• Title, Summary, Keyword: Newcastle disease virus

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Immunohistochemical identification of newcastle disease virus with indirect immunoperoxidase technique (Indirect Immunoperoxidase 법을 이용한 조직내 뉴켓슬병 바이러스 항원동정)

  • Nho, Whan-goog;Sur, Jung-hyang;Kim, Soon-bok
    • Korean Journal of Veterinary Research
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    • v.30 no.3
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    • pp.309-315
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    • 1990
  • The present experiment was done to identify newcastle disease virus(NDV) antigens in frozen sections of various oragns from experimentally NDV-infected with indirect immunoperoxidase method. Section were incubated with rabbit anti-NDV polyclonal as first antibody, followed by incubation with goat anti-rabbit or protein A peroxidase conjugate. Positive reactions were often detected in the epithelium of trachea and in the lymphocyte of spleen at 24 hours after virus inoculation. the viral antigen was localized mainly in the cytoplasm of infected cells. The method approved to be highly specific for the identification of NDV and allowed a precise localization of the viral antigens in infected cells.

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Molecular Biological Characterization of the First Newcastle Disease Virus Isolated in Mongolia (몽골에서 최초로 분리된 뉴캣슬병 바이러스의 분자생물학적 특성)

  • Choi, Kang-Seuk;Lee, Eun-Kyoung;Jeon, Woo-Jin;Batchuulon, D.;Sodnomdarjaa, R.;Park, Mi-Ja;Yoo, Ye-Nah;Kwon, Jun-Hun
    • Korean Journal of Poultry Science
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    • v.38 no.2
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    • pp.89-96
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    • 2011
  • The outbreak of Newcastle disease occurred for the first time at a commercial chicken farm near Ulaanbaatar, Mongolia in August 2010. Newcastle disease virus (NDV) obtained from infected chickens in Mongolia was characterized by biological and molecular biological approches. Mongolian NDV isolate killed all of chicken embryos within 60 h in the mean death time assay, indicating virulent for chicken. A genomic region of 695 nts between nts 1055 of the M gene and 508 of the F gene was amplified by RT-PCR and sequenced. The deduced amino acid sequence of the F protein cleavage site was $^{112}RRQKRF^{117}$, which is a typical sequence of velogenic strains of NDV and is agreement with the result of the MDT assay. The sequence of the partial F gene (nts 47 to 435) was used for genotyping by phylogenetic analysis. The phylogenetic analysis showed that the Mongolian isolate was of genotype VII within class II of NDV. Further phylogenetic analysis on the genotype VII strains revealed that the isolates placed in a genetic sublineage of VIId and most closely related with velogenic strains of NDV circulating in Far-east Asian region especially China, suggesting the introduction of velogenic NDV into Mongolia from neighboring countries.

Coinfected cases with adenovirus, chicken infectious anemia virus and Newcastle disease in broiler chickens (육계에서 아데노바이러스, 전염성빈혈 및 뉴캣슬병 복합감염 증례)

  • Chu, Keum-Suk;Kang, Mi-Seon;Rim, Sang-Hyun;Lee, Jeong-Won
    • Korean Journal of Veterinary Service
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    • v.33 no.1
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    • pp.7-12
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    • 2010
  • There are several immunosuppressive viral diseases in chickens such as avian adenovirus (AAV), chicken anemia virus (CAV), infectious bursal disease (IBD) and Marek's disease (MD). In this study, we have investigated two broiler chicken farms suffered from high mortality in Jeonbuk in July to August 2009. Clinically high fever and growth retardation were observed in the diseased chicken. In necropsy, the hemorrhages in thigh leg and thymus, hemorrhages and enlargement of liver, kidney and proventriculus, and yellowish fluid in heart were seen. Histologically, necrotic foci and basophilic intranuclear inclusion bodies of hepatocytes, hemorrhages and infiltrated lymphocytes in kidney and proventriculus were observed. By using polymerase chain reaction (PCR), the genes of avian adenovirus, CAV and ND virus were detected in specimens. We suggested that these coinfection cases with high mortality were due to primarily infection of immunosuppressive diseases such as avian adenovirus, CAV, followed by secondary infection of Newcastle disease (ND) virus.

