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Complete genome sequence of a velogenic newcastle disease virus isolated from an apparently healthy village chicken in South India.

ABSTRACT: We report the complete genome sequence of a Newcastle disease virus (NDV) isolate, NDV-D1/1998, from an apparently healthy village chicken in South India. This class II, genotype II virus is 15,186 nucleotides in length with unique amino acid variations and was found to be a velogenic pathotype by standard pathogenicity tests.

SUBMITTER: Uthrakumar A

PROVIDER: S-EPMC4064031 | biostudies-literature | 2014 Jun

REPOSITORIES: biostudies-literature

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Complete genome sequence of a velogenic newcastle disease virus isolated from an apparently healthy village chicken in South India.

Uthrakumar Arumugam A Vijayarani Kumanan K Kumanan Kathaperumal K Bhuvaneswari Srinivasan S Kuchipudi Suresh Varma SV Elankumaran Subbiah S

Genome announcements 20140619 3

We report the complete genome sequence of a Newcastle disease virus (NDV) isolate, NDV-D1/1998, from an apparently healthy village chicken in South India. This class II, genotype II virus is 15,186 nucleotides in length with unique amino acid variations and was found to be a velogenic pathotype by standard pathogenicity tests. ...[more]

PMID: 24948766

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Project description:BACKGROUND: Newcastle disease virus (NDV) causes severe and economically important disease in poultry around the globe. None of NDV strains in Pakistan have been completely characterized and the role of rural poultry in harbouring NDV is unclear. Since they have a very important role for long-term circulation of the virus, samples were collected from apparently healthy backyard poultry (BYP) flocks. These samples were biologically analyzed using mean death time (MDT) and intracerebral pathogenicity index (ICPI), whereas genotypically characterized by the real-time PCRs coupled with sequencing of the complete genome. FINDINGS: Despite of being non-pathogenic for BYP, the isolate exhibited MDT of 49.6 h in embryonated chicken eggs and an ICPI value of 1.5. The F gene based real-time PCR was positive, whereas M-gene based was negative due to substantial changes in the probe-binding site. The entire genome of the isolate was found to be 15192 nucleotides long and encodes for six genes with an order of 3'-NP-P-M-F-HN-L-5'. The F protein cleavage site, an indicative of pathogenicity, was 112RRQKRF117. Complete genome comparison indicated that the RNA dependent RNA polymerase gene was the most and the phosphoprotein was least conserved gene, among all the genes. The isolate showed an Y526Q substitution in the HN protein, which determines neuraminidase receptor binding and fusion activity of NDV. Phylogenetic analysis, based on F and HN genes, classified this isolate into genotype VII, a predominant genotype responsible for ND outbreaks in Asian countries. However, it clustered well apart from other isolates in this genotype to be considered a new subgenotype (VII-f). CONCLUSIONS: These results revealed that this isolate was similar to virulent strains of NDV and was avirulent in BYP either due to resistance of local breeds or due to other factors such as substantial mutations in the HN protein. Furthermore, we have characterized the first isolate of NDV, which could act as domestic reference strain and could help in development and selection of appropriate strain of NDV for vaccine in the country.

Project description:Newcastle disease virus (NDV) belongs to genus Avulavirus and family Paramyxoviridae. There are thirteen serotypes named APMV-I (Avian Paramyxovirus-I) to APMV-13 of which NDV has been designated as APMV-1. The disease has been reported worldwide affecting both domestic and wild avian species. Morbidity and mortality rates up to 100% have been reported in cases of unvaccinated flocks. Stringent vaccination schedule is practiced in endemic/disease prone areas in order to prevent the disease. Despite this, NDV outbreaks have been reported even in cases of vaccinated populations. In this study we describe detailed pathological and molecular investigation that were undertaken in an organized poultry farm from Bareilly region, Uttar Pradesh, India, involving layer flocks which succumbed to ND outbreak in spite of following strict vaccination protocol. The mortality rate ranged from 76.80 to 84.41% in different flocks with an average mortality of 79.50%. Necropsied birds had gross lesions suggestive of viscerotropic ND including petechial hemorrhages on the proventricular tips, intestinal lumen with necrotic areas covered with hemorrhages, hemorrhagic cecal tonsils, para-tracheal edema and mottling of spleen. The characteristic histopathological lesions were mainly seen in the blood vessels and lymphoid tissues. Vascular changes characterized by congestion, edema, and hemorrhage were found in majority of the organs. Lymphocytolysis in spleen and cecal tonsils was evident. Immunohistochemical studies revealed positive signals mostly in macrophage and lymphocytes. PCR assay was done to confirm the NDV genome, which revealed an amplicon size of 356 bp. The phylogenetic analysis revealed the resemblance of the present isolate (ADS01) with class II genotype NDV XIIIA. The isolate belonged to velogenic NDV as the Minimum Lethal Dose (MLD) and Mean Death Time (MDT) for the present isolate were 10-8 and 41 h, respectively. Thus this study clearly demonstrates that in spite of strict vaccination regime and biosecurity procedures, ND continues to be rampant. Hence it is important to effectively administer the present vaccine in addition to strains matching to the field isolates to provide longer and optimal protection against spreading of virus by means of reducing the extent of viral shedding.

Project description:Newcastle disease virus (NDV) remains a constant threat to the poultry industry. There is scarce information concerning the pathogenicity and genetic characteristics of the circulating velogenic Newcastle disease virus (NDV) in Egypt. In the present work, NDV was screened from tracheal swabs collected from several broiler chicken farms (N = 12) in Dakahlia Governorate, Egypt. Real-time reverse transcriptase polymerase chain reaction (RRT-PCR) was used for screening of velogenic and mesogenic NDV strains through targeting F gene fragment amplification, followed by sequencing of the resulting PCR products. The identified strain, namely, NDV-CH-EGYPT-F42-DAKAHLIA-2019, was isolated and titrated in the allantoic cavity of 10 day old specific pathogen-free (SPF) embryonated chicken eggs (ECEs), and then their virulence was determined by mean death time (MDT) and intracerebral pathogenicity index (ICPI). The pathogenicity of the identified velogenic NDV strain was also assessed in 28 day old chickens using different inoculation routes as follows: intraocular, choanal slit, intranasal routes, and a combination of both intranasal and intraocular routes. In addition, sera were collected 5 and 10 days post inoculation (pi) for the detection of NDV antibodies by hemagglutination inhibition test (HI), and tissue samples from different organs were collected for histopathological and immunohistochemical examination. A series of different clinical signs and postmortem lesions were recorded with the various routes. Interestingly, histopathology and immunohistochemistry for NDV nucleoprotein displayed widespread systemic distribution. The intensity of viral nucleoprotein immunolabeling was detected within different cells including the epithelial and endothelium lining, as well as macrophages. The onset, distribution, and severity of the observed lesions were remarkably different between various inoculation routes. Collectively, a time-course comparative pathogenesis study of NDV infection demonstrated the role of different routes in the pathogenicity of NDV. The intranasal challenge was associated with a prominent increase in NDV lesions, whereas the choanal slit route was the route least accompanied by severe NDV pathological findings. Clearly, the present findings might be helpful for implementation of proper vaccination strategies against NDV.

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Complete genome sequence of a velogenic newcastle disease virus isolated from an apparently healthy village chicken in South India.

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Complete genome sequence of a velogenic newcastle disease virus isolated from an apparently healthy village chicken in South India.

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