the possibility that a single active site on the HN molecule may be responsible for both activities has not been excluded.

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Influence of Ligustrum lucidum and Schisandra chinensis Fruits on Antioxidative Metabolism and Immunological Parameters of Layer Chicks

  • Ma, Deying;Liu, Yuqin;Liu, Shengwang;Li, Qundao;Shan, Anshan
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.9
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    • pp.1438-1443
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    • 2007
  • The experiment was conducted to evaluate the effects of Ligustrum lucidum (LL) and Schisandra chinensis (SC) on the growth, antioxidative metabolism and immunity of laying strain male chicks. The results showed that diets supplemented with 1% of either LL or SC had no effects on the growth performance of chicks compared with the control. Furthermore, both LL and SC significantly reduced malondialdehyde (MDA) concentration of serum and heart of chicks (p<0.05). In addition, superoxide dismutase (SOD) activity of serum of the birds was significantly elevated by supplementation with SC (p<0.05). Glutathione reductase (GR) activity of heart and serum of the birds was significantly elevated by supplementation with LL or SC (p<0.05). LL supplementation significantly elevated antibody values against Newcastle Disease virus (NDV)(p<0.05) and lymphoblastogenesis (p<0.05) of the birds. The results suggest that diets supplemented with 1% of either LL or SC may improve immune function and antioxidant status of chicks.

Characteristics of a NDV isolated from apparently healthy wild spot-billed ducks (Anas poecilorhyncha) (흰뺨검둥오리(Anas poecilorhyncha)에서 분리된 뉴캣슬병 바이러스의 특성)

  • Choi, Kang-Seuk;Lee, Eun-Kyoung;Jeon, Woo-Jin;Kwon, Jun-Hun;Yang, Chang-Bum
    • Korean Journal of Veterinary Research
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    • v.48 no.2
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    • pp.153-159
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    • 2008
  • Newcastle disease virus (NDV) is the causative agent of a highly contagious and devastating Newcastle disease of poultry. A NDV (isolate DK1/07) was isolated from apparently healthy wild spot-billed ducks (Anas poecilorhyncha) captured at upper branch of the SapGyo Creek in Chungbuk province, Korea during early 2007. The DK1/07 isolate of minimum chicken embryo lethal dose killed all SPF chicken embryos within 60 h. The cleavage site of the F protein possessed the amino acid sequence $^{112}R-R-Q-K-R-F^{117}$, which is a motif characteristic of virulent NDV strains. The F protein-based phylogenetic analysis revealed that the DK1/07 duck isolate was included in the cluster of genotype VIId and most closely related to recent NDV isolates obtained from chicken farms in Korea. Epidemiological importance of virulent NDV from wild duck is discussed.

Comparative Evaluation of Three Purification Methods for the Nucleocapsid Protein of Newcastle Disease Virus from Escherichia coli Homogenates

  • Tan Yan Peng;Ling Tau Chuan;Yusoff Khatijah;Tan Wen Siang;Tey Beng Ti
    • Journal of Microbiology
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    • v.43 no.3
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    • pp.295-300
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    • 2005
  • In the present study, the performances of conventional purification methods, packed bed adsorption (PBA), and expanded bed adsorption (EBA) for the purification of the nucleocapsid protein (NP) of Newcastle disease virus (NDV) from Escherichia coli homogenates were evaluated. The conventional methods for the recovery of NP proteins involved multiple steps, such as centrifugation, precipitation, dialysis, and sucrose gradient ultracentrifugation. For the PBA, clarified feedstock was used for column loading, while in EBA, unclarified feedstock was used. Streamline chelating immobilized with $Ni^{2+}$ ion was used as an affinity ligand for both PBA and EBA. The final protein yield obtained in conventional and PBA methods was $1.26\%$ and $5.56\%$, respectively. It was demonstrated that EBA achieved the highest final protein yield of $9.6\%$ with a purification factor of 7. Additionally, the total processing time of the EBA process has been shortened by 8 times compared to that of the conventional method.