Molecular differentiation of Korean Newcastle disease virus (NDV) by restriction enzyme analysis and pathotype-specific RT-PCR

  • Kwon, Hyuk-Joon;Cho, Sun-Hee;Kim, Sun-Joong
    • Korean Journal of Veterinary Research
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    • v.46 no.4
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    • pp.371-379
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    • 2006
  • Newcastle disease virus (NDV) is a single-stranded negative sense RNA virus, which has been classified as a member of the Avulavirus genus of the Paramyxoviridae family. It is also one of the most important pathogens in the poultry industry. The glycoproteins, fusion (F) and hemagglutinin-neuraminidase (HN), determine the virulence of NDV, and the relevant molecular structures have already been determined. NDV isolates differ in terms of virulence, and at least 2 of 9 genotypes (I-IX) have been shown to co-circulate. Therefore, it is clearly important to differentiate between vaccine strains and field isolates. In vivo pathogenicity tests have been the standard protocol for some time, but molecular methods appear preferable in terms of the rapidity of diagnosis, as well as animal welfare concerns. In this study, we have designed primer sets from HN gene for phylogenetic analysis and restriction enzyme analysis, and from F gene for pathotype-specific RT-PCR. Via the combination of 2 methods, 106 Korean NDV isolates obtained from 1980 to 2005 were differentiated into vaccine strains, and virulent genotypes VI and VII. The genotype VI viruses were only rarely isolated after 1999, and genotype VII, after it was initially isolated from poultry in 1995, recurred in 2000, and then became the main NDV constituting a threat to the Korean poultry industry.

Molecular Cloning and Nucleotide Sequence of the Gene Encoding Fusion(F) Protein of the Thermostable Newcastle Disease Virus Isolated from a Diseased Pheasant (꿩에서 분리된 Newcastle Disease Virus 내열성주 (CBP)의 Fusion(F) 유전자 클론닝과 염기서열 분석)

  • Chang, Kyung-Soo;Jun, Moo-Hyung;Song, Hee-Jong;Kim, Kui-Hyun;Park, Jong-Hyeon
    • The Journal of Korean Society of Virology
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    • v.28 no.3
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    • pp.233-245
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    • 1998
  • The gene encoding F protein of CBP-1 strain, a heat-stable Newcastle disease virus (NDV) isolated from the diseased pheasants in Korea, was characterized by reverse transcription-polymerase chain reaction (RT-PCR), nucleotide and amino acid sequences. Virus RNA was prepared from the chorioallatoic fluid infected with NDV CBP-1 virus and cDNA was amplified by RT-PCR, cloned and sequenced to analyze. The PCR was sensitive as to detect the virus titer above $2^5$ hemagglutination unit. 1.7kb (1,707bp) size of the cDNA was amplified and cloned into BamHI site of pVL1393 Baculo transfer vector. The nucleotide sequences for F protein were determined by dye terminator cyclic sequencing using four pairs of primers, and 553 amino acid sequences were predicted. In comparison of the nucleotide sequence of F gene of CBP-1 with those of other NDV strains, the homology revealed 88.8%, 98.5% and 98.7% with Kyojungwon (KJW), Texas GB and Beaudette C strains, respectively. As the deduced 553 amino acid sequences of F protein of CBP-1 were compared with those of other NDV strains, the homology appeared 89.9%, 98.7% and 98.9% with KJW, Texas GB and Beaudette C strains, respectively. The putative protease cleavage site (112-116) was R-R-Q-K-R, indicating that CBP-1 strain is velogenic type. The amino acid sequences include 6 sites of N-asparagine-linked glycosylation and 13 cysteine residues. These data indicate that the genotype of CBP-1 strain is more closely associated with the strains of Texas GB and Beaudette C than KJW strain.

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Immunohistochemistry and RT-PCR for pathogenesis of Newcastle disease in chickens (닭 뉴캣슬병의 발병기전 규명을 위한 RT-PCR 및 면역조직화학적 연구)

  • 이민권;진영배;문운경;김순복
    • Korean Journal of Veterinary Service
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    • v.27 no.1
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    • pp.63-73
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    • 2004
  • The present experiment was carried out to study the pathogenesis of Newcastle disease(ND), ND virus (NDV) antigens and genes in various organs from NDV inoculated chickens were detected by immunohistochemistry and RT-PCR. Immunohistochemically, NDV antigens were detected in the spleen, thymus, cecal tonsil, proventriculus, trachea and lungs at 12 hour post-inoculation (hpi). Viral antigens were localized mainly in the cytoplasm of lymphocytes and macrophages. After 48 hpi, clinical findings of the affected chickens were open-mouth breathing, conjunctivitis, watery diarrhea and edema around the eye and neck. After 72 hpi, chickens showed muscular tremor, paralysis of the legs and wings, and coma. Histopathological results consist of multi-focal necrosis with hemorrhages in lymphoid aggregates of the intestinal tracts, necrosis of the lymphoid tissues, neuronal degeneration and necrosis, and perivascular cuffing. Using RT-PCR, virus genes were detected in the spleen and proventriculus at 48 hpi, and in the brain at 60 hpi.