Characterization and comparison of the pathogenicity of viscerotropic velogenic Newcastle disease virus isolates in Korea

  • Kim, Jae-Hong;Sung, Haan-Woo;Kim, Il-Hwan;Lee, Eun-Kyoung;Choi, Kang-Seuk;King, Daniel Jack
    • Korean Journal of Veterinary Research
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    • v.52 no.4
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    • pp.213-221
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    • 2012
  • A total of 18 Newcastle disease virus (NDV) isolates that were recovered from 1949 through 1997 were characterized and pathotyped. All viruses were highly virulent as determined by intracerebral pathogenicity indices ${\geq}1.81$ in day-old. These pathotypes are typical for viscerotropic velogenic NDV (VVNDV) pathotype viruses. Some differences were observed for the chicken red blood cell elution rate and thermostability of the hemagglutinin at $56^{\circ}C$. Three antigenic groups were identified by a hemagglutination-inhibition assay using NDV monoclonal antibodies. And the predominant gross lesions were as follows: discharge from the nasal cavity, tracheal mucus, petechial hemorrhage in the heart fat, kidney urates and hemorrhage with or without necrosis in the gastrointestinal tract. Severe hemorrhagic or necrotic lesions were also noted in the lymphoid organs and were localized primarily in the spleen and cecal tonsil. However, differences in the occurrence and frequency of the gross lesions were observed between the virus strains. Among them, NDV strains that induced neurological symptoms belonged only to genotype VI. This strain had spread throughout Korea during the late 1980s to the 1990s, which suggests that specific VVNDVs genotypes might result in neurological symptoms.

Production of Monoclonal Antibody to Newcastle Disease Virus and its Diagnostic Use (뉴켓슬병 바이러스에 대한 단크론성 항체생산 및 진단에 이용)

  • Chung Ok Choi;Chung Gil Lee;Sung Man Cho;Jin Soo Na;Soo Hwan An;Joon Hun Kwon
    • Korean Journal of Poultry Science
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    • v.15 no.3
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    • pp.207-210
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    • 1988
  • A total of 3 hybridoma clones producting menoclonal antibody (MCA) against Newcastle disease virus(NDV) was raised by cell fusion method. The MCAs did not cross react against other avian or mammalian viruses tested. However, these antibodies reacted with all strains of velogenic and lentogenic NDVs tested indicating that they are unable to discriminate the possible antigenic differences among NDVs. All. the MCAs were classified as IgG type and did not show neutralizing and hemagglutination inhibition (HAI) activity except one clone which has low HAI activity. One of these MCA raised in mouse ascites revealed the titer of $10^6$ by indirect immunofluorescent antibody (IFA) test Using the MCA, virulent NDV could easily be detected from tracheal and conjunctival smears made 2 to 3 days after experimental infection.

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육계에서 멜라토닌의 주기적인 변화와 면역성 및 생산성에 미치는 영향에 대한 고찰

  • 류명선;김상호;류경선
    • Proceedings of the Korea Society of Poultry Science Conference
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    • pp.81-84
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    • 2000
  • Effects of different photoperiod regimens on the cellular and humoral immunity in broiler chickens were studied(Exp 1). Total one hundred ninety two one-day-old commercial broiler chicks(Cobb$\times$Cobb) were raised between constant lighting(CL) and intermittent lighting (1h light: 3h darkness(IL; 1l; 3D) Body weight, feed intake and feed conversion were measured for seven week. Peripheral blood and splenic lymphocyte activities were tested at 3 and 5 wk of age by performing a mitogen cellproliferation assay with a polyclonal T-cell mitogen, concanavalin A (Con A), and B-cell mitogen, lipopolysaccharide (LPS). To investigate the effect of photoperiod on the humoral immunity, chicks were immunized with sheep red blood cell(SRBC) and iinactivated Newcastle disease virus(NDV) vaccine. Total immunoglobulin G(IgG) concentration was also determined. Diurnal change of melatonin was tested in sera. In experiment 2, 0.1ml melatonin were subcutaneously injected from three to five weeks old if immunomodulation effect of lighting regimen was due to the melatonin or not. Injections of melatonin were made at 0700h and the dosage was 10ng (M2), 100ng(M3), 1$\mu\textrm{g}$(M4) per bird daily, respectively. control were quivalent injections of vehicle(M1). Lymphocyte activities were tested and humoral immunities were examined at 5 weeks of age. Blood melatonin concentration was determined at 0h, 1, h, 2h, and 3h posterior to injection at five weeks old. It was higher in CL chicks than IL chickens during the subsequent period of 3 to 5 wk of age. However, weight gain of chicks raised IL were significantly higher at 6 wk of age than CL(P<0.05). Antibody response to NDV was not affected by both photoperiod regimens and melatonin injection, whereas anti-SRMB titer and IgG concentration were enhanced. Lymphocyte activity of chickens raised under IL was sighificantly higher than those of chickens raised under CL. Melatonin injection also increased lymphocyte activity. When peripheral blood lymphocytes were used, proliferation response to LPS and Con A were significantly increased in M2 and respectively. The results of this experiments suggest that IL improved host immune response and melatonin have immunomodulatory roles.

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