Validation of a Real-Time RT-PCR Method to Quantify Newcastle Disease Virus (NDV) Titer and Comparison with Other Quantifiable Methods

  • Jang, Juno;Hong, Sung-Hwan;Kim, Ik-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.1
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    • pp.100-108
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    • 2011
  • A method for the rapid detection and quantification of Newcastle disease virus (NDV) produced in an animal cell culture-based production system was developed to enhance the speed of the NDV vaccine manufacturing process. A SYBR Green I-based real-time RT-PCR was designed with a conventional, inexpensive RT-PCR kit targeting the F gene of the NDV LaSota strain. The method developed in this study was validated for specificity, accuracy, precision, linearity, limit of detection (LOD), limit of quantification (LOQ), and robustness. The validation results satisfied the predetermined acceptance criteria. The validated method was used to quantify virus samples produced in an animal cell culture-based production system. The method was able to quantify the NDV samples from mid- or late-production phases, but not effective on samples from the early-production phase. For comparison with other quantifiable methods, immunoblotting, plaque assay, and tissue culture infectious dose 50 ($TCID_{50}$) assay were also performed with the NDV samples. The results demonstrated that the real-time RT-PCR method is suitable for the rapid quantification of virus particles produced in an animal cell-culture-based production system irrespective of viral infectivity.

A Novel Role of Classical Swine Fever Virus Erns Glycoprotein in Counteracting the Newcastle Disease Virus (NDV)-mediated IFN-β Induction

  • Xia, Yan-Hua;Chen, Liu;Pan, Zi-Shu;Zhang, Chu-Yu
    • BMB Reports
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    • v.40 no.5
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    • pp.611-616
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    • 2007
  • $E^{rns}$ is an envelope glycoprotein of classical swine fever virus (CSFV) and has an unusual feature of RNase activity. In the present study, we demonstrate that $E^{rns}$ counteracts Newcastle disease virus (NDV)-mediated induction of IFN-$\beta$. For this purpose, $E^{rns}$ fused to the enhanced green fluorescent protein (EGFP) was transiently expressed in porcine kidney 15 (PK15) cells. In luciferase activity assay, $E^{rns}$-EGFP was found to prevent IFN-$\beta$ promoter-driven luciferase expression and block the induction of IFN-$\beta$ promoter mediated by NDV in a dose-dependent manner. Through IFN-specific semi-quantitative RT-PCR detection, obvious decrease of IFN-$\beta$ mRNA in NDV-infected PK15 cells was observed in the presence of $E^{rns}$-EGFP. In contrast, EGFP alone showed none of this block capacity. In addition, $E^{rns}$-EGFP mutations with RNase inactivation were also found to block NDV-mediated induction of IFN-$\beta$. These evidences establish a novel function for CSFV $E^{rns}$ glycoprotein in counteraction of the IFN-$\beta$ induction pathway.

Newcastle Disease in the Ostriches reared in Korea (타조의 뉴캣슬병 증례보고)

  • Kwon, Yong-Kuk
    • Korean Journal of Veterinary Pathology
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    • v.7 no.1
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    • pp.23-26
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    • 2003
  • Newcastle disease (ND) is acute respiratory disease with high mortality in chicken and pet birds. Two ostriches aged on 51 days old submitted had showed clinical signs of severe torticolis and poor locomotion. In gross, fibrinous air-sacculitis and hemorrhagic enteritis were present. The significant lesions were histopathologically lymphocytic encephalitis such as perivascular cuffings and gliosis in neuropil. The velogenic ND virus was isolated ftom 10 day-old embryonated eggs inoculated with hemogenized cecal tonsils. These results suggest that NO occurred in the ostrich in Korea.

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Partial nucleotide sequencing and phylogenetic analyses of Newcastle disease virus and infectious bursal disease virus isolated in South Korea

  • Son So-Youn;Kim Duk-Soon;Kim Hyun-Soo;Kim Won-Seol;Park Jae-Myoung;Shin Hyun-Jin
    • Korean Journal of Veterinary Service
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    • v.28 no.4
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    • pp.375-385
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    • 2005
  • The present study was conducted to investigate the genetic profile of two prevalent avian pathogens in Korea namely, Newcastle disease virus (NDV) and infectious bursal disease virus (IBDV). Two farms located in Yeongi-gun, Chungnam were selected for this study. The two viruses were isolated from various organs (spleen, trachea, bursa of Fabricius) of deceased chickens that showed clinical symptoms of Newcastle Disease or Infectious bursal disease like swelling and congestion of the F bursa, facial edema, lacrimation, greenish yellow diarrhea as well as pathological signs like airsacculitis, haemorrhages in the intestines and so on. For analysis of NDV and IBDV, a 466 and 435 base pair fragments corresponding to the HN and VP2 regions which are highly conserved among related strains of NDV and IBDV, respectively, were amplified by RT-PCR and analyzed by sequencing. Comparison of the VP2 region showed a $99.3\%$ homology between the Korean IBDV isolate and the BJ836-attenuated vaccine strain. In contrast, the HN region of the Korean NDV isolate only has an 83 to $84\%$ homology with the vaccine strains LaSota, B1 and VGGA. Our findings reveal that the prevalent NDV strain in Korea is genetically different from the vaccine strains and may explain the recent outbreaks of Newcastle disease in the region.